CN101563461A - Plants having improved characteristics and a method for making the same - Google Patents

Abstract

The present invention relates generally to the field of molecular biology and concerns a method for enhancing various economically important characteristics in plants. More specifically, the present invention concerns a method for improving yield-related traits, such as enhanced yield and/or enhanced growth, or modified content of storage compounds in plants by modulating expression in a plant of a nucleic acid encoding a GRP (Growht Related Protein) polypeptide. The present invention also concerns plants having modulated expression of a nucleic acid encoding a GRP polypeptide, which plants have improved characterisitics relative to control plants. The invention also provides hitherto unknown GRP-encoding nucleic acids, and constructs comprising the same, useful in performing the methods of the invention.

Description

Has plant of improved characteristics and preparation method thereof

Relate generally to biology field of the present invention also relates to the method that is used for improving in the plant expression by the nucleic acid of regulating coding GRP (growth associated protein) the various plants feature.The invention still further relates to the plant of being regulated expression of the nucleic acid with coding GRP polypeptide, wherein said plant has the feature of improvement with respect to corresponding wild-type plant or other control plants.The present invention also is provided for the construct in the inventive method.

The world population of sustainable growth is supplied the research that atrophy has stimulated relevant increase farm efficiency with agricultural with the arable land.The plant that conventional crop and the utilization of Horticulture improved means select breeding technique to have welcome characteristic with evaluation.Yet this type of selects breeding technique to have several defectives, and promptly these technology generally expend a lot of work and produce such plant, and it often contains the heterology hereditary component, and it may always not cause the desired proterties transmitted from the parental generation plant.Recent advances in molecular biology has allowed the germplasm of human improvement animal and plant.The genetic engineering of plant makes and can separate and operate genetic material (generally being in DNA or rna form) and introduce this genetic material subsequently to plant.This type of technology has generation and possesses diversified economy, agronomy or the crop of Horticulture improvement proterties or the ability of plant.

Proterties with special economic meaning is the output that increases.But output is normally defined the measuring result from the economic worth of crop.This result can define with regard to quantity and/or quality aspect.Output directly depends on Several Factors, for example the number of organ and size, plant structure (for example Zhi number), seed generation, leaf aging etc.The important factor that root development, nutrition intake, stress tolerance and early stage vigor (early vigor) also can be decision output.Optimize aforementioned factor thereby can contribution be arranged increasing crop yield.

Seed production is the proterties of particularly important, because the seed of numerous plants is important to people and Animal nutrition.Crop such as corn, rice, wheat, OK a karaoke club promise rape and soybean account for above the human total heat of half and take in, no matter by direct consumption seed itself or by consuming the meat product that produces based on the seed of processing.Crop also is the source of used numerous type metabolites in sugar, oil and the industrial processes.Seed contains embryo (origin of new talent and Xin Gen) and endosperm (source of nutrition that is used for embryonic development during duration of germination and the seedling early growth).Seed development relates to several genes and needs metabolite to be transferred to the seed of growing from root, leaf and stem.Endosperm especially assimilates carbohydrate, oil and proteinic metabolic precursor thereof and they is synthesized the storage macromole to fill seed.

Another important character for numerous crops is early stage vigor.Improving early stage vigor is the important goal of modern rice breeding plan on temperate zone and tropical rice varieties.It is important that long root is planted in the rice for correct soil fixing at water.In that direct sowing is to the situation that is submerged the field with rice, and under the situation that plant must emerge rapidly from water, long seedling is relevant with vigor.Under the situation of implementing drilling, long mesocotyl and coleoptile are important for well emerging.With early stage vigor artificial reconstructed will be extremely important in agricultural to endophytic ability.For example, bad early stage vigor has limited based on corn (the Zea mayes L.) hybrid of Corn Belt germplasm (Corn Belt germplasm) and has introduced a fine variety European Atlantic ocean region.

Another important character is improved abiotic stress tolerance.Abiotic stress is the major cause of world wide crop loss, reduces mean yield and surpass 50% (Wang etc., Planta (2003) 218:1-14) for most of staple crop plants.Abiotic stress can be caused by arid, salinity, extreme temperature, chemical toxicity and oxidative stress.Improving plant will be very big economic advantages to the peasant and can allow during unfavourable condition and in arable farming otherwise be impossible land raise crop the ability of abiotic stress tolerance at world wide.

Crop yield thereby can increase by optimizing one of aforementioned factor.

Depend on end-use, may have precedence over other yield traitses the improvement of some yield traits.For example for use as feed or timber production or biofuel resource for, increasing the plant nutrition body portion may wish, and for use as flour, starch or oil production for, increase is planted a subparameter and may especially be wished.Even if in the middle of kind of subparameter, some parameter can be more preferably in other parameters, and this depends on application.Number of mechanisms can have contribution to increasing seed production, and no matter form is the seed size of increase or the number seeds of increase.

A kind of method that increases output (seed production and/or biomass) in the plant can be the multiple signal pathway by the inherent growth mechanism of regulating plant such as cell cycle or involved in plant growth or participation defense mechanism.

Have now found that and in plant, to express the multiple growth characteristics that improves in the plant by the nucleic acid of regulating coding GRP polypeptide in the plant.The GRP polypeptide can be one of following albumen: anchorin-zinc finger polypeptide (AZ), SYT polypeptide, chloroplast(id) fructose-1 (cpFBPase) polypeptide, little inducibility kinases (SIK), II class homeodomain leucine zipper (HD Zip) transcription factor and SYB1 polypeptide.The feature of improvement comprises the output correlated character, as output and/or the growth of increase or the storage compound thing content of improvement that increases.

Background technology

Anchorin-zinc finger polypeptide

Phytomass is the output of fodder crop such as clover, ensiling cereal and hay.In bread crop, use many alternate parameter of output.Wherein primary is estimation plant size.Difference according to species and etap, can measure the plant sizes by many methods, but comprise plant gross dry weight, dry weight, long-pending, the plant height of fresh weight, leaf area, caulome, lotus throne plant diameter, leaf length, root length, root biomass, tiller number and the number of sheets on the ground on the ground.Many species are kept conservative ratio between plant different piece size in the given etap.Utilize these allometry relation and measuring results of these relevant sizes are carried out from one to the other extrapolation (as 2005Agric Ecosys﹠amp such as Tittonell; Environ 105:213).The plant size of early development stage is usually with relevant with the plant size of etap in late period.Plant usually can be smaller greatly more light of plant absorbing and carbonic acid gas with bigger leaf area are therefore probably at the more (Fasoula﹠amp of the weightening finish same period; Tollenaar 2005Maydica50:39).Except reaching at first than big or small microenvironment or prepotent potential continuity that plant had, this is its additive effect.Plant size and growth velocity exist strong genetic module (as 2005 Plant Physiology 139:1078 such as terSteege), and up to now, all diversified genotype plants are at the size under a kind of envrionment conditions relevant with the size under the another kind of envrionment conditions probably (2003 Theoretical Applied Genetics 107:679 such as Hittalmani).By this way, use the alternate parameter of the diversified dynamic environment that standard environment met with in different time and place as crop in the field.

Harvest index is the ratio of seed production and ground dry weight, it is relatively stable under many envrionment conditionss, therefore can obtain more firm dependency (as 2002 Crop Science 42:739 such as Rebetzke) between plant size and grain yield usually.These methods link together inherently, because the photosynthesis productivity that most of cereal biomasss depend on leaf and stem is current or store (1985 Physiology of Crop Plants.Iowa State University Press such as Gardener, pp68-73).Therefore, to the selection of plant size, or even in the selection of growing commitment, the index that has been used as following potential production is (as 2005Agric Ecosys﹠amp such as Tittonell; Environ 105:213).When test hereditary difference during to the influencing of stress tolerance, greenhouse or plant culturing chamber are compared with the field has inherent advantages: promptly can make the operability and the light intensity stdn of soil function, temperature, water and nutrition.But, for want of wind-force or insect cause bad pollination, or since insufficient space allowing matured root or canopy growth or the like, the sex-limited meeting of these manual offices that output is caused limits these application in the test volume variance of controling environment.Therefore, under culturing room or greenhouse standard conditions, measure the plant size of early development stage, provide the standard method of potential hereditary yield heterosis index.

Transcribe by RNA polymerase and undertaken.These polysaccharases are relevant with decision specific other albumen of transcription (transcription factor) usually.Transcription factor combines with the cis-regulating element of gene and also can mediate other regulates proteic combination.Stegmaier etc. propose the classification based on the transcription factor of transcription factor DNA binding domains.Based on following existence, distinguish 5 super classes (supperclass): the β supporting structure territory and 0 that 1) foundation structure territory, 2) zinc synergetic structure territory, 3) spiral corner spirane structure territory, 4) has the ditch contact) other structural domains (Stegmaier etc., Genome informatics 15,276-286,2004).Comprise that the transcription factor group in zinc synergetic structure territory is extremely various and can further classify, comprise that WRKY structural domain, C6 zinc cluster, DM and GCM structural domain according to its conservative halfcystine and histidine residues.

Except the DNA binding motif, transcription factor also can comprise the protein-protein interaction motif.A kind of this motif is the ankyrin motif.It appears in the various protein families, arrives the repetition that surpasses 20 units as 2 usually.Each unit comprises two antiparallel spirals and beta hairpin.

Although many plant proteins with Zinc finger domain are fully characterized, to known to the plant protein that comprises the C3H1 zinc-finger motif seldom.It is reported that PEI1 is a kind of transcription factor that fetal development is worked, it has the zinc-finger motif similar to the C3H1 motif, but lacks ankyrin motif (Li and Thomas, Plant Cell 10,383-398,1998).WO02/44389 has described AtSIZ, a kind of transcription factor of separating from Arabidopis thaliana (Arabidopsis).The AtSIZ of discovery under the control of CaMV35S promotor expresses and promotes the stress-inducing gene transcription, and it is reported that plant that AtSIZ with increase expresses compares with control plant and have higher survival rate under salt stress, but the analysis of relevant seed production is not provided.Infer that AtSIZ can be used for increasing the resistance of plant to osmotic stress.

SYT

Abiotic stress, as drought stress, salt stress, heat stress and cold coerce or these coerce in one or more combination, be plant-growth and bathmic key constraints (Boyer (1982) Science 218:443-448).Arid, heat, cold-peace salt stress have for the very important common theme of plant-growth, the i.e. availability of water.Because the high salt concentration in some soil will cause the less water that can be taken in by cell, its effect is to observed similar under drought condition.In addition, under freezing temperature, to cause water to flow out owing to start from the formation of the ice in the apoplast (apoplast) from synplasm, cause vegetable cell forfeiture moisture content (McKersie and Leshem thus, 1994.Stress andStress Coping in Cultivated Plants, Kluwer Academic Publishers).In heat stress, influence air vent aperture regulating cooling by evapotranspiration, thereby influence the moisture content of plant.Usually, plant is similar to the molecular reaction mechanism of each condition of these stress conditions.

Plant can be exposed to the condition of the environment water content of minimizing in its whole life history.Most of plants evolved out protection they oneself avoid the strategy of the injury of these conditions.Yet,, will be far-reaching to the influence of development of plants, growth and the output of most of crop plants if the severity of drought condition and time length are too big.Continue to be exposed to the great change that causes plant metabolism under the ambient water operability of minimizing.Metabolic these great changes cause necrocytosis the most at last, therefore cause production loss.For example the crop loss of rice, corn (corn) and wheat and crop yield loss are that important economy and political factor and many in the world areas have been caused and be short of food to coerce the staple crops that cause by these.

Another example that abiotic environment is coerced is plant-growth and the operability of growing one or more nutraceutical minimizings that need absorption.Because nutritional utilization efficient is to the brute force influence of plant biomass and quality product, a large amount of fertilizer are administered to the field, to optimize plant-growth and quality.Plant production power is limited by three kinds of main nutrient: phosphorus, potassium and nitrogen, they are the rate limiting factor in the plant-growth normally.The required main nutrient elements of plant-growth is nitrogen (N).It is the component of finding in the life cells that comprises amino acid, protein (enzyme), nucleic acid and chlorophyllous a large amount of important compound.What plant do to be divided 1.5% to 2% is nitrogen, and approximately all plant proteins 16% be nitrogen.Therefore, the operability of nitrogen is key constraints (Frink etc. (1999) the Proc Natl Acad Sci U.S. 96 (4): 1175-1180), and protein accumulation and amino acid formed have main influence of plant-growth and production.Therefore, under the nitrogen restricted condition growth time to have the cereal of output of increase highly beneficial.

Phytomass is the output of fodder crop such as clover, ensiling cereal and hay.In bread crop, use many alternate parameter of output.Wherein primary is estimation plant size.Difference according to species and etap, can measure the plant sizes by many methods, but comprise plant gross dry weight, dry weight, long-pending, the plant height of fresh weight, leaf area, caulome, lotus throne plant diameter, leaf length, root length, root biomass, tiller number and the number of sheets on the ground on the ground.Many species are kept conservative ratio between plant different piece size in the given etap.Utilize these allometry relation and measuring results of these relevant sizes are carried out from one to the other extrapolation (as (2005) Agric Ecosys﹠amp such as Tittonell; Environ105:213).The plant size of early development stage is usually with relevant with the plant size of etap in late period.Plant usually can be smaller greatly more light of plant absorbing and carbonic acid gas with bigger leaf area are therefore probably at the more (Fasoula﹠amp of the weightening finish same period; Tollenaar (2005) Maydica50:39).Except reaching at first than big or small microenvironment or prepotent potential continuity that plant had, this is its additive effect.Plant size and growth velocity exist strong genetic module (as (2005) Plant Physiology 139:1078 such as terSteege), and up to now, all diversified genotype plants are at the size under a kind of envrionment conditions relevant with the size under the another kind of envrionment conditions probably (Hittalmani etc. (2003) Theoretical Applied Genetics 107:679).By this way, use the alternate parameter of the diversified dynamic environment that standard environment met with in different time and place as crop in the field.

The plant of development stress tolerant is the strategy (McKersie and Leshem, (1994) Stress and Stress Coping in CultivatedPlants, Kluwer Academic Publishers) that possible solve or reconcile at least some these type of problems.Yet, development to this type of conventional plant breeding strategy of coercing the new plant strain that presents resistance (tolerance) be relatively slowly and the specific resistance of needs be with hope be to hybridize.Cross-incompatibility between limited stress tolerance germ plasm resource and the source far away plant species is the significant problem that runs in the conventional breeding.Yet these technology are normally labor-intensive, and produce and to contain usually from the plant of the allogeneic heredity component of mother plant heredity, and these hereditary components are not always to produce paternal anticipant character.Molecular biological progress has allowed the idioplasm (germplasm) of human reconstruction animal and plant.Genetically engineered plant need separate and operate genetic material (being generally the form of DNA or RNA) and reach subsequently with described genetic material introduced plant.Such technology can provide the crop or the plant of economy, agricultural or gardening proterties with multiple improvement.

SYT is for transcribing coactivator, and in plant, the transcriptional activation agent of itself and GRF (growth regulatory factor) family protein forms function mixture (Kim HJ, Kende H (2004) Proc Nat AcadSc 101:13374-9).SYT is also referred to as GIF in this article, for the GRF interaction factor ( GRF- iNteracting fActor) abbreviation and AN3 are the abbreviation of angustifolia3, in (2005) Plant J 43:68-78 such as Horiguchi.The SWI/SNF protein shared structure territory (in the N-terminal zone) of agent of GRF transcriptional activation and yeast Chromatin Remodeling mixture (van der Knaap E etc., (2000) Plant Phys 122:695-704).Think that the coactivator of transcribing of these mixtures participates in the SWI/SNF mixture is raised to enhanser and promoter region, with realize local Chromatin Remodeling (

Summaries such as AM, (2001) Annu Rev Biochem 70:475-501).Transcriptional activation is regulated in the change of local chromatin Structure.More precisely, think that SYT and plant SWI/SNF mixture interact, to realize the transcriptional activation (Kim HJ, Kende H (2004) Proc NatAcad Sc 101:13374-9) of GRF target gene.

SYT belongs in the Arabidopis thaliana gene family by three member compositions.Described SYT polypeptide and people SYT have homology.Shown that people SYT polypeptide is for transcribing coactivator (Thaete etc. (1999) Hum Molec Genet 8:585-591).Following three structural domains have characterized Mammals SYT polypeptide:

(i) N-terminal SNH (the N-terminal homology of SYT) structural domain is guarded in Mammals, plant, nematode and fish;

(ii) C-terminal QPGY is rich in structural domain, mainly is made up of glycine, proline(Pro), glutamine and tyrosine, occurs with indefinite interval;

(iii) the methionine(Met) between aforementioned two structural domains is rich in (being rich in Met) structural domain.

In plant SYT polypeptide, the SNH structural domain is fully conservative.The C-terminal structural domain is rich in glycine and glutamine, but not proline rich or tyrosine.Therefore it is named as and is rich in the QG structural domain, forms contrast with mammiferous QPGY structural domain.SYT is the same with Mammals, can identify at QG structural domain N-terminal and be rich in the Met structural domain.Being rich in that the QG structural domain can be considered is the remainder (removing the SHN structural domain) of peptide C end basically; Be rich in the Met structural domain be generally comprised within be rich in the QG structural domain first half within (direction from the N-terminal to the C-terminal).Having second is rich in the Met structural domain and is positioned at (see figure 1) before the plant SYT polypeptide SNH structural domain.

It is reported that the SYT mutant of loss of function and SYT express the transgenic plant that reduce and grow little and narrow leaf and petal, they have less cell (Kim HJ, Kende H (2004) Proc Nat Acad Sc 101:13374-9).

The leaf that the plant that the AN3 overexpression produces in the Arabidopis thaliana has is than the big 20-30% of leaf (Horiguchi etc., (2005) Plant J 43:68-78) of wild-type.

The method of coming leaf size on the controlled levels direction by control AN3 genetic expression has been described among the Japanese patent application 2004-350553.

cpFBPase

Harvest index is the ratio of seed production and ground dry weight, and it is relatively stable under many envrionment conditionss, therefore can obtain more firm dependency (as 2002Crop Science 42:739 such as Rebetzke) between plant size and grain yield usually.These methods itself link together, because the photosynthesis productivity that most of cereal biomasss depend on leaf and stem is current or store (1985Physiology of Crop Plants.Iowa State University Press such as Gardener, pp68-73).Therefore, to the selection of plant size, or even in the selection of growing commitment, the index that has been used as following potential production is (as 2005Agric Ecosys﹠amp such as Tittonell; Environ 105:213).When test hereditary difference during to the influencing of stress tolerance, greenhouse or plant culturing chamber are compared with the field has inherent advantages: promptly can make the operability and the light intensity stdn of soil function, temperature, water and nutrition.But, for want of wind-force or insect cause bad pollination, or since insufficient space allowing matured root or canopy growth or the like, the sex-limited meeting of these manual offices that output is caused limits these application in the test volume variance of controling environment.Therefore, under culturing room or greenhouse standard conditions, measure the plant size of early development stage, provide the standard method of potential hereditary yield heterosis index.

Photosynthetic carbon metabolism is the main process of plant-growth and grain yield in the higher plant.(Calvin, Calvin) the approach fix atmospheric carbon dioxide produces carbohydrate in higher plant by the reduction pentose phosphate.This process occurs in the chloroplast(id), and new synthetic triose phosphate can be as in the starch synthetic matrix compartment, or is output in the cell colloid that sucrose forms.In photosynthesis, new synthetic carbohydrate forms (channel) a kind of or other forms according to needs and the envrionment conditions of plant.

Calvin cycle is complicated approach, comprises the three phases by 11 kinds of enzymatic 13 reactions.A kind of prior enzyme is chloroplast(id) fructose-1 (cpFBPase), and its catalysis fructose-1,6-diphosphate is to the irreversible conversion of fructose-6-phosphate and Pi (inorganic P).Compare with other enzymes of Calvin cycle, the level of cpFBPase polypeptide in chloroplast(id) is very low.

By redox potential regulation and control cpFBPase activity, described system is according to illumination/dark condition and pH value and Mg by Triphosphopyridine nucleotide photoreductase-thioredoxin system ²⁺Light dependent change regulatory enzyme activity (Chiadmi etc., (1999) EMBO 18 (23): 6809-6815) on the level.More specifically, the cpFBPase polypeptide is activated under illumination, and is non-activity in the dark, and this is by redox interaction between the enzyme molecule SH group of thioredoxin mediation and also by pH value and Mg in the chloroplast stroma ²⁺Concentration is subjected to light to induce rising and (Buchanan (1980) AnnuRev Plant Physiol 31:341-374 takes place; Jacquot (1984) Bot Acta 103:323-334).

Higher plant and algae cpFBPase polypeptide are carried out same enzyme catalysis step, but slightly different in enzymatic activity is regulated.More specifically, algae cpFBPase polypeptide is strict with reduction and is had activity, but the reductive agent specificity is required not too strict, that is, and they can activate by homoplasmon thioredoxin not (Huppe and Buchanan (1989) Z Naturforsch 44 (5-6): 487-94).

In photosynthetic (autotrophy) cell,, be arranged in cytosol (cyFBPase) in addition and participate in the 2nd FBPase polypeptide (isotype) of the synthetic and glyconeogenesis of sucrose except the cpFBPase polypeptide.Compare with the cpFBPase polypeptide, the cyFBPase polypeptide has very different modulating properties: it is suppressed by excessive substrate, and by AMP and fructose-2, the 6-bisphosphate shows that allosteric suppresses, and presents the neutral pH optimum.In heterotrophism system (as zooblast), find the cyFBPase polypeptide.The key distinction between cpFBPase polypeptide and cyFBPase polypeptide is to have aminoacid insertion among the former, and described aminoacid insertion has two conservative cysteine residues at least, and described cysteine residues is the target of thioredoxin regulation and control.

Under the regulation and control of stem tuber specificity promoter, express the existing report of transgenic Rhizoma Solani tuber osi plant (Thorbjornsen etc., (2002) Planta 214:616-624) of potato cyFBPase nucleotide sequence.The author find the transgenosis stem tuber aspect starch content or neutral sugar and phosphorylation hexoses aspect and wild-type stem tuber as broad as long.

Effectively be connected in tomato rbcS transit peptides encoding sequence (Miyagawa etc., (2001) Nature Biotech 19:965-969 of chloroplast(id) ubcellular target chimeric protein from two cyFBPase encoding genes of cyanobacteria (cyanobacterium Synechococcus) (FBPase/SBPase and FBPaseII); Tamoi etc., (2006) Plant Cell Physiol 47 (3): 380-390).Do not find of the redox condition adjusting of the activity of FBPase/SBPase and FBPaseII polypeptide, because their lack conservative cysteine residues by Triphosphopyridine nucleotide photoreductase/thioredoxin system.Use the expression of two mosaic genes of tomato rbcS promoter regulation.Reported the rotaring gene tobacco plant that transforms with the chimeric construct body obviously faster and bigger than the wild-type plant growth under atmospheric condition.

SIK

The plant cultivation person is interested according to the part of the plant of the particular aspects of the improvement output of discussing crop or plant and economically valuable or crop usually.For example, to specified plant or specific end-use, the plant cultivation person can seek improvement especially aspect the phytomass (weight) of the one or more parts of plant, and described part comprises (can gather in the crops) part and/or underground (can gather in the crops) part on the ground.What especially relevant was for consumption is that plant shoot divides or underground part.For many crops, cereal especially, the improvement of seed production extremely needs.The seed production itself that increases can show as multiple mode, depends on institute crop of discussing or plant and its end-use, and each independent aspects of seed production has the importance of variation to the plant cultivation person.For example seed production can show as or from: a) seed biomass (seed gross weight) increases, and this can be based on single seed and/or every strain plant and/or every square metre; B) every strain plant increases spends number; C) (full) seed number of Zeng Jiaing; D) the full rate of the seed of Zeng Jiaing (it is expressed as the ratio between full seed number and the seed sum usually); E) harvest index of Zeng Jiaing, it is expressed as the ratio that can gather in the crops part (as seed) output and total biomass usually; And f) thousand seed weight of Zeng Jiaing (TKW), this extrapolates from the full seed number and the gross weight thereof of counting usually.

The increase of seed production also can show as the increase of seed size and/or seed volume.In addition, the increase of seed production itself can self-expression be the increase of seed area and/or seed length and/or seed width and/or seed girth also.The output that increases also can produce the structure of improvement, or can occur because of the structure of improvement.

With the corn is example, and the output increase can show as following one or more indexs: the increase of the increase of the increase of every square metre of plant number, every strain plant spike number, the increase of line number, every row grain number, grain weight, thousand seed weight, fringe length/diameter, the full rate of seed (wherein the full rate of seed is that the full seed number is total and multiply by 100 divided by seed) and other.With the rice is example, and itself can show as the increase of following one or more indexs the output increase: the increase of every square metre of plant number, every strain plant panicle number, every panicle spikelet number, every panicle flower (Xiao Hua) number (it is expressed as the ratio of full seed number to former panicle number), the full rate of seed (wherein the full rate of seed be the full seed number divided by the seed sum and multiply by 100), the increase of thousand seed weight and other.

Concerning the plant cultivation person, can select and want the aspect of reformed seed production highly beneficial.What very expectation can be non-customized chooses, and promptly chooses to be fit to change the particular aspects of seed production or the gene of component.For example the increase with the full rate of the thousand seed weight combination that increases is to expect very much crop as corn.The combination of the thousand seed weight of full rate, harvest index and the increase that rice and wheat are increased is expected very much.

The disclosed International Patent Application WO 02/074801 of Genomine Inc. has been described from the AtSIK protein of Arabidopis thaliana (Arabidopsis thaliana) and the gene of code for said proteins.It is mentioned and can make plant have resistance to osmotic stress by the expression that suppresses AtSIK.Yet the expression of not mentioning by suppressing AtSIK can improve those output aspects.

HD Zip

Study on plants and genetically manipulated have long-range history, itself in addition at famous Gregor

Just begun before the Mendel research.In improving this science, scientist has finished the improvement of specific trait in the plant, and described proterties contains the potato tuber of the starch content with increase and has oilseeds plant such as the canola oil dish and the Sunflower Receptacle of fatty acid content increase or that change.Along with the vegetables oil consumption that increases with use, the improvement of seed oil content and seed oil level more prevalent (as

Deng,, 1995, Science 268:681-686).Handling biosynthetic pathway in transgenic plant provides a large amount of chances to remove to influence plant metabolism for molecular biologist and plant biochemistry scholar, to cause the generation of specific more high-value product.The generation of seed oil or synthetic (composition) a large amount of traditional oilseeds plants such as soybean (United States Patent (USP) the 5th, 955, No. 650), canola oil dish (United States Patent (USP) the 5th, 955, No. 650), Sunflower Receptacle (United States Patent (USP) the 6th, 084, No. 164), rape (

Deng, 1995, Science 268:681-686) and non-traditional foxy old hand plant such as the middle change of tobacco (Cahoon etc., 1992, Proc.Natl.Acad.Sci. U.S. 89:11184-11188).

Plant seed oils comprises neutral and polar lipid (seeing Table 1).Neutral lipid mainly contains triacylglycerol, and it is the main storage lipid that accumulates in the seed oil body.Polar lipid is mainly found in the various films of seed cell, as microsome, plastid and mitochondrial membrane and cytolemma.Neutral and polar lipid contains several frequently seen lipid acid (seeing Table 2) and a series of not too common lipid acid.The lipid acid of membrane lipid synthesizes by altitude mixture control and has only a spot of lipid acid to find in membrane lipid.On the other hand, the neutrality that a large amount of unconventional lipid acid can be incorporated in many plant species seeds is stored (Van de Loo F.J. etc., 1993 in the lipid, Unusual Fatty Acids in Lipid Metabolism inPlants, the 91-126 page or leaf, TS Moore writes, and Jr.CRC publishes; Millar etc., 2000, Trends Plant Sci.5:95-101).

The classification of table 1 vegetable lipid

Neutral lipid	Triacylglycerol (TAG)
			Diacylglycerol (DAG)
	Monoacylglycerol (MAG)
		Polar lipid	Single semi-lactosi diacylglycerol (MGDG)
	Two semi-lactosi diacylglycerols (DGDG)
			Phosphatidyl glycerol (PG)
	Phosphatidylcholine (PC)
			Phosphatidylethanolamine (PE)
	Phosphatidylinositols (PI)
			Phosphatidylserine (PS)
	Sulfo-quinovose diacylglycerol

Table 2 conventional plant lipid acid

16:0	Palmitinic acid
		16:1	Zoomeric acid
16:3	hiragonic acid
		18:0	Stearic acid
18:1	Oleic acid
		18:2	Linolic acid
18:3	Linolenic acid
		γ-18:3	Gamma-linolenic acid *
20:0	Eicosanoic acid
		20:1	20 carbon monoenoic acids
22:6	Docosahexenoic acid (DHA) ＊
		20:2	Eicosadienoic acid
20:4	Arachidonic acid (AA) ＊
		20:5	Timnodonic acid (EPA) ＊
22:1	Sinapinic acid

In the table 2, the lipid acid that has asterisk does not occur in plant seed oils usually, but their production in transgenic plant seed oil has importance in Plant Biotechnology.

The main site of fatty acid biological synthetic is a plastid in the plant.Fatty acid biological is synthetic to begin by acetyl-CoA carboxylase (ACCase) acetyl-CoA is converted into malonyl CoA.Propanedioic acid partly passes through malonyl CoA then: the ACP acyl transferase is transferred to acyl carrier protein (ACP).Enzyme β-ketoacyl-initial the condensation reaction of ACP-synthase III (KAS III) catalysis fatty acid biological synthetic, wherein after propanedioic acid-ACP decarboxylation, the carbanion that obtains is transferred to acetyl-CoA by the nucleophillic attack of carbonyl-carbon, generates 3-batanone acid-ACP (3-ketobutyryl-ACP).This reaction cycle is by reducing, dewater and reducing butyro-generation once more and finish.This reaction cycle repeats (having the reaction of KAS I or KAS II catalyzing and condensing) reaches a common 16-18 carbon atom to carboxyl groups chain length.These acyl-acps, as the substrate of the plastid acyltransferase of lipid formation in the so-called protokaryon approach or output in the cytosol behind the effect excision ACP by thioesterase by stearic acid-ACP desaturase desaturation.They enter acetyl-CoA storehouse (acyl-CoA pool) and are used in so-called eucaryon approach in cytosol, synthetic lipid in endoplasmic reticulum.

Lipid by protokaryon and eucaryon approach is synthetic all take place glycerol-3-phosphate acyltransferase (GPAT) and lysophosphatidate acyltransferase (LPAAT) continuous acylation (Browse etc., 1986, Biochemical is J.235:25-31; Ohlrogge﹠amp; Browse, 1995,5Plant Ce117:957-970).The phosphatidic acid that obtains (PA) is the precursor of other polarity film fat, described polarity film fat for example is phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylinositols (PI) and the phosphatidylserine (PS) in single semi-lactosi diacylglycerol (MGD), two semi-lactosi diacylglycerol (DGD), phosphatidyl glycerol (PG) and sulfo-quinovose diacylglycerol (SQD) and the endoplasmic reticulum in the plastid.Polar lipid also is that acyl chain is further modified the site of (for example desaturation, ethinylation (acetylenation) and hydroxylation).In endoplasmic reticulum, PA also is the intermediate product of biosynthesizing triacylglycerol (TAG), the important component of neutral lipid and the important component of seed oil herein.In addition, exist the biosynthetic alternative route of TAGS (that is, pass through phosphatidylcholine: the transacylation that Diacrylglycerol acyl transferase plays a role) (Voelker, 1996, Genetic Engineering, Setlow writes, 18:111-113; Shanklin﹠amp; Cahoon, 1998, Annu.Rev.Plant Physiol.PlantMol.Biol.49:611-641; Frentzen, 1998, Lipids 100:161-166; Millar etc., 2000, Trends Plant Sci.5:95-101).Reversed reaction, triacylglycerol to the conversion (breakdown) of diacylglycerol and lipid acid by lipase-catalyzed.This type of catalysis can be regarded the terminal point of seed development as, causes certain reduction (Buchanan etc., 2000) of seed oil.

Storage fat in the seed is synthetic from carbohydrate-deutero-precursor.Plant has complete glycolytic pathway (Plaxton, 1996, Annu.Rev.Plant Physiol.Plant Mol.Biol.47:185-214) in cytosol, and has shown also have complete approach (Kang﹠amp in the plastid of rape; Rawsthorne, 1994, Plant is J.6:795-805).Sucrose is the main source of carbon and energy, is transported to from leaf in the seed of growth.In the storage stage of seed, sucrose transforms in cytosol, so that metabolic precursor thereof fructose-6-phosphate and pyruvic acid to be provided.These precursors are transported to plastid and are converted into acetyl-CoA, and acetyl-CoA is the main precursor of lipid acid synthetic.Acetyl-CoA in the plastid is the biosynthetic center of a lipid precursor.Acetyl-CoA can form by different being reflected in the plastid, and the accurate contribution of each reaction still has arguement (Ohlrogge﹠amp; Browse, 1995, Plant Ce117:957-970).Yet, accepted be most of acetyl-CoA derived from fructose-6-phosphate and pyruvic acid, described fructose-6-phosphate and pyruvic acid are imported plastid from tenuigenin.Sucrose is also referred to as the storehouse organ at the seed that comes to produce and be transported among source organ's (leaf, or the place of any photosynthesis generation) growth.In the seed of growing, sucrose is the precursor of all stocks, and described stock is starch, lipid and part lipid storage protein.Therefore, be clear that the metabolism that sucrose plays the carbohydrate of central role is very important to the accumulation of seed storage thing.

Though the lipid of seed oil and fatty acid content can change by the traditional method of plant breeding, the appearance of recombinant DNA technology allows the content of easier manipulation oil of plant, and in some cases, the permission inaccessiable seed oil of dependence breeding institute change separately (referring to, for example

Deng, 1995, Science 268:681-686).For example, in transgene tobacco, introduce Δ ¹²-hydroxylase nucleotide sequence causes the generation (Van de Loo etc., 1995, Proc.Natl.Acad.Sci U.S. 92:6743-6747) of new fats acid (ricinolic acid) in tobacco-seed oil.Tobacco plant has also passed through introducing and the expression acyl-acp desaturase from coriandrum (coriander), transform as and produces low-level petroselinic acid (Cahoon etc., 1992, Proc.Natl.Acad.Sci U.S. 89:11 184-1 1 188).

The modification of oil of plant content has important medical, nutrition and economic consequence.About economic consequence, the clinical disease that the longer chain fatty acid of finding in some seed oil (C18 and longer) is relevant with other coronary heart disease with hypercholesterolemia related (Brenner, 1976, Adv.Exp.Med.Biol.83:85-101).Therefore, the plant that eats the lipid acid of these types with level increase can reduce cardiopathic risk.The extensive generation that the level of raising seed oil-contg also can be used for improving seed oil reduces these oily costs thus.

As previously mentioned, such as Δ ⁶-desaturase nucleic acid, Δ ¹²Some desaturase nucleic acid of-desaturase nucleic acid and acyl-acp desaturase nucleic acid are cloned, and are proved to be the synthetic required enzyme of lipid acid in the coding various plants species.Cloned from oleosin nucleotide sequence, and be proved to be the relevant protein of oil body phosphatide unitary film in coding and these plants such as the species of Btassica (Brassica), soybean, Radix Dauci Sativae, pine tree and Arabidopis thaliana.

Have now found that the nucleotide sequence of coding II class homeodomain leucine zipper (HD-Zip) transcription factor is used for the content of improved seed stock.

The protein that transcription factor is normally defined the combination of display sequence specific DNA and can activates and/or suppress to transcribe.At least 1533 transcriptional regulatory of arabidopsis gene group coding, this account for its estimation gene number～5.9%.Report these transcription factors of about 45% from plant specificity family (Riechmann etc., 2000 (Science the 290th volumes, 2105-2109)).An example of this type of special family of plant of transcription factor is the HD-Zip transcription factor family.

Homoeobox gene is the gene family that appears at the transcription factor in all eukaryotes and constitute at least 89 members in the Arabidopis thaliana.It is characterized in that having the homology frame, described homology frame generally includes 60 conservative amino acid residues, forms the helix-loop-helix-corner-spirane structure in conjunction with DNA.The normally false palindrome of this DNA binding site.Homoeobox gene grow many aspect have important and various effect, comprise in the early development, yeast of animal embryo that the initial sum of bud apical meristem is kept (Sakakibara etc. in the cell type specialization and flowering plant, Mol.Biol.Evol.18 (4): 491-502,2001).

Based on its amino acid similarity, separated a large amount of angiosperm homoeobox genes and be divided into seven not on the same group.These groups comprise that KNOX, BELL, HD-PHD-refer to, HAT1, HAT 2, GL2 and ATHB8 group.The leucine zipper motif of the contiguous homeodomain C-terminal of the gene in back four groups-coding, and form homeodomain leucine zipper (HD-Zip) gene family together.Aso etc., Mol.Biol.Evol.16 (4): 544-552,1999.Comprise in the Arabidopis thaliana among at least 47 among at least 89 members of homoeobox gene family and comprise homeodomain and leucine zipper.Although the combination of homeodomain and leucine zipper motif is a plant specific, except vascular plant, also in moss (moss), run into (Sakakibara etc., (2001) Mol Biol Evol 18 (4): 491-502).

Homeodomain leucine zipper (HDZip) albumen constitutes transcription factor family, it is characterized in that existing DNA binding domains (HD) and contiguous leucine zipper (Zip) motif.Leucine zipper is adjacent to the C-end of homeodomain, comprises several seven peptide repeating units (at least 4), wherein leucine (being Xie Ansuan or Isoleucine once in a while) just occurs every 6 amino acid usually.Leucine zipper is extremely important for protein dimerization.Dimerization is DNA bonded precondition (Sessa etc. (1993) EMBO J12 (9): 3507-3517), and can occur between two identical HDZip albumen (homodimer), perhaps between two different HDZip albumen (heterodimer).

The group RNTO HD-Zip I-IV subfamily of angiosperm homoeobox gene HAT1, HAT 2, GL2 and ATHB8 group.The combination of homeodomain and leucine zipper motif is that higher plant is distinctive, and this shows the adjusting that the HD-Zip gene can the distinctive growth course of involved in plant.Aso etc., Mol.Biol.Evol.16 (4): 544-552,1999.The function difference of HD-Zip gene in the different subfamilies.HD-Zip I and II gene participate in light, dehydration inductive ABA or growth hormone signal transduction network similarly.These signal transduction networks are relevant with the whole adjusting and controlling growth of plant.The overexpression of justice or antisense HD-Zip I or II mRNA changes growth velocity and growth usually.The big logarithm member of HD-Zip III subfamily plays a role in the cytodifferentiation of center pillar.HD-Zip IV gene is relevant with the differentiation of outermost cellular layer.Sakakibara etc., Mol.Biol.Evol.18 (4): 491-502,2001.

Proved different HDZip albumen activation or check and transcribe.Utilized reporter gene luciferase (Henriksson etc. (2005) Plant Phys 139:509-518) to prove in Arabidopis thaliana: I class HDZip ATHB1 ,-5 ,-6 and-16 works as activating transcription factor in the transient expression of Arabidopis thaliana leaf is measured.Utilize another reporter gene (glycuronidase; Meijer etc. (2000) MolGen Genet 263:12-21), prove that two kinds of rice I class HDZip albumen are that Oshox4 and Oshox5 work as incitant in the transient expression of rice cell suspension culture is measured.On the contrary, two kinds of rice II class HDZip albumen be Oshox1 with Oshox3 in identical test as transcription repressor (Meijer etc. (1997) the Plant J 11:263-276 that works; Meijer etc. (2000), the same).

From the II class HD-Zip gene of Arabidopis thaliana, HAT4 is also referred to as ATHB-2, has been in the news as keeping away shady reaction instrumentality (Morelli and Ruberti TIPS, the 7th volume, the 9th phase, in September, 2002).

SYB1

Ran is the little signal conduction GTP enzyme (gtp binding protein matter) in the zooblast, and it participates in nucleo-cytoplasmic transport.Nucleo-cytoplasmic transport is mainly studied in animal system.Several Ran conjugated proteins are known, and wherein RanBP2 is also referred to as Nup358.Suppose that RanBP2 links to each other with the proteinic kytoplasm side of nuclear pore complex, and the supposition function be as SUMO E3 ligase enzyme, though its structure is not typical E3 ligase enzyme.SUMO (ubiquitin be correlated with little modified protein) is an eukaryotic protein, and is covalently bound with other protein, regulates and control a large amount of cell processes thus.Relevant with Ubc9 (as the E2 conjugated protein), RanBP2 SUMO-1 labeled substrate, described substrate is connected with nuclear import receptor, and this is similar with proteinic ubiquitinization.Pass through the nucleopore input nucleus after the substrate of SUMOization.Comprise NFX1-p15 (chaperone in the mRNA output) and histone deacetylase HDAC-4 based on the proteinic example of the input of SUMOization.SUMO modifies and RanBP2 afterwards also shows in genetic expression, cell cycle (in nuclear envelope disintegration process) and the subcellular structure such as the promyelocytic leukemia corpusculum and works.

Major part in these researchs is carried out in the model animal system, and the plant protein and the process of correspondence are known little.The RanBP2 related protein of identifying in the Arabidopis thaliana has Zinc finger domain, but inequality in structure.

Summary of the invention

Surprisingly, the expression that has now found that the coding nucleic acid of regulating the GRP polypeptide has produced the plant that has the feature of improvement with respect to control plant.

One aspect of the present invention has now found that regulating coding has produced the plant that has the output of increase with respect to control plant from the AZ polypeptide of Arabidopis thaliana (AtAZ) or the expression of nucleic acid in plant of its homologue.According to one embodiment of the invention, the method that increases plant biomass is provided, it comprises regulates coding AZ polypeptide or the expression of its homologue in plant.Advantageously, implement method of the present invention and produced the plant that has the output of increase with respect to corresponding wild plant, particularly the plant of the seed production of Zeng Jiaing.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.Another aspect of the present invention has now found that the expression of nucleotide sequence in plant of regulating coding SYT polypeptide produced the plant that has the output of increase under abiotic stress with respect to control plant.Therefore, the present invention also is provided at the method that increases plant biomass under the abiotic stress with respect to control plant, and it comprises the expression of nucleotide sequence in plant of regulating coding SYT polypeptide.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.

Another aspect of the present invention has now found that the expression of nucleotide sequence in plant shoot divides that increases coding chloroplast(id) fructose-1 (cpFBPase) polypeptide increased plant biomass with respect to control plant.Therefore, according to the present invention, provide the method that increases plant biomass with respect to control plant, it comprises the expression of nucleotide sequence in plant shoot divides that increases coding cpFBPase polypeptide.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.

Another aspect of the present invention, have now found that adjusting coding SIK nucleic acid and/or the expression of SIK polypeptide in plant have produced the plant that has the output correlated character of multiple improvement with respect to control plant, wherein the overexpression of SIK coding nucleic acid in plant produced the every strain plant flowers number that increases with respect to control plant, and wherein reduces or remove thousand seed weight, the harvest index of increase and the full rate of increase that SIK nucleic acid has produced to be increased with respect to the corresponding wild-type plant substantially.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.

Another aspect of the present invention, the nucleic acid that has now found that coding II class HD-Zip transcription factor can be used for changing on the content of repertory in the seed useful.Therefore the present invention provides by regulating the expression of coding II class HD-Zip transcription factor in plant to change the method for the content of repertory in the seed with respect to control plant.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.The present invention also provides the seed that has the repertory content of change with respect to control plant, and described seed has the adjusting that is subjected to of the nucleic acid of coding II class HD-Zip transcription factor expresses.The invention provides the method that changes repertory content in the seed with respect to control plant, it comprises regulates the expression of coding II class HD-Zip transcription factor in plant.

Another aspect of the present invention has now found that the expression of nucleic acid in plant of regulating coding SYB1 polypeptide produced the plant that has the enhanced yield associated shape with respect to control plant.When plant (non-stress conditions) under no stress conditions was cultivated, being surprised to find this output increased.Therefore, the present invention also provides with respect to control plant increases the method that plant strengthens the output correlated character, and it comprises the expression of nucleic acid in plant of regulating coding SYB1 polypeptide.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.

Definition

Polypeptides

Term " polypeptide " is used interchangeably in this article with " protein " and refers to be in the amino acid that is connected by peptide bond in the random length polymerized form.

Polynucleotide/nucleic acid/nucleotide sequence/nucleotide sequence

Term " polynucleotide ", " nucleotide sequence ", " nucleotide sequence ", " nucleic acid ", " nucleic acid molecule " are used interchangeably in this article and refer to be in Nucleotide in the non-branch of the random length polymerized form, i.e. ribonucleotide or deoxyribonucleotide or these two combination.

Control plant

To select suitable control plant be the habitual part that is provided with of experiment and can comprise the corresponding wild-type plant or not have the corresponding plant of goal gene.Control plant generally is plant species or or even the identical mutation identical with plant to be assessed.Control plant also can be the inefficacy zygote of plant to be assessed.The inefficacy zygote is because genetically modified individuality is lost in separation." control plant " not only refers to whole strain plant as used in this article, also refers to plant part, comprises seed and plants subdivision.

Homologue

Proteinic " homologue " comprises such peptide, oligopeptides, polypeptide, protein and enzyme, and they have aminoacid replacement, disappearance and/or insertion and have similar biologic activity and functionally active to the non-modifying protein of described peptide, oligopeptides, polypeptide, protein and enzyme source with respect to the above-mentioned protein of non-modification.

Disappearance refers to remove one or more amino acid from protein.

Insertion refers to the introducing in the predetermined site in protein of one or more amino-acid residues.Insertion can comprise single or multiple amino acid whose aminoterminals fusions and/or carboxyl terminal merges and the interior insertion of sequence.Usually, littler in the insertion meeting of aminoacid sequence inside than aminoterminal fusion or carboxyl terminal fusion, about 1-10 residue rank.The example of aminoterminal or carboxyl terminal fusion rotein or fusogenic peptide comprise as the binding domains of used transcriptional activator in the yeast two-hybrid system or activation structure territory, bacteriophage coat protein, (Histidine)-6-label, glutathione S-transferase-label, albumin A, maltose binding protein, Tetrahydrofolate dehydrogenase, Tag100 epi-position, c-myc epi-position,

-epi-position, lacZ, CMP (calmodulin binding peptide), HA epi-position, PROTEIN C epi-position and VSV epi-position.

Replace other amino acid that refer to have similar characteristics (as the tendency of similar hydrophobicity, wetting ability, antigenicity, formation or destruction α-Luo Xuanjiegou or beta sheet structure) and replace proteinic amino acid.Aminoacid replacement generally is single residue, but can be a bunch collection property, and this depends on the functional constraint that places polypeptide; Inserting can be about 1-10 amino-acid residue rank usually.Aminoacid replacement preferably conservative amino acid replaces.Conservative property replacement table is (seeing for example Creighton (1984) Proteins.W.H.Freeman and Company (writing) and following table 3) well-known in the art.

Table 3: the example that conservative amino acid replaces

Residue	Conservative property replaces	Residue	Conservative property replaces
				Ala	Ser	Leu	Ile；Val
Arg	Lys	Lys	Arg；Gln
				Asn	Gln；His	Met	Leu；Ile
Asp	Glu	Phe	Met；Leu；Tyr
				Gln	Asn	Ser	Thr；Gly
Cys	Ser	Thr	Ser；Val
				Glu	Asp	Trp	Tyr
Gly	Pro	Tyr	Trp；Phe
				His	Asn；Gln	Val	Ile；Leu
Ile	Leu，Val

Aminoacid replacement, disappearance and/or insert and to use the peptide synthetic technology well-known in the art such as the solid phase method of peptide synthesis etc. or by the recombinant DNA operation and carry out easily.Being used to operate dna sequence dna is well-known in the art with the method that produces proteinic replacement, insertion or disappearance variant.For example, it is well-known and comprise M13 mutagenesis, T7-Gen vitro mutagenesis method (USB to be used for producing at the predetermined site place of DNA the technology that replaces sudden change and to be those skilled in the art, Clevelaand, OH), the site-directed mutagenesis (Stratagene of QuickChange, San Diego, CA), site-directed mutagenesis or other site-directed mutagenesiss of PCR-mediation.

Derivative

" derivative " comprises such peptide, oligopeptides, polypeptide, wherein compare with the aminoacid sequence of the protein (as target protein) of natural generation form, they comprise the interpolation of the amino-acid residue that the amino-acid residue that takes place with non-natural takes place amino acid whose replacement or non-natural.Proteinic " derivative " also comprises such peptide, oligopeptides, polypeptide; wherein compare with the aminoacid sequence of the natural generation form of polypeptide, they comprise change (glycosylation, acidylate, isoprenylation, phosphorylation, Semen Myristicae acidylate, sulphating etc.) amino-acid residue or non-natural change amino-acid residue of natural generation.Compare with the aminoacid sequence that derivative is originated, this derivative can also comprise and covalently or non-covalently one or more non-aminoacid replacement bases of bonded or the interpolation (for example reporter molecule or other parts) of described aminoacid sequence, as amino-acid residue for promote to detect this derivative bonded reporter molecule and take place with non-natural that the proteinic aminoacid sequence of natural generation compares.In addition, " derivative " also comprises proteinic natural generation form and such as the fusions of the labelled peptide of FLAG, HIS6 or Trx (summary of labelled peptide referring to Terpe, Appl.Microbiol.Biotechnol.60,523-533,2003).

Directly to homologue/collateral line homologue

Directly comprise the evolution notion that is used for describing the gene ancestral relationship to homologue and collateral line homologue.The collateral line homologue is the gene of same species endogenous origin in my late grandfather's gene replication, is from the different biological genes that originate from species formation to homologue directly, also is derived from identical my late grandfather's gene.

Structural domain

Term " structural domain " refers to along the sequence alignment result of evolution related protein and at one group of conservative amino acid of specific location.Although the amino acid in other positions can change between homologue, yet may be essential amino acid in proteinic structure, stability or function aspects in the amino acid indication of the high conservative of specific location.Structural domain is because of being identified by the conservative degree of the height in the aligned sequences of protein homology thing family, and they can be as identifying that thing is to determine whether the polypeptide of being discussed belongs to the peptide family of before having identified arbitrarily.

Motif/consensus sequence/label

Term " motif " or " consensus sequence " or " label " refer to short conserved regions in the sequence of evolution related protein.Motif is the high conservative part of structural domain often, but also can only comprise the part of structural domain, maybe can be positioned at (if whole amino acid of motif are positioned at outside the structural domain of definition) outside the conserved domain.

Hybridization

Term as defined herein " hybridization " is the process of the mutual renaturation of homologous complementary nucleotide sequence basically wherein.Crossover process can be carried out in solution fully, and promptly two kinds of complementary nucleic acid all are in the solution.Crossover process also can take place under one of complementary nucleic acid is fixed to the situation of matrix such as magnetic bead, agarose (Sepharose) pearl or any other resin.Crossover process also can be fixed on solid support such as nitrocellulose filter or the nylon membrane or be fixed to by for example photolithography under the situation on the silicate glasses upholder (latter is called nucleic acid array or microarray or is called nucleic acid chip) for example at one of complementary nucleic acid carries out.For hybridization is taken place, usually with nucleic acid molecule thermally denature or chemical modification so that double-stranded unwinding become two strands and/or remove hair clip or other secondary structures from single-chain nucleic acid.

Term " severity " refer to the condition of hybridizing therein.The severity of hybridization is formed by condition such as temperature, salt concn, ionic strength and hybridization buffer to be influenced.Usually, low stringency is chosen as when ionic strength of determining and pH, is lower than particular sequence pyrolysis chain temperature (T _m) about 30 ℃.Medium stringency is that temperature is lower than T at this moment _mAbout 20 ℃ and high stringency be this moment temperature be lower than T _mAbout 10 ℃.High stringency hybridization condition generally is used to separate the hybridization sequences that has high sequence similarity with target nucleic acid sequence.Yet nucleic acid can depart from sequence and because of the degeneracy of the genetic codon substantially the same polypeptide of still encoding.Thereby sometimes may need medium stringency hybridization condition to identify this type of nucleic acid molecule.

T _mBe the temperature when ionic strength of determining and pH, 50% target sequence and the probe hybridization that mates fully under described temperature.T _mThe based composition and the length that depend on solution condition and probe.For example, long sequence is hybridized under comparatively high temps specifically.From being lower than T _mAbout 16 ℃ obtain maximum hybridization speed until 32 ℃.The existence of monovalent cation in solution reduced the Coulomb repulsion of two nucleic acid interchain, thereby promotes hybrid molecule to form; This effect is tangible (for greater concn, this effect can be ignored) for the na concn up to 0.4M.Methane amide reduces the melting temperature(Tm) of DNA-DNA and DNA-RNA duplex, and every percentage ratio methane amide reduces 0.6-0.7 ℃, and adds 50% methane amide and allow to hybridize at 30-45 ℃, though hybridization speed can reduce.Base-pair mismatch has reduced the thermostability of hybridization speed and duplex.On average and for big probe, every % base mispairing T _mDescend about 1 ℃.The type that depends on hybrid molecule, T _mCan use following equation to calculate:

1) DNA-DNA crossbred (Meinkoth and Wahl, Anal.Biochem., 138:267-284,1984):

T _m=81.5 ℃+16.6xlog ₁₀[Na ⁺] ^a+ 0.41x%[G/C ^b]-500x[L ^c] ^-1-0.61x% methane amide

2) DNA-RNA or RNA-RNA crossbred:

T _m＝79.8+18.5(log ₁₀[Na ⁺] ^a)+0.58(％G/C ^bb)+11.8(％G/C ^b) ²-820/L ^c

3) few DNA or few RNA ^dCrossbred:

For＜20 Nucleotide: T _m=2 (l _n)

For 20-35 Nucleotide: T _m=22+1.46 (l _n)

^aOr for other monovalent cations, but only be accurate in the 0.01-0.4M scope.

^bBe accurate in the 30%-75% scope only for %GC.

^cThe length of L=duplex (in base pair).

^dOligo, oligonucleotide; l _n, the useful length of=primer=2 * (G/C number)+(A/T number).

Can for example handle to hybridization buffer and with the RNA enzyme with any control non-specific binding of numerous known technologies with proteinaceous solution closed film, interpolation heterology RNA, heterology DNA and SDS.For the non-homology probe, a series of hybridization can be undertaken by changing one of following condition: (i) reduce renaturation temperature (for example from 68 ℃ to 42 ℃) progressively or (ii) reduce methane amide concentration (for example from 50% to 0%) progressively.The technician understands during the hybridization can change and will keep or change the multiple parameter of stringency.

Except that the hybridization condition, the hybridization specificity generally also depends on the function of post-hybridization washing.For removing because of the background due to the non-specific hybridization, sample is with the salts solution washing of dilution.The key factor of this type of washing comprises the ionic strength and the temperature of final washing soln: salt concn is low more and wash temperature is high more, and then Xi Di severity is high more.Wash conditions is generally on the hybridization severity or be lower than hybridization severity and carrying out.Positive hybridization produces the signal that doubles background signal at least.Usually, the suitable stringency that is used for nucleic acid hybridization analysis method or gene amplification detection method as mentioned above.Also can select stricter or more undemanding condition.The technician understands during the washing can change and will keep or change the multiple parameter of stringency.

For example, be used for length and be included in 65 ℃ greater than the common high stringency hybridization condition of the DNA hybrid molecule of 50 Nucleotide and in 1 * SSC and 50% methane amide, hybridize, wash in 0.3 * SSC at 65 ℃ subsequently in 1 * SSC or at 42 ℃.Be used for length and be included in 55 ℃ greater than the example of the medium stringency hybridization condition of the DNA hybrid molecule of 50 Nucleotide and in 6 * SSC and 50% methane amide, hybridize, wash in 2 * SSC at 50 ℃ subsequently in 4 * SSC or at 40 ℃.The length of hybrid molecule is the expection length of hybrid nucleic acid.When the known nucleic acid hybridization of sequence, can determine hybrid molecule length herein by aligned sequences and the described conserved regions of evaluation.1 * SSC is 0.15M NaCl and 15mM Trisodium Citrate; Hybridization solution and washing soln can comprise 5 * Denhardt reagent, 0.5-1.0%SDS, the fragmentation salmon sperm DNA of 100 μ g/ml sex change, 0.5% trisodium phosphate extraly.

In order to define the purpose of severity level, can be with reference to (2001) MolecularCloning:a laboratory manual such as Sambrook, the third edition, Cold Spring Harbor LaboratoryPress, CSH, New York or with reference to Current Protocols in Molecular Biology, John Wiley﹠amp; Sons, N.Y. (1989 and annual upgrade version).

Splice variant

Term as used in this article " splice variant " comprise wherein excise, replace, be shifted or add selected intron and/or exon or wherein intron shortened or the variant of the nucleotide sequence that extends.This type of variant will be a kind of variant that has wherein kept proteinic biologic activity basically; This can realize by the proteinic functional fragment of selective retention.This type of splice variant can find or can manually make at occurring in nature.Be used to predict that with the method for separating this type of splice variant be (seeing for example Foissac and Schiex (2005), BMC Bioinformatics.6:25) well-known in the art.

Allelic variant

Allelotrope or allelic variant are the alternative forms of given gene, are positioned at identical chromosome position.Allelic variant comprises single nucleotide polymorphism (SNP) and little insertion/deletion polymorphism (INDEL).The size of INDEL is usually less than 100bp.SNP and INDEL are formed on the maximum set of sequence variants in the most of biological natural generation sexual polymorphism strain system.

Gene reorganization/orthogenesis

Consisting of of gene reorganization or orthogenesis: DNA reorganization repeatedly, suitably screening and/or selection have the proteinic nucleic acid of improvement biologic activity or variant (Castle etc., (2004) Science 304 (5674): 1151-4 of its part to produce coding subsequently; United States Patent (USP) 5,811,238 and 6,395,547).

Regulatory element/regulating and controlling sequence/promotor

Term " regulatory element ", " regulating and controlling sequence " and " promotor " all are used interchangeably and mean in a broad sense the modulability nucleotide sequence that can realize that the sequence that is attached thereto is expressed in this article.Term " promotor " refer generally to be positioned at genetic transcription starting point upstream and participate in identification and in conjunction with RNA polymerase and other protein, thereby instruct the nucleic acid regulating and controlling sequence of the transcribed nucleic acid that effectively connects.Aforementioned term comprises from typical eukaryotic gene group gene and (comprising for the required TATA frame of accurate transcripting starting, have or do not have the CCAAT box sequence) in deutero-transcriptional regulatory sequences and replying grow stimulation and/or outside stimulus or with the tissue specificity mode change genetic expression the additional adjustment element (as, upstream activating sequence, enhanser and silencer).This term also comprises the transcriptional regulatory sequences of typical prokaryotic gene, and it can comprise-35 frame sequences and/or-10 frame transcriptional regulatory sequences in the case.Term " regulatory element " also comprises gives, activates or strengthen synthetic fusion molecule or the derivative that nucleic acid molecule is expressed in cell, tissue or organ.

" plant promoter " comprises the regulatory element that mediation encoding sequence section is expressed in vegetable cell.Therefore, plant promoter needs not be plant origin, but can be derived from virus or microorganism, for example from the virus of invasion and attack vegetable cell." plant promoter " also can plant-derived cell, for example comes to use by oneself to treat the nucleotide sequence institute plant transformed expressing and describe in this article in the inventive method.This also is applicable to other " plant " modulability signals, as " plant " terminator.The promotor upstream that is used for the nucleotide sequence of the inventive method can be replaced, be inserted and/or disappearance and being modified by one or more Nucleotide, but do not disturb promotor, open reading-frame (ORF) (ORF) or 3 ' regulatory region such as terminator or functional or active away from other 3 ' regulatory regions of ORF.The activity of promotor also might be because of the sequence of modifying this promotor or by more active promotor even thoroughly replace this promotor from the promotor of allos biology and increase.For expressing in plant, as mentioned above, nucleic acid molecule must effectively be connected to or comprise suitable promotor, and wherein said promotor is on orthochronous point and with needed space expression pattern expressing gene.

For identifying functional equivalent promotor, the promotor intensity of candidate's promotor and/or expression pattern can be by effectively being connected this promotor with reporter gene and analyzing this report gene and analyze in the expression level and the pattern of the multiple tissue of plant.Suitable known reporter gene comprises for example β-glucuronidase or beta-galactosidase enzymes.Promoter activity is analyzed by the enzymic activity of measuring β-glucuronidase or beta-galactosidase enzymes.Promotor intensity and/or expression pattern can compare with the promotor intensity and/or the expression pattern of reference promotor (as a kind of promotor used in the inventive method) subsequently.Alternatively, promotor intensity can be used the densitometric analysis method of means known in the art such as Northern blotting and autoradiogram(ARGM), quantitative PCR in real time or RT-PCR (Heid etc., 1996GenomeMethods 6:986-994), by quantitative mRNA or by the mRNA level of used nucleic acid in the inventive method and the mRNA level comparison of housekeeping gene (as 18S rRNA) are analyzed.Usually " weak promoter " means and drives encoding sequence expression promoter on low-level." low-level " means at about 1/10,000 transcript of each cell to about 1/100,000 transcript, to the level of about 1/500,0000 transcript.On the contrary, " strong promoter " drive encoding sequence high level or at about 1/10 transcript of each cell to about 1/100 transcript, to about 1/1,000 transcript, express.

Effectively connect

Term as used in this article " effectively connect " refer to functionally be connected between promoter sequence and the goal gene, to such an extent as to can starting goal gene, promoter sequence transcribes.

Constitutive promoter

" constitutive promoter " refers in the major part of g and D but all during the stage and in the promotor that transcriptional activity is arranged at least one cell, tissue or organ under most of envrionment conditions.Following table 4a provides the example of constitutive promoter.

Table 4a: the example of constitutive promoter

Gene source	Reference
		Actin muscle	McElroy etc., Plant Cell, 2:163-171,1990
HMGP	WO 2004/070039
		CAMV 35S	Odell etc., Nature, 313:810-812,1985
CaMV 19S	Nilsson etc., Physiol.Plant.100:456-462,1997
		GOS2	De Pater etc., Plant J Nov; 2 (6): 837-44,1992, WO 2004/065596
Ubiquitin	Christensen etc., Plant Mol.Biol.18:675-689,1992
		The rice cyclophilin	Buchholz etc., Plant Mol Biol.25 (5): 837-43,1994
Corn H3 histone	Lepetit etc., Mol.Gen.Genet.231:276-285,1992

Clover H3 histone	Plant Mol.Biol.11:641-649 such as Wu, 1988
		Actin muscle 2	An etc., Plant are (1) J.10; 107-121,1996
34S FMV	Sanger etc., Plant.Mol.Biol., 14,1990:433-443
		The Rubisco small subunit	US 4,962,028
OCS	Leisner (1988) the Proc Natl Acad Sci U.S. 85 (5): 2553
		SAD1	Jain etc., Crop Science, 39 (6), 1999:1696
SAD2	Jain etc., Crop Science, 39 (6), 1999:1696
		nos	Shaw etc. (1984) Nucleic Acids Res.12 (20): 7831-7846
The V-ATP enzyme	WO 01/14572
		Super promotor	WO 95/14098
G box protein matter	WO 94/12015

The omnipresence promotor

Institute is in a organized way or activity arranged in the cell basically at biology for the omnipresence promotor.

Grow the modulability promotor

Grow the modulability promotor and during certain growth period or in experience is grown the plant part that changes activity is being arranged.

Inducible promoter

(summary is seen Gatz 1997 to inducible promoter replying chemical, Annu.Rev.PlantPhysiol.Plant Mol.Biol., 48:89-108), the transcripting starting that has induced or increase when environmental stimulus or physical stimulation, maybe can be " stress-inducing ", promptly when being exposed to multiple stress conditions, plant activated, or " pathogen-inducible ", promptly when being exposed to multiple pathogenic agent, plant activated.

Organ specificity/tissue-specific promoter

Organ specificity or tissue-specific promoter can be preferentially start the promotor of transcribing in some organ or tissue such as leaf, root, seed tissue etc.For example, " root-specific promoter " is that advantage ground has the promotor of transcriptional activity in roots of plants, and essentially no activity in any other part of plant is although allow any leakage to express in these other parts of plant.Can only in some cell, start the promotor of transcribing and be called " cell-specific " in this article.

Seed specific promoters is transcriptional activation in seed tissue mainly, but needn't only activate (under the situation of leakage expression) in seed tissue.Seed specific promoters can activate in seed development and/or duration of germination.Seed specific promoters can be endosperm, aleuron, embryo-specific.The example of seed specific promoters is shown in following table 4b-4c.Other examples of seed specific promoters are provided by QingQu and Takaiwa (Plant Biotechnol.J.2,113-125,2004), the document complete being incorporated herein by reference disclosed.

Table 4b: the example of seed specific promoters

Table 4c: the example of endosperm specificity promoter

Gene source	Reference
		Gluten (rice)	Takaiwa etc., (1986) Mol Gen Genet 208:15-22 Takaiwa etc., (1987) FEBS Letts.221:43-47
Zein	Matzke etc., (1990) Plant Mol Biol 14 (3): 323-32
		Wheat LMW and HMW glutenin-1	Colot etc., (1989) Mol Gen Genet 216:81-90 Anderson etc., (1989) NAR 17:461-2
Wheat SPA	Albani etc., (1997) Plant Cell 9:171-184
		The wheat gliadine	Rafalski etc., (1984) EMBO 3:1409-15
Barley Itr1 promotor	Diaz etc., (1995) Mol Gen Genet 248 (5): 592-8
		Barley B1, C, D, hordein	Cho etc., (1999) Theor Appl Genet 98:1253-62 Muller etc., (1993) Plant J 4:343-55 Sorenson etc., (1996) Mol Gen Genet 250:750-60
Barley DOF	Mena etc., (1998) Plant J 116 (1): 53-62

blz2	Onate etc., (1999) J Biol Chem 274 (14): 9175-82
		Synthetic promoter	Vicente-Carbajosa etc., (1998) Plant J 13:629-640
Paddy prolamine NRP33	Wu etc., (1998) Plant Cell Physiol 39 (8) 885-889
		Rice Lysozyme lb-1	Wu etc., (1998) Plant Cell Physiol 39 (8) 885-889
Rice sphaeroprotein REB/OHP-1	Nakase etc., (1997) Plant Molec Biol 33:513-522
		Rice ADP-glucose pyrophosphorylase	Russell etc., (1997) Trans Res 6:157-68
Corn ESR gene family	Opsahl-Ferstad etc., (1997) Plant J 12:235-46
		Kafirin	DeRose etc., (1996) Plant Mol Biol 32:1029-35

Table 4d: the example of embryo-specific promoter

Gene source	Reference
		Rice OSH1	Sato etc., the Proc.Natl.Acad.Sci. U.S., 93:8117-8122,1996
KNOX	Postma-Haarsma etc., Plant Mol.Biol.39:257-71,1999
		PRO0151	WO 2004/070039
PRO0175	WO 2004/070039
		PRO005	WO 2004/070039
PRO0095	WO 2004/070039

Table 4e: the example of aleuron specificity promoter

Gene source	Reference
		α-Dian Fenmei (Amy32b)	Lanahan etc., Plant Cell 4:203-211,1992; Skriver etc., Proc Natl Acad Sci U.S. 88:7266-7270,1991
Kethepsin β-sample gene	Cejudo etc., Plant Mol Biol 20:849-856,1992
		Barley Ltp2	Kalla etc., Plant J.6:849-60,1994
Chi26	Leah etc., Plant J.4:579-89,1994
		Corn B-Peru	Selinger etc., Genetics 149; 1125-38,1998

Chlorenchyma specificity promoter as defined herein is mainly to have the promotor of transcriptional activity in chlorenchyma, and essentially no activity in any other part of plant is although allow any leakage to express in these other parts of plant.

Another example of tissue-specific promoter is the meristematic tissue specificity promoter, it mainly has transcriptional activity in the merism tissue, essentially no activity in any other part of plant is although allow any leakage to express in these other parts of plant.

Terminator

Term " terminator " comprise such regulating and controlling sequence, it is the dna sequence dna at transcription unit's end, sends primary transcript is carried out the signal that 3 ' processing and poly-adenosine and termination are transcribed.Terminator can be derived from natural gene, from multiple other plant gene or from T-DNA.Terminator to be added can be from for example nopaline synthase or octopine synthase gene or alternatively from the other plant gene or more preferably from any other eukaryotic gene.

Regulate

Term " adjusting " with regard to expression or genetic expression, mean such process, wherein expression level is compared with control plant because of described expression of gene changes, and expression level can increase or reduce.Any kind that original expression of being regulated can be structure RNA (rRNA, tRNA) or mRNA is expressed, and is translation subsequently.Term " adjusting is active " should mean any variation of nucleotide sequence of the present invention or coded protein expression, and this causes the output of plant increase and/or the growth of increase.

Express

Term " expression " or " genetic expression " mean transcribing of one or more genes of Yin Teding or gene construct.Term " expression " or " genetic expression " especially mean one or more genes or gene construct is transcribed into structure RNA (rRNA, tRNA) or mRNA, and being with or without subsequently, mRNA translates into protein.This process comprises that DNA transcribes, processes the mRNA product that obtains.

Expression/the overexpression that increases

Term as used in this article " expression that increases " or " overexpression " to mean for original wild-type expression level be extra any formal representation.

In this area write up be used to increase the method for gene or gene product expression and they for example comprise, by the overexpression of suitable promoters driven, use transcriptional enhancer or translational enhancer.Can in the suitable location (generally being the upstream) of the polynucleotide of non-allos form, introduce isolating nucleic acid, so that go up the expression of nucleic acids of tone coded desired polypeptides as promotor or enhancer element.For example, the endogenous promotor can and/or replace and changes in vivo and (see Kmiec, the U.S. 5,565,350 by sudden change, disappearance; Zarling etc., WO9322443), maybe can be with isolating promotor with respect to the correct direction of gene of the present invention and apart from the introduced plant cell, so that controlling gene is expressed.

If need expression of polypeptides, wish that usually 3 ' end in the polynucleotide encoding district comprises the poly-adenosine district.The poly-adenosine district can be from natural gene, from multiple other plant gene or from T-DNA.3 ' end sequence to be added can be from for example nopaline synthase or octopine synthase gene or alternatively from another plant gene or more not preferably from any other eukaryotic gene.

Intron sequences also can be added on the encoding sequence of 5 ' non-translational region (UTR) or part coding property sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.But verified montage intron being included in the transcription unit in expression of plants construct and animal expression construct increases genetic expression to reaching 1000 times of (Buchman and Berg (1988) Mol.Cell biol.8:4395-4405 on mRNA level and the protein level; Callis etc. (1987) Gens Dev 1:1183-1200).This type of intron enhancement of genetic expression is the strongest generally near being positioned at transcription unit 5 ' end the time.It is known in the art using corn intron A dh1- S introne 1,2 and 6, Bronze-1 intron.For general information, see: The Maize Handbook, the 116th chapter, editor Freeling and Walbot, Springer, N.Y. (1994).

Native gene

" endogenous " gene of mentioning herein not only refers to the gene of being discussed that exists with its natural form (promptly without any the mankind intervene) as in plant, and also refers to be in the homologous genes (or homologous nucleic acid/gene) basically of (again) the subsequently introduced plant (transgenosis) of unpack format.For example, contain this genetically modified transgenic plant and can meet with the significantly reduction that transgene expression significantly reduces and/or native gene is expressed.Isolating gene is separable from organism, or synthetical, for example by chemosynthesis.

The expression that reduces

" expression of reduction " mentioned herein or " the reducing or basic the removal " of expressing mean native gene expression and/or polypeptide level and/or the polypeptide active reduction with respect to control plant.Compare with control plant, reducing or removing to increase progressively preferred sequence substantially is at least 10%, 20%, 30%, 40% or 50%, 60%, 70%, 80%, 85%, 90%, or 95%, 96%, 97%, 98%, 99% or more reduction.

In order to reduce or to remove the expression of native gene in plant substantially, need the Nucleotide of successive basically of the sufficient length of nucleotide sequence.In order to carry out gene silencing, this length can be few to 20,19,18,17,16,15,14,13,12,11,10 or still less Nucleotide, and perhaps this length can the whole gene of as many as (comprising 5 ' and/or 3 ' UTR, in part or total length).Basically the successive nucleotide fragments can come the own coding target protein nucleic acid (target gene) or from the target protein of can encoding directly to any nucleic acid of homologue, collateral line homologue or homologue.Preferably, basically the successive nucleotide fragments can form hydrogen bond with target gene (sense strand or antisense strand), more preferably, the successive nucleotide fragments has 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity to increase progressively preferred sequence and target gene (sense strand or antisense strand) basically.The nucleotide sequence of coding (functional) polypeptide is not that discussed herein to be used to reduce or to remove substantially the several different methods that native gene expresses required.

This reduction of expressing or basic removal can use conventional instrument and technology to finish.Being used for reducing or removing substantially the preferred method that native gene expresses is to introduce and express such genetic constructs plant, its amplifying nucleic acid (be from goal gene or any nucleic acid one section successive nucleotide sequence basically in the case, wherein said any nucleic acid can encode any target protein directly to homologue, collateral line homologue or homologue) be cloned in the described genetic constructs as (partially or completely) inverted repeats that separates by transcribed spacer (non-coding DNA).

In this preferable methods, use nucleic acid or its part (be in the case from goal gene or from any nucleic acid one section of deutero-successive nucleotide sequence basically, wherein said any nucleic acid can encode target protein directly to homologue, collateral line homologue or homologue) inverted repeats (preferably can form hairpin structure), the expression that the silence effect by RNA mediation reduces or remove basically native gene.Inverted repeats is cloned in comprising the expression vector of regulating and controlling sequence.Non-coding DNA nucleotide sequence (intervening sequence, for example matrix attachment regions fragment (MAR), intron, polylinker etc.) is between two reverse nucleic acid that form inverted repeats.After inverted repeats is transcribed, form chimeric RNA with (partially or completely) self complementary structure.This double-stranded RNA structure is called hairpin RNA (hpRNA).HpRNA is processed into siRNA by plant, and it is impregnated in the reticent mixture of RNA inducibility (RISC).RISC further cuts the mRNA transcript, thereby reduces the number of the mRNA transcript of one-tenth polypeptide to be translated significantly.For other general details, see for example (1998) WO 98/53083 such as Grierson; Waterhouse etc. (1999) WO 99/53050).

The enforcement of the inventive method does not rely in the plant to introduce and to express and wherein is cloned into the genetic constructs of nucleic acid as inverted repeats, but several known " gene silencing " method any or multiplely can be used for realizing identical effect.

It is a kind of that to be used to reduce these class methods that native gene expresses are genetic expression silences (downward modulation) of RNA mediation.Silence acts in this case and is triggered in plant by similar to the endogenous target gene basically double-stranded RNA sequence (dsRNA).This dsRNA is arrived about 26 Nucleotide by the further processing of plant into about 20, is called short interferential RNA (siRNA).SiRNA is impregnated in the reticent mixture of RNA inducibility (RISC), and wherein said RISC further cuts the mRNA transcript of endogenous target gene, thereby reduces the number of the mRNA transcript of one-tenth polypeptide to be translated significantly.Preferably, the double-stranded RNA sequence is corresponding to target gene.

Another example of RNA silencing methods comprise with sense orientation introduce nucleotide sequence or its part (be in the case from goal gene or from any nucleic acid one section of deutero-successive Nucleotide basically, wherein said any nucleic acid can encode target protein directly to homologue, collateral line homologue or homologue) to plant." sense orientation " refers to and its mRNA transcript homologous dna sequence dna.Thereby will in plant, introduce at least one copy of this nucleotide sequence.This extra nucleotide sequence can reduce native gene expresses, and produces and is known as inhibiting altogether phenomenon.When the several additional copy introduced plant of nucleotide sequence, the reduction of genetic expression will be more obvious, because have positive correlation between the inhibiting together triggering of high transcript level.

Another example of RNA silencing methods comprises the use anti sense nucleotide sequence." antisense " nucleotide sequence comprises " justice is arranged " nucleic acid array complementation with coded protein, promptly with the coding strand complementation of double-stranded cDNA molecule, or with mRNA transcript sequence complementary nucleotide sequence.Anti sense nucleotide sequence preferably with treat reticent native gene complementation.Complementary " coding region " that can be positioned at gene and/or " non-coding region ".Term " coding region " refers to comprise the nucleotide sequence district of the codon that is translated into amino-acid residue.Term " non-coding region " refers to be distributed in being transcribed but not translating into amino acid whose 5 ' and 3 ' sequence (be also referred to as 5 ' and 3 ' non-translational region) of both sides, coding region.

Anti sense nucleotide sequence can be according to Watson and the design of Crick base pairing rules.Anti sense nucleotide sequence can with whole nucleic acid array complementation (be in the case from goal gene or from any nucleic acid one section of deutero-successive Nucleotide basically, wherein said any nucleic acid can encode target protein directly to homologue, collateral line homologue or homologue), but also can be only with the oligonucleotide of a part (comprising mRNA 5 ' and the 3 ' UTR) antisense of nucleotide sequence.For example, Antisensedigonucleotsequence sequence can with the regional complementarity around the translation starting point of the mRNA transcript of coded polypeptide.The length of suitable Antisensedigonucleotsequence sequence is known in the art and can be from about 50,45,40,35,30,25,20,15 or 10 Nucleotide of length or Nucleotide still less.Anti sense nucleotide sequence of the present invention can utilize means known in the art, uses chemosynthesis and enzyme ligation and makes up.For example, anti sense nucleotide sequence (for example Antisensedigonucleotsequence sequence) can use the Nucleotide of natural generation or the Nucleotide of multiple modification to synthesize chemically, the Nucleotide of wherein said modification is designed to be intended to increase the biological stability of molecule or increases anti sense nucleotide sequence and the physical stability of the duplex that forms between the phosphorothioate odn sequence is arranged, for example, the Nucleotide that can use phosphorothioate derivative and acridine to replace.The example that can be used for producing the modified nucleotide of anti sense nucleotide sequence is well-known in the art.Known nucleotide modification comprise methylate, cyclisation and ' add cap ' and replace the Nucleotide of one or more natural generations with analogue (as inosine).Other nucleotide modification is well-known in the art.

This anti sense nucleotide sequence can use nucleotide sequence wherein with antisense orientation in addition the expression vector of subclone (promptly the RNA that transcribes from the nucleic acid that inserts will be antisense orientation with the purpose target nucleic acid) produce in the biology mode.Preferably, the generation of anti sense nucleotide sequence in plant undertaken by the nucleic acid construct of stable integration, antisense oligonucleotide and terminator that wherein said nucleic acid construct comprises promotor, effectively connects.

The nucleic acid molecule (no matter introduce in plant or produce in position) that is used for the reticent effect of the inventive method is with the mRNA transcript and/or the genomic dna hybridization of coded polypeptide or combine, so that for example by suppressing to transcribe and/or translation and the expression of arrestin matter.Hybridization can be passed through to form due to the conventional Nucleotide complementarity of stablizing duplex, or under the situation of the anti sense nucleotide sequence that is incorporated into DNA duplex, due to the interaction of duplex major groove internal specific.Anti sense nucleotide sequence can be by transforming or at particular organization's position direct injection and introduced plant.Alternatively, anti sense nucleotide sequence can be modified for the selected cell of target and systemic administration subsequently.For example, for systemic administration, anti sense nucleotide sequence can be modified so that their specific combination are expressed in acceptor or the antigen on the selected cell surface, for example by connect anti sense nucleotide sequence to cell surface receptor or antigen bonded peptide or antibody.Anti sense nucleotide sequence also can use described carrier to send herein and pass to cell.

According to another aspect, anti sense nucleotide sequence is a α-different nucleotide sequence.Different nucleotide sequence of α and complementary RNA form specific double-stranded hybrid molecule, and be wherein opposite with usual b-unit, described chain be parallel to each other (Gaultier etc. (1987) Nucl Ac Res 15:6625-6641).Anti sense nucleotide sequence also can comprise 2 '-the o-methyl ribonucleotides (Inoue etc. (1987) Nucl Ac Res 15,6131-6148) or chimeric RNA-DNA analogue (Inoue etc. (1987) FEBS Lett.215,327-330).

Reduction that native gene is expressed or basic removal also can be used ribozyme and carry out.Ribozyme is the catalytic RNA molecule with ribonuclease activity, can cut the single-chain nucleic acid sequence that has complementary region with it, as mRNA.Therefore, (for example hammerhead ribozyme is (at Haselhoff and Gerlach (1988) Nature 334 for ribozyme, describe among the 585-591) can be used for the mRNA transcript of catalytic ground cutting coded polypeptide, thereby reduce the number of the mRNA transcript of one-tenth polypeptide to be translated significantly.Can design the specific ribozyme of nucleotide sequence tool (is for example seen: U.S. Patent numbers such as Cech 4,987,071; With U.S. Patent numbers 5,116,742 such as Cech).Alternatively, corresponding to the mRNA transcript of nucleotide sequence can be used for from the RNA library of molecules, selecting catalytic RNA with specific ribonucleic acid enzymic activity (Bartel and Szostak (1993) Science 261,1411-1418).The purposes that ribozyme is used for the plant gene silencing is ((1994) WO94/00012 such as Atkins for example known in the art; Lenne etc. (1995) WO 95/03404; Lutziger etc. (2000) WO 00/00619; (1997) WO 97/38116 such as Prinsen etc. (1997) WO 97/13865 and Scott).

Gene silencing also can be by inserting mutagenesis (for example T-DNA inserts or transposon inserts) or by ((1999) Plant is (3) J.20: 357-62), (Amplicon VIGSWO 98/36083) or Baulcombe (WO 99/15682) and other people strategy of description realize as Angell and Baulcombe.

When having sudden change on having sudden change and/or the isolating gene/nucleic acid introduced plant subsequently on the native gene, gene silencing also can take place.Reduction or basic removal can be caused by non-functional polypeptide.For example, polypeptide can with multiple interaction protein bound; One or more sudden changes and/or brachymemma thereby can provide still can binding interactions protein (as receptor protein) but can not show the polypeptide (as playing the part of signal effect) of its normal function.

The method of another kind of gene silencing is the triple-helix structure that target is fixed and generegulation district (for example promotor and/or enhanser) complementary nucleotide sequence stops gene to be transcribed in target cell with formation.See Helene, C., Anticancer Drug Res.6,569-84,1991; Helene etc., Ann.N.Y.Acad.Sci.660,27-361992 and Maher, L.J.Bioassays 14,807-15,1992.

Additive method, as using at the antibody of endogenous polypeptide suppressing the function of this polypeptide in plant, or the signal pathway that disturbs described polypeptide to participate in, will be well-known for the technician.Especially, what can conceive is the biological function that artificial molecule can be used to suppress the target polypeptide, or is used to the signal pathway that disturbs the target polypeptide to participate.

Alternatively, can set up screening procedure to identify the natural variant of gene in plant population, wherein said variant coding has the active polypeptide of reduction.This type of natural variant also can be used for for example carrying out homologous recombination.

Artificial and/or natural microRNA (miRNA) can be used for knocking out genetic expression and/or mRNA translation.Endogenous miRNA is the little RNA of strand of a common 19-24 length of nucleotides.Their major function is that regulatory gene is expressed and/or the mRNA translation.Most plant micrornas (miRNA) has completely with its target sequence or is approaching complementary completely.Yet, exist to have the nearly natural target of 5 mispairing.They by the double-stranded specific RNA enzyme of cutting enzyme family from having the characteristic processing the long non-coding RNA of structure of turning back.Adding man-hour, they are by mixing this complex body with the main component Argonaute protein bound of the reticent mixture of RNA inducibility (RISC).MiRNA serves as the specific component of RISC, so target nucleic acid (the being mRNA mostly) base pairing in they and the tenuigenin.Follow-up adjusting incident comprises the said target mrna cutting and destroys and/or the translation inhibition.Therefore the effect of miRNA overexpression obtains reflection on the mRNA level that target gene reduces.

The artificial microRNA (amiRNAs) of common 21 length of nucleotides can genetic modification with the negative genetic expression of regulating single or multiple goal gene specifically.The determinative of the selection of plant micrornas target is well-known in the art.The empirical parameter that is used for target identification has been determined and can be used for the specific amiRNA of aided design, (Schwab etc., Dev.Cell 8,517-527,2005).The convenient tool that is used to design and produce amiRNA and precursor thereof also is the public obtainable (Schwab etc., Plant Cell 18,1121-1133,2006).

Be optimum performance, be used for reducing gene silent technology that native gene expresses plant and need use from monocotyledonous nucleotide sequence with transforming monocots with use nucleotide sequence from dicotyledons to transform dicotyledons.Preferably, will introduce in the same species from the nucleotide sequence of any given plant species.For example, will be converted into rice plant from the nucleotide sequence of rice.Yet, be not absolute requirement nucleotide sequence to be introduced originate from this nucleotide sequence will the identical plant species of exotic plant.As long as exist sizable homology just enough between endogenous target gene and the nucleic acid to be introduced.

Above-described is the example that is used for reducing or removes substantially the several different methods that native gene expresses plant.To such an extent as to those skilled in the art can adjust easily and aforementionedly be used for reticent method for example by utilizing suitable promotor to realize to reduce native gene whole strain plant or in the expression of its part.

Selected marker (gene)/reporter gene

" selected marker ", " selected marker " or " reporter gene " comprise any gene from phenotype to cell that give, wherein at the described gene of described cell inner expression promote to identify and/or to select with nucleic acid construct institute's transfection of the present invention or cell transformed.These marker gene can be identified the successful transfer of nucleic acid molecule by a series of different principle.Suitable mark can be selected from the mark of giving antibiotic resistance or Herbicid resistant, the new metabolism proterties of introducing or allowing visual selection.Selected marker's example comprise the gene of giving antibiotic resistance (as make the nptII of Xin Meisu and kantlex phosphorylation or make the hpt of Totomycin phosphorylation or give to for example bleomycin, Streptomycin sulphate, tsiklomitsin, paraxin, penbritin, gentamicin, Geneticin (Geneticin) (G418), the gene of the resistance of spectinomycin or blasticidin), the gene of conferring herbicide resistance (for example provides

The bar of resistance; The aroA or the gox of glyphosate resistance be provided or give for example gene of the resistance of imidazolone, phosphinothricin or sulfourea) or provide the metabolism proterties gene (as allow plant use seminose as the manA of sole carbon source or utilize xylose isomerase or the anti-nutrition mark such as the 2-deoxyglucose resistance of wood sugar).The expression of visual marker gene causes forming color (for example β-glucuronidase, GUS or beta-galactosidase enzymes substrate coloured with it for example X-Gal), luminous (as luciferin/luciferase system) or fluorescence (green fluorescent protein GFP and derivative thereof).This list is only represented the possible mark of minority.The technician is familiar with this type of mark.Depend on biology and system of selection, preferred different mark.

Known to nucleic acid stability or integration,temporal during to vegetable cell, the cellular uptake foreign DNA of small portion and as required it is integrated into cellular genome only, this depends on the rotaring dyeing technology of used expression vector and use.For identifying and select these integrons, the gene of the selected marker of will encoding usually one of (as indicated above) is introduced host cell together with goal gene.These marks therein these genes because of using in the non-functional mutant of disappearance due to the ordinary method for example.In addition, the nucleic acid molecule of coding selected marker can be introduced in the host cell, with the sequence of used polypeptide in comprising code book invention polypeptide or the inventive method on identical carrier, or on independent carrier.Can be by having selected to identify (for example having the cell survival of selected marker of integration and other necrocytosiss) with the nucleic acid stability cells transfected of introducing.

Because in case successfully introduced nucleic acid, then no longer need in the genetically modified host cell or do not wish underlined gene, especially antibiotic resistance gene and herbicide resistance gene, the inventive method that therefore is used to introduce nucleic acid is advantageously used the technology that can remove or excise these marker gene.A kind ofly be called the cotransformation method as this method.The cotransformation method is used two kinds of carriers being used to simultaneously transform, and a kind of carrier carries nucleic acid of the present invention and another kind of carrier carries marker gene.A high proportion of transformant is accepted, or under the situation of plant, comprise (up to 40% or more transformant) these two kinds of carriers.Under situation about transforming with Agrobacterium (Agrobacterium), transformant is only accepted the part of carrier usually, and promptly flank has the sequence of T-DNA, and it represents expression cassette usually.Marker gene can be removed from plant transformed by hybridizing subsequently.In another approach, the marker gene that is integrated into transposon is used for transforming (being called the Ac/Ds technology) with the nucleic acid of wanting.Transformant can be instantaneous or stably transform with the nucleic acid construct that causes transposase to be expressed with originate plant hybridization or transformant of transposase.(about 10%) in some cases, transposon is jumped out the genome of host cell and is lost when successfully taking place to transform.Under other more susceptible conditions, transposon skips to different positions.In these cases, marker gene must be removed by hybridizing.In microbiology, developed the technology that realizes or promote to detect this class incident.Another advantageous method depends on recombination system; The advantage of this method is and needn't removes by hybridization.The most well-known system of the type is called the Cre/lox system.Cre1 is the recombinase that removes sequence between the loxP sequence.If marker gene is integrated between the loxP sequence, when then having expressed successfully generation conversion by recombinase, marker gene is removed.Other recombination systems are HIN/HIX, FLP/FRT and REP/STB system (Tribble etc., J.Biol.Chem., 275,2000:22255-22267; Velmurugan etc., J.Cell Biol., 149,2000:553-566).Nucleotide sequence of the present invention might be integrated into Plant Genome in the locus specificity mode.These methods also can be applied to microorganism such as yeast, fungi or bacterium naturally.

Genetically modified/transgenosis/reorganization

Be the object of the invention, " genetically modified ", " transgenosis " or " reorganization " biology of with regard to nucleotide sequence, meaning the expression cassette, gene construct or the carrier that comprise this nucleotide sequence or transforming with nucleotide sequence of the present invention, expression cassette or carrier, these make up all and produce by recombination method, wherein

(a) coding is used for the nucleic acid sequences to proteins of the inventive method, or

(b) the genetic regulation sequence that effectively is connected with nucleotide sequence of the present invention, promotor for example, or

(c) (a) and (b)

Be not in its natural genotypic environment or modify by genetic manipulation method, be modified with may for example adopt replace, interpolation, disappearance, inversion or insert the form of one or more nucleotide residues.Natural genotypic environment be interpreted as mean the source in the plant natural gene group locus or chromogene seat or in genomic library, exist.Under the situation of genomic library, the natural genotypic environment of nucleotide sequence preferably obtains keeping, and is kept at least in part.This environment is distributed at least one side of nucleotide sequence and has 50bp at least, preferred 500bp at least, especially preferred 1000bp at least, the most preferably sequence length of 5000bp at least.The combination of the natural generation of the corresponding nucleotide sequence of used polypeptide in the natural promoter of the nucleotide sequence of the expression cassette of natural generation-for example and the code book inventive method, as hereinbefore defined-when being subjected to modifying, become transgene expression cassette by non-natural synthetic (" manually ") method (as mutagenic treatment) at this expression cassette.Appropriate method is for example at US 5,565,350 or WO00/15815 in describe.

Be the object of the invention, therefore transgenic plant are interpreted as above and mean the natural gene seat that nucleic acid used in the inventive method is not arranged in described this nucleic acid of Plant Genome that described nucleic acid might homology or the expression of allos ground.Yet as mentioned, although transgenosis also means nucleic acid of the present invention or used in the methods of the invention nucleic acid is in the natural place of this nucleic acid in the Plant Genome, yet its sequence is modified for native sequences, and/or the adjusting sequence of described native sequences is modified.Transgenosis is interpreted as preferably to mean in the non-natural locus of nucleic acid of the present invention in genome and expresses that the homology that nucleic acid promptly takes place is expressed or preferred heterogenous expression.Preferred transgenic plant have been mentioned in this article.

Transform

Term " introducing " or " conversion " comprise that exogenous polynucleotide are transferred in the host cell as mentioned in this article, and what the method that no matter is used to transform is.Can follow-up clone's property propagation the plant tissue of (no matter take place or the embryo is taken place) by organ can transform and the whole strain plant that can therefrom regenerate with genetic constructs of the present invention.The concrete tissue of selecting will depend on clone's property proliferating system of the concrete species that can be used for and be suitable for just transforming most.The example organization target comprises leaf dish, pollen, embryo, cotyledon, hypocotyl, megagametophyte, callus, existing meristematic tissue (for example apical meristem, axillalry bud and root meristematic tissue) and inductive meristematic tissue (for example cotyledon meristematic tissue and hypocotyl meristematic tissue).Polynucleotide can instantaneous or stably be introduced host cell and can keep to nonconformity, for example as plasmid.Alternatively, polynucleotide can be integrated in the host genome.The transformed plant cells that produces can be used for regenerating in the manner known to persons skilled in the art the conversion plant subsequently.

Alien gene is converted into and is called conversion in the Plant Genome.The conversion of plant species is quite conventional technology now.Advantageously, the arbitrary method in several method for transformation can be used for goal gene is introduced suitable ancester cell.Be used for transforming and the described method of the plant that regenerates can be used for instantaneous conversion or be used for stable conversion from plant tissue or vegetable cell.Method for transformation comprises that the chemical, the dna direct that use liposome, electroporation, increase dissociative DNA to take in are injected to the conversion method and the micro-projective method (microprojection) of plant, particle gun blast technique, use virus or pollen.Method for transformation can be selected from calcium/polyoxyethylene glycol method (Krens, F.A. etc., (1982) Nature296, the 72-74 that is used for protoplastis; (1987) Plant Mol Biol 8:363-373 such as Negrutiu I); The electroporation of protoplastis ((1985) Bio/Technol 3 such as Shillito R.D., 1099-1102); Micro-injection (Crossway A etc., (1986) Mol.Gen Genet 202:179-185) to vegetable material; The particle bombardment method (Klein TM etc., (1987) Nature 327:70) of DNA or RNA coating, (nonconformity) virus infection method etc.Transgenic plant comprise the genetically modified crops plant, preferably produce by agriculture bacillus mediated conversion method.Favourable method for transformation is the conversion method of in plant (in planta).For this purpose, for example might make Agrobacterium act on the meristematic tissue that plant seed maybe might be inoculated plant with Agrobacterium.To act on complete plant or act on flower primordium at least be particularly advantageous to the verified Agrobacterium suspension that makes conversion according to the present invention.Plant continues subsequently to cultivate that (Clough and Bent, Plant J. (1998) 16,735-743) until the seed that obtains the plant of handling.Be used for method that agriculture bacillus mediated rice transforms and comprise and be used for the known method that rice transforms, as those methods of in arbitrary following document, describing: European patent application EP 1198985 A1, Aldemita and Hodges (Planta199:612-617,1996); Chan etc. (Plant Mol Biol 22 (3): 491-506,1993), Hiei etc. (Plant J 6 (2): 271-282,1994), its disclosure is incorporated herein by reference in this article, as providing fully.Under the situation that corn transforms, (Nat.Biotechnol 14 (6): 745-50 for preferable methods such as Ishida etc., 1996) or Frame etc. (Plant Physiol 129 (1): 13-22,2002) describe, its disclosure is incorporated herein by reference as fully in this article.Described method by way of example mode further by B.Jenes etc., Techniques for Gene Transfer,: Transgenic Plants, the 1st volume, Engineering and Utilization, editor S.D.Kung and R.Wu, Academic Press (1993) 128-143 and at Potrykus Annu.Rev.PlantPhysiol.Plant Molec.Biol.42 (1991) 205-225) in the description.Nucleic acid to be expressed or construct preferably are cloned into the carrier that is suitable for transforming agrobacterium tumefaciens (Agrobacterium tumefaciens), for example pBin19 (Bevan etc., Nucl.Acids Res.12 (1984) 8711).The Agrobacterium that is transformed by this carrier can be used to transform plant according to known way subsequently, the plant of using for example as model, (Arabidopsis is in scope of the present invention as Arabidopis thaliana, be not considered as crop plants) or crop plants, for example tobacco plant is also cultivated them subsequently by the leaf that soaks abrasive leaf or chopping in Agrobacterium solution in suitable medium.The conversion of plant by agrobacterium tumefaciens for example by

With Willmitzer at Nucl.Acid Res. (1988) 16, Vectors for Gene Transfer in Higher Plants is described in 9877 or especially from F.F.White; At Transgenic Plants, the 1st volume, Engineering and Utilization, editor S.D.Kung and R.Wu, AcademicPress is known in 1993, the 15-38 pages or leaves.

Except transformant cell (its necessary subsequently complete plant of regeneration), also might transform the merismatic cell of plant and special those cells that develop into gamete that transform.In this case, the gamete of conversion is followed natural development of plants process, produces transgenic plant.Therefore, for example the Arabidopis thaliana seed is handled with Agrobacterium and obtain seed from is grown plant, and wherein a certain proportion of described plant is transformed and is genetically modified [Feldman, KA and Marks MD (1987) Mol GenGenet 208:274-289 therefore; Feldmann K (1992).: editor C Koncz, N-H Chua and J Shell, Methods in Arabidopsis Research.Word Scientific, Singapore, 274-289 page or leaf].Alternative method based on remove inflorescence repeatedly and make in the rosette in the heart the excision position and the Agrobacterium incubation of conversion, thereby the seed that transforms can obtain at later time point equally, and (Chang (1994) Plant is J.5:551-558; Katavic (1994) Mol Gen Genet, 245:363-370).Yet especially effective means is the vacuum infiltration method of improvement, as " flower is contaminated " method.Under the situation of Arabidopis thaliana vacuum infiltration method, complete plant is under reduced pressure handled [Bechthold, N (1993) with the Agrobacterium suspension.C R Acad Sci Paris Life Sci, 316:1194-1199], and under the situation of " flower dip method ", of short duration incubation [the Clough of Agrobacterium suspension that the flower tissue and the tensio-active agent of growing handled, SJ and Bent, AF (1998) The Plant J.16,735-743].Gathered in the crops a certain proportion of transgenic seed in both cases, and these seeds can be distinguished by under aforesaid selection condition, cultivating with the non-transgenic seed.In addition, the stable conversion of plastid is favourable because plastid in most of crop with the heredity of parent mode, reduce or eliminated transgenosis through the pollen flow risk.The conversion of chloroplast gene group generally by at Klaus etc., 2004[Nature Biotechnology 22 (2), 225-229] in the exemplary method realization of being showed.In brief, sequence to be transformed be cloned into together with the selected marker and chloroplast gene group homologous flanking sequence between.These homologous flanking sequences instruct locus specificity to be integrated in the plastom(e).Numerous different plant species having been described plastid transforms and summarizes and can come from Bock (2001) transgenosis plastid (Transgenicplastids in basic research and plant biotechnology) .J Mol Biol.2001 September 21 in fundamental research and Plant Biotechnology; 312 (3): 425-38 or Maliga, P (2003) plastid transformation technology commercialization progress (Progress towards commercialization of plastid transformationtechnology) .Trends Biotechnol.21,20-28.Further the biotechnology progress has been made report with the form of unmarked plastid transformant recently, described unmarked plastid transformant can produce (Klaus etc. by the instantaneous marker gene of integrating altogether, 2004, Nature Biotechnology 22 (2), 225-229).

T-DNA activates mark

T-DNA activates mark (Hayashi etc., Science (1992) 1350-1353) relates in the genome area of goal gene or gene coding region upstream or downstream 10kb sentence structure like this and insert T-DNA (containing promotor (also can be translational enhancer or intron) usually), make promotor instruct and decided expression of gene by target.Usually, the promotor control that the natural promoter of deciding gene by target regulating effect that described target is decided genetic expression is destroyed and this gene is in new introducing down.Promotor generally is embedded among the T-DNA.This T-DNA inserts Plant Genome randomly, for example by agroinfection, and causes near the improvement of the gene insertion T-DNA to be expressed.Cause is expressed near the improvement of the gene of the promotor of introducing, the transgenic plant performance dominant phenotype of generation.

TILLING

Term " TILLING " is that the abbreviation of " genome interior orientation inductive local damage " and meaning is used for producing and/or identifying the nucleic acid induced-mutation technique, and wherein said nucleic acid encoding has to modify expresses and/or active protein.TILLING also allows to select to carry the plant of this type of mutation variants.These mutation variants may be displayed on the intensity aspect or aspect the position or in the expression (if for example sudden change influence promotor) of improvement aspect the time.These mutation variants can show than showed active higher activity by the gene that is in its natural form.TILLING is with high-density mutagenesis and high-throughput screening method combination.The general step of following in TILLING is: (Redei GP and Koncz C (1992) are at Methods in Arabidopsis Research in (a) EMS mutagenesis, Koncz C, Chua NH, Schell J, Singapore writes, World Scientific Publishing Co, the 16-82 page or leaf; Feldmann etc., at Meyerowitz EM, Somerville CR writes (1994), Arabidopsis.ColdSpring Harbor Laboratory Press, Cold Spring Harbor, NY, 137-172 page or leaf; Lightner J and Caspar T (1998) be at J Martinez-Zapater, J Salinas editor, Methods on Molecular Biology the 82nd volume .Humana Press, Totowa, NJ, 91-104 page or leaf); (b) DNA of individual prepares and compiles; (c) pcr amplification purpose district; (d) sex change and renaturation are to allow to form heteroduplex; (e) DHPLC, wherein with heteroduplex whether the existence in compiling thing detect and be an extra peak in the color atlas; (f) identify mutated individual; (g) to the order-checking of sudden change PCR product.The method that is used for TILLING is (McCallum etc., (2000) Nat Biotechnol 18:455-457 well-known in the art; Summary is seen Stemple (2004) Nat Rev Genet5 (2): 145-50).

Homologous recombination

The nucleic acid that homologous recombination allows to select is introduced in the genome on the selected location of determining.Homologous recombination is the standard technique that is used for unicellular lower eukaryote such as yeast or liver moss sword-like leave moss (Physcomitrella) in bio-science routinely.The method that is used for carrying out homologous recombination plant is not only to model plant (Offringa etc. (1990) EMBO J 9 (10): 3077-84) but also to crop plants rice (Terada etc. (2002) Nat Biotech 20 (10): 1030-4 for example; Iida and Terada (2004) Curr Opin Biotech 15 (2): 132-8) be described.

Output

Term " output " but mean the measuring result of economic worth usually, general with specify crop, and area and relevant with the time period.Single plant part based on they number, size and/or weight and directly output is had contribution, or actual output is every square metre of output for certain crop and 1 year, and this determines divided by square metre number of plantation by ultimate production (comprise results with output assessment)." output " of term plant relates to nutrients biological amount (root and/or branch biomass), reproductive organ and/or the propagulum (for example seed) of this plant.

Early stage vigor

" early stage vigor " refers to especially growth active, healthy, well balanced during plant-growth is early stage, and can produce because of plant adaptability increases, its reason is that for example plant adapts to its environment (promptly optimizing the use of the energy and the distribution between the Miao Yugen) better.Plant with early stage vigor also shows the seedling survival of increase and better crop foundation, this often causes field piece (crop fitly grows, and promptly most of plants reach each stage of growth on the substantially the same time) and often better and higher output highly uniformly.Thereby early stage vigor can be by the multiple factor such as thousand seed weight, sprout percentage ratio, the percentage ratio of emerging, growth of seedling, seedling height, root length, root and seedling biomass and numerous other factors are determined.

Increase/improvement/enhancing

Term " increase ", " improvement " or " enhancing " be interchangeable and should on the application's implication, refer to compare at least 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%, preferably at least 15% or 20%, more preferably 25%, 30%, 35% or 40% more output and/or growth with control plant as defined herein.

Seed production

The seed production itself that increases can show as following one or more indexs: a) seed biomass (seed gross weight) increases, and this can be based on single seed and/or every strain plant and/or every square metre; B) every strain plant increases spends number; C) (full) seed number of Zeng Jiaing; D) the full rate of the seed of Zeng Jiaing (it is expressed as the ratio between full seed number and the seed sum); E) harvest index of Zeng Jiaing, it is expressed as the ratio that can gather in the crops part (as seed) output and total biomass; And f) thousand seed weight of Zeng Jiaing (TKW), this is from the full seed number and the gross weight extrapolation thereof of counting.The TKW that increases can be because of due to the seed size and/or seed weight that increase, and also can be because of due to the increase of embryo and/or endosperm size.

The increase of seed production also can show as the increase of seed size and/or seed volume.In addition, the increase of seed production itself can self-expression be the increase of seed area and/or seed length and/or seed width and/or seed girth also.The output that increases also can produce the structure of improvement, or can occur because of the structure of improvement.

Green degree index

Used " green degree index " calculates the digital picture from plant in the literary composition.For each pixel that belongs to the plant target of correspondence on the image, calculate the ratio (in the RGB of encoded colors pattern) of greenness index and red scale value.Green degree exponential representation is the pixel per-cent that green and red ratio surpasses given threshold value.Under the normal growth condition,, measure during the last capture of the green degree index of plant before blooming under the salt stress growth conditions and can obtain under the growth conditions that nutrition reduces.In contrast, under the drought stress growth conditions, the green degree index of plant the arid before first capture the time measure.

Plant

Term as used in this article " plant " comprise ancestors and offspring and the plant part of whole strain plant, plant, comprise seed, branch, stem, leaf, root (comprising stem tuber), flower and tissue and organ, wherein every kind of mentioned object comprises goal gene/nucleic acid.Term " plant " also comprise vegetable cell, suspension culture, callus, embryo, meristem zone, gametophyte, sporophyte, pollen and sporule, same every kind of object of mentioning comprises goal gene/nucleic acid.

The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia (Viridiplantae) superfamily, especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub: maple species (Acer spp.), Actinidia species (Actinidia spp.), Abelmoschus species (Abelmoschus spp.), sisal hemp (Agavesisalana), Agropyron species (Agropyron spp.), the bent grass (Agrostisstolonifera) of crawling, allium species (Allium spp.), Amaranthus species (Amaranthus spp.), Europe beach grass (Ammophila arenaria), pineapple (Ananas comosus), Anona species (Annona spp.), celery (Apium graveolens), Arachis species (Arachis spp.), Artocarpus Forst species (Artocarpus spp.), officinalis (Asparagus officinalis), Avena species (Avena spp.) (oat (Avena sativa) for example, wild avena sativa (Avena fatua), than praising oat (Avena byzantina), Avena fatua var.sativa, hybrid oat (Avena hybrida), carambola (Averrhoa carambola), Ce Sinobambusa (Bambusa sp.), wax gourd (Benincasahispida), Brazil's chestnut (Bertholletia excelsea), beet (Beta vulgaris), Btassica species (Brassica spp.) (colea (Brassica napus) for example, overgrown with weeds blue or green species (Brassica rapassp.) [canola oil dish, rape (oilseed rape), turnip (turnip rape)]), Cadabafarinosa, tea (Camellia sinensis), Canna generalis Bailey (Canna indica), hemp (Cannabissativa), Capsicum species (Capsicum spp.), Carex elata, papaya (Carica papaya), carissa macrocarpa (Carissa macrocarpa), hickory species (Carya spp.), safflower (Carthamus tinctorius), Castanea species (Castanea spp.), America kapok (Ceibapentandra), hare's-lettuce (Cichorium endivia), Cinnamomum species (Cinnamomum spp.), watermelon (Citrullus lanatus), Citrus species (Citrus spp.), cocoanut species (Cocos spp.), Coffea species (Coffea spp.), taro (Colocasia esculenta), Africa Firmiana species (Colaspp.), Corchorus (Corchorus sp.), coriander (Coriandrum sativum), Corylus species (Corylus spp.), hawthorn species (Crataegus spp.), Stigma Croci (Crocus sativus), Cucurbita species (Cucurbita spp.), Cucumis species (Cucumis spp.), cynara scolymus belongs to (Cynara spp. species), Radix Dauci Sativae (Daucus carota), acutifoliate podocarpium herb species (Desmodium spp.), longan (Dimocarpus longan), Wild yam species (Dioscorea spp.), Diospyros species (Diospyrosspp.), Echinochloa species (Echinochloa spp.), oil palm belongs to (Elaeis) (oil palm (Elaeisguineensis) for example, America oil palm Elaeis (oleifera)) Finger-millet (Eleusine coracana), Plumegrass (Erianthus sp.), loquat (Eriobotrya japonica), eucalyptus belongs to (Eucalyptus sp.), red young fruit (Eugenia uniflora), Fagopyrum species (Fagopyrum spp.), Fagus species (Fagusspp.), alta fascue (Festuca arundinacea), Fructus Fici (Ficus carica), cumquat species (Fortunella spp.), Fragaria species (Fragaria spp.), ginkgo (Ginkgo biloba), Glycine (Glycine spp.) (soybean for example, soybean (Soja hispida) or soybean (Soja max)), upland cotton (Gossypium hirstum), Helianthus (Helianthus spp.) (for example Sunflower Receptacle (Helianthusannuus)), long tube tawny daylily (Hemerocallis fulva), hibiscus species (Hibiscus spp.), Hordeum (Hordeum spp.) (for example barley (Hordeum vulgare)), sweet potato (Ipomoeabatatas), Juglans species (Juglans spp.), lettuce (Lactuca sativa), Lathyrus species (Lathyrus spp.), Lens culinaris (Lens culinaris), flax (Linum usitatissimum), lichee (Litchi chinensis), Lotus species (Lotus spp.), patola (Luffa acutangula), lupinus species (Lupinus spp.), Luzula sylvatica, tomato belongs to (Lycopersicon spp.) (tomato (Lycopersicon esculentum for example, Lycopersicon lycopersicum, Lycopersicon pyriforme)), sclerderm Macroptilium species (Macrotyloma spp.), Malus species (Malus spp.), recessed edge Malpighia coccigera (Malpighia emarginata), shea (Mammeaamericana), mango (Mangifera indica), cassava species (Manihot spp.), sapota (Manilkara zapota), clover (Medicago sativa), Melilotus suaveolens Ledeb. species (Melilotus spp.), Mentha species (Mentha spp.), awns (Miscanthus sinensis), Momordica species (Momordica spp.), black mulberry (Morus nigra), Musa species (Musa spp.), Nicotiana species (Nicotiana spp.), Olea species (Olea spp.), Opuntia species (Opuntiaspp.), bird foot Macroptilium species (Ornithopus spp.), Oryza (Oryza spp.) (rice for example, read leaf rice (Oryza latifolia)), millet (Panicum miliaceum), switchgrass (Panicum virgatum), Purple Granadilla (Passiflora edulis), Selinum pastinaca (Pastinaca sativa), Pennisetum species (Pennisetum sp.), Persea species (Persea spp.), celery (Petroselinum crispum), Phalaris grass (Phalaris arundinacea), Phaseolus species (Phaseolus spp.), timothy grass (Phleumpratense), thorn certain herbaceous plants with big flowers species (Phoenix spp.), south reed (Phragmites australis), Physalis species (Physalis spp.), Pinus species (Pinus spp.), Pistacia vera (Pistacia vera), Pisum species (Pisum spp.), annual bluegrass species (Poa spp.), Populus species (Populusspp.), mesquite grass species (Prosopis spp.), Prunus species (Prunus spp.), Psidium species (Psidium spp.), pomegranate (Punica granatum), European pear (Pyrus communis), oak species (Quercus spp.), radish (Raphanus sativus), rheum rhabarbarum (Rheumrhabarbarum), currant species (Ribes spp.), castor-oil plant (Ricinus communis), rubus species (Rubus spp.), saccharum species (Saccharum spp.), Salix species (Salixsp.), Sambucus species (Sambucus spp.), rye (Secale cereale), flax species (Sesamum spp.), sinapsis alba species (Sinapis sp.), Solanum (Solanum spp.) (potato (Solanum tuberosum) for example, red eggplant (Solanum integrifolium) or tomato (Solanumlycopersicum)), dichromatism chinese sorghum (Sorghum bicolor), spinach species (Spinacia spp.), Syzygium species (Syzygium spp.), Tagetes species (Tagetes spp.), tamarind (Tamarindus indica), cocoa tree (Theobroma cacao), Clover species (Trifoliumspp.), Triticosecale rimpaui, Triticum (Triticum spp.) (common wheat (Triticum aestivum) for example, durum wheat (Triticum durum), cylinder wheat (Triticumturgidum), Triticum hybernum, Macha wheat (Triticum macha) (Triticum macha), common wheat (Triticum sativum) or common wheat (Triticum vulgare)), little Flower of Chinese Globeflower (Tropaeolumminus), Flower of Chinese Globeflower (Tropaeolum majus), genus vaccinium species (Vaccinium spp.), tare species (Vicia spp.), Vigna species (Vigna spp.), sweet violet (Viola odorata), Vitis species (Vitis spp.), Zea mays, Zizania palustris, zizyphus species (Ziziphus spp.) or the like.

Detailed Description Of The Invention

Ankyrin-Zn refers to the detailed description of polypeptide

Now be surprised to find the adjusting coding and produced the plant that has the output of increase with respect to control plant from the AZ polypeptide of Arabidopis thaliana (AtAZ) or the expression of nucleic acid in plant of its homologue.According to one embodiment of the invention, the method that increases plant biomass is provided, it comprises regulates coding AZ polypeptide or the expression of its homologue in plant.Advantageously, implement the method according to this invention and produced the plant that has the output of increase with respect to corresponding wild plant, particularly seed production.

" reference ", " with reference to plant ", " contrast ", " control plant ", " wild-type " or " wild-type plant " especially such cell, tissue, organ, plant or its part, it does not produce according to the inventive method.Therefore, term " wild-type ", " contrast " or " reference " be the cell or the part that can exchange mutually and can be the plant of not modified or handling according to described the inventive method herein, as organoid or tissue or plant.Therefore, as the cell of the plant of wild-type, contrast or reference or part as organoid or plant as much as possible corresponding to cell, plant or its part, and identical as far as possible with theme of the present invention on other any characteristics except that the result of the inventive method.Therefore, wild-type, contrast or reference are handled in the same manner or as far as possible in the same manner, also promptly only so the conditioned disjunction characteristic can be different, and wherein said conditioned disjunction characteristic does not influence the quality of institute's test characteristic.In other words, this meaning wild-type refers to such plant (1), and it carries gene or allelic change or does not adjust form, or refers to that (2) are used to obtain parent material/plant of plant that the inventive method produces.

Preferably, under similar condition, carry out to any comparison between the plant that produces by the inventive method at wild-type plant.Term " similar condition " mean full terms, for example culture condition or growth conditions, analysis condition (as damping fluid composition, temperature, substrate, pathogenic agent strain system, concentration etc.) keep identical between experiment to be compared.

" reference ", " contrast " or " wild-type " preferably such object, for example organoid, cell, tissue, plant, it is not adjusted, is modified or handled and be similar to theme of the present invention as far as possible on any other characteristic according to described the inventive method herein.Reference, contrast or wild-type are similar to theme of the present invention as far as possible on its genome, transcript group, protein group or metabolite group.Preferably, term " with reference to-", the organoid of " contrast-" or " wild-type-", cell, tissue or plant relate to such organoid, cell, tissue or plant, itself and organoid of the present invention, cell, in tissue or plant or its part heredity much at one, preferred 95%, more preferably 98%, even more preferably 99.00%, especially 99.10%, 99.30%, 99.50%, 99.70%, 99.90%, 99.99%, 99.999% or higher, most preferably, " reference ", " contrast " or " wild-type " is such object, organoid for example, cell, tissue or plant, itself and the plant of using according to the inventive method, cell, organoid is identical in heredity, removes nucleic acid molecule or is changed according to the inventive method by the gene product of described nucleic acid molecule encoding, outside adjusting or modifying.

Preferably term " expression " or " genetic expression " cause the phenotypic character due to one or more genetic transcriptions of Yin Teding to occur as herein defined.

The increase of the activity that relates to polypeptide in cell, tissue, organoid, organ or biological or its part compared with contrast, reference or wild-type and preferably reached at least 5%, preferably at least 10% or at least 15%, especially preferably at least 20%, 25%, 30% or higher, extremely especially preferably at least 40%, 50% or 60%, most preferably be at least 70% or higher.

The biomass (weight) that term defined herein " output of increase " means one or more parts of plant increases, and described part can comprise (can gather in the crops) part and/or underground (can gather in the crops) part on the ground.In preferred embodiments, the output of increase is the seed production that increases.

Therefore, this type of can gather in the crops preferably seed of part, and the enforcement of the inventive method produces the plant that has the seed production of increase with respect to the seed production of control plant.

The increase of seed production also can show as the increase of seed size and/or seed volume, and it also can influence the component (comprising oil, albumen and carbohydrate total content and component) of seed.

With the corn is example, and the output increase can show as following one or more indexs: the increase of the increase of the increase of every square metre of plant number, every strain plant spike number, the increase of line number, every row grain number, grain weight, thousand seed weight, fringe length/diameter, the full rate of seed (wherein the full rate of seed is that the full seed number is total and multiply by 100 divided by seed) and other.With the rice is example, and itself can show as the increase of following one or more indexs the output increase: the increase of every square metre of plant number, every strain plant panicle number, every panicle spikelet number, every panicle flower (Xiao Hua) number (it is expressed as the ratio of full seed number to former panicle number), the full rate of seed (wherein the full rate of seed be the full seed number divided by the seed sum and multiply by 100), the increase of thousand seed weight and other.The output that increases also can produce the structure of improvement, or can occur because of the structure of improvement.

According to preferred feature of the present invention, the enforcement of the inventive method produces the plant of the output with increase, the preferably plant of the seed production of Zeng Jiaing.Thereby, providing the method that increases plant biomass according to the present invention, described method comprises the expression of nucleic acid in plant of regulating coding AZ polypeptide or its homologue.

Because transgenic plant of the present invention have the output of increase, thereby with respect to the growth velocity of control plant, these plants might show on the corresponding stage in its life cycle increase growth velocity (its life cycle during the small part).The growth velocity that increases can be specific for one or more parts (comprising seed) of plant, or can spread all over whole strain plant basically.Plant with growth velocity of increase can possess short life cycle.The life cycle of plant can be considered as meaning from dry mature seed and grow to the needed time in stage that plant has produced the dry mature seed similar to parent material.This life cycle can be influenced by following factors, as early stage vigor, growth velocity, green degree index, flowering time and seed maturity speed.The increase of growth velocity can take place during life cycle on the one or more stages in life cycle or in whole plants basically plant.The growth velocity that increases during plant early stage in life cycle can reflect the enhanced vigor.The increase of growth velocity can change the harvest cycle of plant, allows the later sowing of plant and/or than early harvest, otherwise this can not (similar effect can obtain with flowering time early).If growth velocity increases fully, can allow further to sow the seed (for example sow and gather in the crops rice plant, sow and gather in the crops other rice plants subsequently, all rice plant is all in a conventional growth period) of identical plant species.Similarly, if growth velocity sufficiently increases, can allow further to sow the seed (for example sowing and harvesting corn plant are for example sowed and optional results soybean, potato or any other suitable plant subsequently) of different plant species.The results additional times also is possible in the situation of some crop plants from identical rhizome.The harvest cycle that changes plant can cause the increase of every acre year biomass yield (number of times (as in a year) that can grow and gather in the crops because of any specified plant increases).The increase of growth velocity also can allow cultivating transgenic plant in the geographic area widely than its wild type counterparts, because the region limits of cultivating crop is often determined by the plantation time (season early) or in the adverse environment condition of results period (season in evening).If shorten harvest cycle, then can avoid this class unfavourable condition.Growth velocity can determine that this type of parameter can be by obtain multiple parameter from growth curve: T-Mid (plant reaches the time that its 50% overall dimension is spent) and T-90 (plant reaches the time that its 90% overall dimension is spent), or the like.

According to preferred feature of the present invention, the plant of the output of growth velocity with increase or increase is compared in the enforcement generation of the inventive method with control plant.Thereby, providing the method that increases plant biomass and/or growth velocity according to the present invention, described method comprises the expression of nucleic acid in plant of regulating coding AZ polypeptide or its homologue.Compare with control plant, no matter plant is under the non-stress conditions still is that plant is exposed to multiple coercing down, and the increase of output and/or growth velocity all takes place.Plant is generally replied being exposed to coerce to make by growing slowlyer.Under the condition of serious stress of soil condition, plant even can stop growing fully.On the other hand, slightly coerce and be defined as plant in this article any of its exposure coerced, the wherein said ability that does not cause plant to stop growing fully and recover growth of coercing.Compare with the control plant under the non-stress conditions, slightly coerce and in meaning of the present invention, cause being coerced the plant-growth reduction less than 40%, 35% or 30%, preferably, be more preferably less than 14%, 13%, 12%, 11% or 10% or lower less than 25%, 20% or 15%.Because the progress on the agricultural practice (irrigation, fertilising, pesticide treatments) does not often run into condition of serious stress of soil in the raise crop plant.Therefore, by the agriculture often undesirable characteristic that goes up of the impaired growth of slight stress-inducing.Slightly coerce is that the common biological and/or inanimate (environment) that plant exposes is coerced.Abiotic stress can because of arid or waterlogging, anaerobism are coerced, due to salt stress, chemical toxicity, oxidative stress and heat, cold or the freezing temperature.Abiotic stress can be to coerce (especially because arid), salt stress, oxidative stress or ion by water to coerce the osmotic stress that causes.It generally is that those that caused by pathogenic agent such as bacterium, virus, fungi and insect are coerced that biology is coerced.

Especially, method of the present invention can implemented the plant that has the output of increase with respect to control plant to produce under the non-stress conditions or under slight drought condition.As report in (Planta (2003) 218:1-14) such as Wang, abiotic stress causes influencing unfriendly a series of morphological change of plant-growth and productivity, physiology to change, biological chemistry changes and molecule changes.Known arid, salinity, extreme temperature and oxidative stress are also can damaging and primary cellular defect by induced growth by similar mechanism of connecting each other.Rabbani etc. (Plant Physiol (2003) 133:1755-1767) have described " cross-talk " that drought stress and high salinity are coerced a very high degree.For example, arid and/or salinification mainly show as osmotic stress, cause the destruction of cell homeostasis and ion distribution.Often follow the oxidative stress of high temperature or low temperature, salinity or drought stress can cause functional protein and structural protein sex change.Therefore, these various environment-stress usually activate similar cell signal approach and cell response, as producing stress protein matter, raising antioxidant, accumulation compatible solute and growth-inhibiting.Term used herein " non-coercing " condition is plant not to be applied as the above-mentioned envrionment conditions of coercing.Non-stress conditions allows the plant optimum growh.Those skilled in the art know that normal edaphic condition and weather condition for given place.

The enforcement of the inventive method is with respect to the control plant of cultivating under comparable conditions, gives under the non-stress conditions or the output that increases of the plant of cultivating under slight drought condition.Thereby according to the present invention, be provided under the non-stress conditions or increase the method for output in the plant of under slight drought condition, cultivating, described method comprises that the nucleic acid that increases coding AZ polypeptide expresses in plant.

In the preferred embodiment of the invention, occur according to being increased under the non-stress conditions of the output of the inventive method and/or growth velocity.

The enforcement of the inventive method produces with respect to the appropriate control plant of cultivating under comparable conditions, the plant of under the nutritive deficiency condition, especially cultivating under nitrogen shortage condition that increases output that has.Thereby, increasing the method for output in the plant that is provided under the nutritive deficiency condition, cultivating according to the present invention, described method comprises the expression of nucleic acid in plant that increases coding AZ polypeptide.Nutritive deficiency can be because of due to the nutraceutical shortage, and described nutrition for example is nitrogen, phosphoric acid salt and other P contained compounds, potassium, calcium, cadmium, magnesium, manganese, iron and boron etc.

The inventive method advantageously is applicable to any plant.Growth characteristics above-mentioned can advantageously change in any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.

Other favourable plants be selected from composite family (Asteraceae) as Helianthus (Helianthus), Tagetes for example species Sunflower Receptacle [Sunflower Receptacle (sunflower)], spiceleaf Flower of Aztec Marigold (Tagetes lucida), Flower of Aztec Marigold (Tagetes erecta) or Tagetes signata (Tagetes tenuifolia) [Marigold], Cruciferae (Brassicaceae) as Btassica (Brassica), for example species colea, overgrown with weeds blue or green [canola oil dish, rape, turnip]; Pulse family (Fabaceae) is as Glycine for example species soybean, soybean (Sojahispida) or soybean (Soja max) [soybean]; Flax family (Linaceae) (Linaceae) is as linum (Linum) species flax [flax (flax, linseed)] for example; Gramineae such as Hordeum (Hordeum), Secale (Secale), Avena (Avena), sorghum (Sorghum), Oryza (Oryza), Zea (Zea), Triticum (Triticum) is species barley [barley] for example, rye [rye], oat, wild avena sativa, than praising oat, Avenafatua var.sativa, hybrid oat [oat], dichromatism chinese sorghum [Chinese sorghum, grain], rice, broad-leaved rice [rice], Zea mays [corn, corn], common wheat, durum wheat, the cylinder wheat, Triticum hybernum, Macha wheat (Triticum macha), common wheat (Triticum sativum) or common wheat (Triticum vulgare) [wheat, bread wheat, common wheat]; Solanaceae (Solanaceae) belongs to (Lycopersicon) for example potato [potato], tomato (Lycopersicon esculentum, Lycopersicon lycopersicum, Lycopersicon pyriforme, Solanumintegrifolium or Solanum lycopersicum) [tomato (tomato)] as Solanum (Solanum), tomato.

Term " AtAZ polypeptide or its homologue " refers to comprise that at least one ankyrin repeats to refer to at least one zinc the protein of C3H1 structural domain as herein defined, and described ankyrin repeats to be positioned at the upstream of C3H1 structural domain.Preferably, AZ protein comprises that two ankyrins repeat and two C3H1 structural domains the protein of described AZ protein shown in SEQ ID NO:2.Also preferably, two ankyrins repeat close to each other.More preferably, zinc refers to that the C3H1 structural domain is also close to each other and is positioned at ankyrin multiple C-end.Among the SEQ ID NO:2, ankyrin repeat to be positioned at position D90 to R120 and D125 to L157, two Zinc finger domains be positioned at position H301 to V327 and Q336 to P359 (Fig. 1).

Also preferably, AtAZ protein comprises at least one following consensus sequence:

(P/A) CSRAY (S/T) HDWTEC (motif 1, SEQ ID NO:3)

HPGENARRRDPR (motif 2, SEQ ID NO:4)

HG (V/I) FE (C/S) WLHP (A/S) QY (R/K) TRLCK (motif 3, SEQ ID NO:5)

CFFAH (motif 4, SEQ ID NO:6)

Preferred motif 1 is PCSRAYSHDWTEC, and preferred motif 3 is HGVFECWLHPAQYRTRLCK.

More preferably, AtAZ protein comprises two above-mentioned motifs, especially preferably comprises three above-mentioned motifs, most preferably comprises all four motifs.

As described in the Interpro database, (SMART SM00248 InterproIPR002110) is one of prevailing protein-protein interaction motif in essence in the ankyrin repetition.It is usually about 33 amino acid whose repetitive components that ankyrin repeats (series connection usually).They appear at a large amount of diverse in function mainly from the Eukaryotic protein.The seldom known example from prokaryotic organism and virus may be the result that horizontal gene transmits.Find that described repetition has multiple function in protein, as transcripting promoter, Cycle Regulation, cytoskeleton, ion transporter and signal transmitter.As if ankyrin folding by its organization definition rather than its function, because it is not by universally recognized special sequence or structure.Known ankyrin repeating unit is conservative folding from several crystal and solution structure.Each repeats to be folded into the helix-loop-helix structure in the β-hair clip/ring district that has to go out with an angle of 90 degrees projection from spiral.Described repeat to be deposited in form L shaped structure together.

As described in the Interpro database, think that Zinc finger domain ZnF_C3H1 (is also referred to as Znf_CCCH, SMART SM00356; Interpro IPR000571) participated in the DNA combination.Zinc refers to exist with multi-form, depends on the position of halfcystine.(wherein x represents any amino acid to contain the C-x8-C-x5-C-x3-H type, and the amino acid number between numeral conservative C of 8,5 and 3 representatives or the H residue) protein of Zinc finger domain comprises the zinc finger protein matter from Eukaryotic participation cell cycle or growth phase associated adjustment, as human TIS11B (butyrates response factor 1), participate in regulating may regulate protein and having the derivable nucleoprotein of mouse TTP somatomedin of said function the response of somatomedin.Mouse TTP protein is by growth factor-induced.Another protein that contains this structural domain is human splicing factor U2AF 35kD subunit, it plays a crucial role in the montage of composing type and enhanser dependence, and it selects the main protein-protein interaction and the protein-RNA interaction of requirement by mediation 3 ' splice site.Shown that different CCCH zinc finger protein matter interact with 3 ' non-translational region of different mRNA.This class zinc refers to occur with two copies very at large.

Fig. 2 has described the consensus sequence that the SMART database that is used for ankyrin structural domain and Zinc finger domain defines; Yet, should notice that these consensus sequences may be partial to the animal proteinum sequence.

Term " structural domain " and " motif " such as this paper " definition " part definition.There is the specialized database that is used to identify structural domain.C3H1 in the AZ protein or ankyrin structural domain can use (Schultz etc. (1998) the Proc.Natl.Acad.Sci. U.S. 95,5857-5864 as SMART; Letunic etc. (2002) Nucleic Acids Res 30,242-244), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318), Prosite (Bucher and Bairoch (1994), be used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and in the function of automatization sequence interpretation, () ISMB-94; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002)) evaluation.The one group of instrument that is used for analysing protein sequence on the computer chip can obtain (resident (Gasteiger etc. on Swiss Institute of Bioinformatics on the ExPASY proteomics server, ExPASy: be used for the protein science server of deep understanding and analysing protein, Nucleic AcidsRes.31:3784-3788 (2003)).With SMART instrument (edition 4 .1; Schultz etc. (1998) Proc.Natl.Acad.Sci. U.S. 95,5857-5864; Letunic etc. (2002) NucleicAcids Res 30,242-244) analyze the AZ protein sequence, and use AZ protein sequence screening Pfam (version in March, 17.0,2005; Bateman etc. (2004) Nucl.Acids Res.32, D138-141) (Release 11.0, and 26July 2005 with the InterPro database; Mulder etc. (2005) Nucl.Acids.Res.33, D201-205).

By with other protein sequences and SEQ ID NO:2 comparison, can easily identify corresponding consensus sequence, C3H1 structural domain, ankyrin structural domain or other sequence motifs.Like this, AZ polypeptide or its homologue (comprising directly to homologue and collateral line homologue) use routine techniques well known in the art easily to identify, for example identify by sequence alignment.It is well-known in the art being used for the method that aligned sequences is used for comparison, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.GAP uses Needleman and Wunsch algorithm ((1970) J Mol Biol48:443-453) to find the comparison that makes the maximization of coupling number and make minimized two complete sequence of room number.BLAST algorithm (Altschul etc. (1990) J Mol Biol 215:403-10) sequence of calculation identity percentage ratio and execution are to the statistical study of similarity between two sequences.Being used to carry out software that BLAST analyzes and being the public, to pass through NCBI (NCBI) obtainable.Homologue can use for example ClustalW multiple sequence alignment algorithm (1.83 version) evaluation easily to give tacit consent to pairing comparison parameter and methods of marking (with percentage ratio).The overall percentage ratio of similarity and identity also can use one of obtainable method in the MatGAT software package and determine (Campanella etc., BMCBioinformatics.2003 July 10; 4:29.MatGAT:an application thatgenerates similarity/identity matrices using protein or DNA sequences.).Can carry out trickle craft and write to optimize the comparison between the conservative motif, apparent as those skilled in the art.Except using full length sequence, also can use ad hoc structure territory (as C3H1 or one of ankyrin structural domain or motif defined above) in addition to identify homologue.The sequence identity value of describing with per-cent is used said procedure below, utilizes default parameters, determines on complete conserved domain or nucleic acid or aminoacid sequence.

The example of AZ protein or its homologue comprises protein sequence listed in the table 1 of embodiment 1.

Be to be understood that the sequence in the definition that falls into " AZ polypeptide or its homologue " is not limited to execute the listed peptide sequence of table 1 of example 1, or SEQ ID NO:262 or the listed homologue of SEQ ID NO:275, but any comprise that at least one ankyrin repeats and at least one C3H1 Zinc finger domain and also preferred at least one as defined above the polypeptide of SEQ ID NO:3,4,5 or 6 consensus sequences applicable in the method for the present invention.Preferably, this polypeptide is the polypeptide from Arabidopis thaliana.

Term " homologue " comprises directly to (orthologous) homologous sequence and collateral line (paralogous) homologous sequence.Can easily find by carrying out so-called mutual blast research directly to homologue and collateral line homologue.This can be undertaken by a blast (comprising at for example BLAST analysis of the obtainable ncbi database enforcement search sequence of the public (for example, SEQ ID NO:1 or SEQ ID NO:2) of any sequence library).When starting from nucleotide sequence, can use BLASTN or TBLASTX (use standard default), when starting from protein sequence, can use BLASTP or TBLASTN (use standard default).Can randomly filter BLAST result.Then filtering result or unfiltered result's full length sequence is analyzed (the 2nd BLAST) (when search sequence is SEQ ID NO:1 or SEQ ID NO:2, the 2nd blast will carry out at arabidopsis thaliana sequence) at return BLAST from the sequence of the biology that search sequence derived from.The result who compares first and second BLAST then.If from identical species, be accredited as the collateral line homologue so from the high-level hit event of the 2nd blast and search sequence; If high-level hit event and search sequence from different species, are accredited as so directly to homologue.Be that SEQ ID NO:1 or SEQID NO:2 are directly to homologue directly preferably to homologue.High-level hit event is the hit event with low E value.The E value is low more, mark remarkable more (or in other words, the probability that chances on this hit event is low more).The calculating of E value is known in this area.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Preferred score value is greater than 50, more preferably greater than 100; And preferred E-value is more preferably less than e-6 less than e-5.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the generation of tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.The straight example to homologous sequence of SEQ ID NO:2 comprises SEQ ID NO:11, SEQ ID NO:15 and SEQ ID NO:17.SEQ ID NO:19 is the example of the collateral line homologue of SEQ ID NO:2.

Preferably, the AZ polypeptide that is used for the inventive method repeats and at least one C3H1 structural domain except having at least one ankyrin, and also the protein with SEQ ID NO:2 has certain sequence identity, and described sequence identity is at least 26% according to the preferred sequence that increases, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%.Matrix shown in Fig. 3 (matrix A) shows different AZ protein total length similaritys and identity (runic).If only compare the ad hoc structure territory, identity or similarity may higher (matrix B: the comparison of Zinc finger domain sequence) between different proteins.

Can implement assay method to determine the AZ activity.Dna binding activity and protein-protein interaction can use technology well-known in the art and easily external or determine in vivo.The example that is used for the external test method of dna binding activity comprises: gel retardation assay or yeast one-hybrid analysis.The example that is used for the external test method of protein-protein interaction is yeast two-hybrid analysis (Fields and Song (1989) Nature 340:245-6).

In addition, AZ protein or its homologue in plant and the expression in rice especially have the effect that increases transgenic plant output, compare with corresponding wild-type plant, wherein the output of Zeng Jiaing comprises one of following at least: seed gross weight, seed sum and full seed number.

AZ polypeptide or its homologue are by AZ nucleic acid/genes encoding.Therefore term " AZ nucleic acid/gene " is encode the AZ polypeptide of definition as mentioned or any nucleic acid/gene of its homologue as herein defined.The example of AZ nucleic acid includes but not limited to those nucleic acid of representing in the Table A of embodiment 1.AZ nucleic acid/gene and its variant can be suitable for implementing method of the present invention.Preferably, the variant of AZ gene is derived from Arabidopis thaliana.The variant of AZ nucleic acid/gene comprise AZ nucleic acid/gene part, splice variant, allelic variant and/or can with the nucleic acid of AZ nucleic acid/gene recombination.

Quoting of " nucleotide sequence " meant the two strands of arbitrary length of polymerized form or the deoxyribonucleotide polymkeric substance or the ribonucleoside acid polymer of strand herein, or its analogue, it has the essential characteristic of natural nucleus sugar nucleotide, and promptly it can be with mode and the nucleic acid array hybridizing that is similar to naturally occurring polynucleotide.

Term is meant that partly coding comprises the dna fragmentation of the polypeptide of at least one ankyrin repetition and at least one C3H1 structural domain as herein defined.For example, can prepare part by AZ nucleic acid is carried out one or more disappearances.Part can be used with isolating form, perhaps itself and other coding (or non-coding) sequence can be merged, so that for example, produce and made up some active protein.When merging with other encoding sequences, the polypeptide that is produced after translating may be bigger than the AZ fragment of prediction.The part normal length is at least 500,700 or 900 Nucleotide, and preferred length is at least 1100,1300 or 1500 Nucleotide, and more preferably length is at least 1700,1900 or 2100 Nucleotide, and most preferably length is at least 2300 or 2400 Nucleotide.Preferably, part is the part of the nucleic acid of expression in the table 1 of embodiment 1.Most preferred part is the part of the AZ nucleic acid represented of SEQ ID NO:1 or SEQID NO:53.

Term " fragment ", " sequence fragment " or " part of sequence ", " part " or " its part " be meant with reference to the truncated sequence of original series.Truncated sequence (nucleic acid or protein sequence) variable-length; Minimum length is such sequence, its size be enough to provide have with reference to original series suitable function and/or active sequence, perhaps under stringent condition with making nucleic acid molecular hybridization of the present invention or that the inventive method is used, and the highest length is unimportant.In some applications, how much the highest length grows unlike original series activity that expectation is provided and/or the required size of function usually.Suitable functional representation be at least original series 40%, 45% or 50%, preferably at least 60%, 70%, 80% 90% or more than.

Other variants of AZ nucleic acid/gene are under the stringent condition that reduces, preferably under stringent condition, can with defined AZ nucleic acid/gene above or with the nucleic acid of defined part hybridization above.The hybridization sequences normal length is at least 300 Nucleotide, and preferred length is at least 400 Nucleotide, and more preferably length is at least 500 Nucleotide, and most preferably length is at least 600 Nucleotide.

Preferred hybridization sequences can with SEQ ID NO:1, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16, SEQ ID NO:18, SEQ ID NO:20, SEQ ID NO:22, SEQ ID NO:24, SEQ ID NO:26, SEQ ID NO:28, SEQ ID NO:30, SEQ ID NO:32, SEQ ID NO:34, SEQ ID NO:36, SEQ ID NO:38, SEQ ID NO:40, nucleic acid hybridization shown in SEQ ID NO:42 or the SEQ ID NO:53, or with the above defined part hybridization of arbitrary aforementioned sequence.Most preferably hybridization sequences can be hybridized with SEQ ID NO:1, SEQ ID NO:53 or its part (or probe).The method of designing probe is well known in the art.Probe usually on length less than 1000bp, 900bp, 800bp, 700bp, 600bp, preferably less than 500bp, 400bp, 300bp, 200bp or 100bp.Usually, hybridize for DNA-DNA, Southern trace for example, probe length changes between 100-500bp, yet at DNA-DNA hybridization, for example in pcr amplification, the hybridization region in the probe is shorter than 50 Nucleotide usually, but be longer than 10 Nucleotide, preferably they are 15,20,25,30,35,40,45 or 50bp on length.

The coding nucleic acid of the homologue of the aminoacid sequence that SEQ ID NO:2 or derivatives thereof is represented (comprise directly to or collateral line) also is useful in the methods of the invention.

Another nucleic acid variant that is used for the inventive method is the splice variant of the AZ polypeptide of definition as mentioned of encoding.Preferred splice variant is that coding comprises that at least one ankyrin repeats and the splice variant of the nucleic acid of the polypeptide of at least one C3H1 structural domain.Preferably, has 26%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity at least by the AZ polypeptide of splice variant coding or its homologue and SEQ ID NO:2.The more preferably splice variant represented of the nucleic acid of representing in the Table A by embodiment 1.For example, SEQ ID NO:25 and SEQ ID NO:47 are encoded by the splice variant of same gene.The splice variant of representing by SEQ ID NO:1 or SEQ ID NO:53 most preferably.

Another nucleic acid variant that is used for the inventive method is the nucleic acid allelic variant of the AZ polypeptide of definition as mentioned of encoding.Preferably, by in the Table A of allelic variant encoded polypeptides by embodiment 1 shown in the listed peptide sequence.Most preferably, the allelic variant of coding AZ polypeptide is by shown in the SEQ ID NO:1.

Another nucleic acid variant that is used for method of the present invention is the nucleic acid variant that obtains by gene reorganization.In addition, can use site-directed mutagenesis to produce the variant of AZ nucleic acid.Available several method carries out site-directed mutagenesis; The most frequently used method that is based on PCR (Current Protocols in MolecularBiology.Wiley writes).

Therefore, the invention provides the method that increases output and/or plant growth rate, it comprises the expression of variant in plant of regulating AZ nucleic acid, and described AZ nucleic acid is selected from:

(i) part of AZ nucleic acid;

(ii) with the nucleic acid of AZ nucleic acid hybridization;

(iii) the encode splice variant of nucleic acid of AZ polypeptide;

(iv) the encode allelic variant of nucleic acid of AZ polypeptide; With

(the v) nucleic acid variant of the coding AZ polypeptide that obtains by gene reorganization or site-directed mutagenesis.

AZ nucleic acid or its variant can be from any natural or artificial sources.Can on composition and/or genome environment, modify the natural form of described nucleic acid by careful manual operation.Nucleic acid preferred plant source, preferred source is from dicotyledonous species again, and preferred source more preferably is derived from Arabidopis thaliana from Cruciferae (Brassicaceae) again.Most preferably, AZ nucleic acid is the arabidopsis thaliana sequence that SEQ ID NO:1 represents, and the AZ aminoacid sequence is shown in SEQ ID NO:2.Alternatively, the AZ aminoacid sequence shown in AZ nucleic acid shown in the SEQ ID NO:53 or the SEQ ID NO:47 also can be used for method of the present invention.

Therefore any AZ polypeptide of addressing of this paper is interpreted as the above defined AZ protein of expression.The proteinic any nucleic acid of such AZ of encoding all is applicable to the method for the present invention of implementing.

According to a preferred aspect of the present invention, consider the expression that AZ nucleic acid or its variant are regulated, the preferred expression that increases.The method that increases genetic expression or gene product has sufficient record in this area.Can regulate the expression of the nucleotide sequence of coding AZ polypeptide by introducing genetic modification, can introduce described genetic modification by arbitrary (or multiple) following method: T-DNA activation, TILLING, site-directed mutagenesis, orthogenesis and homologous recombination, or by in plant, introducing and express the nucleic acid of coding AZ polypeptide or its homologue.Step after the introducing genetic modification is the expression that the coding nucleic acid of selection AZ polypeptide or its homologue is regulated, and the described expression of being regulated increases plant biomass.The method that reduces genetic expression or gene product also is well known in the art.

The T-DNA activation has above been described.Also can use TILLING (the local sudden change of the genome of targeted induction) technology that genetic modification is introduced the AZ locus.The locus of this paper definition means the genome district, and described genome district comprises goal gene and upstream of coding region or downstream 10kb.Can obtain several method and carry out site-directed mutagenesis; The most frequently used method that is based on PCR (Current Protocols inMolecular Biology.Wiley writes .).Also can use homologous recombination to produce effect of the present invention.

The preferred method that is used for introducing genetic modification be with coding as mentioned the AZ polypeptide of definition or its homologue the nucleic acid introduced plant and express therein.The nucleic acid that is introduced in the plant can be total length nucleic acid, maybe can be part or hybridization sequences as the preamble definition.

The present invention also provides genetic constructs and carrier to promote to introduce and/or express the nucleotide sequence that is used for the inventive method.

Thereby, gene construct is provided, it comprises:

(i) AZ nucleic acid as hereinbefore defined or its variant;

(ii) with effective one or more regulating and controlling sequences that are connected of the nucleic acid of (i).

Used construct can use the well-known recombinant DNA technology of those skilled in the art to make up in the inventive method.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for expressing goal gene in cell transformed.The present invention also provides as defined gene construct purposes in the methods of the invention in the literary composition.

Plant transforms with the carrier that comprises aim sequence (nucleic acid of promptly encode AZ polypeptide or its homologue).Those skilled in the art are appreciated that successfully and transform, select and breed the host cell that contains aim sequence and the genetic elements that must exist very much on carrier.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.Term " regulatory element ", " regulating and controlling sequence " and " promotor " have been defined above.Advantageously, the promotor of any kind can be used for driving the expression of nucleotide sequence.

The common known suitable promotor that function is arranged in plant.They can take the form of composing type or inducible promoter.Suitable promotor can make development-specific and/or the tissue specific expression in the many cells eukaryote become possibility; Therefore can in plant, advantageously use leaf-, root-, flower-, seed-, pore-, stem tuber-or fruit-specific promoter.

The different plant promoters that can be used in the plant are such promotors, for example USP, LegB4-, DC3 promotor or from the ubiquitin promotor of parsley (parsley).

" plant " promotor comprises controlling element, its mediation encoding sequence section expression in vegetable cell.Thereby plant promoter need not to plant origin, but can derive from virus or microorganism, for example, particularly derives from the virus of attacking vegetable cell.

" plant " promotor also can derive from vegetable cell, for example, derives from the nucleotide sequence plant transformed of expressing in the methods of the invention through desire as herein described.This is suitable equally for other " plant " adjustment signals, for example the situation of " plant " terminator.

For expressing in plant, as mentioned above, nucleic acid molecule must effectively be connected or comprise this promotor with suitable promotor, and this promotor is expressed gene on correct time point and in cell or tissue specificity mode.The available promotor is constitutive promoter (Benfey etc., EMBO is (1989) 2195-2202 J.8), those constitutive promoters as plant-derived virus, as 35SCAMV (Franck etc., Cell 21 (1980) 285-294), 19S CaMV (seeing also US 5352605 and WO 84/02913), 34S FMV (Sanger etc., Plant.Mol.Biol., 14,1990:433-443), parsley ubiquitin promotor or as at US 4,962, the plant promoter such as Rubisco small subunit promotor or the plant promoter PRP1[Ward etc. that describe in 028, Plant.Mol.Biol.22 (1993)], SSU, PGEL1, OCS[Leisner (1988) the Proc Natl Acad Sci U.S. 85 (5): 2553-2557], lib4, usp, mas[Comai (1990) Plant Mol Biol 15 (3): 373-381], STLS1, ScBV (Schenk (1999) Plant Mol Biol 39 (6): 1221-1230), B33, SAD1 or SAD2 (flax promotor, ain etc., Crop Science, 39 (6), 1999:1696-1701) or (1984) Nucleic Acids Res.12 (20): 7831-7846 such as nos[Shaw].Other examples of constitutive plant promoters are beet V-ATP enzyme promotors (WO 01/14572).Synthetic constitutive promoter example be super promotor (WO 95/14098) and from the G-box deutero-promotor (WO 94/12015).As required, compare, can also further use the chemical induced promotor with EP-A 388186, EP-A 335528, WO 97/06268.The stable constitutive expression of protein of the present invention in plant can be favourable.Yet, be favourable if for example express the late period before results, the inducible expression of polypeptide of the present invention is favourable, because the metabolic operation can cause plant-growth slow.

Also can promote the expression of plant gene by the chemical induced promotor.When needs during with temporal mode expressing gene, the chemical induced promotor is a particularly suitable.The example of this type of promotor is Induced by Salicylic Acid promotor (WO 95/19443) and dormin-inducible promoter (EP 335 528), tsiklomitsin-inducible promoter (Gatz etc. (1992) Plant J.2,397-404), hexalin-or ethanol-inducible promoter (WO 93/21334) or other chemical induced promotors as described herein.

Other suitable promotor is to coerce or those promotors of inanimate stress conditions reaction biological, pathogen-inducible PRP1 gene promoter (Ward etc. for example, Plant.Mol.Biol.22 (1993) 361-366), (US 5 for tomato thermal induction hsp80 promotor, 187,267), cold inducibility α-Dian Fenmei promotor (WO 96/12814) of potato or wound inducibility pinII promotor (EP-A-0375 091) or other promotors as described herein.

Preferred promotor especially in tissue and organ, in the protoblast of seed cell such as albuminous cell and growth, cause those promotors of genetic expression.Suitable promotor is that (US 5 for the rapeseed protein gene promoter of rape, 608,152), broad bean (Vicia faba) USP promotor (Baeumlein etc., Mol Gen Genet, 1991,225 (3): 459-67), Arabidopis thaliana oleosin promotor (WO98/45461), (US 5 for Kidney bean (Phaseolus vulgaris) phaseolin promoter, 504,200), Brassica plants (Brassica) Bce4 promotor (WO 91/13980), beans arc5 promotor, Radix Dauci Sativae DcG3 promotor or leguminous plants (Legumin) B4 promotor (LeB4; Baeumlein etc., 1992, PlantJournal, 2 (2): 233-9) and cause the promotor of the seed-specific expression in monocotyledons such as corn, barley, wheat, rye, rice etc.Favourable seed specific promoters is sucrose-binding proteins promotor (WO 00/26388), phaseolin promoter and rapeseed protein promotor.The suitable promotor that must take in is that barley lpt2 or lpt1 gene promoter (WO 95/15389 and WO 95/23230) and the promotor described in WO 99/16890 are (from the hordein gene of barley, the glutenin gene of rice, the paddy rice plain gene of rice, the prolamine gene of rice, the gliadine gene of wheat, the glutenin gene of wheat, the zein spirit-soluble gene of corn, the avenaceous glutenin gene, the promotor of the secalin gene of Chinese sorghum kasirin gene and rye).Other suitable promotors are Amy32b, Amy 6-6 and Aleurain[US 5,677,474], [US 5 for Bce4 (rape), 530,149], glycinin (soybean) [EP 571 741], phosphoric acid enol pyruvic acid carboxylase (soybean) [JP 06/62870], ADR12-2 (soybean) [WO 98/08962], [US 5 for isocitrate lyase (rape), 689,040] or α-Dian Fenmei (barley) [EP 781 849].Be used in that other promotors of expressing gene are leaf specificity promoters in the plant, regulate promotor as, pea petE promotor for example as those promotors of in DE-A 19644478, describing or light.

Other suitable plant promoters are cytosol FBP enzyme promotor or potato ST-LSI promotor (Stockhaus etc., EMBO J.8,1989,2445), the joint specificity promoter of soybean Phosphoribosyl tetra-sodium amide transferase promotor (GenBank accession number U87999) or description in EP-A-0 249 676.

According to a preferred feature of the present invention, AZ nucleic acid or its variant effectively are connected in seed specific promoters.Seed specific promoters can activate in seed development and/or duration of germination.It is well-known that seed-specific is enabled in this area.The preferred seed specificity promoter is embryo-specific, endosperm-specific, aleuron specificity promoter.Also the preferred seed specificity promoter drives the expression in one of embryo, endosperm, aleuron at least.More preferably promotor is the promotor of WSI18 or its functional equivalent.Most preferably promoter sequence such as SEQ ID NO:9 or SEQ ID NO:55 are represented.Should be understood that practicality of the present invention is not limited to the AZ nucleic acid shown in SEQ ID NO:1 or the SEQ ID NO:53, practicality of the present invention also is not limited to the AZ expression of nucleic acids that driven by seed specific promoters.The example (embryo-specific, endosperm-specific, aleuron specificity promoter) of other seed specific promoters that can be used for driving the AZ expression of nucleic acid equally above is provided.

According to a preferred feature of the present invention, AZ nucleic acid or its variant effectively are connected in constitutive promoter.Preferred constitutive promoter is the also constitutive promoter of omnipresence ground expression basically.Also preferred promoter is from plant, more preferably from monocotyledons.An example of this promotor is the GOS2 promotor (SEQ ID NO:54 or SEQ ID NO:56) from rice.Should be understood that practicality of the present invention is not limited to the AZ nucleic acid shown in the SEQ ID NO:1, practicality of the present invention also is not limited to the AZ expression of nucleic acids that is subjected to constitutive promoter and especially is subjected to the GOS2 promoters driven.The example that has above shown other constitutive promoters that can be used for driving the AZ expression of nucleic acid equally.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.Extra regulatory element can comprise transcriptional enhancer and translational enhancer.One skilled in the art will recognize that the terminator sequence and the enhancer sequence that can be suitable in the embodiment of this invention.As described in definitional part, intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also be included in keeps and/or duplicates required replication orgin sequence in the particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to f1-ori and colE 1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Thereby genetic constructs can randomly comprise the selected marker.The more detailed selective marker of having described of " definition " part in the literary composition.This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top " definition " part.

The present invention also comprises plant, plant part or the vegetable cell that available method of the present invention obtains.The plant that therefore the present invention provides available method of the present invention to obtain is introduced as top defined AZ nucleic acid or its variant in the described plant.

The present invention also is provided for producing the method for the transgenic plant of the output with increase, its be included in introduce in the plant and express as mentioned in defined AZ nucleic acid or its variant.

Host plant advantageously can synthesize whole plants of used polypeptide in the inventive method in principle for used nucleic acid or carrier, expression cassette or construct or carrier in the inventive method.

More specifically, the invention provides the method that is used to produce the transgenic plant with increase output, described method comprises:

(i) in plant or vegetable cell, introduce and express AZ nucleic acid or its variant; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

Nucleic acid is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleic acid is preferably by transforming introduced plant.Definitional part is more detailed has described term " introducing " or " conversion ".Usually after conversion, vegetable cell or the cell colony of selecting one or more marks to exist, wherein said mark become whole strain plant with the material regeneration that transforms subsequently by the effable genes encoding of the plant that moves with the goal gene corotation.As mentioned, the Agrobacterium that transforms with expression vector of the present invention also can transform plant with himself known method, as being used as model plant, as Arabidopis thaliana (Arabidopis thaliana is not considered as crop plants in the scope of the invention) or crop plants, for example as cereal, corn, oat, rye, barley, wheat, soybean, rice, cotton, beet, rape, Sunflower Receptacle, flax, hemp, potato, tobacco, tomato, Radix Dauci Sativae, pimento, the Semen Brassicae campestris rape, tapioca (flour), cassava, arrowroot, Flower of Aztec Marigold, clover, romaine lettuce and multiple trees, nut and grape vine species, oil-containing crop plants particularly, as soybean, peanut, the Viscotrol C plant, Sunflower Receptacle, corn, cotton, flax, the Semen Brassicae campestris rape, coconut, oil palm, safflower (Carthamus tinctorius), cocoa beans, for example the leaf or the leaf segment of scratching by water-bath in Agrobacterium solution cultivated it subsequently in suitable medium.

The vegetable cell of genetic modification can be regenerated by all methods that those skilled in the art are familiar with.Suitable method be found in S.D.Kung and R.Wu, Potrykus or

Above-mentioned publication with Willmitzer.

Usually after conversion, vegetable cell or the cell colony of selecting one or more marks to exist, wherein said mark become whole strain plant with the material regeneration that transforms subsequently by the effable genes encoding of the plant that moves with the goal gene corotation.In order to select plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example, can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying.Another kind of possibility is included in the seed of growing on the agar plate that uses suitable selective agent (as required after sterilization), makes the seed that only transforms can grow into plant.Perhaps, plant transformed is by the existence screening of above-mentioned those selective markers.

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition can for example use Southern to analyze goal gene were estimated.Alternative or extraly, the expression level of newly introducing DNA can use Northern and/or Western to analyze and monitor, and these two kinds of technology are that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clonal expansion method or classical breeding technique.For example, the first-generation (or T1) transforms plant can carry out selfing, the s-generation (or T2) transformant that selection is isozygotied, and the T2 plant can further breed by classical breeding technique subsequently.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's transformant (for example being transformed) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the root stock of the conversion of unconverted grafting of tender branch.

The present invention extends to any vegetable cell or the plant that produces by any means described in the literary composition clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by any preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical one or more yielding characteristicses and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.The present invention also comprises host cell, and it contains separative AZ nucleic acid or its variant.The preferred host cell of the present invention is a vegetable cell.The present invention also extend to plant the part gathered in the crops as, but be not limited to seed, leaf, fruit, flower, stem, root, root stock, stem tuber and bulb.The invention further relates to directly from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, oil, fat and lipid acid, starch or protein.

The present invention also comprises the purposes of AZ nucleic acid or its variant and the purposes of AZ polypeptide or its homologue.

This purposes relates to improvement plant-growth feature, particularly improvement output, especially seed production.The seed production that increases preferably includes the thousand seed weight of increase.

The nucleic acid of coding AZ polypeptide can be used for the procedure of breeding, wherein identifies the dna marker that can be connected with the AZ gene hereditarily.Described nucleic acid/gene can be used for defining molecule marker.This DNA or protein labeling can be used for selecting to have the plant of enhanced yield subsequently in the procedure of breeding.

The allelic variant of AZ nucleic acid/gene also can be used for the auxiliary procedure of breeding of mark.This procedure of breeding needs to introduce allelic variation by for example using the EMS mutagenesis that plant is carried out mutagenic treatment sometimes; Alternatively, this program can be from one group of allelic variant of the non-artificial what is called that causes " nature " origin.Carry out the evaluation of allelic variant subsequently, for example by the PCR method.After this be used to select discuss and cause increasing the step of excellent allelic variant of the sequence of output.Generally the growth performance that contains the plant of the different allelic variants that sequence is discussed to some extent by monitoring is implemented to select, as the different allelic variants of listed arbitrary nucleic acid in the table 1 of embodiment 1.Can be in the greenhouse or field monitoring growth performance.Other optional step comprise and will identify the plant and the another kind of plant hybridization of excellent allelic variant.This can be used for for example producing target phenotype combination of features.

AZ nucleic acid also can be as probe so that carry out genetic mapping or physical mapping to gene, and described probe reaches the mark of the proterties related with these genes as the part of described gene.This type of information can be used for plant breeding, so that exploitation has the strain system that wants phenotype.This purposes of AZ nucleic acid or its variant only needs to have the nucleotide sequence of at least 15 length of nucleotides.AZ nucleic acid or its variant can be used as restriction fragment length polymorphism (RFLP) mark.The Southern trace of the plant genome DNA of restrictive diges-tion (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) can be used the AZ nuclei acid probe.What produce carries out genetic analysis to make up genetic map in conjunction with graphic can use a computer subsequently program such as MapMaker (Lander etc. (1987) Genomics 1:174-181).In addition, this nucleic acid can be used for surveying the Southern trace of the genomic dna that contains one group of individuality handling through restriction endonuclease, and wherein said one group of individual representative has the parental generation and the offspring of definite genetic cross.The separation of dna polymorphism is marked and is used for calculating the position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) of AZ nucleic acid in using the previous genetic map that obtains of this colony.

Generation and its purposes in genetic mapping of plant gene deutero-probe have been described in Bernatzky and Tanksley (1986) Plant Mol.Biol.Reporter 4:37-41.Numerous publications have been described the genetic mapping that uses methodology mentioned above or its modification method that specific cDNA is cloned.For example, to hand over group, the group that backcrosses, panmictic population, contiguous isozygotying mutually be can be used for mapping with other population of individuals to F2.This type of methodology is that those skilled in the art are well-known.

It (is the arrangement of sequence on physical map that described nucleic acid probe also can be used for physical mapping; See that Hoheisel etc. exists: Non-mammalian Genomic Analyasis:A Practical Guide, Academic press 1996, the 319-346 pages or leaves and the reference of wherein quoting).

In another embodiment, nucleic acid probe can directly use in fluorescence in situ hybridization (FISH) graphing method (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although large-scale clone is used in current FISH graphing method support; See (1995) Genome Res.5:13-20 such as Laan), however the improvement of sensitivity can allow to use shorter probe to carry out the FISH mapping.

The multiple method based on nucleic acid amplification that is used for genetic mapping and physical mapping can be used described nucleic acid and implement.Example is found in " definition " part in the literary composition.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med 11:95-96), pcr amplified fragment; Sheffield etc., (1993) Genomics 16:325-332), allele-specific connects (Landegren etc., (1988) Science 241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc., Nat.Genet.7:22-28) and Happy mapping (Dear and Cook, (1989) Nucleic Acid Res.17:6795-6807) (1997).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The inventive method produces has the plant that has enhanced yield as previously described.These favourable growth characteristics also can make up with other favourable economically proterties, as other output increase proterties, to multiple tolerance of coercing, regulate the proterties of multiple constructivity feature and/or biochemical characteristics and/or physiologic character.

The detailed description of SYT polypeptide

Now be surprised to find being expressed under the abiotic stress of nucleotide sequence of regulating coding SYT polypeptide and increased plant biomass and/or early stage vigor with respect to control plant.Thereby, the invention provides the method that under abiotic stress, increases plant biomass and/or early stage vigor with respect to control plant, it comprises the expression of nucleotide sequence in plant of regulating coding SYT polypeptide.

One or more control plants or the plant that quoting of " control plant " meant any appropriate in this article.

Term " wild-type ", " contrast " or " reference " use interchangeably at this paper, and preferably such object, for example organoid, cell, tissue, plant, it is similar to theme of the present invention as far as possible.Reference, contrast or wild-type are similar to theme of the present invention as far as possible on its genome, transcript group, protein group or metabolite group.Preferably, term " reference-", " contrast-" or organoid, cell, tissue or the plant of " wild-type-" relate to such organoid, cell, tissue or plant, much at one, have 95%, 98%, 99.00%, 99.10%, 99.30%, 99.50%, 99.70%, 99.90%, 99.99%, 99.999% or higher genetic identity in itself and organoid of the present invention, cell, tissue or plant or its part heredity with the preferred sequence that increases and organoid of the present invention, cell, tissue or plant.Most preferably, " reference ", " contrast " or " wild-type " preferably such object, for example organoid, cell, tissue or plant, it is identical in heredity with the plant, tissue, cell, the organoid that use according to the inventive method, remove nucleotide sequence the gene product discussed is subjected to changing, adjusts according to the inventive method or modification.

Preferably, under similar condition, carry out to any comparison between the plant that produces by the inventive method at control plant.Term " similar condition " mean full terms, for example culture condition or growth conditions, analysis condition (as damping fluid composition, temperature, substrate, pathogenic agent strain system, concentration etc.) keep identical between experiment to be compared.

Any SYT polypeptide that means as defined herein of quoting of " protein that is used for the inventive method " after this.Any nucleic acid that means this type of SYT polypeptide of to encode of quoting of " nucleic acid that is used for the inventive method " after this.The nucleic acid for the treatment of introduced plant (and thereby be used to implement the inventive method) is any nucleic acid of the present described protein type of coding, hereinafter is called " SYT nucleic acid " or " SYT gene " again.

Term " sequence " relates to polynucleotide, nucleic acid, nucleic acid molecule, peptide, polypeptide and protein, depends on the context that uses term " sequence "." encoding sequence " is nucleotide sequence, when under the control that is placed on suitable regulating and controlling sequence, is transcribed into mRNA and/or is translated as polypeptide.The border of encoding sequence is defined as translation stop codon of translation initiation codon and the 3 '-end of 5 '-end.Encoding sequence can be including, but not limited to mRNA, cDNA, recombinant nucleic acid sequence or genomic dna, and also can have intron under certain situation.Term " expression " or " genetic expression " define as mentioned.Above define term " adjusting " and meant the increase on expressing with regard to gene or genetic expression.

The term " SYT polypeptide " of this paper definition refers to such polypeptide, it comprises from the N-terminal to the C-terminal: (i) according to the preferred sequence that increases, and the SNH structural domain that has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity with the SNH structural domain of SEQ ID NO:58; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.Preferably, the SNH structural domain comprises the residue that is shown as black among Fig. 6.Further preferably, the SNH structural domain is a SEQ ID NO:57 representative.

In addition, the SYT polypeptide can comprise following one or more sequence: (a) SEQ ID NO:146; (b) SEQ ID NO:147; What (c) be positioned at N-terminal before the SNH structural domain is rich in the Met structural domain.

The SYT polypeptide is by the SYT nucleic acid sequence encoding.Therefore the term " SYT nucleotide sequence " that defines in the literary composition is any nucleotide sequence of above defined SYT polypeptide of encoding.

Term " motif " and " structural domain " define as the definition part branch.There is the specialized database that is used to identify structural domain, as SMART (Schultz etc. (1998) the Proc.Natl.Acad.Sci. U.S. 95,5857-5864; Letunic etc. (2002) Nucleic Acids Res 30,242-244), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318), Prosite (Bucher and Bairoch (1994), be used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and in the function of automatization sequence interpretation, () ISMB-94; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002)) evaluation.The one group of instrument that is used for analysing protein sequence on the computer chip can obtain on ExPASY proteomics server (at Swiss Institute of Bioinformatics (Gasteiger etc., ExPASy: be used for the protein science server of deep understanding and analysing protein, Nucleic Acids Res.31:3784-3788 (2003)).Also can use routine techniques to identify structural domain and motif, as pass through sequence alignment.Can utilize well-known routine techniques in this area,, easily identify the SYT polypeptide as by sequence alignment.The method of sequence alignment is well-known in the art, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.The algorithm ((1970) J.Mol.Biol.48:443-453) of GAP application Needleman and Wunsch is sought the comparison of two complete sequence, and maximization of coupling number and room number are minimized.BLAST algorithm (Basic Local Alignment Search Tool; Altschul etc. (1990) J Mol Biol215:403-10) per-cent (%) of sequence of calculation identity, and the similarity between two sequences (E-value) carried out statistical study.Identity per-cent refers to two relatively numbers of the identical Nucleotide (or amino acid) on length-specific between nucleic acid (or polypeptide) sequence.Similarity between two sequences is high more, E-value low more (or in other words, it is low more to chance on the chance of hitting).The calculating of E-value is well known in the art.The software of carrying out the BLAST analysis can obtain publicly from biotechnology infonation center.Can identify like this to comprise the SYT polypeptide that has the SNH structural domain of certain sequence identity with the SNH structural domain of SEQ ID NO:58 that described sequence identity is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% according to the preferred sequence that increases.Alternatively, the SYT polypeptide and the polypeptide SEQ ID NO:60 that are used for the inventive method have certain sequence identity, and described sequence identity is at least 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% according to the preferred sequence that increases.Can carry out trickle craft and write to optimize the comparison between the conservative motif, apparent as those skilled in the art.In some cases, can adjust default parameters to regulate the severity of search.For example use BLAST, can improve the statistical significance threshold value (being called " expectation " value) that is used to report at the coupling of database sequence to show the lower coupling of severity.Like this, can identify almost accurate short coupling.Can identify the SEQ ID NO:146 that all is included in the SYT polypeptide that is used for the inventive method and the existence of SEQ ID NO:147 like this.

In addition, the existence of being rich in the Met structural domain or being rich in the QG structural domain also can easily be identified.As shown in Figure 7, be rich in Met structural domain and be rich in the QG structural domain and be positioned at after the SNH structural domain.Being rich in that the QG structural domain can be considered is the terminal remainder (removing the SNH structural domain) of peptide C basically; Be rich in the Met structural domain be generally comprised within be rich in the QG structural domain first half within (direction from the N-terminal to the C-terminal).The one-level amino acid whether decision polypeptide structure territory is rich in specific amino acids is formed (representing with %) and can be used to software program, particularly ProtParam instrument from the ExPASy server and calculate (2003) ExPASy:the proteomics server for in-depth proteinknowledge and analysis.Nucleic Acids Res 31:3784-3788 such as () Gasteiger E.The composition of desired polypeptides and the average amino acid composition (representing with %) in the Swiss-Prot protein sequence database can be compared (table 5) then.In this database, average Met (M) content is 2.37%, and average Gln (Q) content is 3.93%, and average Gly (G) content is 6.93% (table 5).Such as herein defined, be rich in the Met structural domain or be rich in the QG structural domain and have and be higher than Met content (representing) or Gln and the Gly content (representing) that the average amino acid of Swiss-Prot protein sequence database is formed (representing with %) with % with %.For example in SEQ ID NO:60, the Met structural domain (from the 1st to the 24th in amino acid) that is rich in that is positioned at N-terminal before the SNH structural domain has 20.8% Met content, is rich in QG structural domain (from the 71st to the 200th in amino acid) and has 18.6% Gln (Q) content and 21.4% Gly (G) content.Preferably, the Met structural domain of this paper definition has Met content (with %), described Met content is to form with the average amino acid of described protein sequence class to count 1.25 at least, 1.5,1.75,2.0,2.25,2.5,2.75,3.0,3.25,3.5,3.75,4.0,4.25,4.5,4.75,5.0,5.25,5.0,5.75,6.0,6.25,6.5,6.75,7.0,7.25,7.5,7.75,8.0,8.25,8.5,8.75,9.0,9.25,9.5,9.75,10 or higher (with %), described average amino acid is formed and is included in the Swiss-Prot protein sequence database.Preferably, the QG structural domain of this paper definition has Gln (Q) content and/or Gly (G) content, described Gln (Q) content and/or Gly (G) content are to form with the average amino acid of described protein sequence class to count 1.25 at least, 1.5,1.75,2.0,2.25,2.5,2.75,3.0,3.25,3.5,3.75,4.0,4.25,4.5,4.75,5.0,5.25,5.0,5.75,6.0,6.25,6.5,6.75,7.0,7.25,7.5,7.75,8.0,8.25,8.5,8.75,9.0,9.25,9.5,9.75,10 or higher (with %), described average amino acid is formed and is included in the Swiss-Prot protein sequence database.

Average amino acid is formed (%) (in July, 2004) in the table 5:Swiss-Prot protein sequence database:

Residue	％	Residue	％
				A＝Ala	7.80	M＝Met	2.37
C＝Cys	1.57	N＝Asn	4.22
				D＝Asp	5.30	P＝Pro	4.85
E＝Glu	6.59	Q＝Gln	3.93
				F＝Phe	4.02	R＝Arg	5.29
G＝Gly	6.93	S＝Ser	6.89

H＝His	2.27	T＝Thr	5.46
				I＝Ile	5.91	V＝Val	6.69
K＝Lys	5.93	W＝Trp	1.16
				L＝Leu	9.62	Y＝Tyr	3.09

The example of SYT polypeptide comprises (by the polynucleotide sequence coding shown in the accession number in the bracket, referring to what mentioned in table 6 and the sequence table): Arabidopis thaliana Arath_SYT1 (AY102639.1) SEQ IDNO:60, Arabidopis thaliana Arath_SYT2 (AY102640.1) SEQ ID NO:62, Arabidopis thaliana Arath_SYT3 (AY102641.1) SEQ ID NO:64, Aspergillus offcinalisAspof_SYT (CV287542) SEQ ID NO:66, colea (Brassica napus) Brana_SYT (CD823592) SEQ ID NO:68, sweet orange (Citrus sinensis) Citsi_SYT (CB290588) SEQ ID NO:70, tree cotton (Gossypium arboreum) Gosar_SYT (BM359324) SEQ ID NO:72, puncture vine shape clover (Medicago trunculata) Medtr_SYT (CA858507.1) SEQ ID NO:74, rice (Oryza sativa) Orysa_SYT1 (AK058575) SEQ ID NO:76, rice Orysa_SYT2 (AK105366) SEQ ID NO:78, rice Orysa_SYT3 (BP185008) SEQ ID NO:80, potato Soltu_SYT (BG590990) SEQ ID NO:82, Zea mays (Zea mays) Zeama_SYT1 (BG874129.1, CA409022.1) SEQ ID NO:84, Zea mays Zeama_SYT2 (AY106697) SEQ IDNO:86, homo sapiens (Homo sapiens) Homsa_SYT (CAG46900) SEQ ID NO:88, onion (Allium cepa) Allce_SYT2 (CF437485) SEQ ID NO:90, Aquilegiaformosa x Aquilegia pubescens Aqufo_SYT1 (DT758802) SEQ ID NO:92, two fringe false bromegrasses (Brachypodium distachyon) Bradi_SYT3 (DV480064) SEQ IDNO:94, colea Brana_SYT2 (CN732814) SEQ ID NO:96, sweet orange Citsi_SYT2 (CV717501) SEQ ID NO:98, Euphorbia esula L (Euphorbia esula) Eupes_SYT2 (DV144834) SEQ ID NO:100, soybean (Glycine max) Glyma_SYT2 (BQ612648) SEQ ID NO:102, wild soybean (Glycine soya) Glyso_SYT2 (CA799921) SEQ ID NO:104, upland cotton (Gossypium hirsutum) Goshi_SYT1 (DT558852) SEQ ID NO:106, upland cotton Goshi_SYT2 (DT563805) SEQ ID NO:108, barley (Hordeum vulgare) Horvu_SYT2 (CA032350) SEQID NO:110, fireweed (Lactuca serriola) Lacse_SYT2 (DW110765) SEQ IDNO:112, tomato (Lycopersicon esculentum) Lyces_SYT1 (AW934450, BP893155) SEQ ID NO:114, domestication apple (Malus domestica) Maldo_SYT2 (CV084230, DR997566) SEQ ID NO:116, puncture vine shape clover Medtr_SYT2 (CA858743, BI310799, AL382135) SEQ ID NO:118, switchgrass (Panicumvirgatum) Panvi_SYT3 (DN152517) SEQ ID NO:120, picea sitchensis (Piceasitchensis) Picsi_SYT1 (DR484100, DR478464) SEQ ID NO:122, torch pine (Pinus taeda) Pinta_SYT1 (DT625916) SEQ ID NO:124, trembling poplar (Populustremula) Poptr_SYT1 (DT476906) SEQ ID NO:126, sugarcane (Saccharumofficinarum) Sacof_SYT1 (CA078249, CA078630, CA082679, CA234526, CA239244, CA083312) SEQ ID NO:128, sugarcane Sacof_SYT2 (CA110367) SEQ ID NO:130, sugarcane Sacof_SYT3 (CA161933, CA265085) SEQ ID NO:132, potato Soltu_SYT1 (CK265597) SEQ ID NO:134, dichromatism chinese sorghum (Sorghumbicolor) Sorbi_SYT3 (CX611128) SEQ ID NO:136, common wheat (Triticumaestivum) Triae_SYT2 (CD901951) SEQ ID NO:138, common wheat Triae_SYT3 (BJ246754, BJ252709) SEQ ID NO:140, grape (Vitis vinifera) Vitvi_SYT1 (DV219834) SEQ ID NO:142, Zea mays Zeama_SYT3 (CO468901) SEQ IDNO:144, colea (Brassica napus) Brana_SYT SEQ ID NO:151, soybean (Glycine max) Glyma_SYT SEQ ID NO:153.

Table 6: the example of the nucleotide sequence of coding SYT polypeptide

Title	NCBI Nucleotide accession number	Nucleic acid sequence SEQ ID NO	Translation polypeptide SEQ ID NO	The source
					Arath_SYT1	AY102639.1	59	60	Arabidopis thaliana
Arath_SYT2	AY102640.1	61	62	Arabidopis thaliana

Arath_SYT3	AY102641.1	63	64	Arabidopis thaliana
					Aspof_SYT1	CV287542
	65	66	Aspergillus officinalis
				Brana_SYT1	CD823592		67	68	Colea
Citsi_SYT1	CB290588
			69	70	Sweet orange
Gosar_SYT1	BM359324
			71	72	Tree cotton
Medtr_SYT1	CA858507.1					73	74	Puncture vine shape clover
		Orysa_SYT1	AK058575
	75			76	Rice
		Orysa_SYT2	AK105366
	77			78	Rice
		Orysa_SYT3	BP185008
	79			80	Rice
		Soltu_SYT2	BG590990
	81			82	Potato
		Zeama_SYT1	BG874129.1 CA409022.1 ＊			83	84	Zea mays
Zeama_SYT2	AY106697
			85	86	Zea mays
Homsa_SYT	CR542103
			87	88	The homo sapiens
Allce_SYT2	CF437485						89	90	Onion
		Aqufo_SYT1	DT758802.1	91	92					Aquilegia formosa x Aquilegia pubescens
Bradi_SYT3	DV480064.1					93	94	Two fringe false bromegrasses
		Brana_SYT2	CN732814	95	96				Colea
Citsi_SYT2	CV717501
			97	98	Sweet orange
Eupes_SYT2	DV144834
			99	100	Euphorbia esula L
Glyma_SYT2	BQ612648
			101	102	Soybean
Glyso_SYT2	CA799921
			103	104	Wild soybean
Goshi_SYT1	DT558852
			105	106	Upland cotton

Goshi_SYT2	DT563805		107	108	Upland cotton
						Horvu_SYT2	CA032350
	109	110	Barley
				Lacse_SYT2	DW110765
	111	112	Fireweed
				Lyces_SYT1	AW934450.1 BP893155.1 ＊	113	114	Tomato
Maldo_SYT2	CV084230 DR997566	＊	115						116	The domestication apple
				Medtr_SYT2	CA858743 BI310799.1 AL382135.1 ＊	117	118	Puncture vine shape clover
Panvi_SYT3	DN152517
			119	120	Switchgrass
Picsi_SYT1	DR484100 DR478464.1					121	122	Picea sitchensis
		Pinta_SYT1	DT625916
	123			124	Torch pine
		Poptr_SYT1	DT476906
	125			126	Trembling poplar
		Sacof_SYT1	CA078249.1 CA078630 CA082679 CA234526 CA239244 CA083312 ＊			127	128	Sugarcane
Sacof_SYT2	CA110367
			129	130	Sugarcane
Sacof_SYT3	CA161933.1 CA265085 ＊					131	132	Sugarcane
		Soltu_SYT1	CK265597
	133			134	Potato
		Sorbi_SYT3	CX611128
	135			136	Dichromatism chinese sorghum

Triae_SYT2	CD901951		137	138	Common wheat
						Triae_SYT3	BJ246754 BJ252709	＊	139	140	Common wheat
Vitvi_SYT1	DV219834
			141	142	Grape
Zeama_SYT3	CO468901
			143	144	Zea mays
Brana_SYT	NA
			150	151	Colea
Glyma_SYT	NA
			152	153	Soybean

^*By the accession number splicing of being drawn

NA: non-availability (individual has)

The example of the nucleic acid of coding SYT polypeptide provides in last table 6.This class nucleic acid can be used for implementing method of the present invention.The aminoacid sequence that provides in the last table 6 be the SYT polypeptide shown in the SEQ ID NO:58 directly to the example of homologue and collateral line homologue, term " directly to homologue " and " collateral line homologue " such as in the literary composition definition.Other directly can easily be identified by carrying out so-called interactivity blast search to homologue and collateral line homologue.This is usually directed to a BLAST, and a wherein said BLAST comprises that submission search sequence (for example use and go up arbitrary sequence of listing in the table 6) is used for the blast search at arbitrary sequence library (as public's available ncbi database).When nucleotide sequence begins, generally use BLASTN or TBLASTX (using the standard default value), and, can use BLASTP or TBLASTN (use standard default value) when when protein sequence begins.Randomly can screen BLAST result.The full length sequence of submitting The selection result and non-The selection result subsequently to is with at carry out reverse BLAST (the 2nd BLAST) from the sequence of biology, wherein search sequence is from described biology (be under the situation of SEQ ID NO:59 or SEQ ID NO:60 in search sequence wherein, the 2nd BLAST thereby will be at arabidopsis thaliana sequence).The result who compares first and second blast searches subsequently.If from the high-order position of a BLAST hit be derived from search sequence from identical species of deutero-species wherein, oppositely BLAST produces in the highest search sequence of hitting subsequently ideally, then identifies the collateral line homologue; If the high-order position among the BLAST hit be not derived from search sequence from identical species of deutero-species wherein, and preferably when reverse BLAST, produce the search sequence that belongs to the highest row that hit, then identify directly to homologue.

It is that with low E-value those hit that high-order position is hit.The E-value is low more, mark remarkable more (or in other words, this hit because of the chance odds low more).The calculating of E-value is well-known in the art.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.

Be to be understood that the sequence in the definition that falls into " SYT polypeptide " is not limited to table 6 listed polypeptide (mentioning with sequence table), also be not limited to any polypeptide, described polypeptide comprises from the N-terminal to the C-terminal: (i) the SNH structural domain that has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity with the SNH structural domain of SEQ ID NO:58; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.Preferably, the SNH structural domain comprises the residue that is shown as black among Fig. 6, all can be used for carrying out the inventive method.In addition, the SYT polypeptide can comprise following one or more sequence: (a) SEQ ID NO:146; (b) SEQ ID NO:147; What (c) be positioned at N-terminal before the SNH structural domain is rich in the Met structural domain.Most preferred SYT polypeptide is shown in SEQ ID NO:60, SEQ ID NO:151 or SEQ ID NO:153.

In yeast two-hybrid system, common and GRF (growth regulatory factor) the polypeptide interaction of SYT polypeptide.Yeast two-hybrid interacts and to be determined at that this area is well-known (sees (1989) Nature 340 (6230) such as Field: 245-246).For example, the SYT polypeptide of SEQ ID NO:4 representative can interact with AtGRF5 and with AtGRF9.

Another embodiment of the present invention is an isolated nucleic acid sequences, and it comprises the nucleotide sequence that is selected from down group:

(a) isolated nucleic acid sequences is shown in SEQ ID NO:150 and SEQ ID NO:152;

(b) isolated nucleic acid sequences, the polypeptide of coding shown in SEQ ID NO:151 and SEQ ID NO:153;

(c) isolated nucleic acid sequences, as the result of genetic code degeneracy, its sequence can be inferred out from the polypeptide shown in SEQ ID NO:151 and SEQ ID NO:153;

(d) isolated nucleic acid sequences, its encoded polypeptides with (a) have at least 70% identity to (c) amplifying nucleic acid sequence encoded polypeptide;

(e) isolated nucleic acid sequences, homologue, derivative or the active fragments of the polypeptide of coding shown in SEQ ID NO:151 and SEQ ID NO:153, described homologue, derivative or fragment are plant origin, and advantageously comprise from N-terminal to C-terminal:

(i) the SNH structural domain that has certain sequence identity with the SNH structural domain of SEQ ID NO:58, described sequence identity is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% according to the preferred sequence that increases.

(ii) be rich in the Met structural domain;

(iii) be rich in the QG structural domain;

(f) isolated nucleic acid sequences or its complement, can with above-mentioned (a) to (c) amplifying nucleic acid molecular hybridization, the plant protein in wherein said hybridization sequences or its complement coding (a) to (e); Wherein, compare with control plant, being expressed in of the change of this nucleotide sequence increases output and/or early stage vigor in the plant under abiotic stress.

The nucleotide sequence of the coding SYT polypeptide that provides in the table 6, or any aforementioned SEQ ID NO directly need not be the total length nucleotide sequence to homologue or collateral line homologue because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.

SYT nucleic acid variant can be suitable for implementing method of the present invention.Variant SYT nucleotide sequence is generally those nucleotide sequences that have identical function with naturally occurring SYT nucleotide sequence, it can be identical biological function, or under abiotic stress, when being subjected to regulating in the expression plant of described nucleotide sequence, increase the function of output and/or early stage vigor with respect to control plant.The variant of the SYT nucleotide sequence that such variant comprises the allelic variant of splice variant, the SYT nucleotide sequence of part, the SYT nucleotide sequence of SYT nucleotide sequence hereinafter described, obtain by gene reorganization and/or can with the nucleotide sequence of SYT nucleic acid array hybridizing.Preferably, the nucleic acid variant is the variant as the represented nucleotide sequence of SEQ ID NO:59, SEQ IDNO:150 or SEQ ID NO:152.Term hybridization sequences, splice variant, allelic variant and gene reorganization are as described herein.

The nucleic acid of coding SYT polypeptide needs not be total length nucleic acid, because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.The invention provides the method that in plant, under abiotic stress, strengthens output and/or early stage vigor, it is included in the plant in the nucleotide sequence that provides in the table 6 of introducing and expressing any part, or in the aminoacid sequence that provides in the table 6 any directly to the part of the coding nucleic acid of homologue, collateral line homologue or homologue.

Term as used herein " part " refer to the to encode dna fragmentation of such polypeptide, described polypeptide comprises from the N-terminal to the C-terminal: (i) according to the preferred sequence that increases, and the SNH structural domain that has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity with the SNH structural domain of SEQ ID NO:58; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.Of course, for example, prepare part by in the SYT nucleotide sequence, producing one or more disappearances.Part can be used with isolating form, perhaps they can encode with other (or non-coding) sequence merge with, for example, produce several active polypeptide of combination.When merging with other encoding sequences, the polypeptide that translation produces can be greater than the SYT segment of prediction.Preferably, part is the straight part to homologue or the represented nucleotide sequence of collateral line homologue by the arbitrary or any aforementioned SEQ ID NO that provides in the table 6.Most preferred part is the part as SEQ ID NO:59, SEQID NO:150 or the represented nucleic acid of SEQ ID NO:152.

Another variant of SYT nucleotide sequence is under the stringent condition that reduces, preferably under stringent condition, can with the nucleic acid of defined SYT nucleic acid array hybridizing above, this hybridization sequences coding SYT polypeptide or the part of definition as mentioned.

The invention provides the method that in plant, under abiotic stress, strengthens output and/or early stage vigor, its be included in the plant introduce and express can with the nucleic acid of any nucleic acid hybridization of providing in the table 6, or be included in the plant and introduce and to express nucleic acid like this, its can with any nucleotide sequence of providing in the table 6 directly to the nucleic acid of the coding nucleic acid hybridization of homologue, collateral line homologue or homologue.

The such SYT polypeptide of hybridization sequences coding that is used for the inventive method, described polypeptide comprises from the N-terminal to the C-terminal: (i) according to the preferred sequence that increases, and the SNH structural domain that has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% sequence identity with the SNH structural domain of SEQ ID NO:58; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.Preferably, hybridization sequences be can with the nucleotide sequence that provides in the table 6 or any aforementioned SEQ ID NO directly to homologue or collateral line homologue or the hybridization sequences of the part hybridization of any aforementioned sequence of definition as mentioned.Most preferred hybridization sequences be can with as the represented nucleotide sequence of SEQ ID NO:59, SEQ ID NO:150 or SEQ ID NO:152 or the hybridization sequences of its part (or probe) hybridization.The method of designing probe is well known in the art.Probe usually on length less than 1000bp, preferably on length less than 500bp.Usually, for DNA-DNA hybridization, Southern trace for example, probe length changes between 100-500bp, however at DNA-DNA hybridization, for example in pcr amplification, the hybridization region in the probe is shorter than 50 Nucleotide usually, but be longer than 10 Nucleotide.Hybridization sequences is 100,125,150,175,200 or 225 Nucleotide usually at least on length, preferably be 250,275,300,325,350,375,400,425,450 or 475 Nucleotide at least on length, also preferably be 500,525,550,575,600,625,650,675,700 or 725 Nucleotide at least on length, or the same long with total length SYT cDNA.

Another nucleic acid variant that is used for the inventive method is the splice variant of the SYT polypeptide of definition as mentioned of encoding.Preferred splice variant be the SYT nucleotide sequence that provides in the table 6 or any aforementioned SEQIDNO directly to the splice variant of homologue or collateral line homologue.Most preferred splice variant is the splice variant as the represented SYT nucleotide sequence of SEQ ID NO:59, SEQ ID NO:150 or SEQ ID NO:152.

The invention provides the method that in plant, under abiotic stress, strengthens output and/or early stage vigor, its be included in the plant introduce and expression table 6 in the splice variant of any nucleotide sequence of providing, or the arbitrary amino acid sequence that provides in the table 6 directly to the splice variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

The another kind of nucleic acid variant that is used to implement the inventive method is the allelic variant of coding nucleic acid of SYT polypeptide as hereinbefore defined of encoding.The natural existence of allelic variant, and comprise in the methods of the invention be these natural allelic purposes.Preferred allelic variant be the SYT nucleotide sequence that provides in the table 6 or any aforementioned SEQ ID NO directly to the allelic variant of homologue or collateral line homologue.Most preferably by the splice variant of the SYT nucleotide sequence shown in the SEQ ID NO:59.

The invention provides the method that in plant, under abiotic stress, strengthens output and/or early stage vigor, its be included in the plant introduce and expression table 6 in the allelic variant of any nucleic acid of providing, or be included in introduce in the plant and expression table 6 in the arbitrary amino acid sequence that provides directly to the allelic variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant of the coding SYT polypeptide that obtains by gene reorganization (or orthogenesis).

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant of the coding SYT polypeptide that obtains by site-directed mutagenesis.Site-directed mutagenesis can be used for producing the variant of SYT nucleotide sequence.Several method can be used for realizing site-directed mutagenesis, the method for the modal PCR of being based on (CurrentProtocols in Molecular Biology.Wiley writes).

The invention provides the method that in plant, under abiotic stress, strengthens output and/or early stage vigor, its be included in the plant introduce and expression table 6 in the variant of any nucleotide sequence of providing, or be included in the plant introduce and expression table 6 in the arbitrary amino acid sequence that provides directly to the variant of the coding nucleic acid of homologue, collateral line homologue or homologue, by gene reorganization or site-directed mutagenic obtained this variant nucleic acid.

Following SYT nucleic acid variant is the variant example that is fit to implement the inventive method:

(i) part of SYT nucleotide sequence;

(ii) can with the nucleotide sequence of SYT nucleic acid array hybridizing;

The (iii) splice variant of SYT nucleotide sequence;

The (iv) allelic variant of SYT nucleotide sequence;

(v) reorganize the SYT nucleotide sequence that obtains by gene;

(vi) by site-directed mutagenic obtained SYT nucleotide sequence.

What also be used for the inventive method is such nucleic acid, and it is encoded as the straight homologue to homologue or collateral line homologue of the homologue of the SYT polypeptide that provides in the table 6 or any aforementioned SEQ ID NO.

Also can be used in the inventive method is such nucleic acid, the derivative of the arbitrary SYT nucleotide sequence that provides in its coding schedule 6 or any aforementioned SEQ ID NO directly to the derivative of homologue or collateral line homologue.The straight derivative to homologue or collateral line homologue of any aforementioned SEQ ID NO is another embodiment that can be suitable for the inventive method.The SYT nucleotide sequence can be from any artificial or natural sources, as plant, algae, fungi or animal.Can on composition and/or genome environment, modify the natural form of described nucleotide sequence by careful manual operation.The nucleotide sequence that preferably has the coding SYT polypeptide of plant origin.Can preferably from Cruciferae (Brassicaceae), more preferably separate described nucleotide sequence from the dicotyledons species from Arabidopis thaliana or colea (Brassica napus).Alternatively, can be from pulse family (Fabaceae), preferably from soybean (Glycine max) separated nucleic acid sequence.More preferably, the SYT nucleotide sequence separates certainly:

(a) the SYT polypeptide with shown in SEQ ID NO:60 shown in SEQ ID NO:59 separates from Arabidopis thaliana;

(b) the SYT polypeptide with shown in SEQ ID NO:151 shown in SEQ ID NO:150 separates from colea;

(c) the SYT polypeptide with shown in SEQ ID NO:153 shown in SEQ ID NO:152 separates from soybean.

Term " output ", " seed production " and " early stage vigor " have above been defined.Term " increase ", " improvement ", " enhanced ", " amplification ", " extension " or " rising " can exchange mutually.With the corn is example, the output increase can show as following one or more indexs: the increase of every square metre of plant number, the increase of the increase of every strain plant spike number, the increase of line number, every row grain number, grain weight, thousand seed weight, fringe length/diameter, the full rate of seed (wherein the full rate of seed is that the full seed number is total and multiply by 100 divided by seed) and other.With the rice is example, and the output increase can be by the increase of following one or more indexs performance: the increase of every square metre of plant number, every strain plant panicle number, every panicle spikelet number, every panicle flower (Xiao Hua) number (it is expressed as the ratio of full seed number to former panicle number), the full rate of seed (wherein the full rate of seed be the full seed number divided by the seed sum and multiply by 100), the increase of thousand seed weight and other.

The output that increases also can produce the structure of improvement, or can occur because of the structure of improvement.

According to preferred feature of the present invention, the enforcement of the inventive method is created in the biomass that has increase under the abiotic stress condition with respect to control plant, the seed production of increase and/or the plant of early stage vigor.Thereby, being provided at the method that increases biomass in the plant, seed production and/or early stage vigor under the abiotic stress condition with respect to control plant according to the present invention, described method comprises the expression of nucleotide sequence in plant of regulating coding SYT polypeptide.Over-ground part (leaf biomass) when preferably, the biomass of the increase that this paper quoted means development of plants phase and maturation.Preferably, the biomass of the increase that this paper quoted mean following each: seed ultimate production, full seed number, the full rate of seed, TKW and harvest index.

Because transgenic plant of the present invention have the output of increase under abiotic stress, thereby with respect to the growth velocity of control plant, these plants might be on the corresponding stage in its life cycle performance increases under abiotic stress growth velocity (its life cycle during the small part, the emerging the stage of early stage vigor).The growth velocity that increases can be specific for one or more parts (comprising seed) of plant, or can spread all over whole strain plant basically.The increase of growth velocity can take place during life cycle on the one or more stages in life cycle or in whole plants basically plant.The growth velocity that increases during plant early stage in life cycle can reflect the enhanced vigor.The increase of growth velocity can change the harvest cycle of plant, allows the later sowing of plant and/or than early harvest, otherwise this is with impossible.If growth velocity increases fully, can allow further to sow the seed (for example plant and gather in the crops rice plant, plant and gather in the crops other rice plants subsequently, all rice plant is all in a conventional growth period) of identical plant species.Similarly, if growth velocity sufficiently increases, can allow further to sow the seed (for example sowing and harvesting corn plant are for example sowed and optional results soybean, potato or any other suitable plant subsequently) of different plant species.The results additional times also is possible in the situation of some crop plants from identical rhizome.The harvest cycle that changes plant can cause the increase of every square metre year biomass yield (number of times (as in a year) that can grow and gather in the crops because of any specified plant increases).The increase of growth velocity also can allow cultivating transgenic plant in the geographic area widely than its wild type counterparts, because the region limits of cultivating crop is often determined by the plantation time (season early) or in the adverse environment condition of results period (season in evening).If shorten harvest cycle, then can avoid this class unfavourable condition.Growth velocity can determine that this type of parameter can be by obtain multiple parameter from growth curve: T-Mid (plant reaches the time that its 50% overall dimension is spent) and T-90 (plant reaches the time that its 90% overall dimension is spent), or the like.Under abiotic stress, measure plant growth rate, described abiotic stress such as salt stress; Water is coerced (arid or waterlogging); The nutrition availability that reduces is coerced; Unusual heat or the temperature that cold/freezing temperature causes are coerced; Oxidative stress; Metal is coerced; Chemical toxicity is coerced; Or its combination.

The enforcement of the inventive method is created in the plant that has the growth velocity of increase under the abiotic stress with respect to control plant.Thereby, being provided at the method that increases plant growth rate under the abiotic stress with respect to control plant according to the present invention, described method comprises the expression of nucleotide sequence in plant of regulating coding SYT polypeptide.

Plant is generally replied being exposed to coerce to make by growing slowlyer.Under the condition of serious stress of soil condition, plant even can stop growing fully.Compare with the control plant under the non-stress conditions, slightly coerce and in meaning of the present invention, cause being coerced the plant-growth reduction less than 40%, 35% or 30%, preferably, be more preferably less than 14%, 13%, 12%, 11% or 10% or lower less than 25%, 20% or 15%.Because the progress on the agricultural practice (irrigation, fertilising, pesticide treatments) does not often run into condition of serious stress of soil in the raise crop plant.Therefore, by the agriculture often undesirable characteristic that goes up of the impaired growth of slight stress-inducing.Slightly coercing is coercing of may being exposed to of typical plant.These are coerced is that the common biological and/or inanimate (environment) that plant exposed is coerced.It generally is that those that caused by pathogenic agent such as bacterium, virus, fungi, nematode and insect are coerced that biology is coerced.Typical abiotic stress comprises appoints one or more following coercing: the temperature that the nutrition availability that salt stress, water coerces (arid or waterlogging), reduce coerces, unusual heat or cold/freezing temperature causes is coerced, oxidative stress, metal is coerced or chemical toxicity is coerced.

The enforcement of the inventive method is created under the abiotic stress has the output of increase and/or the plant of early stage vigor with respect to control plant.As report in (Planta (2003) 218:1-14) such as Wang, abiotic stress causes influencing unfriendly a series of morphological change of plant-growth and productivity, physiology to change, biological chemistry changes and molecule changes.What arid, salinity, extreme temperature and oxidative stress normally connected each other also can damage and primary cellular defect by induced growth by similar mechanism.For example, arid and/or salinification mainly show as osmotic stress, cause the destruction of cell homeostasis and ion distribution.Often follow the oxidative stress of high temperature or low temperature, salinity or drought stress can cause functional protein and structural protein sex change.The nutrition availability that reduces, especially the nitrogen availability of Jian Shaoing is the key constraints of plant-growth, for example is used for the amino acid whose availability of albumen synthetic by minimizing.Therefore, these various environment-stress usually activate similar cell signal approach and cell response, as producing stress protein matter, incremental adjustments antioxidant, accumulation compatible solute and growth-inhibiting.

Since diversified environment-stress activates similar path, the present invention should not be considered as being confined to salt stress about illustrating of salt stress, and should be considered as more showing that the SYT polypeptide participates in the screen display information of abiotic stress (screen) in general.Summary among the Plant Science among the TRENDS (Jian-Kang Zhu, the 6th volume, the 2nd phase, February calendar year 2001) confirms that under salt stress better transgenic plant of performance are comprising that quenching, severe cold, heat and arid other coerce down also performance better usually.(Rabbani etc. (Plant Physiol (2003) 133:1755-1767) have described " cross-talk " that drought stress and high salinity are coerced a very high degree.Therefore, obviously SYT polypeptide (as one's own department or unit definition) increases purposes aspect plant biomass and/or the early stage vigor together with it under salt stress, also will increase plant biomass and/or find a place where one can give full play to one's talent aspect the vigor in early days under multiple other abiotic stress.

Term defined herein " abiotic stress " means following one or more aspect: the nutrition availability that salt stress, water coerces (arid or waterlogging), reduce coerces, by unusual heat, cold/temperature that freezing temperature causes coerce, oxidative stress, metal is coerced or chemical toxicity is coerced.The term salt stress is not limited to common salt (NaCl), and can be following any or multiple: NaCl, KCl, LiCl, MgCl ₂, CaCl ₂Or the like.

The enforcement of the inventive method is created in the plant of the output that has increase under the abiotic stress condition with respect to control plant and/or early stage vigor.Thereby, being provided at the method that increases plant biomass and/or early stage vigor under the abiotic stress condition with respect to control plant according to the present invention, described method comprises the expression of nucleotide sequence in plant of regulating coding SYT polypeptide.The abiotic stress of quoting means following one or more aspect: the nutrition availability that salt stress, water coerces (arid or waterlogging), reduce coerces, by unusual heat, cold/temperature that freezing temperature causes coerce, oxidative stress, metal is coerced or chemical toxicity is coerced.

The inventive method advantageously is applicable to any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.

The preferred method that is used for introducing genetic modification is the nucleotide sequence of introducing and expressing coding SYT polypeptide plant.Definition SYT polypeptide is such polypeptide, it comprises from the N-terminal to the C-terminal: (i) according to the preferred sequence that increases, and the SNH structural domain that has at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93,94%, 95%, 96,97%, 98%, 99% sequence identity with the SNH structural domain of SEQ ID NO:58; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.Preferably, the SNH structural domain comprises the residue that is shown as black among Fig. 6.Further preferably, the SNH structural domain is a SEQ ID NO:57 representative.

According to a preferred aspect of the present invention, the expression regulated of SYT nucleotide sequence is the expression that increases.The expression that increases can cause that SYT mRNA or polypeptide level improve, and this equates the activity that improves the SYT polypeptide; Perhaps, perhaps even when the polypeptide level descends also can improve activity when the not variation of polypeptide level.When this situation appears at SYT polypeptide natural characteristics and changes, for example, have more active mutant forms than wild type peptide by preparation.The method that increases or reduce gene or gene product expression is known in the art.

The present invention also provides genetic constructs and carrier to promote to introduce and/or express the nucleotide sequence that is used for the inventive method in plant.

Thereby, gene construct is provided, it comprises:

The nucleotide sequence of (i) nucleotide sequence of coding as top defined SYT polypeptide, or SEQ ID NO:150 or SEQ ID NO:152 representative;

(ii) one or more control sequences that can drive the expression of (i) amplifying nucleic acid sequence; Randomly

(iii) transcription termination sequence.

Preferred construct is a kind of such construct, and wherein regulating and controlling sequence is the promotor from plant, if monocotyledons is transformed preferably from monocotyledons.Used construct can use the well-known recombinant DNA technology of those skilled in the art to make up in the inventive method.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for expressing goal gene in cell transformed.The present invention also provides as defined gene construct purposes in the methods of the invention in the literary composition.

Plant transforms (as the nucleotide sequence of coding SYT polypeptide) with the carrier that comprises aim sequence.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.

Advantageously, can drive the expression of nucleotide sequence with the promotor of any kind.

Useful promotor has constitutive promoter (Benfey etc., EMBO is (1989) 2195-2202 J.8), as derive from those promotors of plant virus, as 35S CAMV (Franck etc., Cell 21 (1980) 285-294), 19S CaMV (also referring to US 5352605 and WO84/02913), 34S FMV (Sanger etc., Plant.Mol.Biol., 14,1990:433-443), the parsley ubiquitin promoter, or plant promoter such as US 4,962, contracting of ribulose diphosphate enzyme-oxygenase (Rubisco) small subunit promotor described in 028 or plant promoter PRP1[Ward etc., Plant.Mol.Biol.22 (1993)], SSU, PGEL1, OCS[Leisner (1988) the Proc NatlAcad Sci U.S. 85 (5): 2553-2557], lib4, usp, mas[Comai (1990) PlantMol Biol 15 (3): 373-381], STLS1, ScBV (Schenk (1999) Plant Mol Biol39 (6): 1221-1230), B33, SAD1 or SAD2 (flax promotor, Jain etc., CropScience, 39 (6), 1999:1696-1701) or (1984) Nucleic Acids Res.12 (20): 7831-7846 such as nos[Shaw].The example of other constitutive plant promoters is beet V-ATPase promotor (WO 01/14572).The example of synthetic constitutive promoter has super promotor (Superpromoter) (WO 95/14098) and derives from the promotor (WO 94/12015) of G-box.In addition, under the suitable situation,, can also use the chemical induced promotor with respect to EP-A 388186, EP-A 335528, WO 97/06268.It is favourable stablizing constitutive expression polypeptide of the present invention in plant.Yet favourable if express the late period before the results, preferably inducible expression's polypeptide of the present invention causes plant production to postpone because metabolism is operated possibly.

Also can promote the expression of plant gene (about summary, referring to Gatz 1997.Annu.Rev.Plant Physiol.Plant Mol.Biol., 48:89-108) by the chemical induced promotor.When expecting with temporal mode expressing gene, the chemical induced promotor is especially suitable.The example of this type of promotor has Induced by Salicylic Acid promotor (WO 95/19443), dormin inducible promoter (EP 335 528), tsiklomitsin inducible promoter, and (Gatz etc. (1992) Plant J.2,397-404), hexalin or alcohol induced property promotor (WO 93/21334), other promotors perhaps as herein described.

Other suitable promotors promotor that to be those react at biology or abiotic stress condition, for example pathogeny evoked PRP1 gene promoter (Ward etc., Plant.Mol.Biol.22 (1993) 361-366), (US 5 for tomato thermal induction hsp80 promotor, 187,267), the cold inducibility α-Dian Fenmei of potato promotor (WO 96/12814) or wound-induced pinII promotor (EP-A-0375091), other promotors perhaps as herein described.

Especially those cause gene in tissue and organ, expression promoter in seed cell such as albuminous cell and developmental protoblast to preferred promotor.Suitable promotor has Semen Brassicae campestris rape napin gene promoter, and (US 5,608,152), broad bean (Vicia faba) USP promotor (Baeumlein etc., Mol Gen Genet, 1991,225 (3): 459-67), Arabidopis thaliana oleosin promotor (WO98/45461), (US 5 for Kidney bean (Phaseolus vulgaris) phaseolin promoter, 504,200), rape Bce4 promotor (WO 91/13980), beans arc5 promotor, Radix Dauci Sativae DcG3 promotor or legumin B4 promotor (LeB4; Baeumlein etc., 1992, Plant Journal, 2 (2): 233-9), and the promotor of in monocotyledons, bringing seed-specific expression, as corn, barley, wheat, rye, rice etc.Favourable seed specific promoters has sucrose-binding proteins promotor (WO 00/26388), phaseolin promoter and napin promotor.The suitable promotor that must consider has barley lpt2 or lpt1 gene promoter (WO 95/15389 and WO 95/23230), and the promotor described in the WO 99/16890 (from the promotor of barley hordein gene, paddy protein gene, rice oryzin gene, rice prolamin gene, wheat gliadin gene, wheat gluten gene, corn zein spirit-soluble gene, avenin gene, Chinese sorghum kasirin gene, rye secaline gene).Other suitable promotors are Amy32b, Amy 6-6 and Aleurain[US 5,677,474], [US 5 for Bce4 (Semen Brassicae campestris rape), 530,149], glycinin (soybean) [EP 571 741], phosphoric acid enol pyruvic acid carboxylase (soybean) [JP 06/62870], ADR12-2 (soybean) [WO 98/08962], isocitrate lyase (Semen Brassicae campestris rape) [US 5,689,040] or α-Dian Fenmei (barley) [EP 781849].Be used in that other promotors of expressing gene have the leaf specificity promoter in the plant, those described in DE-A 19644478; The perhaps promotor of light regulation and control is as pea petE promotor.

Other suitable plant promoters have kytoplasm FBPase promotor or potato ST-LSI promotor (Stockhaus etc., EMBO J.8,1989,2445), the knurl specificity promoter of describing among soybean phosphoribosylpyrophosphate amidotransferase promotor (GenBank accession number U87999) or the EP-A-0 249 676 (node-specific promoter).

In one embodiment, the SYT nucleotide sequence effectively is connected with constitutive promoter.Constitutive promoter is at the great majority of its g and D but must not be all stages transcriptional activations all, and is generally to express basically.Preferred promoter derives from plant, is transformed if also preferably derive from the monocotyledons monocotyledons.Preferred again constitutive promoter is by the GOS2 promotor shown in the nucleotide sequence that is substantially similar to SEQ ID NO:145 or SEQ ID NO:56.Most preferably the GOS2 promotor is shown in SEQ ID NO:56 or SEQ ID NO:145.Be understood that suitability of the present invention is not limited to the SYT nucleotide sequence by SEQ ID NO:59 representative, SYT expression of nucleic acids when suitability of the present invention also is not limited to by the GOS2 promoters driven simultaneously.The example that also can be used for driving other constitutive promoters that coding SYT nucleotide sequence expresses shows in definitional part.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.The additional adjustment element can comprise transcribes and translates reinforcement.One skilled in the art will recognize that and go for implementing terminator of the present invention and strengthen subsequence.Intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also comprise need be used for the replication sequence starting point of keeping and/or duplicating in particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to f1-ori and colE1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Thereby genetic constructs can randomly comprise the selected marker.This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top definitional part.

The present invention also comprises the plant that available the inventive method obtains.Plant, plant part and vegetable cell that therefore the present invention provides available the inventive method to obtain are introduced the SYT nucleotide sequence in the described plant and described plant, plant part and vegetable cell are preferably selected from crop, more preferably are selected from monocotyledons.

The present invention also is provided for being created in the method for the transgenic plant that have enhanced yield and/or early stage vigor under the abiotic stress, and it is included in introduces and express the SYT nucleotide sequence in the plant.

More specifically, the invention provides and be used to be created in the transgenic plant that have enhanced yield and/or early stage vigor under the abiotic stress, preferred monocotyledonous method, described method comprises:

(i) nucleotide sequence of introducing and expression coding SYT polypeptide in plant or vegetable cell; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

(i) nucleic acid can be can encode as the nucleic acid of SYT polypeptide defined above arbitrarily, or the nucleotide sequence shown in SEQ ID NO:150 or SEQ ID NO:152.

The plant offspring who obtains (ii) from incubation step can be bred by several different methods, as by clonal propagation method or classical breeding technique.For example, the first-generation (or T1) transforms plant can carry out selfing, produce the s-generation (or T2) transformant of isozygotying, and the T2 plant breeds further by classical breeding technique.

Nucleotide sequence is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleotide sequence is preferably by transforming introduced plant.

Usually after conversion, vegetable cell or cell colony are selected the existence of one or more marks, wherein said mark becomes whole strain plant with the material regeneration that transforms subsequently by the expressive gene of plant coding that moves with the goal gene corotation.For selecting plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example.Can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying and handles.Another kind of possibility is included in cultivates seed (as required after sterilization) on the agar plate that uses suitable selective agent, to such an extent as to only the seed of Zhuan Huaing can grow into plant.Alternatively, plant transformed is screened the existence of selected marker (those selected markers as indicated above).

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition also can for example use Southern to analyze goal gene were estimated.Alternative or extraly, the expression level of newly introducing DNA can use Northern and/or Western to analyze, and these two kinds of technology are that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clone's property method of proliferating or classical breeding technique.For example, the conversion plant of the first-generation (or T1) can self-pollination produces the s-generation (or T2) transformant of purifying, and the T2 plant further breeds by classical breeding technique.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's property transformant (for example conversion) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the conversion rhizome of unconverted grafting of tender branch.

The present invention extends to any vegetable cell or the plant by described arbitrary method generation herein clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by arbitrary preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical yielding characteristics and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.The present invention also comprises host cell, and it contains separative SYT nucleotide sequence.The preferred host cell of the present invention is a vegetable cell.The present invention also extend to plant the part gathered in the crops as, but be not limited to seed, leaf, fruit, flower, stem, root, root stock, stem tuber and bulb.The invention further relates to from oneself, preferably directly from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, break chop (meal), oil, fat and lipid acid, starch or protein.

According to preferred feature of the present invention, the expression of being regulated is the expression that increases.The method that increases the expression of nucleic acid, gene or gene product is fully put down in writing in this area, and example provides at definitional part.

Alternatively, can regulate the expression of the nucleotide sequence of coding SYT polypeptide by introducing genetic modification, for example can introduce described genetic modification: T-DNA activation, TILLING, homologous recombination or by in plant, introducing and express the nucleotide sequence of the SYT polypeptide of encoding by arbitrary (or multiple) following method.Introducing genetic modification step afterwards is the expression of selecting SYT peptide coding nucleotide sequence to be regulated, and the described expression of being regulated makes plant have the output and/or the early stage vigor of increase under abiotic stress.

A kind of this type of technology is that T-DNA activates mark.The promotor of introducing can be arbitrarily can be in the expectation organism (being plant in this case) drive the promotor of genetic expression.For example, composing type, that organize preference, cell type preference is applicable to all that with epigamic promotor T-DNA activates.Can also use TILLING (the local sudden change of the genome of directional induction) technology to reproduce effect of the present invention.Can also use the homologue reorganization to reproduce effect of the present invention.

The present invention also comprises purposes and the purposes of SYT polypeptide and construct purposes in increasing plant biomass and/or early stage vigor under abiotic stress of definition as mentioned of SYT nucleotide sequence.

Can in the procedure of breeding, use SYT nucleotide sequence or SYT polypeptide, wherein identify the dna marker that can be connected in SYT hereditarily.Can use SYT nucleotide sequence or SYT polypeptide to define molecule marker.Then this DNA or polypeptide marker can be used in the procedure of breeding, have the plant of the output of increase with selection.For example, the SYT gene can be the nucleotide sequence of the SYT nucleotide sequence that provides in the table 6 or any aforementioned SEQ ID NO straight arbitrary representative in homologue or collateral line homologue.

The allelic variant of SYT nucleotide sequence also can be used for the auxiliary procedure of breeding of mark.This class procedure of breeding needs to use sometimes, and for example EMS mutagenesis is introduced allele variant by the plant mutagenic treatment; Alternative, this program can begin with the allele variant of collecting what is called " natural " origin that is not intended to generation.Identify allele variant by for example PCR then.Be to select step subsequently, in order to select the better allele variant of the sequence of discussing, described allele variant is given the output that plant increases.Generally the growth behavior that contains the different allele variant plants of studying sequence to some extent by monitoring is selected, the SYT nucleotide sequence that for example provides in the table 6 or aforementioned any SEQ ID NO straight any different allele variants in homologue or collateral line homologue.Can in greenhouse or field, monitor growth behavior.More optional step comprise, will contain plant and another plant hybridization of better allele variant through evaluation.For example, can make the combination that produces phenotypic characteristic interested in this way.

The SYT nucleotide sequence can also be as probe, is used for for the part of those gene linkage proterties and carry out the mapping of heredity and physics as the gene of its mark.Such information can be used in plant breeding, to obtain having the strain of desired phenotype.This class of SYT nucleotide sequence is used the nucleotide sequence that only needs to grow to few 15 Nucleotide.The SYT nucleotide sequence can be used as restriction fragment length polymorphism (RFLP) mark.Available SYT nucleotide sequence is surveyed the Southern trace (Sambrook J, Fritsch EF and Maniatis T (1989) " molecular cloning: laboratory manual ") of the plant genome DNA of restriction digest.The program that uses a computer subsequently such as MapMaker (Lander etc. (1987) Genomics 1:174-181) carry out genetic analysis to the banding pattern that produces, to make up genetic map.In addition, can use nucleotide sequence to survey the Southern trace in the genomic dna that the restriction enzyme that contains one group of individuality is handled, described one group of individuality is the parent of the clear and definite genetic cross of representative and one group of individuality of filial generation.The separation of record dna polymorphism also is used for calculating formerly position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) with the genetic map SYT nucleotide sequence of this colony's acquisition.

The derive generation and the purposes of probe of the plant gene that uses in genetic mapping is described among Bematzky and Tanksley (1986) the Plant Mol.Biol.Reporter 4:37-41.Described in numerous publications with aforesaid method or its flexible form specific cDNA clone was carried out genetic mapping.For example, can use F2 hybridization colony, backcross population, panmictic population, the homogenic strain of close relative and the mapping of other group of individuals.These class methods are that those skilled in the art are well-known.

Nucleic acid probe also can be used for physical mapping and (promptly settle sequence on physical map; See In:Non-mammalian Genomic Analysis:A Practical Guide such as Hoheisel, Academicpress 1996, the 319-346 pages or leaves, and the reference of wherein quoting).

In another embodiment, nucleic acid probe can be used for direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although the method inclination of FISH mapping at present is used for big clone; See (1995) Genome Res.5:13-20 such as Laan), but the raising of susceptibility allows to use short probe in the FISH mapping.

The multiple method based on nucleic acid amplification that is used for heredity and physical mapping can use described nucleotide sequence to carry out.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med11:95-96), pcr amplified fragment; Sheffield etc. (1993) Genomics16:325-332), allele-specific connects (Landegren etc. (1988) Science 241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc. (1997) Nat.Genet.7:22-28) and Happy mapping (Dear and Cook (1989) Nucleic Acid Res.17:6795-6807).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The method according to this invention obtains having as previously mentioned the plant of the output of increase under abiotic stress.The proterties that these output improve can also make up other favourable economically proterties, as the proterties of more raising output, to the proterties of multiple tolerance of coercing, the multiple structural attitude of improvement and/or biochemistry and/or physiologic character.

The detailed description of cpFBPase polypeptide

Now be surprised to find, with respect to control plant, the expression of nucleotide sequence in plant shoot divides that increases coding chloroplast(id) fructose-1 (cpFBPase) polypeptide increases plant biomass.Thereby, the invention provides the method that increases plant biomass with respect to control plant, it comprises the expression of nucleotide sequence in plant shoot divides that increases coding cpFBPase polypeptide.

One or more control plants that quoting of " control plant " meant any appropriate in this article.Term " reference ", " contrast " or " wild-type " use interchangeably at this paper, and preferably such object, for example organoid, cell, tissue, plant, it is similar to theme of the present invention as far as possible.Reference, contrast or wild-type are similar to theme of the present invention as far as possible on its genome, transcript group, protein group or metabolite group.Preferably, term " reference-", " contrast-" or organoid, cell, tissue or the plant of " wild-type-" relate to such organoid, cell, tissue or plant, in itself and organoid of the present invention, cell, tissue or plant or its part heredity much at one, preferred 95%, 98%, 99.00%, 99.10%, 99.30%, 99.50%, 99.70%, 99.90%, 99.99%, 99.999% or higher identity.Most preferably, " reference ", " contrast " or " wild-type " preferably such object, for example organoid, cell, tissue or plant, it is identical in heredity with the plant, tissue, cell, the organoid that use according to the inventive method, remove nucleotide sequence or its coded gene product be subjected to changing, adjust according to the inventive method or modification.

Unless point out separately, term " polynucleotide ", " nucleic acid " reach " nucleic acid molecule " and exchange use in the text.Unless point out separately, term " peptide ", " polypeptide " reach " protein " and exchange use in this article.Term " sequence " may relate to polynucleotide, nucleic acid, nucleic acid molecule, peptide, polypeptide and protein, depends on the context that uses term " sequence ".Term " gene ", " polynucleotide ", " nucleotide sequence " " nucleotide sequence " or " nucleic acid molecule " are used in reference to the Nucleotide of the polymer form of any length in the text, perhaps are ribonucleotide or are deoxyribonucleotide.The primary structure of molecule only addressed in these terms.

Therefore, term " nucleotide sequence ", " gene ", " polynucleotide ", " nucleotide sequence " or " nucleic acid molecule " comprise double-stranded and single stranded DNA and RNA as used herein.They also comprise the modification of known type, for example methylate, " adding cap ", replace one or more naturally occurring Nucleotide with analogue.Preferred DNA of the present invention or RNA sequence comprise the encoding sequence that coding this paper defines polypeptide.

" encoding sequence " is nucleotide sequence, when under the control that is placed on suitable regulating and controlling sequence, is transcribed into mRNA and/or is translated as polypeptide.The border of encoding sequence is defined as translation stop codon of translation initiation codon and the 3 '-end of 5 '-end.Encoding sequence can be including, but not limited to mRNA, cDNA, recombinant nucleic acid sequence or genomic dna, and also can have intron under certain situation.

Term as defined herein " chloroplast(id) fructose-1 (cpFBPase) polypeptide " refers in chloroplast(id) the performance function and comprises: (i) at least one FBPase structural domain; (ii) the polypeptide of fragment (redox regulatory insertion) is inserted at least one redox regulation and control.

As shown in SEQ ID NO:155, described cpFBPase brings into play function and comprises in chloroplast(id): (i) at least one FBPase structural domain as an example of cpFBPase polypeptide defined above; (ii) fragment is inserted at least one redox regulation and control.Also having this type of example is by SEQ ID NO:157, SEQ ID NO:159, SEQ ID NO:161, SEQ ID NO:163, SEQ ID NO:165, SEQ ID NO:167, SEQ ID NO:169, SEQ ID NO:171, SEQ ID NO:173, SEQ ID NO:175, SEQ ID NO:177, SEQ ID NO:179, SEQ ID NO:181, SEQ ID NO:183, SEQ ID NO:185, SEQ ID NO:187, SEQ ID NO:189, SEQ ID NO:191, SEQ ID NO:193, SEQ ID NO:195 and SEQ ID NO:197, or any aforementioned SEQ ID NO directly in homologue or collateral line homologue shown in any.The present invention is by illustrating from Chlamydomonas reinhardtii sequence plant transformed with the coded polypeptide SEQ ID NO:155 shown in the SEQ ID NO:154.SEQ ID NO:157 (is encoded by SEQ ID NO:156, from Bigelowiella natans), SEQ ID NO:159 (is encoded by SEQ ID NO:158, from Aquilegia formosa x Aquilegia pubescens), SEQ ID NO:161 (is encoded by SEQ IDNO:160, from Arabidopis thaliana), SEQ ID NO:163 (is encoded by SEQ ID NO:162, from colea (Brassica napus)), SEQ ID NO:165 (is encoded by SEQ ID NO:164, from Cyanidioschyzon merolae), SEQ ID NO:167 (is encoded by SEQ ID NO:166, from soybean (Glycine max)), SEQ ID NO:169 (is encoded by SEQ ID NO:168, from tomato (Lycopersicon esculentum)), SEQ ID NO:171 (is encoded by SEQ ID NO:170, from puncture vine clover (Medicago truncatula)), SEQ ID NO:173 (is encoded by SEQ IDNO:172, from tobacco (Nicotiana tabacum)), SEQ ID NO:175 (is encoded by SEQID NO:174, from rice (Oryza sativa)), SEQ ID NO:177 (is encoded by SEQ ID NO:176, from Ostreococcus lucimarinus), SEQ ID NO:179 (is encoded by SEQ ID NO:178, from Ostreococcus tauri), SEQ ID NO:181 (is encoded by SEQ ID NO:180, from Phaeodactylum tricornutum (Phaeodactylum tricornutum)), SEQ ID NO:183 (is encoded by SEQ ID NO:182, from ripe pea), SEQ ID NO:185 (is encoded by SEQ ID NO:184, from trifoliate orange (Poncirus trifoliata)), SEQ ID NO:187 (is encoded by SEQ ID NO:186, from Populus tremuloides (Populus tremuloides)), SEQ ID NO:189 (is encoded by SEQID NO:188, from potato), SEQ ID NO:191 (is encoded by SEQ ID NO:190, from spinach (Spinacia oleracea)), SEQ ID NO:193 (is encoded by SEQ ID NO:192, from wheat), SEQ ID NO:195 (is encoded by SEQ ID NO:194, from Zea mays (Zeamays)) and SEQ ID NO:197 (by SEQ ID NO:196 coding, from small liwan moss (Physicomitrella patens)) be polypeptide SEQ ID NO:155 directly to homologue.

Be to be understood that the sequence in the definition that falls into " cpFBPase polypeptide " is not limited to the given polypeptide of table 7 (mentioning with sequence table), also be not limited to any polypeptide, described polypeptide is brought into play function and is comprised in chloroplast(id): (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control, all can be used for carrying out the inventive method.Preferably, the cpFBPase polypeptide is represented as SEQ ID NO:155.

Yet enforcement of the present invention is not limited to these sequences; Can use as defined herein any cpFBPase coding nucleic acid or cpFBPase polypeptide advantageously to implement the inventive method.

Provided among the table C of this paper embodiment 12 coding cpFBPase polypeptide the nucleic acid example.This type of nucleic acid can be used for implementing method of the present invention.The aminoacid sequence that provides among the table C of embodiment 12 be the cpFBPase polypeptide represented by SEQ ID NO:155 directly to the exemplary sequences of homologue and collateral line homologue, term " directly to homologue " and " collateral line homologue " are as defined herein.Directly can easily find by carrying out so-called interactivity blast search to homologue and collateral line homologue.This can be finished by a BLAST, and a described BLAST participates in submitting to search sequence (for example SEQ ID NO:154or SEQ ID NO:155) to be used for the blast search at arbitrary sequence library (as public's available ncbi database).When nucleotide sequence begins, can use BLASTN or TBLASTX (using the standard default value), and, can use BLASTP or TBLASTN (use standard default value) when when peptide sequence begins.Randomly can screen BLAST result.The full length sequence of submitting The selection result and non-The selection result subsequently to is with at carry out reverse BLAST (the 2nd BLAST) from the sequence of biology, wherein search sequence is from described biology (be under the situation of SEQ ID NO:154 or SEQ ID NO:155 in search sequence wherein, the 2nd BLAST thereby will be at from chlamydomonas (chlamydomonas) sequence).The result who compares first and second blast searches subsequently.If from the high-order position of a BLAST hit be derived from search sequence from identical species of deutero-species wherein, oppositely BLAST produces in the search sequence with the highest hitting (besides itself) subsequently ideally, then identifies the collateral line homologue; If the high-order position among the BLAST hit be not derived from search sequence from identical species of deutero-species wherein, and preferably when reverse BLAST, produce the search sequence that belongs to the highest row that hit, then identify directly to homologue.It is that with low E-value those hit that high-order position is hit.The E-value is low more, mark remarkable more (or in other words, this hit because of the chance odds low more).The calculating of E-value is well-known in the art.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Directly the detailed example to homologue and the evaluation of collateral line homologue provides in embodiment 12.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.Preferably, the cpFBPase polypeptide that is used for the inventive method is in chloroplast(id) performance function and comprise: (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control; (iii), has at least 40%, 45%, 50%, 55%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% sequence identity (calculating is shown in embodiment 14) with SEQ ID NO:155 by the preferred sequence that increases.

Determine to evolve chain (evolutionary linkage) and the improved prediction of secondary structure and tertiary structure in, use the comparison of a plurality of peptide sequences to find conserved domain and feature motif in the protein families.Those skilled in the art can use many programs to implement this alanysis, for example, and the program that the ExPASy protein group instrument cases that Switzerland information biology institute has are provided.It is well-known in the art being used for the method that aligned sequences compares, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.GAP uses Needleman and Wunsch algorithm ((1970) J Mol Biol 48:443-453) to find the overall comparison that makes the maximization of coupling number and make minimized two sequences of room number (the covering whole sequence).BLAST algorithm (Altschul etc. (1990) J Mol Biol 215:403-10) sequence of calculation identity percentage ratio and execution are to the statistical study of similarity between two sequences.Being used to carry out software that BLAST analyzes and being the public, to pass through NCBI (NCBI) obtainable.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.The overall percentage ratio of similarity and identity also can use one of obtainable method in the MatGAT software package and determine (Campanella etc., BMC Bioinformatics.2003 July 10; 4:29.MatGAT: with protein or dna sequence dna and the application that produces similarity/identity matrix).Can carry out trickle edit to optimize the comparison between the conservative motif, apparent as those skilled in the art.As using full length sequence, also can use the ad hoc structure territory in addition to identify substituting of homologue.Use program mentioned above, use default parameters, measured sequence identity value at whole nucleotide sequence or aminoacid sequence and/or at structural domain or the one or more conservative motif selected.

Term " structural domain " and " motif " define as the definition part branch.There is the specialized database that is used to identify structural domain.FBPase structural domain in the cpFBPase polypeptide can use for example SMART ((1998) Proc.Natl.Acad.Sci. U.S. 95 such as Sch ultz, 5857-5864; Letunic etc. (2002) Nucleic Acids Res 30,242-244; EMBL by Heidelberg, Germany has), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318; European bioinformation institute (EBI) by Britain has), (Bucher and Bairoch (1994) are used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and at the function of automatization sequence interpretation, (In) ISMB-94 to Prosite; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)), ExPASy protein group server that provides to scientific circles as a service (the Switzerland information biology institute (SIB) by Switzerland has) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002) is had by the Sanger institute of Britain) identified.For example, in the InterPro database, the FBPase structural domain that is included in the cpFBPase polypeptide is named as IPR000146.Relate to the cpFBPase polypeptide of representing by SEQ ID NO:155, the accession number that identifies in the mass data storehouse is listed among the embodiment 15.

An important motif that is included in the FBPase structural domain is that fragment is inserted in the redox regulation and control, and described motif only is present in the cpFBPase polypeptide, and is not present in the cyFBPase polypeptide.Compare all FBPases polypeptide by the method for using (and Figure 12) among mentioned above and the embodiment 13, identify to comprise that disulfide linkage forms the insertion fragment amino acid of (being redox regulation) necessary at least two cysteine residues.Conservative halfcystine is with them the position name in ripe pea (Pisum sativa) polypeptide, i.e. Cys153, Cys173 and Cys178.Cys153 and Cys173 normally participate in two mating partners that disulfide linkage forms (people such as Chiadmi. (1999) EMBO J 18 (23): 6809-6815).Cys153 is encircled with Cys173 to be separated, and the length of described ring is 10,11,12,13,14,15,16,17,18,19,20,21,22,23,24 or 25 amino-acid residues, is preferably 14,15,16,17,18 or 19 amino-acid residues.

The task of Prediction of Protein Subcellular Location is important and is fully studied.Know that proteinic being positioned with helps illustrate its function.The localized experimental technique of albumen is contained from immunolocalization to the protein labeling that uses green fluorescent protein (GFP).These class methods are accurately, are labour-intensive although compare with method of calculation.Recently, localized calculating prediction has obtained very much progress from the albumen of sequence data.Algorithm well-known to those skilled in the art can obtain in the ExPASy protein group instrument that Switzerland information biology institute has such as Psort, TargetP, ChloroP, Predotar, LipoP, MITOPROT, PATS, PTS1, SignalP etc.The evaluation of the Subcellular Localization of polypeptide of the present invention is shown in embodiment 16.Particularly, in plastid (chloroplast(id)) compartment of SEQ ID NO:155 of the present invention photosynthetic by arranging (autotrophy) cell.

The method that is targeted to plastid is well known in the art and comprises the use transit peptides.Following table 7 has shown can be used for any FBPase polypeptide is targeted to the transit peptides example of plastid, and the FBPase polypeptide of its natural form normally is not targeted to plastid, and perhaps the FBPase polypeptide of its natural form is targeted to plastid by the effect of different transit peptides (for example its natural transit peptides).The nucleotide sequence of cyFBPase of for example encoding also can be suitable for method of the present invention, as long as described nucleic acid is targeted to plastid, preferably arrive chloroplast(id), and described nucleic acid comprises at least one regulation and control redox insertion fragment.

Table 7: the example that amino acid is targeted to the transit peptide sequence of plastid

The NCBI accession number	The source is biological	Protein function	Transit peptide sequence
				P07839	Chlamydomonas	Ferredoxin	MAMAMRSTFAARVGAKPAVRGARPAS RMSCMA
AAR23425	Chlamydomonas	The rubisco activating enzyme	MQVTMKSSAVSGQRVGGARVATRSVR RAQLQV
				CAA56932	Arabidopis thaliana	The Asp transaminase	MASLMLSLGSTSLLPREINKDKLKLGTS ASNPFLKAKSFSRVTMTVAVKPSR
CAA31991	Arabidopis thaliana	Acyl carrier protein 1	MATQFSASVSLQTSCLATTRISFQKPALI SNHGKTNLSFNLRRSIPSRRLSVSC
				CAB63798	Arabidopis thaliana	Acyl carrier protein 2	MASIAASASISLQARPRQLAIAASQVKSF SNGRRSSLSFNLRQLPTRLTVSCAAKP ETVDKVCAVVRKQL
CAB63799	Arabidopis thaliana	Acyl carrier protein 3	MASIATSASTSLQARPRQLVIGAKQVKS FSYGSRSNLSFNLRQLPTRLTVYCAAK PETVDKVCAVVRKQLSLKE

CpFBPase polypeptide shown in SEQ ID NO:155 is to have (the EC of EC; Enzyme classification based on enzymatic reaction) numbering EC 3.1.3.11 is the enzyme of fructose-diphosphatase.CpFBPase polypeptide catalysis fructose-1,6-diphosphate is to the irreversible conversion of fructose-6-phosphate and Pi.This functional assays can be based on the mensuration of the cpFBPase determination of activity of Pi colorimetric, as Huppe and Buchanan (1989), described in Naturforsch.44c:487-494.Other methods of measuring enzymic activity are described in Plant Physiol 70:728-734 by Alscher-Herman (1982).

" bring into play function in chloroplast(id) " herein meaning the cpFBPase polypeptide is activated in chloroplast(id), and promptly the cpFBPase polypeptide carries out being hydrolyzed to the enzymatic reaction that fructose-6-phosphate and Pi form by fructose-bisphosphate in chloroplast(id).

The straight nucleotide sequence to homologue or collateral line homologue of the cpFBpase polypeptide that provides in the coding schedule 7 or aforementioned any SEQ ID NO needs not be total length nucleic acid, because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.

The nucleic acid variant of cyFBPase also can be suitable for implementing method of the present invention.Variant cyFBPase nucleotide sequence is generally those nucleotide sequences that have identical function with naturally occurring cyFBPase nucleotide sequence, it can be identical biological function, or when the expression of described nucleotide sequence in plant shoot divides is increased, increase the function of output with respect to control plant.Such cyFBPase variant example comprises the part of nucleotide sequence naturally occurring or that obtain by DNA operation, the cyFBPase nucleotide sequence that can obtain with the nucleotide sequence of cyFBPase hybridization, splice variant, allelic variant, by gene reorganization or by site-directed mutagenic obtained cyFBPase nucleotide sequence.

As used herein term " part " is meant the dna fragmentation of coded polypeptide, and described dna fragmentation is brought into play function and comprised in chloroplast(id): (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control.

For example, can prepare part by the nucleic acid of coding cpFBPase polypeptide of definition is as mentioned carried out one or more disappearances.Part can be used with isolating form, perhaps itself and other coding (or non-coding) sequence can be merged, so that for example, produce and made up some active polypeptide.In another embodiment, the transit peptide sequence of natural generation can be replaced by the transit peptide sequence from another photosynthetic organ, or is synthesized the transit peptide sequence replacement.Transform if consider chloroplast(id), can shift transit peptide sequence in the lump.When merging with other encoding sequences, the polypeptide that is produced after translating may be bigger than the cpFBPase part of prediction.The part normal length that is used for the inventive method is at least 900 Nucleotide, and preferred length is at least 1000 Nucleotide, more preferably length be at least 1100 Nucleotide and most preferably length be at least 1200 Nucleotide.Preferably, part is the part of the nucleotide sequence that provides in the table 7 or aforementioned any SEQ ID NO straight any represented nucleotide sequence in the nucleic acid sequence encoding of homologue or collateral line homologue.Most preferred part is the part of the nucleotide sequence represented of SEQ IDNO:154.

According to the present invention, the method that increases plant biomass is provided, it comprise in the nucleotide sequence that provides among the table C that increases embodiment 12 any part or the table C of embodiment 12 in the expression of part in plant of straight coding nucleic acid to homologue, collateral line homologue or homologue of any aminoacid sequence of providing.

Another nucleic acid variant that is used for the inventive method is under the stringent condition that reduces, preferably under stringent condition, can with the nucleic acid sequence encoding of defined cpFBPase polypeptide above or with the nucleic acid of defined part hybridization above.The present invention of root a tree name, the method that increases plant biomass is provided, it comprise increase can with the nucleic acid of any hybridization in the nucleotide sequence that provides among the table C of embodiment 12 in plant expression or comprise can with any nucleotide sequence of providing among the table C of embodiment 12 directly to the expression of nucleic acid in plant of homologue, collateral line homologue or homologue hybridization.

Be used for the hybridization sequences coding cpFBPase polypeptide of the inventive method, described cpFBPase polypeptide is brought into play function and is comprised in chloroplast(id): (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control, and has and the essentially identical biologic activity of cpFBPase polypeptide shown in the SEQ ID NO:155.The method of designing probe is well known in the art.Hybridization sequences usually on length less than 1000bp, preferably on length less than 900,800,700,600 or 500bp.Usually, for DNA-DNA hybridization, Southern trace for example, probe length changes between 100-500bp, however at DNA-DNA hybridization, for example in pcr amplification, probe is shorter than 50 Nucleotide usually, but be longer than 10 Nucleotide.Preferably, hybridization sequences is such hybridization sequences, its can with the straight nucleic acid array hybridizing of arbitrarily represented nucleotide sequence (or deutero-probe) or any aforementioned SEQ ID NO that encodes to homologue or collateral line homologue as the nucleotide sequence that provides in the table 7, or with the hybridization of the part of any aforementioned sequence, part is definition as mentioned.Most preferably hybridization sequences can be hybridized with SEQ ID NO:154 or with its part (or probe).

Another nucleic acid variant that is used for the inventive method is the splice variant of the cpFBPase polypeptide of definition as mentioned of encoding.This type of variant will be the variant that has wherein kept proteinic biologic activity basically; This can realize by the proteinic functional sections of selective retention.Preferred splice variant be the cpFBPase nucleotide sequence that provides in the table 7 or any aforementioned SEQ ID NO that encodes directly to the splice variant of the nucleotide sequence of homologue or collateral line homologue.Most preferred splice variant is the splice variant as the represented cpFBPase nucleotide sequence of SEQ IDNO:154.

The another kind of nucleic acid variant that is used to implement the inventive method is the allelic variant of nucleotide sequence of cpFBPase polypeptide as hereinbefore defined of encoding.The natural existence of allelic variant, and comprise in the methods of the invention be these natural allelic purposes.Preferred allelic variant be the cpFBPase nucleotide sequence that provides in the table 7 or any aforementioned SEQ ID NO that encodes directly to the allelic variant of the nucleotide sequence of homologue or collateral line homologue.Most preferably by the splice variant of the cpFBPase nucleotide sequence shown in the SEQ ID NO:154.

The invention provides in plant the method that increases output, its be included in the arbitrary amino acid sequence that provides among the table C of the splice variant that increases any nucleotide sequence that provides among the table C that expresses embodiment 12 in the plant or embodiment 12 directly to the splice variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant of the coding cpFBPase polypeptide that obtains by gene reorganization (or orthogenesis).Most preferably reorganize the nucleic acid variant that obtains by the gene of the cpFBPase nucleotide sequence shown in SEQ ID NO:155.

The invention provides the method that in plant, increases output, its be included in the plant variant that increases any nucleotide sequence that provides among the table C that expresses embodiment 12 or be included in increase in the plant arbitrary amino acid sequence that provides among the table C that expresses embodiment 12 directly to the variant of the coding nucleic acid of homologue, collateral line homologue or homologue, described variant nucleic acid obtains by gene reorganization.

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant of the coding cpFBPase polypeptide that obtains by site-directed mutagenesis.Site-directed mutagenesis also can be used for producing the variant of cpFBPase nucleotide sequence.Several method can be used for realizing site-directed mutagenesis, the method for the modal PCR of being based on (Current Protocols in Molecular Biology.Wiley writes).For example, change into Serine for one that influences sudden change that disulfide linkage forms and be in will conservative halfcystine, produce thus the cpFBPase constitutive activity (Chiadmi etc., (1999) EMBO J 18 (23): 6809-6815). the site-directed mutagenic obtained nucleic acid variant of the cpFBPase nucleotide sequence by shown in SEQ ID NO:154 most preferably.

Following cpFBPase nucleic acid variant is the variant example that is fit to implement the inventive method:

(i) part of cpFBPase nucleotide sequence;

(ii) can with the nucleotide sequence of cpFBPase nucleic acid array hybridizing;

The (iii) splice variant of cpFBPase nucleotide sequence;

The (iv) allelic variant of cpFBPase nucleotide sequence;

(v) reorganize the cpFBPase nucleotide sequence that obtains by gene;

(vi) by site-directed mutagenic obtained cpFBPase nucleotide sequence

Also can be used in the inventive method is such nucleotide sequence, the homologue of its coding as the cpFBPase polypeptide that provides in the table 7 or the straight homologue to homologue or collateral line homologue of any aforementioned SEQ ID NO that encodes.

Also can be used in the inventive method is such nucleotide sequence, the derivative of the arbitrary cpFBPase nucleotide sequence that provides in its coding schedule 7 or any aforementioned SEQ ID NO directly to the derivative of homologue or collateral line homologue.Provide in the table 7 arbitrary shown in the cpFBPase polypeptide or any aforementioned SEQ ID NO be another example that can be suitable for the inventive method directly to the derivative of homologue or collateral line homologue.

The nucleotide sequence of coding cpFBPase polypeptide can be from any artificial or natural sources, as plant, algae, diatom or animal.Can on composition and/or genome environment, modify the natural form of described nucleotide sequence by careful manual operation.The nucleotide sequence of optimized encoding cpFBPase polypeptide is from photosynthetic cells (vegitabilia). and the nucleotide sequence of going back optimized encoding cpFBPase polypeptide is from vegetable cell.More preferably encode the nucleotide sequence of cpFBPase polypeptide from frustule.Most preferably encode the nucleotide sequence of cpFBPase polypeptide from algae (red algae, brown alga or green alga) cell.Can be subordinated to the green alga of Chlorophyta (Chlorophyta) or charophyta (Charophyta) or separate described nucleotide sequence from terrestrial plant (non-vascular plant or vascular plant).For example, the nucleotide sequence of coding cpFBPase polypeptide separates certainly: the Chlamydomonas species, chlorella species (Chlorella sp.), Bigelowiella natans, Cyanidioschyzon merolae, Ostreococcus lucimarinus, Ostreococcus tauri, Galderia sulphuraria, small liwan moss, Phaeodactylum tricornutum, Aquilegia formosa x Aquilegia pubescen, Arabidopis thaliana, colea, soybean, tomato, the puncture vine clover, tobacco, rice, ripe pea, trifoliate orange (Poncirus trifoliate), Populus tremuloides, potato, spinach, wheat, Zea mays etc.The nucleotide sequence of cpFBPase polypeptide of most preferably encoding separates from Chlamydomonas reinhardtii.

The enforcement of the inventive method produces has the plant that increases output.Term " output ", " increase ", " improvement ", " enhanced ", " amplification ", " extension ", " reinforcement " or " raising " be can exchange mutually and above define.The biomass that increases can show as the root biomass of increase.The root biomass that increases can be owing to the radical amount that increases, the root thickness and/or the root length of increase.The output that increases can show as following one or more indexs:

(i) the one or more parts of plant, particularly the root biomass of the biomass (weight) of (can gather in the crops) part increase on the ground, increase or any other can be gathered in the crops the biomass that part increases; (ii) the early stage vigor of Zeng Jiaing is defined as the ground area of sprouting back three all seedling in the literary composition;

The (iii) seed ultimate production of Zeng Jiaing, this comprises the increase of seed biomass (seed weight), and it can be that seed weight on every plant or the single seed basis increases;

(iv) every strain plant panicle number of Zeng Jiaing;

(the v) paniculiform flower of each of Zeng Jiaing (" small ear floret ") quantity;

(the vi) full rate of the seed of Zeng Jiaing;

(vii) (full) seed amount of Zeng Jiaing;

(the viii) seed size of Zeng Jiaing (length and width area, girth), this also can influence the composition of seed;

(ix) the seed volume of Zeng Jiaing, this also can influence the composition of seed;

(x) harvest index of Zeng Jiaing, it is expressed as can gather in the crops the part output of [as seed] and the ratio of total biomass; With

(xi) thousand seed weight of Zeng Jiaing (TKW), this is by counting full seed quantity and gross weight thereof, and extrapolating obtains.TKW increases the increase that can come from seed size and/or seed weight.TKW increases the increase that can come from embryo size and/or endosperm size.

Increase aspect seed size, seed volume, seed area, seed girth, seed width and seed length can be owing to the increase of seed specific part, as the increase owing to the size of other parts of embryo and/or endosperm and/or aleuron and/or scultellum or seed.

Especially, the output of increase is meant the seed production of increase and is selected from one or more following index: (i) seed weight of Zeng Jiaing; The (ii) full seed quantity of Zeng Jiaing; The (iii) full rate of the seed of Zeng Jiaing; The (iv) harvest index of Zeng Jiaing.

With the corn is example, and the output increase can show as following one or more indexs: the increase of the increase of the increase of every square metre of plant number, every strain plant spike number, line number, every row grain number, grain weight, thousand seed weight, fringe length/diameter, the increase of the full rate of seed (wherein the full rate of seed is that the full seed number is total and multiply by 100 divided by seed) and other.

With the rice is example, and itself can show as the increase of following one or more indexs the output increase: the increase of every square metre of plant number, every strain plant panicle number, every panicle spikelet number, every panicle flower (Xiao Hua) number (it is expressed as the ratio of full seed number to former panicle number), the full rate of seed (wherein the full rate of seed be the full seed number divided by the seed sum and multiply by 100), the increase of thousand seed weight and other.

The output that increases also can produce the structure of improvement, or can occur because of the structure of improvement.

According to preferred feature of the present invention, the enforcement of the inventive method produces the plant that has the increase seed production with respect to control plant.Thereby, providing the method that increases seed production according to the present invention, described method comprises the expression of nucleotide sequence in the over-ground part of plant that increases coding cpFBPase polypeptide.

Because transgenic plant of the present invention have the output of increase, thereby with respect to the growth velocity of control plant, these plants might show on the corresponding stage in its life cycle increase growth velocity (its life cycle during the small part).

The growth velocity that increases can be specific for one or more parts (comprising seed) of plant, or can spread all over whole strain plant basically.Plant with growth velocity of increase can possess short life cycle.The life cycle of plant can be considered as meaning from dry mature seed and grow to the needed time in stage that plant has produced the dry mature seed similar to parent material.This life cycle can be influenced by following factors, as early stage vigor, growth velocity, green degree index, flowering time and seed maturity speed.The increase of growth velocity can take place during life cycle on the one or more stages in life cycle or in whole plants basically plant.The growth velocity that increases during plant early stage in life cycle can reflect the enhanced vigor.The increase of growth velocity can change the harvest cycle of plant, allows the later sowing of plant and/or than early harvest, otherwise this can not (similar effect can obtain with flowering time early).If growth velocity increases fully, can allow further to sow the seed (for example sow and gather in the crops rice plant, sow and gather in the crops other rice plants subsequently, all rice plant is all in a conventional growth period) of identical plant species.Similarly, if growth velocity sufficiently increases, can allow further to sow the seed (for example sowing and harvesting corn plant are for example sowed and optional results soybean, potato or any other suitable plant subsequently) of different plant species.The results additional times also is possible in the situation of some crop plants from identical rhizome.The harvest cycle that changes plant can cause the increase of every acre year biomass yield (number of times (as in a year) that can grow and gather in the crops because of any specified plant increases).The increase of growth velocity also can allow cultivating transgenic plant in the geographic area widely than its wild type counterparts, because the region limits of cultivating crop is often determined by the plantation time (season early) or in the adverse environment condition of results period (season in evening).If shorten harvest cycle, then can avoid this class unfavourable condition.Growth velocity can determine that this type of parameter can be by obtain multiple parameter from growth curve: T-Mid (plant reaches the time that its 50% overall dimension is spent) and T-90 (plant reaches the time that its 90% overall dimension is spent), or the like.

According to preferred feature of the present invention, the plant of the growth velocity with increase is compared in the enforcement generation of the inventive method with control plant.Thereby, providing the method that increases plant growth rate according to the present invention, described method comprises adjusting, the preferred expression of nucleic acid in plant that increases coding cpFBPase polypeptide as defined herein.

Compare with control plant, no matter plant is under the non-stress conditions still is that plant is exposed to multiple coercing down, and the increase of output and/or growth velocity all takes place.Plant is generally replied being exposed to coerce to make by growing slowlyer.Under the condition of serious stress of soil condition, plant even can stop growing fully.On the other hand, slightly coerce and be defined as plant in this article any of its exposure coerced, the wherein said ability that does not cause plant to stop growing fully and recover growth of coercing.Compare with the control plant under the non-stress conditions, slightly coerce and in meaning of the present invention, cause being coerced the plant-growth reduction less than 40%, 35% or 30%, preferably, be more preferably less than 14%, 13%, 12%, 11% or 10% or lower less than 25%, 20% or 15%.Because the progress on the agricultural practice (irrigation, fertilising, pesticide treatments) does not often run into condition of serious stress of soil in the raise crop plant.Therefore, by the agriculture often undesirable characteristic that goes up of the impaired growth of slight stress-inducing.Slightly coerce is that the common biological and/or inanimate (environment) that plant exposes is coerced.Abiotic stress can because of arid or waterlogging, anaerobism are coerced, due to salt stress, chemical toxicity, oxidative stress and heat, cold or the freezing temperature.Abiotic stress can be to coerce (especially because arid), salt stress, oxidative stress or ion by water to coerce the osmotic stress that causes.It generally is that those that caused by pathogenic agent such as bacterium, virus, fungi and insect are coerced that biology is coerced.

Especially, method of the present invention can implemented the plant that has the output of increase with respect to control plant to produce under the non-stress conditions or under slight drought condition.As report in (Planta (2003) 218:1-14) such as Wang, abiotic stress causes influencing unfriendly a series of morphological change of plant-growth and productivity, physiology to change, biological chemistry changes and molecule changes.Known arid, salinity, extreme temperature and oxidative stress are also can damaging and primary cellular defect by induced growth by similar mechanism of connecting each other.Rabbani etc. (Plant Physiol (2003) 133:1755-1767) have described " cross-talk " that drought stress and high salinity are coerced a very high degree.For example, arid and/or salinification mainly show as osmotic stress, cause the destruction of cell homeostasis and ion distribution.Often follow the oxidative stress of high temperature or low temperature, salinity or drought stress can cause functional protein and structural protein sex change.Therefore, these various environment-stress usually activate similar cell signal approach and cell response, as producing stress protein matter, raising antioxidant, accumulation compatible solute and growth-inhibiting.Term as used in this article " non-coercing " condition is the envrionment conditions that allows the plant optimum growh.Those skilled in the art know that normal edaphic condition and weather condition for given place.

The enforcement of the inventive method is with respect to the appropriate control plant of cultivating under comparable conditions, gives under the non-stress conditions or the output that increases of the plant of cultivating under slight drought condition.Thereby according to the present invention, be provided under the non-stress conditions or increase the method for output in the plant of under slight drought condition, cultivating, described method comprises that the nucleic acid that increases coding cpFBPase polypeptide expresses in plant.

The enforcement of the inventive method produces with respect to the appropriate control plant of cultivating under comparable conditions, the plant of under the nutritive deficiency condition, especially cultivating under nitrogen shortage condition that increases output that has.Thereby, increasing the method for output in the plant that is provided under the nutritive deficiency condition, cultivating according to the present invention, described method comprises the expression of nucleic acid in plant that increases coding cpFBPase polypeptide.Nutritive deficiency can be because of due to the nutraceutical shortage, and described nutrition for example is nitrogen, phosphoric acid salt and other P contained compounds, potassium, calcium, cadmium, magnesium, manganese, iron and boron etc.

The increase of preferred output and/or growth velocity occurs under non-stress conditions or slight abiotic or biological stress conditions according to the inventive method.

Term " expression " or " genetic expression " mean transcribing of one or more genes of Yin Teding or gene construct.Term " expression " or " genetic expression " especially mean one or more genes or gene construct is transcribed into structure RNA (rRNA, tRNA) or mRNA, and being with or without subsequently, mRNA translates into protein.This process comprises that DNA transcribes, processes the mRNA product that obtains.

Above defined term " expression of increase ".The nucleotide sequence that increases coding cpFBPase polypeptide is the expression of the part output that causes plant to increase with respect to control plant on the ground.Preferably, expression of nucleic acid increase to endogenous plant cpFBPase expression of polypeptides 1.25,1.5,1.75,2,5,7.5,10,15,20,25,30,35,40,45,50,55,60,65,70,75,80,85,90,95,100 or more times.

This paper quotes term " plant shoot branch " and means plant part, and described plant part does not comprise root, Gen Mao and any other plant part that (promptly directly is not exposed under the illumination) in soil.

The expression (in plastid) of the nucleotide sequence by increasing coding cpFBPase polypeptide obtains the increase of cpFBPase polypeptide quantity.The increase of this cpFBPase polypeptide (in plastid) quantity causes the active increase of cpFBPase.Alternatively, when cpFBPase polypeptide quantity does not change or when cpFBPase polypeptide quantity has minimizing, also can increase activity.When this situation appears at cpFBPase polypeptide natural characteristics and changes, for example, have more active mutant forms than wild type peptide by preparation.

Use technology known in the art to increase the expression of nucleotide sequence in plastid of coding cpFBPase polypeptide, as by using transit peptide sequence that the cpFBPase polypeptide is targeted to plastid or directly being transformed in the plastid by the cpFBPase polypeptide that does not have transit peptide sequence.Can in any plastid, increase expression, express but preferably in chloroplast(id), preferably increase.

Can regulate the expression of the nucleotide sequence of coding cpFBPase polypeptide by introducing genetic modification, can introduce described genetic modification by arbitrary (or multiple) following method: T-DNA activation, TILLING, homologous recombination, or by in plant, introducing and express the nucleotide sequence of coding cpFBPase polypeptide.Introduce step after the genetic modification and be the expression of the increase of the nucleotide sequence of selecting coding cpFBPase polypeptide, the expression of described increase produces the plant that has the output of increase with respect to control plant.

A kind of this type of technology is that T-DNA activates mark.The promotor of introducing can be arbitrarily can be in the expectation organism (being plant in this case) drive the promotor of genetic expression.For example, composing type, over-ground part, underground part, organize preference, the cell type preference be applicable to all that with epigamic promotor T-DNA activates.Can also use the reorganization of TILLING or homologue to reproduce effect of the present invention.

The preferred method that is used to introduce genetic modification is to divide the nucleotide sequence of introducing and expressing coding cpFBPase polypeptide at plant shoot.CpFBPase as defined herein refers to polypeptide, and described polypeptide is brought into play function and comprised in chloroplast(id): (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control.

In one embodiment of the invention, the expression of the nucleotide sequence of coding cpFBPase polypeptide is the expression (in the over-ground part of plant) that increases.The expression that increases can cause that cpFBPasemRNA or polypeptide level improve, and this equates the activity that improves the cpFBPase polypeptide; Perhaps, perhaps even when the polypeptide level descends also can improve activity when the not variation of polypeptide level.When this situation appears at cpFBPase polypeptide natural characteristics and changes, for example, have more active mutant forms than wild type peptide by preparation.The method that increases or reduce genetic expression or gene product is known in the art.

The present invention also provides genetic constructs and carrier to promote to introduce and/or express the nucleotide sequence that is used for the inventive method in plant.

Thereby, gene construct is provided, it comprises:

(i) nucleotide sequence of coding as top defined cpFBPase polypeptide;

(ii) one or more control sequences that can drive the expression of (i) amplifying nucleic acid sequence in plant shoot divides; Alternatively

(iii) transcription termination sequence.

Preferred construct is a kind of such framework body, and wherein regulating and controlling sequence is the promotor from plant, if monocotyledons is transformed preferably from monocotyledons.

Used construct can use the well-known recombinant DNA technology of those skilled in the art to make up in the inventive method.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for expressing goal gene in cell transformed.The present invention also provides as defined gene construct purposes in the methods of the invention in the literary composition.

Plant transforms (as the nucleotide sequence of coding cpFBPase polypeptide) with the carrier that comprises aim sequence.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.Term " regulatory element ", " regulating and controlling sequence " and " promotor " all are used interchangeably at this paper, and in above definition.

Advantageously, the expression promoter that is used to drive nucleotide sequence is to organize the promotor of preference, promptly be can be preferentially in particular organization such as leaf, stem, seed tissue etc. the preferential promotor of transcribing that starts.Only can in particular organization, start the promotor this paper that transcribes and be called " tissue-specific ".Similarly, only in specific cells, start the promotor this paper that transcribes and be called " cell-specific ".In addition or alternatively, promotor can produce naturally or be synthetic.Preferably, the nucleic acid that promotor can drive coding cpFBPase polypeptide promptly is exposed to expression in the part of illumination at the over-ground part of plant, is used for suitable cpFBPase redox regulation and control.

Other suitable promotors have constitutive promoter (Benfey etc., EMBO is (1989) 2195-2202 J.8), as derive from those promotors of plant virus, as 35S CAMV (Franck etc., Cell 21 (1980) 285-294), 19S CaMV (also referring to US 5352605 and WO84/02913), 34S FMV (Sanger etc., Plant.Mol.Biol., 14,1990:433-443), the parsley ubiquitin promoter, or plant promoter such as US 4,962, contracting of ribulose diphosphate enzyme-oxygenase (Rubisco) small subunit promotor described in 028 or plant promoter PRP1[Ward etc., Plant.Mol.Biol.22 (1993)], SSU, PGEL1, OCS[Leisner (1988) the Proc NatlAcad Sci U.S. 85 (5): 2553-2557], lib4, usp, mas[Comai (1990) PlantMol Biol 15 (3): 373-381], STLS1, ScBV (Schenk (1999) Plant Mol Biol39 (6): 1221-1230), B33, SAD1 or SAD2 (flax promotor, Jain etc., CropScience, 39 (6), 1999:1696-1701) or (1984) Nucleic Acids Res.12 (20): 7831-7846 such as nos[Shaw].The example of other constitutive plant promoters is beet V-ATPase promotor (WO 01/14572).The example of synthetic constitutive promoter has super promotor (Superpromoter) (WO 95/14098) and derives from the promotor (WO 94/12015) of G-box.In addition, under the suitable situation,, can also use the chemical induced promotor with respect to EP-A 388186, EP-A 335528, WO 97/06268.It is favourable stablizing constitutive expression polypeptide of the present invention in plant.Yet favourable if express the late period before the results, preferably inducible expression's polypeptide of the present invention causes plant production to postpone because metabolism is operated possibly.

Also can promote the expression of plant gene by the chemical induced promotor.The example of this type of promotor has Induced by Salicylic Acid promotor (WO 95/19443), dormin inducible promoter (EP 335 528), tsiklomitsin inducible promoter, and (Gatz etc. (1992) Plant J.2,397-404), hexalin or alcohol induced property promotor (WO 93/21334), other promotors perhaps as herein described.

Other suitable promotors promotor that to be those react at biology or abiotic stress, for example pathogeny evoked PRP1 gene promoter (Ward etc., Plant.Mol.Biol.22 (1993) 361-366), (US 5 for tomato thermal induction hsp80 promotor, 187,267), the cold inducibility α-Dian Fenmei of potato promotor (WO 96/12814) or wound-induced pinII promotor (EP-A-0 375 091), other promotors perhaps as herein described.

Especially those influence gene in tissue and organ, expression promoter in seed cell such as albuminous cell and developmental protoblast to preferred promotor.Suitable promotor has Semen Brassicae campestris rape napin gene promoter, and (US 5,608,152), broad bean (Vicia faba) USP promotor (Baeumlein etc., Mol Gen Genet, 1991,225 (3): 459-67), Arabidopis thaliana oleosin promotor (WO98/45461), (US 5 for Kidney bean (Phaseolus vulgaris) phaseolin promoter, 504,200), rape Bce4 promotor (WO 91/13980), beans arc5 promotor, Radix Dauci Sativae DcG3 promotor or legumin B4 promotor (LeB4; Baeumlein etc., 1992, Plant Journal, 2 (2): 233-9), and the promotor of in monocotyledons, bringing seed-specific expression, as corn, barley, wheat, rye, rice etc.Favourable seed specific promoters has sucrose-binding proteins promotor (WO 00/26388), phaseolin promoter and napin promotor.The suitable promotor that must consider has barley lpt2 or lpt1 gene promoter (WO 95/15389 and WO 95/23230), and the promotor described in the WO 99/16890 (from the promotor of barley hordein gene, paddy protein gene, rice oryzin gene, rice prolamin gene, wheat gliadin gene, wheat gluten gene, corn zein spirit-soluble gene, avenin gene, Chinese sorghum kasirin gene, rye secaline gene).Other suitable promotors are Amy32b, Amy 6-6 and Aleurain[US 5,677,474], [US 5 for Bce4 (Semen Brassicae campestris rape), 530,149], glycinin (soybean) [EP 571741], phosphoric acid enol pyruvic acid carboxylase (soybean) [JP 06/62870], ADR12-2 (soybean) [WO 98/08962], isocitrate lyase (Semen Brassicae campestris rape) [US 5,689,040] or α-Dian Fenmei (barley) [EP 781849].

Be used in that other promotors of expressing gene have the leaf specificity promoter, those described in DE-A19644478 in the plant; The perhaps promotor of light regulation and control is as pea petE promotor.

In one embodiment, nucleotide sequence effectively is connected with constitutive promoter.Constitutive promoter is at the great majority of its g and D but must not be all stages transcriptional activations all, and is generally to express (over-ground part that is included in plant) basically.Preferred promoter derives from plant, is transformed if also preferably derive from the monocotyledons monocotyledons.Preferred again constitutive promoter is the GOS2 promotor of being represented by the nucleotide sequence that is substantially similar to SEQ ID NO:208 or SEQ ID NO:56.Most preferably GOS2 promotor such as SEQ ID NO:56 or SEQ ID NO:208 are represented.Be understood that suitability of the present invention is not limited to the cpFBPASE nucleotide sequence by SEQ ID NO:154 representative, cpFBPASE expression of nucleic acids when suitability of the present invention also is not limited to by the GOS2 promoters driven simultaneously.The example that also can be used for driving other constitutive promoters that coding cpFBPASE nucleotide sequence expresses shows as mentioned.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.The additional adjustment element can comprise transcribes and translates reinforcement.One skilled in the art will recognize that and go for implementing terminator of the present invention and strengthen subsequence.Intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also comprise need be used for the replication sequence starting point of keeping and/or duplicating in particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to fl-ori and colE1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Thereby genetic constructs can randomly comprise the selected marker.This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top " definition " part.Preferred different mark depends on tissue and system of selection.

The present invention also comprises with the obtainable plant of the method according to this invention (comprising seed).Therefore the present invention provides with the obtainable plant of the method according to this invention, plant part and vegetable cell, introduces cpFBPASE nucleotide sequence and described plant, plant part and vegetable cell in the described plant preferably from crop, more preferably from monocotyledons.

The present invention also is provided for producing the method that has the transgenic plant of enhanced yield with respect to control plant, and it is included in the nucleotide sequence of introducing in the plant and expressing coding cpFBPase polypeptide.

More specifically, the invention provides and be used to produce the preferred monocotyledonous method of transgenic plant that has enhanced yield with respect to control plant, described method comprises:

(i) nucleotide sequence of introducing and expression coding cpFBPase polypeptide in plant shoot branch or vegetable cell; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

(i) nucleic acid can be the nucleic acid of the cpFBPase polypeptide of encoding of any this paper definition.

Nucleotide sequence is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleotide sequence is preferably by transforming introduced plant.

Can be by the regenerate vegetable cell of genetic modification of all methods that the technician was familiar with.S.D.Kung that can mention in the above and R.Wu, Potrykus or

With find suitable method in the publication of Willmitzer.

Usually after conversion, vegetable cell or cell colony are selected the existence of one or more marks, wherein said mark becomes whole strain plant with the material regeneration that transforms subsequently by the expressive gene of plant coding that moves with the goal gene corotation.For selecting plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example.Can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying and handles.Another kind of possibility is included in cultivates seed (as required after sterilization) on the agar plate that uses suitable selective agent, to such an extent as to only the seed of Zhuan Huaing can grow into plant.Alternatively, plant transformed is screened the existence of selected marker (those selected markers as indicated above).

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition also can for example use Southern to analyze goal gene were estimated.Alternative or extraly, the expression level of newly introducing DNA can use that Northern and/or Western analyze, quantitative PCR, and this type of technology is that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clone's property method of proliferating or classical breeding technique.For example, the conversion plant of the first-generation (or T1) can self-pollination produces the s-generation (or T2) transformant of purifying, and the T2 plant further breeds by classical breeding technique.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's property transformant (for example conversion) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the conversion rhizome of unconverted grafting of tender branch.

The inventive method advantageously is applicable to any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.

Other favourable plants be selected from composite family (Asteraceae) as Helianthus (Helianthus), Tagetes for example species Sunflower Receptacle [Sunflower Receptacle (sunflower)], spiceleaf Flower of Aztec Marigold (Tagetes lucida), Flower of Aztec Marigold (Tagetes erecta) or Tagetes signata (Tagetes tenuifolia) [Marigold], Cruciferae (Brassicaceae) as Btassica (Brassica), Arabidopsis, for example species colea, overgrown with weeds overgrown with weeds blue or green [canola oil dish, rape, turnip] or Arabidopis thaliana; Pulse family (Fabaceae) is as Glycine for example species soybean, soybean (Soja hispida) or soybean (Soja max) [soybean]; Flax family (Linaceae) (Linaceae) is as linum (Linum) species flax [flax (flax, linseed)] for example; Gramineae such as Hordeum (Hordeum), Secale (Secale), Avena (Avena), sorghum (Sorghum), Oryza (Oryza), Zea (Zea), Triticum (Triticum) is species barley [barley] for example, rye [rye], oat, oat (Avena fatua), wild avena sativa, than praising oat, Avena fatua var.sativa, hybrid oat [oat], dichromatism chinese sorghum [Chinese sorghum, grain], rice, read leaf rice [rice], Zea mays [corn, corn], common wheat, durum wheat, the cylinder wheat, Triticum hybernum, Macha wheat (Triticum macha), common wheat (Triticum sativum) or common wheat (Triticum vulgare) [wheat, bread wheat, common wheat]; Solanaceae (Solanaceae) belongs to (Lycopersicon) for example potato [potato], tomato (Lycopersicon esculentum, Lycopersiconlycopersicum, Lycopersicon pyriforme, Solanum integrifolium or Solanumlycopersicum) [tomato (tomato)] as Solanum (Solanum), tomato.

The present invention extends to any vegetable cell or the plant by described arbitrary method generation herein clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by arbitrary preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical yielding characteristics and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.The present invention also comprises host cell, and it contains separative cpFBPase nucleotide sequence.The preferred host cell of the present invention is a vegetable cell.The present invention also extend to plant the part gathered in the crops as, but be not limited to seed, leaf, fruit, flower, stem, root stock, stem tuber and bulb.The invention further relates to from oneself, preferably directly from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, oil, fat and lipid acid, starch or protein.

The present invention also comprise the purposes of the purposes of cpFBPase nucleotide sequence and cpFBPase polypeptide and as mentioned definition construct with respect to the purposes of control plant in increasing plant biomass.The plant biomass that increases refers in particular to the seed production of increase.The seed production that this paper quotes increase means following arbitrary index: (i) seed weight of Zeng Jiaing; The (ii) full seed quantity of Zeng Jiaing; The (iii) full rate of the seed of Zeng Jiaing; The (iv) harvest index of Zeng Jiaing.

Can in the procedure of breeding, use cpFBPase nucleotide sequence or cpFBPase polypeptide, wherein identify the dna marker that can be connected in the cpFBPase locus hereditarily.Can use cpFBPase nucleotide sequence or cpFBPase polypeptide to define molecule marker.Then this DNA or polypeptide marker can be used in the procedure of breeding, have the plant of the output of increase with selection.For example, the cpFBPase gene can be the nucleotide sequence of the cpFBPase nucleotide sequence that provides in the table 7 or any aforementioned SEQ ID NO straight arbitrary representative in the nucleic acid sequence encoding of homologue or collateral line homologue.

The allele variant of cpFBPase nucleotide sequence also can be used for the auxiliary procedure of breeding of mark.This class procedure of breeding needs to use sometimes, and for example EMS mutagenesis is introduced allele variant by the plant mutagenic treatment; Alternative, this program can begin with the allele variant of collecting what is called " natural " origin that is not intended to generation.Identify allele variant by for example PCR then.Be to select step subsequently, in order to select the better allele variant of the sequence of discussing, described allele variant is given the output that plant increases.Generally the growth behavior that contains the different allele variant plants of studying sequence to some extent by monitoring is selected the straight different allele variants arbitrary in the nucleic acid sequence encoding of homologue or collateral line homologue of the cpFBPase nucleotide sequence that for example provides in the table 7 or aforementioned any SEQ IDNO.Can in greenhouse or field, monitor growth behavior.More optional step comprise, will contain plant and another plant hybridization of better allele variant through evaluation.For example, can make the combination that produces phenotypic characteristic interested in this way.

The cpFBPase nucleotide sequence can also be as probe, is used for for the part of those gene linkage proterties and carry out the mapping of heredity and physics as the gene of its mark.Such information can be used in plant breeding, to obtain having the strain of desired phenotype.This class of cpFBPase nucleotide sequence is used the nucleotide sequence that only needs to grow to few 15 Nucleotide.The cpFBPase nucleotide sequence can be used as restriction fragment length polymorphism (RFLP) mark.Available cpFBPase nucleotide sequence is surveyed the Southern trace (Sambrook J, Fritsch EF and ManiatisT (1989) " molecular cloning: laboratory manual ") of the plant genome DNA of restriction digest.The program that uses a computer subsequently such as MapMaker (Lander etc. (1987) Genomics 1:174-181) carry out genetic analysis to the banding pattern that produces, to make up genetic map.In addition, can use nucleotide sequence to survey the Southern trace in the genomic dna that the restriction enzyme that contains one group of individuality is handled, described one group of individuality is the parent of the clear and definite genetic cross of representative and one group of individuality of filial generation.The separation of record dna polymorphism also is used for calculating formerly position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) with the genetic map cpFBPase nucleotide sequence of this colony's acquisition.

The derive generation and the purposes of probe of the plant gene that uses in genetic mapping is described among Bematzky and Tanksley (1986) the Plant Mol.Biol.Reporter 4:37-41.Described in numerous publications with aforesaid method or its flexible form specific cDNA clone was carried out genetic mapping.For example, can use F2 hybridization colony, backcross population, panmictic population, the homogenic strain of close relative and the mapping of other group of individuals.These class methods are that those skilled in the art are well-known.

Nucleic acid probe also can be used for physical mapping and (promptly settle sequence on physical map; See In:Non-mammalian Genomic Analysis:A Practical Guide such as Hoheisel, Academicpress 1996, the 319-346 pages or leaves, and the reference of wherein quoting).

In another embodiment, nucleic acid probe can be used for direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although the method inclination of FISH mapping at present is used for big clone; See (1995) Genome Res.5:13-20 such as Laan), but the raising of susceptibility allows to use short probe in the FISH mapping.

The multiple method based on nucleic acid amplification that is used for heredity and physical mapping can use described nucleotide sequence to carry out.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med11:95-96), pcr amplified fragment; Sheffield etc. (1993) Genomics16:325-332), allele-specific connects (Landegren etc. (1988) Science 241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc. (1997) Nat.Genet.7:22-28) and Happy mapping (Dear and Cook (1989) Nucleic Acid Res.17:6795-6807).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The method according to this invention is had the plant of the output of increase as previously mentioned.The proterties that output improves can also make up other favourable economically proterties, as the proterties of more raising output, to the proterties of multiple tolerance of coercing, the multiple structural attitude of improvement and/or biochemistry and/or physiologic character.

The detailed description of SIK polypeptide

Have now found that regulating SIK nucleic acid and/or the expression of SIK polypeptide in plant produces the plant that has the relevant proterties of the output of multiple improvement with respect to control plant, wherein the overexpression of SIK coding nucleic acid in plant produces to have every strain plant flowers number of increase and wherein to reduce or remove SIK nucleic acid substantially with respect to control plant and produces and have the thousand seed weight of increase, the harvest index of increase and the full rate of increase with respect to corresponding wild-type plant.

Therefore the invention provides method with respect to output correlated character in the control plant improvement plant, it comprises regulates SIK nucleic acid and/or the expression of SIK polypeptide in plant, the expression of wherein being regulated is the overexpression of SIK coding nucleic acid in plant, it produces the every strain plant flowers number that increases with respect to control plant, the expression of wherein being regulated is minimizing or removes SIK nucleic acid substantially, the full rate that it produces the thousand seed weight (TKW) that increases with respect to corresponding wild-type plant, the harvest index (HI) of increasing and increases.

Term as defined herein " adjusting " means the change of comparing gene expression dose with control plant.If the SIK coding nucleic acid is used to increase every strain plant flowers number, just increase expression level, if SIK nucleic acid is used to increase TKW, HI or full rate, just reduce expression level.Any kind that original expression of being regulated can be structure RNA (rRNA, tRNA) or mRNA is expressed, and is translation subsequently.

Term " expression " or " genetic expression " mean transcribing of one or more genes of Yin Teding or gene construct.Term " expression " or " genetic expression " especially mean one or more genes or gene construct is transcribed into structure RNA (rRNA, tRNA) or mRNA, and being with or without subsequently, mRNA translates into protein.This process comprises that DNA transcribes, processes the mRNA product that obtains.

The output correlated character of multiple improvement compares modified at least 5%, 6%, 7%, 8%, 9% or 10% with control plant, preferably at least 15% or 20%, more preferably 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65% or 70%.

Improvement can be a kind of in following or more plant: the full rates of seed (it is the ratio between full seed number and the seed sum as defined herein) of spending number, increasing that every strain plant increases; The harvest index that increases, it is the ratio that can gather in the crops part (as seed) output and total biomass as defined herein; With the thousand seed weight (TKW) that increases, this extrapolates from the full seed number and the gross weight thereof of counting as defined herein.The TKW that increases can be because of due to the seed size and/or seed weight that increase, and also can be because of due to the increase of embryo and/or endosperm size.

Compare with control plant, no matter plant is under the non-stress conditions still is that plant is exposed to multiple coercing down, and the increase of thousand seed weight, the harvest index of increase and the full rate of increase all take place.Plant is generally replied being exposed to coerce to make by growing slowlyer.Under the condition of serious stress of soil condition, plant even can stop growing fully.On the other hand, slightly coerce and be defined as plant in this article any of its exposure coerced, the wherein said ability that does not cause plant to stop growing fully and recover growth of coercing.Compare with the control plant under the non-stress conditions, slightly coerce and in meaning of the present invention, cause being coerced the plant-growth reduction less than 40%, 35% or 30%, preferably, be more preferably less than 14%, 13%, 12%, 11% or 10% or lower less than 25%, 20% or 15%.Because the progress on the agricultural practice (irrigation, fertilising, pesticide treatments) does not often run into condition of serious stress of soil in the raise crop plant.Therefore, by the agriculture often undesirable characteristic that goes up of the impaired growth of slight stress-inducing.Slightly coerce is that the common biological and/or inanimate (environment) that plant exposes is coerced.Abiotic stress can because of arid or waterlogging, anaerobism are coerced, due to salt stress, chemical toxicity, oxidative stress and heat, cold or the freezing temperature.Abiotic stress can be to coerce (especially because arid), salt stress, oxidative stress or ion by water to coerce the osmotic stress that causes.It generally is that those that caused by pathogenic agent such as bacterium, virus, fungi and insect are coerced that biology is coerced.

Especially, method of the present invention can implemented the plant that has the full rate of the harvest index of thousand seed weight, increase of increase and increase with respect to control plant to produce under the non-stress conditions or under slight drought condition.As report in (Planta (2003) 218:1-14) such as Wang, abiotic stress causes influencing unfriendly a series of morphological change of plant-growth and productivity, physiology to change, biological chemistry changes and molecule changes.Known arid, salinity, extreme temperature and oxidative stress are also can damaging and primary cellular defect by induced growth by similar mechanism of connecting each other.Rabbani etc. (PlantPhysiol (2003) 133:1755-1767) have described " cross-talk " that drought stress and high salinity are coerced a very high degree.For example, arid and/or salinification mainly show as osmotic stress, cause the destruction of cell homeostasis and ion distribution.Often follow the oxidative stress of high temperature or low temperature, salinity or drought stress can cause functional protein and structural protein sex change.Therefore, these various environment-stress usually activate similar cell signal approach and cell response, as producing stress protein matter, raising antioxidant, accumulation compatible solute and growth-inhibiting.Term as used in this article " non-coercing " condition is the envrionment conditions that allows the plant optimum growh.Those skilled in the art know that normal edaphic condition and weather condition for given place.

The enforcement of the inventive method is with respect to the control plant of cultivating under comparable conditions, gives under the non-stress conditions or the plant of cultivating under slight the drought condition thousand seed weight, the harvest index of increase and the full rate of increase that increase.Thereby according to the present invention, be provided for increasing thousand seed weight under the non-stress conditions or in the plant of under slight drought condition, cultivating, increase harvest index and increasing the method for full rate, described method comprises that the nucleic acid that increases coding SIK polypeptide expresses in plant.

The enforcement of the inventive method produces with respect to the appropriate control plant of cultivating under comparable conditions, the plant of the full rate of the harvest index of the thousand seed weight of under the nutritive deficiency condition, especially cultivating under nitrogen shortage condition with increase, increase and increase.Thereby according to the present invention, increase thousand seed weight in the plant that is provided under the nutritive deficiency condition, cultivating, increase harvest index and increase the method for full rate, described method comprises the expression of nucleic acid in plant that increases coding SIK polypeptide.Nutritive deficiency can be because of due to the nutraceutical shortage, and described nutrition for example is nitrogen, phosphoric acid salt and other P contained compounds, potassium, calcium, cadmium, magnesium, manganese, iron and boron etc.

The present invention includes according to the obtainable plant of the inventive method or its part (comprising seed).Plant or its part comprise the nucleic acid transgenosis of the SIK polypeptide of definition as mentioned.

According to an aspect of the present invention, reduce or remove SIK nucleic acid substantially and produce following one or multinomial: the full rate of the thousand seed weight of increase (TKW), the harvest index (HI) that increases and increase with respect to control plant.

" the SIK nucleic acid " mentioned herein means the two strands of arbitrary length of polymerized form or the deoxyribonucleotide polymkeric substance or the ribonucleoside acid polymer of strand, or its analogue, it has the essential characteristic of natural nucleus sugar nucleotide, because it can be with mode and the SIK nucleic acid array hybridizing that is similar to naturally occurring polynucleotide.

" reduce or remove substantially SIK nucleic acid or gene " that this paper mentions and " endogenous " SIK gene are as described in the definitional part.

In order to reduce or to remove the expression of SIK native gene in plant substantially, need the Nucleotide of successive basically of the sufficient length of SIK nucleotide sequence.Basically the successive nucleotide fragments can be from any SIK nucleic acid, preferably by SEQ ID NO 213 to 225, or arbitrary SIK nucleic acid of explaining in the arbitrary nucleotide sequence that provides in following table 8 or the table 9.The SIK nucleotide sequence of coding (functional) polypeptide is not discussed hereinly to be used to reduce or to remove the several different methods of endogenous SIK genetic expression substantially required.

This reduction or basic removal can use conventional instrument and technology as indicated above to finish.The preferred method that is used for reducing or removes the SIK expression of nucleic acid substantially is to introduce and express such genetic constructs plant, wherein the SIK nucleic acid clone to the described genetic constructs as (partially or completely) inverted repeats that separates by transcribed spacer (noncoding DNA).

According to another aspect of the present invention, the overexpression of SIK coding nucleic acid in plant produces the every strain plant flowers number that increases with respect to control plant.

The preferred method that is used for SIK coding nucleic acid overexpression is by introduce and be expressed as follows the nucleic acid of the coding SIK polypeptide of face definition plant.

As defined herein " SIK coding nucleic acid " coding " SIK polypeptide " or " SIK aminoacid sequence " mean according to SEQ ID NO:210 and its straight polypeptide to homologue and collateral line homologue as defined herein.

SIK polypeptide shown in the SEQ ID NO:210 and its directly also can typically comprise following feature to homologue and collateral line homologue:

The ATP-land

ATP-land vsesltstsykasfrddftdpktieaivsrl (motif 1) or have 40% at least with preferred sequence and the motif I that increases, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53,54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, the ATP-land of 99% identity.

Serine threonine kinases avtive spot mark

Serine threonine kinases avtive spot mark AMYN with conservative D residue DFSTSNIQI (motif 2) or with have 40% at least with preferred sequence and the motif II that increases, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity.

Be rich in the ser structure territory

N-terminal is rich in the ser structure territory.

The myristoylation site

Directly can further include one or more myristoylations site to homologue and collateral line homologue, it anchors to protein on the film.

Kinase activity and then, SIK directly will show kinase activity to homologue and collateral line homologue, can use conventional instrument and technology easily to determine described kinase activity.Several mensuration are available (Current Protocols in Molecular Biology for example, 1 and 2 volumes, people such as Ausubel (1994), Current Protocols; Or online, for example Http:// www.protocol-online.org).

In brief, kinase assays generally includes: (1) contains kinase protein contact to remain the substrate polypeptide of the target site of phosphorylation; (2) under suitable condition, in suitable kinase buffer liquid, carry out the phosphorylation of target site; (3) after the appropriate reaction period, the product of phosphorylation is separated from unphosphorylated substrate.The existence of kinase activity or shortage are by the existence of the target of phosphorylation or lack to determine.In addition, can carry out quantitative measurment.

The SIK protein of purifying, perhaps containing or be rich in the proteinic cell extract of SIK can be as the source of kinase protein.Histone h1 or little peptide especially are suitable as substrate.Peptide must contain one or more Serines, Threonine or tyrosine residues in the phosphorylation site motif.Can be at Biochimica et Biophysica Acta 1314,191-225 finds the compilation of phosphorylation site in (1996).In addition, peptide substrates can advantageously have clean positive charge, is beneficial to be attached on the phosphorylated cotton filter (to allow to the peptide of phosphorylation is separated and detected the peptide of phosphorylation from unphosphorylated peptide).If do not know the phosphorylation site motif, can use general tyrosine kinase substrate.For example, Src related peptides (RRLIEDAEYAARG) is the substrate of many acceptors and nonreceptor tyrosine kinase.In order to determine the kinetic parameter of synthetic peptide phosphorylation, need the scope of peptide concentration.For initial action, can use the peptide concentration of 0.7-1.5mM.

For every kind of kinases, importantly to determine active optimized buffer liquid, ionic strength and pH.Standard 5 * kinase buffer liquid contains 5mg/ml BSA (preventing that kinases is adsorbed in the bovine serum albumin of measuring pipe), 150mM Tris-Cl (pH 7.5), 100mM MgCl usually ₂Most of tyrosine-kinase enzyme require divalent cation is although some Tyrosylprotein kinases (for example Regular Insulin-, IGF-1-and pdgf receptor kinase) need MnCl ₂Rather than MgCl ₂(or except MgCl ₂).Must rule of thumb determine the optimum concn of divalent cation for every kind of protein kinase.

Phosphoryl (phophoryl) group donor commonly used be radiolabeled [γ- ³²P] ATP (normally 0.2mM final concentration).In peptide, mix ³²The amount of P can be active next definite by measuring on the dried pad of soluble cotton in scintillometer.

Alternatively or in addition, can by under constitutive promoter regulation and control in plant the nucleic acid of overexpression coding SIK polypeptide measure with the increase that detects every strain plant flowers number with respect to control plant SIK directly to the activity of homologue and collateral line homologue and also can by under the constitutive promoter regulation and control, reduce or remove substantially SIK nucleic acid with detect in thousand seed weight (TKW), harvest index (HI) and the full rate in the plant or more item measure SIK directly to the activity of homologue and collateral line homologue with respect to the increase of control plant.

The present invention is illustrated by the rice sequence plant transformed of using the coded polypeptide sequence SEQ IDNO:210 that is represented by SEQ ID NO:209, yet enforcement of the present invention is not limited to these sequences.The inventive method can advantageously use coding to carry out any nucleotide sequence that for example provides in table 8 and the table 9 as any nucleic acid of defined SIK polypeptide in the literary composition.The SEQ IDNO:210 that table 8 and table 9 provide directly to the example of homologue and collateral line homologue available from gene studies institute (The Instituteof Genetic research, TIGR).% identity in the table 9 provides on nucleotide level.The accession number with ' TC ' beginning that provides is identified the sequence of finding by TIGR.If local nucleic acid, obtain the total length nucleotide sequence length of (or be enough at least implement the inventive method) and belong to those skilled in the art's limit of power fully.

Table 8: the SIK that infers is directly to the example of homologue and collateral line homologue

#	TC	The function of inferring
			1	Arabidopis thaliana | TC272322	(Q9C9Y3) false albuminoid F17O14.23
2	Barley | TC134680	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
			3	Cotton | TC30779	(Q9C9Y3) false albuminoid F17O14.23
4	Grape | TC43027	Q94CI5) protein kinase A tSIK
			5	Japan perch (L.japonicus) TC17767	(Q94CI5) protein kinase A tSIK
6	Lettuce | TC15801	Protein kinase A tSIK
			7	Corn | TC255367	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
8	Corn | TC272965	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
			9	Clover | TC96175	(Q9C9Y3) false albuminoid F17O14.23
10	Pepper | TC5595	(Q94CI5) protein kinase A tSIK
			11	Potato | TC117743	Infer the protein kinase structural domain
12	Rice | TC268277	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
			13	Sugarcane | TC67974	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
14	Chinese sorghum | TC103780	(Q8RUD7) albuminoid kinase AtSIK (P0485B12.21 protein)
			15	Soybean | TC229774	(Q94CI5) protein kinase A tSIK

16	Tomato | TC158378	68416.m01018 the protein kinase family protein comprises protein kinase structural domain Pfam:PF00069
			17	Tomato | TC166426	68416.m01018 the protein kinase family protein comprises protein kinase structural domain Pfam:PF00069
18	Wheat | TC257477	(Q8RUD7 albuminoid kinase AtSIK (P0485B12.21 protein)

Table 9: the SIK that infers is directly to the example of homologue and collateral line homologue

Sequence 1	Sequence 2	% identity	Matching length	The p-value	Recip. the best is hit
						Barley | TC134680	Arabidopis thaliana | TC272322	63	1030	6.80E-66	＊
Cotton | TC30779	Arabidopis thaliana | TC272322	71	1008	9.40E-103	＊
						Cotton | TC30779	Barley | TC134680	68	971	8.30E-80	＊
Grape | TC43027	Arabidopis thaliana | TC272322	60	619	4.30E-25	＊
						Grape | TC43027	Cotton | TC30779	80	245	5.60E-29	＊
Grape | TC43027	Corn | TC272965	58	483	3.20E-13	＊
						Grape | TC43027	Rice | TC268277	66	318	3.50E-19	＊
Grape | TC43027	Sugarcane | TC67974	63	372	2.40E-17	＊
						Grape | TC43027	Soybean | TC229774	77	276	4.60E-30	＊
Grape | TC43027	Tomato | TC158378	69	420	9.20E-33	＊
						Grape | TC43027	Tomato | TC166426	75	241	1.80E-23
The Japan perch | TC17767	Arabidopis thaliana | TC272322	72	659	1.30E-64	＊
						The Japan perch | TC17767	Barley | TC134680	67	659	4.40E-53	＊
The Japan perch | TC17767	Cotton | TC30779	80	654	9.20E-89	＊
						Lettuce | TC15801	Arabidopis thaliana | TC272322	69	529	8.90E-44	＊
Lettuce | TC15801	Cotton | TC30779	77	501	1.20E-58	＊
						Lettuce | TC15801	The Japan perch | TC17767	75	412	2.20E-45	＊
Corn | TC255367	Arabidopis thaliana | TC272322	62	528	5.50E-24	＊
						Corn | TC255367	Barley | TC134680	81	451	1.10E-60	＊
Corn | TC255367	Cotton | TC30779	63	738	6.90E-44	＊
						Corn | TC255367	The Japan perch | TC17767	66	366	2.20E-24	＊
Corn | TC255367	Lettuce | TC15801	65	455	4.10E-30	＊
						Corn | TC272965	Barley | TC134680	68	623	2.90E-43

Clover | TC96175	Arabidopis thaliana | TC272322	67	1245	2.70E-102	＊
						Clover | TC96175	Barley | TC134680	65	1069	2.50E-75	＊
Clover | TC96175	Cotton | TC30779	76	1041	2.20E-125	＊
						Clover | TC96175	The Japan perch | TC17767	89	666	3.50E-114	＊
Clover | TC96175	Lettuce | TC15801	73	504	1.00E-51	＊
						Clover | TC96175	Corn | TC255367	62	813	3.60E-41	＊
Pepper | TC5595	Arabidopis thaliana | TC272322	66	696	6.40E-49	＊
						Pepper | TC5595	Barley | TC134680	66	510	2.90E-38	＊
Pepper | TC5595	Cotton | TC30779	76	553	1.60E-66	＊
						Pepper | TC5595	The Japan perch | TC17767	75	474	4.50E-53	＊
Pepper | TC5595	Lettuce | TC15801	75	541	3.30E-60	＊
						Pepper | TC5595	Corn | TC255367	66	462	1.00E-29	＊
Pepper | TC5595	Clover | TC96175	67	800	1.30E-60	＊
						Potato | TC117743	Arabidopis thaliana | TC272322	64	1082	8.80E-65	＊
Potato | TC117743	Barley | TC134680	66	565	1.30E-39	＊
						Potato | TC117743	Cotton | TC30779	77	587	1.80E-70	＊
Potato | TC117743	The Japan perch | TC17767	75	496	3.60E-54	＊
						Potato | TC117743	Lettuce | TC15801	77	543	2.30E-65	＊
Potato | TC117743	Corn | TC255367	67	444	1.30E-33	＊
						Potato | TC117743	Clover | TC96175	68	804	4.20E-64	＊
Potato | TC117743	Pepper | TC5595	82	826	1.60E-121	＊
						Rice | TC268277	Arabidopis thaliana | TC272322	63	1019	3.10E-66	＊
Rice | TC268277	Barley | TC134680	78	1371	5.70E-179	＊
						Rice | TC268277	Cotton | TC30779	64	1436	3.40E-101	＊
Rice | TC268277	The Japan perch | TC17767	70	656	1.90E-61	＊
						Rice | TC268277	Corn | TC255367	81	860	4.50E-121	＊
Rice | TC268277	Corn | TC272965	74	646	4.00E-68
						Rice | TC268277	Clover | TC96175	63	1684	3.00E-104	＊
Rice | TC268277	Pepper | TC5595	65	600	5.50E-40	＊
						Rice | TC268277	Potato | TC117743	65	691	3.80E-45	＊
Sugarcane | TC67974	Arabidopis thaliana | TC272322	63	938	1.50E-59	＊
						Sugarcane | TC67974	Barley | TC134680	76	1294	2.60E-151	＊

Sugarcane | TC67974	Cotton | TC30779	66	876	5.10E-70	＊
						Sugarcane | TC67974	The Japan perch | TC17767	69	656	4.70E-57	＊
Sugarcane | TC67974	Lettuce | TC15801	66	390	4.70E-28	＊
						Sugarcane | TC67974	Corn | TC255367	96	367	2.80E-72
Sugarcane | TC67974	Corn | TC272965	90	649	2.20E-112	＊
						Sugarcane | TC67974	Clover | TC96175	64	1140	7.80E-75	＊
Sugarcane | TC67974	Pepper | TC5595	66	471	1.80E-33	＊
						Sugarcane | TC67974	Potato | TC117743	67	460	1.00E-35	＊
Sugarcane | TC67974	Rice | TC268277	82	1338	1.20E-194	＊
						Sugarcane | TC67974	Chinese sorghum | TC103780	95	1359	8.70E-277	＊
Sugarcane | TC67974	Soybean | TC229774	70	750	1.70E-67	＊
						Chinese sorghum | TC103780	Arabidopis thaliana | TC272322	63	1057	1.40E-65	＊
Chinese sorghum | TC103780	Barley | TC134680	75	1421	5.10E-163	＊
						Chinese sorghum | TC103780	Cotton | TC30779	64	1333	8.60E-91	＊
Chinese sorghum | TC103780	The Japan perch | TC17767	69	656	6.40E-57	＊
						Chinese sorghum | TC103780	Lettuce | TC15801	64	493	3.70E-30	＊
Chinese sorghum | TC103780	Corn | TC255367	92	948	1.70E-180	＊
						Chinese sorghum | TC103780	Corn | TC272965	89	651	2.40E-110
Chinese sorghum | TC103780	Clover | TC96175	62	1653	5.10E-96	＊
						Chinese sorghum | TC103780	Pepper | TC5595	64	580	2.60E-36	＊
Chinese sorghum | TC103780	Potato | TC117743	65	602	6.50E-39	＊
						Chinese sorghum | TC103780	Rice | TC268277	80	1852	1.40E-260	＊
Soybean | TC229774	Arabidopis thaliana | TC272322	72	757	1.10E-77	＊
						Soybean | TC229774	Barley | TC134680	69	756	5.70E-67	＊
Soybean | TC229774	Cotton | TC30779	78	737	3.40E-96	＊
						Soybean | TC229774	The Japan perch | TC17767	90	495	4.10E-87	＊
Soybean | TC229774	Lettuce | TC15801	77	249	1.90E-26	＊
						Soybean | TC229774	Corn | TC255367	71	203	2.00E-15	＊
Soybean | TC229774	Clover | TC96175	87	717	2.10E-118	＊
						Soybean | TC229774	Pepper | TC5595	76	308	1.50E-32	＊
Soybean | TC229774	Potato | TC117743	76	332	1.80E-35	＊
						Soybean | TC229774	Rice | TC268277	71	744	4.20E-74	＊

Soybean | TC229774	Chinese sorghum | TC103780	69	757	1.30E-65	＊
						Tomato | TC158378	Arabidopis thaliana | TC272322	71	686	3.10E-64	＊
Tomato | TC158378	Barley | TC134680	65	907	8.80E-63	＊
						Tomato | TC158378	Cotton | TC30779	78	669	2.90E-86	＊
Tomato | TC158378	The Japan perch | TC17767	78	427	1.70E-52	＊
						Tomato | TC158378	Lettuce | TC15801	81	181	3.90E-21
Tomato | TC158378	Corn | TC272965	60	428	4.80E-14	＊
						Tomato | TC158378	Clover | TC96175	68	908	4.60E-76	＊
Tomato | TC158378	Pepper | TC5595	84	243	6.40E-34
						Tomato | TC158378	Potato | TC117743	96	264	8.90E-50	＊
Tomato | TC158378	Rice | TC268277	66	897	1.70E-64	＊
						Tomato | TC158378	Sugarcane | TC67974	65	906	2.40E-60	＊
Tomato | TC158378	Soybean | TC229774	74	700	4.80E-78	＊
						Tomato | TC166426	Arabidopis thaliana | TC272322	70	675	1.80E-62
Tomato | TC166426	Barley | TC134680	67	680	1.40E-54
						Tomato | TC166426	Cotton | TC30779	77	679	3.80E-85
Tomato | TC166426	The Japan perch | TC17767	77	440	1.30E-50
						Tomato | TC166426	Lettuce | TC15801	81	193	6.50E-23	＊
Tomato | TC166426	Clover | TC96175	73	675	1.30E-69
						Tomato | TC166426	Pepper | TC5595	95	255	1.10E-46	＊
Tomato | TC166426	Potato | TC117743	85	276	2.40E-39
						Tomato | TC166426	Rice | TC268277	67	680	2.70E-55
Tomato | TC166426	Sugarcane | TC67974	67	680	4.90E-53
						Tomato | TC166426	Chinese sorghum | TC103780	67	680	1.60E-53	＊
Tomato | TC166426	Soybean | TC229774	73	680	2.30E-72
						Wheat | TC257477	Barley | TC134680	91	672	7.00E-119	＊
Wheat | TC257477	Corn | TC272965	69	639	2.80E-49	＊
						Wheat | TC257477	Rice | TC268277	68	647	4.10E-47	＊
Wheat | TC257477	Sugarcane | TC67974	67	648	4.10E-46	＊
						Wheat | TC257477	Chinese sorghum | TC103780	69	648	4.20E-49	＊
Wheat | TC257477	Tomato | TC158378	61	441	1.20E-18	＊

Can find easily that SIK is directly to homologue and collateral line homologue with encode that this type of directly can be used for implementing method of the present invention to the nucleic acid of homologue and collateral line homologue.

Directly can easily find by carrying out so-called interactivity blast search to homologue and collateral line homologue.This generally comprises a BLAST, a described BLAST participates in submitting to search sequence (for example SEQ ID NO:209 or SEQ ID NO:210) to be used for that (as public's available ncbi database, described database can find in following website: blast search http://www.ncbi.nlm.nih.gov) at arbitrary sequence library.When nucleotide sequence begins, use BLASTN or TBLASTX (using the standard default value) usually, and, can use BLASTP or TBLASTN (use standard default value) when when protein sequence begins.Randomly can screen BLAST result.The full length sequence of submitting The selection result and non-The selection result subsequently to is with at carry out reverse BLAST (the 2nd BLAST) from the sequence of biology, wherein search sequence is from described biology (be under the situation of SEQ ID NO:209 or SEQ ID NO:210 in search sequence wherein, the 2nd BLAST thereby will be at rice (Oryza) sequence).The result who compares first and second blast searches subsequently.If from the high-order position of a BLAST hit be derived from search sequence from identical species of deutero-species wherein, oppositely BLAST then identifies the collateral line homologue subsequently ideally to produce in the highest search sequence of hitting; If the high-order position among the BLAST hit be not derived from search sequence from identical species of deutero-species wherein, and preferably when reverse BLAST, produce the search sequence that belongs to the highest row that hit, then identify directly to homologue.

It is that with low E-value those hit that high-order position is hit.The E-value is low more, mark remarkable more (or in other words, this hit because of the chance odds low more).The calculating of E-value is well-known in the art.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Preferably mark greater than 50, more preferably greater than 100; With preferred E-value less than e-5, be more preferably less than e-6.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.

Also can use following blast program to identify directly to homologue and collateral line homologue.Can use routine techniques well known in the art such as sequence alignment easily to identify homologue (or homologue protein, comprise) directly to homologue and collateral line homologue.It is well-known in the art being used for the method that aligned sequences compares, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.GAP uses Needleman and Wunsch algorithm ((1970) J Mol Biol 48:443-453) to find the comparison that makes the maximization of coupling number and make minimized two complete sequence of room number.BLAST algorithm (Altschul etc. (1990) J Mol Biol 215:403-10) sequence of calculation identity percentage ratio and execution are to the statistical study of similarity between two sequences.Being used to carry out software that BLAST analyzes and being the public, to pass through NCBI obtainable.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.The overall percentage ratio of similarity and identity also can use one of obtainable method in the MatGAT software package and determine (Campanella etc., BMC Bioinformatics.4,29,2003).Can carry out trickle edit to optimize the comparison between the conservative motif, apparent as those skilled in the art.As using full length sequence, also can use the ad hoc structure territory in addition to identify substituting of homologue.Use program mentioned above, use default parameters, at whole SIK nucleotide sequence or determined amino acid sequence sequence identity value, represent described sequence identity value with per-cent below.

Preferably, the SIK polypeptide by the SIK coding nucleic acid coding that is used for the inventive method has at least 40% with preferred sequence and the polypeptide SEQ ID NO:210 that increases, 41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity.

Alternatively, can use special structural domain to determine sequence identity in the homologue.Can use database that is used for identification of proteins and the instrument of listing above and/or be used for supplying the method for sequence alignment relatively to identify and illustrate the structural domain that provides arbitrarily.In some cases, can adjust default parameters to regulate the severity of search.For example use BLAST, can improve the statistical significance threshold value (being called " expectation " value) that is used to report at the coupling of database sequence to show the lower coupling of severity.Like this, can identify almost accurate short coupling.

The SIK polypeptide can be determined owing to the existence that ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory and or many myristoylations site.Term " structural domain " and " motif " are as hereinbefore defined.There is the specialized database that is used to identify structural domain, as SMART (Schultz etc. (1998) the Proc.Natl.Acad.Sci. U.S. 95,5857-5864; Letunic etc. (2002) Nucleic Acids Res 30,242-244), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318), Prosite (Bucher and Bairoch (1994), be used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and in the function of automatization sequence interpretation, () ISMB-94; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002)) evaluation.The one group of instrument that is used for analysing protein sequence on the computer chip can obtain (resident (Gasteiger etc. on Swiss Institute of Bioinformatics on the ExPASY proteomics server, ExPASy: be used for the protein science server of deep understanding and analysing protein, Nucleic AcidsRes.31:3784-3788 (2003)).Yet, also can easily identify described multiple structural domain by the SIK polypeptide of inferring and the sequence alignment of SIK polypeptide known in the art.

What also be used for the inventive method is homologue by the SIK polypeptide shown in the SEQ ID NO:210, or as mentioned its of definition directly to the coding nucleic acid of the homologue of homologue and collateral line homologue.

Also be used for the inventive method be by the coding nucleic acid of the derivative of SEQ ID NO:210 or SEQ ID NO:210 directly to the coding nucleic acid of the derivative of homologue, collateral line homologue or homologue.

The coding nucleic acid of the SIK polypeptide of this paper definition needs not be total length nucleic acid, because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.The nucleic acid example that is suitable for implementing the inventive method comprises the nucleotide sequence that provides in table 8 and the table 9, but is not limited to those sequences.The nucleic acid variant can be used for implementing method of the present invention.Such nucleic acid variant example comprises the variant of the coding nucleic acid of the allelic variant of coding nucleic acid of splice variant, SIK polypeptide as defined herein of the coding nucleic acid of the part of the coding nucleic acid of SIK polypeptide as defined herein, SIK polypeptide as defined herein and the SIK polypeptide as defined herein that obtains by gene reorganization.Term part, splice variant, allelic variant and gene reorganization are as described herein.

The invention provides the method that increases every strain plant flowers number with respect to control plant, its be included in the plant introduce and expression table 8 or table 9 in the arbitrary amino acid sequence that provides in the part of arbitrary nucleotide sequence of providing or table 8 or the table 9 directly to the part of the coding nucleic acid of homologue, collateral line homologue or homologue.

The represented SIK polypeptide of the arbitrary amino acid sequence that provides in the part coded polypeptide that is used for the inventive method, described polypeptide and table 8 or table 9 has substantially the same biologic activity.Preferably, part be the part of arbitrary nucleic acid of providing in table 8 or the table 9, by the part of the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225.The part normal length is at least 300 continuous nucleotides, preferred length is at least 400 continuous nucleotides, more preferably length is at least 500 continuous nucleotides, and described continuous nucleotide is arbitrary nucleotide sequence of providing in table 8 and the table 9 or by the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225.Most preferred part is the part of nucleic acid SEQ ID NO:209.Preferably; the part encoding amino acid sequence; described aminoacid sequence comprises that ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory, one or the many myristoylations site any one or more, and has kinase activity.

The part of the coding nucleic acid of SIK polypeptide can prepare part by nucleic acid is carried out one or more disappearances as herein defined.Part can be used with isolating form, perhaps itself and other coding (or non-coding) sequence can be merged, so that for example, produce and made up some active protein.When merging with other encoding sequences, the polypeptide that is produced after translating may be bigger than the part of prediction.

Be used for another nucleic acid variant of the present invention and be under the stringent condition that reduces, preferably under stringent condition, can with the coding nucleic acid of SIK polypeptide defined herein or with the nucleic acid of part hybridization defined herein.

The represented SIK polypeptide of the arbitrary amino acid sequence that provides in the hybridization sequences coded polypeptide that is used for the inventive method, described polypeptide and table 8 or table 9 has substantially the same biologic activity.The hybridization sequences normal length is at least 300 continuous nucleotides, and preferred length is at least 400 continuous nucleotides, and more preferably length is at least 500 continuous nucleotides, and described continuous nucleotide is the arbitrary nucleotide sequence that provides in table 8 and the table 9.Preferably, hybridization sequences be can with the sequence of any nucleic acid hybridization of providing in table 8 or the table 9 or with the sequence of the part hybridization of these sequences arbitrarily, described part defines as mentioned.Most preferred hybridization sequences be can with represented nucleic acid of SEQ ID NO:209 or the hybridization of its part.Preferably; the hybridization sequences encoding amino acid sequence; described aminoacid sequence comprises that ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory, one or the many myristoylations site any one or more, and has kinase activity.

The invention provides the method that increases every strain plant flowers number, described method be included in introduce in the plant and express can be with the nucleic acid of the represented arbitrary nucleic acid array hybridizing of arbitrary nucleic acid of providing in table 8 or the table 9 or SEQ ID NO:213 to 225 or be included in introduce in the plant and express can with the represented arbitrary nucleotide sequence of any nucleotide sequence of providing in table 1 or the table 2 or SEQ ID NO:213 to 225 directly to the nucleic acid of the coding nucleic acid hybridization of homologue, collateral line homologue or homologue.

Another nucleic acid variant that is used for the inventive method is the splice variant of the SIK polypeptide of definition as mentioned of encoding, and term " splice variant " defines as mentioned.

The invention provides the method that increases every strain plant flowers number, described method be included in introduce in the plant and expression table 8 or table 9 in arbitrary nucleotide sequence of providing splice variant or by the arbitrary amino acid sequence that provides in the splice variant of the represented arbitrary nucleic acid of SEQ ID NO:213 to 225 or table 8 or the table 9 or by the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225 directly to the splice variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Preferred splice variant is the straight splice variant to homologue or collateral line homologue by the splice variant of the represented nucleic acid of SEQ ID NO:209 or coding SEQ ID NO:210.Preferably, splice variant amino acids coding ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory, one or the many myristoylations site any one or more, and have kinase activity.

The another kind of nucleic acid variant that is used to implement the inventive method is the allelic variant of coding nucleic acid of SIK polypeptide as hereinbefore defined of encoding.

The invention provides the method that increases every strain plant flowers number, described method be included in introduce in the plant and expression table 8 or table 9 in arbitrary nucleic acid of providing or introduce by the allelic variant of the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225 or in plant and expression table 8 or table 9 in the arbitrary amino acid sequence that provides or by the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225 directly to the allelic variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Preferred allelic variant be the allelic variant of SEQ ID NO:209 or SEQ ID NO:210 directly to the allelic variant of the coding nucleic acid of homologue or collateral line homologue.Preferably, the allelic variant amino acids coding comprises that ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory, one or the many myristoylations site any one or more, and has kinase activity.

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant that obtains by gene reorganization.Also can use gene reorganization or orthogenesis to produce the nucleic acid variant of coding SIK polypeptide.

The invention provides the method that increases every strain plant flowers number, described method be included in the plant introduce and expression table 8 or table 9 in arbitrary nucleotide sequence of providing or introduce by the variant of the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225 or in plant and expression table 8 or table 9 in the arbitrary amino acid sequence that provides or by the represented arbitrary nucleotide sequence of SEQ ID NO:213 to 225 directly to the variant of the coding nucleic acid of homologue, collateral line homologue or homologue, described variant nucleic acid obtains by gene reorganization.Preferably; obtain variant nucleic acid encoding amino acid by gene reorganization; described amino acid ATP-land, serine threonine kinases avtive spot mark, N-end are rich in ser structure territory, one or the many myristoylations site any one or more, and have kinase activity.

In addition, can pass through site-directed mutagenic obtained nucleic acid variant.Several method can be used for realizing site-directed mutagenesis; The most frequently used method that is based on PCR (Current Protocols in MolecularBiology.Wiley writes).

The SIK nucleic acid of coding SIK sample polypeptide can be from any natural or artificial source.Can on composition and/or genome environment, modify the natural form of described SIK nucleic acid by careful manual operation.Preferred SIK coding nucleic acid is from plant, and preferred source more preferably is derived from Gramineae (Poaceae) from monocotyledons again, and most preferably nucleic acid is from rice.

Therefore quoting of any SIK polypeptide of this paper means the SIK polypeptide of definition as mentioned.Any SIK nucleic acid of this type of SIK polypeptide of encoding is suitable for implementing the inventive method to increase every strain plant flowers number.

The present invention also comprises by the obtainable plant of the inventive method or its part (comprising seed).Described plant or its part comprise the nucleic acid transgenosis of the coding SIK polypeptide of definition as mentioned.

The present invention also provides genetic constructs and carrier to promote to introduce and/or express the SIK nucleotide sequence that is used for the inventive method in plant.

Thereby, gene construct is provided, it comprises:

(i) as top defined SIK nucleic acid or SIK coding nucleic acid;

(ii) one or more regulating and controlling sequences that effectively are connected with the SIK nucleic acid of (i).

Reducing or removing substantially under the situation of SIK nucleic acid with the full rate of the harvest index that produces the thousand seed weight that increases, increase and increase, preferred construct is a kind of such construct, it comprises the reverse repetition of SIK nucleic acid, preferably can form the reverse repetition of hairpin structure, the described regulation and control that oppositely repeat to be subjected to constitutive promoter.

The construct that is used for the inventive method can use the well-known recombinant DNA technology of those skilled in the art to make up.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for transcribing goal gene in cell transformed.The present invention also provides the purposes in the methods of the invention of gene construct as hereinbefore defined.

Plant transforms with the carrier that comprises aim sequence.The technician is appreciated that successfully and transforms, selects and breed the host cell that contains aim sequence and the genetic elements that must exist very much on carrier.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.Term " regulatory element ", " regulating and controlling sequence " and " promotor " are in all commutative use of this paper and in above definition.

Advantageously, can use any type promotor in the inventive method.Term " promotor " refer to be positioned at genetic transcription starting point upstream and participate in identification and in conjunction with RNA polymerase and other protein, thereby instruct the nucleic acid regulating and controlling sequence of the transcribed nucleic acid that effectively connects.Promotor can be a constitutive promoter.Alternatively, promotor can be an inducible promoter.In addition or alternatively, promotor can be a tissue-specific promoter.Only can in particular organization, start the promotor this paper that transcribes and be called " tissue-specific ", similarly, only in specific cells, start the promotor this paper that transcribes and be called " cell-specific ".

Preferably, nucleotide sequence effectively is connected with constitutive promoter.Preferred promoter derives from plant, is transformed if also preferably derive from the monocotyledons monocotyledons.Preferred again constitutive promoter is the GOS2 promotor of being represented by the nucleotide sequence that is substantially similar to SEQ ID NO:56 or 226.Most preferably GOS2 promotor such as SEQ ID NO:56 or 226 represented.Be understood that suitability of the present invention is not limited to the SIK nucleotide sequence by SEQ ID NO:209 representative, SIK expression of nucleic acids when suitability of the present invention also is not limited to by the GOS2 promoters driven simultaneously.The example that also can be used for driving other constitutive promoters that coding SIK nucleotide sequence expresses above shows.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.The additional adjustment element can comprise transcribes and translates reinforcement.One skilled in the art will recognize that and go for implementing terminator of the present invention and strengthen subsequence.Intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also comprise need be used for the replication sequence starting point of keeping and/or duplicating in particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to f1-ori and colE1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Thereby genetic constructs can randomly comprise the selected marker.In this paper " definition " part selected marker has been described in more detail.This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top " definition " part.

Can also regulate SIK nucleic acid or SIK polypeptide expression by genetic modification being introduced the SIK locus.The locus of this paper definition means gene regions, and described gene regions comprises goal gene and gene coding region upstream or downstream 10kb.

For example, can introduce described genetic modification by arbitrary (or multiple) following method: T-DNA activation, TILLING and homologous recombination.Introducing genetic modification step afterwards is to select the suitable expression of being regulated.

The inventive method advantageously is applicable to any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.

Other favourable plants be selected from composite family (Asteraceae) as Helianthus (Helianthus), Tagetes for example species Sunflower Receptacle [Sunflower Receptacle (sunflower)], spiceleaf Flower of Aztec Marigold (Tagetes lucida), Flower of Aztec Marigold (Tagetes erecta) or Tagetes signata (Tagetes tenuifolia) [Marigold], Cruciferae (Brassicaceae) as Btassica (Brassica), Arabidopsis, for example species colea, overgrown with weeds overgrown with weeds blue or green [canola oil dish, rape, turnip] or Arabidopis thaliana; Pulse family (Fabaceae) is as Glycine for example species soybean, soybean (Soja hispida) or soybean (Soja max) [soybean]; Flax family (Linaceae) (Linaceae) is as linum (Linum) species flax [flax (flax, linseed)] for example; Gramineae such as Hordeum (Hordeum), Secale (Secale), Avena (Avena), sorghum (Sorghum), Oryza (Oryza), Zea (Zea), Triticum (Triticum) is species barley [barley] for example, rye [rye], oat, oat (Avena fatua), wild avena sativa, than praising oat, Avena fatua var.sativa, hybrid oat [oat], dichromatism chinese sorghum [Chinese sorghum, grain], rice, broad-leaved rice [rice], Zea mays [corn, corn], common wheat, durum wheat, the cylinder wheat, Triticum hybernum, Macha wheat (Triticum macha), common wheat (Triticum sativum) or common wheat (Triticum vulgare) [wheat, bread wheat, common wheat]; Solanaceae (Solanaceae) belongs to (Lycopersicon) for example potato [potato], tomato (Lycopersicon esculentum, Lycopersiconlycopersicum, Lycopersicon pyriforme, Solanum integrifolium or Solanumlycopersicum) [tomato (tomato)] as Solanum (Solanum), tomato.

But the present invention also comprises the plant that obtains with the method according to this invention.But therefore the present invention provides plant, its plant part or the vegetable cell that obtains with the method according to this invention, and described plant or its part or cell comprise SIK nucleic acid transgenosis (its SIK polypeptide of definition as mentioned of can encoding).

The present invention also is provided for producing the method for the transgenic plant of the output correlated character with various improvement above-mentioned, and it is included in the SIK nucleic acid of introducing and expressing as define in the plant.

Host plant advantageously can synthesize whole plants of used polypeptide in the inventive method in principle for used nucleic acid or carrier, expression cassette or construct or carrier in the inventive method.

More specifically, the invention provides and be used to produce the transgenic plant that have the output correlated character of various improvement with respect to control plant, described method comprises:

(i) in vegetable cell, introduce and express SIK nucleic acid; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth;

(ii) obtain having the plant of every strain plant flowers number of increase.

Also be provided for producing the transgenic plant that have the output correlated character of multiple improvement with respect to control plant, described method comprises:

(i) in vegetable cell, introduce the construct that is used for regulating down SIK genetic expression; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth;

(ii) obtain having one or multinomial plant in the full rate of the harvest index of thousand seed weight, increase of increase and increase.

Nucleic acid is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleic acid is preferably by transforming introduced plant.Also be preferred for decrement regulate SIK genetic expression and be introduced into vegetable cell or plant in construct comprise the reverse repetition of SIK nucleic acid or its part.

Usually after conversion, vegetable cell or the cell colony of selecting one or more marks to exist, wherein said mark become whole strain plant with the material regeneration that transforms subsequently by the effable genes encoding of the plant that moves with the goal gene corotation.

As mentioned, the Agrobacterium that transforms with expression vector of the present invention also can transform plant with himself known method, as test plant, picture Arabidopis thaliana or crop plants, for example as cereal, corn, oat, rye, barley, wheat, soya bean (soya), rice, cotton, beet, rape, Sunflower Receptacle, flax, hemp, potato, tobacco, tomato, Radix Dauci Sativae, pimento, the Semen Brassicae campestris rape, tapioca (flour), cassava, arrowroot, Flower of Aztec Marigold, clover, romaine lettuce and multiple trees, nut and grape vine species, oil-containing crop plants particularly, as soya bean, peanut, the Viscotrol C plant, Sunflower Receptacle, corn, cotton, flax, the Semen Brassicae campestris rape, coconut, oil palm, safflower (Carthamus tinctorius), cocoa beans, for example the leaf or the leaf segment of scratching by water-bath in Agrobacterium solution cultivated it subsequently in suitable medium.

The vegetable cell of genetic modification can be regenerated by all methods that those skilled in the art are familiar with.Suitable method be found in S.D.Kung and R Wu, Potrykus or

Above-mentioned publication with Willmitzer.

In order to select plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example, can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying.Another kind of possibility is included in the seed of growing on the agar plate that uses suitable selective agent (as required after sterilization), makes the seed that only transforms can grow into plant.Perhaps, plant transformed is by the existence screening of above-mentioned those selective markers.

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition can for example use Southern to analyze goal gene were estimated.Alternative or extraly, expression level can use Northern and/or Western or quantitative PCR analysis to monitor, all technology are that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clonal expansion method or classical breeding technique.For example, the first-generation (or T1) transforms plant can carry out selfing, and so that the s-generation (or T2) of isozygotying transformant to be provided, and the T2 plant further breeds by classical breeding technique.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's transformant (for example being transformed) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the root stock of the conversion of unconverted grafting of tender branch.

The present invention extends to any vegetable cell or the plant that produces by any means described in the literary composition clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by any preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical one or more yielding characteristicses and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.

The present invention also comprises host cell, and it contains the separative SIK nucleic acid of definition as mentioned.The preferred host cell of the present invention is a vegetable cell.Host plant advantageously can synthesize whole plants of used polypeptide in the inventive method in principle for used nucleic acid or carrier, expression cassette or construct or carrier in the inventive method.

The present invention also extend to plant the part gathered in the crops as, but be not limited to seed, leaf, fruit, flower, stem, root stock, stem tuber and bulb.The invention further relates to from, preferred directly from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, oil, fat and lipid acid, starch or protein.

The present invention also comprises SIK nucleic acid and the purposes of SIK polypeptide in improvement multiple output correlated character As mentioned above.

The nucleic acid of coding SIK polypeptide can be used for the procedure of breeding, wherein identifies the dna marker that can be connected with the SIK gene hereditarily.Described nucleic acid/gene can be used for defining molecule marker.This dna marker can be used for selecting to have in the methods of the invention the plant of the output of the increase of definition as mentioned subsequently in the procedure of breeding.

The allelic variant of SIK nucleic acid/gene also can be used for the auxiliary procedure of breeding of mark.This procedure of breeding needs to introduce allelic variation by for example using the EMS mutagenesis that plant is carried out mutagenic treatment sometimes; Alternatively, this program can be from one group of allelic variant of the non-artificial what is called that causes " nature " origin.Carry out the evaluation of allelic variant subsequently, for example by the PCR method.After this be used to select discuss and cause increasing the step of excellent allelic variant of the sequence of output.Generally the growth performance that contains the plant of the different allelic variants that sequence is discussed to some extent by monitoring is implemented to select.Can be in the greenhouse or field monitoring growth performance.Other optional step comprise and will identify the plant and the another kind of plant hybridization of excellent allelic variant.This can be used for for example producing target phenotype combination of features.

SIK nucleic acid also can be as probe so that carry out genetic mapping or physical mapping to gene, and described probe reaches the mark of the proterties related with these genes as the part of described gene.This type of information can be used for plant breeding, so that exploitation has the strain system that wants phenotype.This purposes of SIK nucleic acid only needs to have the nucleotide sequence of at least 15 length of nucleotides.SIK nucleic acid can be used as restriction fragment length polymorphism (RFLP) mark.The Southern trace of the plant genome DNA of restrictive diges-tion (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, ALaboratory Manual) can be used the SIK nuclei acid probe.What produce carries out genetic analysis to make up genetic map in conjunction with graphic can use a computer subsequently program such as MapMaker people (1987) Genomics 1:174-181 such as () Lander.In addition, this nucleic acid can be used for surveying the Southern trace of the genomic dna that contains one group of individuality handling through restriction endonuclease, and wherein said one group of individual representative has the parental generation and the offspring of definite genetic cross.The separation of dna polymorphism is marked and is used for calculating the position of SIK nucleic acid in using the previous genetic map that obtains of this colony people (1980) Am.J.Hum.Genet.32:314-331 such as () Botstein.

Generation and its purposes in genetic mapping of plant gene deutero-probe have been described in Bernatzky and Tanksley (Plant Mol.Biol.Reporter 4:37-41,1986).Numerous publications have been described the genetic mapping that uses methodology mentioned above or its modification method that specific cDNA is cloned.For example, to hand over group, the group that backcrosses, panmictic population, contiguous isozygotying mutually be can be used for mapping with other population of individuals to F2.This type of methodology is that those skilled in the art are well-known.

It (is the arrangement of sequence on physical map that described nucleic acid probe also can be used for physical mapping; See that Hoheisel etc. exists: Non-mammalian Genomic Analyasis:A Practical Guide, Academic press 1996, the 319-346 pages or leaves and the reference of wherein quoting).

In another embodiment, nucleic acid probe can directly use in fluorescence in situ hybridization (FISH) graphing method (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although large-scale clone is used in current FISH graphing method support; See people such as Laan (1995) Genome Res.5:13-20), however the improvement of sensitivity can allow to use shorter probe to carry out the FISH mapping.

The multiple method based on nucleic acid amplification that is used for genetic mapping and physical mapping can be used described nucleic acid and implement.Example is found in " definition " part in the literary composition.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med 11:95-96), pcr amplified fragment; Sheffield etc., (1993) Genomics 16:325-332), allele-specific connects (Landegren etc., (1988) Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (people such as Walter Science 241:1077-1080),, Nat.Genet.7:22-28) and Happy mapping (Dear and Cook, (1989) Nucleic Acid Res.17:6795-6807) (1997).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The inventive method produces the plant with the output correlated character that has improvement as previously described.These have proterties also can with favourable other proterties combinations economically, as other output increase proterties, to other abiotic and biological tolerances of coercing, the proterties of regulating various constructivity features and/or biochemical characteristics and/or physiologic character.

The detailed description of II class HD-Zip transcription factor

Have now found that the nucleic acid of coding II class HD-Zip transcription factor is used for the content of improved seed stock.Therefore the present invention also provides by regulating the expression of nucleic acid in plant of coding II class HD-Zip transcription factor, with respect to the method for the content of stock in the control plant improved seed.The present invention also is provided for implementing the nucleotide sequence and the construct of these class methods.The present invention and then the seed that has the stock content of improvement with respect to control plant is provided, described seed has the expression of nucleic acids of the coding II class HD-Zip transcription factor of being regulated.

The invention provides the method with respect to the content of stock in the control plant improved seed, described method comprises the expression of nucleic acid in plant of regulating coding II class HD-Zip transcription factor.

The preferred method that is used for regulating the expression of nucleic acids of (preferred increasing) coding II class HD-Zip transcription factor is by introduce and express the nucleic acid of the II class HD-Zip transcription factor of encoding as defined here plant.

" II class HD-Zip transcription factor " means and comprises following polypeptide: (i) homeodomain frame; (ii) leucine zipper; Motif I, II that provides (iii) and III (with random order).

Motif I (SEQ ID NO:279)

RKKLRL or have motif I that one or more conserved amino acids replace in any site and/or have the motif I of one or two non-conservative change in site arbitrarily; With

Motif II (SEQ ID NO:280)

TKLKQTEVDCEFLRRCCENLTEEN or have motif II that one or more conserved amino acids replace in any site and/or have at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or the motif of higher sequence identity with the preferred sequence that increases with motif II; With

Motif III (SEQ ID NO:281)

TLTMCPSCER or have motif III that one or more conserved amino acids replace in any site and/or have one, the motif III of two or three non-conservative changes in site arbitrarily.

This paper means the nucleic acid of the coding II class HD-Zip transcription factor of definition as mentioned to " nucleic acid of coding II class HD-Zip transcription factor " or quoting arbitrarily of II class HD-Zip coding nucleic acid, and this type of nucleic acid is used to implement the inventive method.

As mentioned, the preferred method that is used for regulating the expression of nucleic acids of (preferred increasing) coding II class HD-Zip transcription factor is by introduce and express the nucleic acid of coding II class HD-Zip transcription factor plant.

The present invention illustrates by the nucleotide sequence plant transformed with the represented coded polypeptide sequence SEQ ID NO:230 of SEQ ID NO:229.Yet enforcement of the present invention is not limited to these sequences; Can advantageously use as defined herein any II class HD-Zip transcription factor coding nucleic acid or II class HD-Zip transcription factor to implement the inventive method.

For example, coding can be used to implement the inventive method by the straight nucleic acid to homologue or collateral line homologue of the represented aminoacid sequence of SEQ ID NO:230.In the Table A of embodiment 1, provide this type of directly to the example of homologue and collateral line homologue.

Directly comprise the evolution notion that is used for describing the gene ancestral relationship to homologue and collateral line homologue.The collateral line homologue is the gene of same species endogenous origin in my late grandfather's gene replication; Be from different biological genes by the origin of species directly to homologue.

Directly can easily find by carrying out so-called interactivity blast search to homologue and collateral line homologue.This generally comprises a BLAST, a described BLAST participates in submitting to search sequence (for example SEQ ID NO:229 or SEQ ID NO:230) to be used for that (as public's available ncbi database, described database can find in following website: blast search http://www.ncbi.nlm.nih.gov) at arbitrary sequence library.When nucleotide sequence begins, use BLASTN or TBLASTX (using the standard default value) usually, and, can use BLASTP or TBLASTN (use standard default value) when when protein sequence begins.Randomly can screen BLAST result.The full length sequence of submitting The selection result and non-The selection result subsequently to is with at carry out reverse BLAST (the 2nd BLAST) from the sequence of biology, wherein search sequence is from described biology (be under the situation of SEQ ID NO:229 or SEQ ID NO:230 in search sequence wherein, the 2nd BLAST thereby will be at the rice sequence).The result who compares first and second blast searches subsequently.If from the high-order position of a BLAST hit be derived from search sequence from identical species of deutero-species wherein, oppositely BLAST then identifies the collateral line homologue subsequently ideally to produce in the highest search sequence of hitting; If the high-order position among the BLAST hit be not derived from search sequence from identical species of deutero-species wherein, and preferably when reverse BLAST, produce the search sequence that belongs to the highest row that hit, then identify directly to homologue.It is that with low E-value those hit that high-order position is hit.The E-value is low more, mark remarkable more (or in other words, this hit because of the chance odds low more).The calculating of E-value is well-known in the art.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.

Coding can be used to implement the inventive method by the nucleic acid of the homologue of the arbitrary amino acid sequence that provides among the nucleic acid of the homologue of the represented aminoacid sequence of SEQ ID NO:230 or the coding schedule H.

Also be used for the inventive method be the arbitrary amino acid sequence that provides among the arbitrary amino acid whose derivative that provides of coding schedule H or the table H directly to the nucleic acid of the derivative of homologue or collateral line homologue." derivative " comprises such peptide, oligopeptides, polypeptide, wherein compare with the aminoacid sequence of the protein (as by the represented albumen of SEQID NO:230) of natural generation form, they comprise the interpolation of the amino-acid residue that the amino-acid residue that takes place with non-natural takes place amino acid whose replacement or non-natural.

Can use routine techniques well known in the art such as sequence alignment easily to identify homologue (or homologue protein, comprise) directly to homologue and collateral line homologue.It is well-known in the art being used for the method that aligned sequences compares, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.GAP uses Needleman and Wunsch algorithm ((1970) J Mol Biol48:443-453) to find the comparison that makes the maximization of coupling number and make minimized two complete sequence of room number.BLAST algorithm (Altschul etc. (1990) J Mol Biol 215:403-10) sequence of calculation identity percentage ratio and execution are to the statistical study of similarity between two sequences.Being used to carry out software that BLAST analyzes and being the public, to pass through NCBI obtainable.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.It is the methods of marking of percentage ratio and identifying easily with acquiescence pairing comparison parameter and unit that homologue can use ClustalW multiple sequence alignment algorithm (1.83 version) for example.The overall percentage ratio of similarity and identity also can use one of obtainable method in the MatGAT software package and determine (Campanella etc., BMC Bioinformatics.4,29,2003).Can carry out trickle edit to optimize the comparison between the conservative motif, apparent as those skilled in the art.As using full length sequence, also can use the ad hoc structure territory in addition to identify substituting of homologue.

Use ClustalW and default parameters, at whole nucleotide sequence or determined amino acid sequence sequence identity value, represent described sequence identity value with per-cent below.

Preferably, the polypeptide (as showing any peptide sequence that H provides) by the nucleic acid encoding that is used for the inventive method has at least 40% with the preferred sequence of increase and the polypeptide of SEQ ID NO:230,41%, 42%, 43%, 44%, 45%, 46%, 47%, 48%, 49%, 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% identity.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprise adjusting by SEQ ID NO:2 or its directly to the expression of coding nucleic acid in plant of the represented II class HD-Zip transcription factor of homologue, collateral line homologue or homologue.

The preferred method of expression that is used for regulating the coding nucleic acid of (preferred increasing) II class HD-Zip transcription factor is by introducing and express coding nucleic acid by the represented II class HD-Zip transcription factor of SEQ ID NO:230 plant, or it is directly to the coding nucleic acid of homologue, collateral line homologue or homologue.

Above-mentioned straight in homologue, collateral line homologue and homologue fall into the definition of II class HD-Zip transcription factor, promptly meaning straight is to comprise following polypeptide to homologue, collateral line homologue and homologue: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.

Can use multiple structural domain and motif to help to identify the sequence that is used for the inventive method.There is the specialized database that is used to identify structural domain, for example, SMART (Schultz etc. (1998) the Proc.Natl.Acad.Sci. U.S. 95,5857-5864; Letunic etc. (2002) Nucleic Acids Res 30,242-244), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318), Prosite (Bucher and Bairoch (1994), be used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and in the function of automatization sequence interpretation, () ISMB-94; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002)) evaluation.Can also use routine techniques, sequence alignment as described herein is identified structural domain and motif.

The nucleic acid that is used for the inventive method needs not be total length nucleic acid, because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.Example comprises the part of the nucleotide sequence of being represented by SEQ ID NO:229 or the part of arbitrary nucleotide sequence that table provides among the H.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprises part the expression in plant of adjusting by the represented nucleotide sequence of SEQ ID NO:229, or comprise the expression of part in plant of the arbitrary nucleotide sequence that provides among the reconciliation statement H, or comprise the arbitrary amino acid sequence that provides among the reconciliation statement H directly to the expression of part in plant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

Be used for regulating (preferred increasing) this type of part the preferred method of the expression of plant be by in plant, introduce and expression table H in the part of the nucleotide sequence that provides, or be included in introduce in the plant and expression table H in any peptide sequence of providing directly to the part of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

The part that is used for the inventive method comprises that length is enough to encode and falls into the part of the polypeptide in the definition of II class HD-Zip transcription factor, promptly means to comprise following polypeptide: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.In addition, this type of part and II class HD-Zip transcription factor have essentially identical biologic activity.

Preferably, partial-length is at least 500 continuous nucleotides, preferred length is at least 750 continuous nucleotides, more preferably length is at least 1250 continuous nucleotides, described continuous nucleotide be the arbitrary amino acid sequence that provides among the arbitrary nucleotide sequence that provides among the H of SEQID NO:229 or table or the table H directly to any coding nucleic acid of homologue, collateral line homologue or homologue.Most preferred part is the part of nucleic acid SEQ ID NO:229.

Part can prepare part by discussion nucleic acid is carried out one or more disappearances as herein defined.Part can be used with isolating form, perhaps itself and other coding (or non-coding) sequence can be merged, so that for example, produce and made up some active protein.When merging with other encoding sequences, the polypeptide that is produced after translating may be bigger than the part of prediction.

Another nucleic acid that is used for the inventive method is such nucleic acid, under the stringent condition that reduces, preferably under medium stringent condition, more preferably under stringent condition, its can with the represented nucleic acid hybridization of SEQ ID NO:229, or under the stringent condition that reduces, preferably under medium stringent condition, more preferably under stringent condition, its can with the nucleic acid array hybridizing that provides of table H, or under the stringent condition that reduces, preferably under medium stringent condition, more preferably under stringent condition, its can with any peptide sequence of providing among the table H directly to homologue, the coding nucleic acid hybridization of collateral line homologue or homologue.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprise adjusting can with the expression of nucleic acid in plant by the represented nucleic acid hybridization of SEQ ID NO:229, or comprise the expression of nucleic acid in plant of the nucleic acid array hybridizing that adjusting can provide among the H with table, or comprise adjusting can with any peptide sequence of providing among the table H directly to the expression of nucleic acid in plant of the nucleic acid sequence encoding hybridization of homologue, collateral line homologue or homologue.Most preferably hybridization sequences can with by the represented nucleic acid hybridization of SEQ ID NO:229.Hybridization sequences preferably can be under the stringent condition that reduces, preferably under medium stringent condition, more preferably hybridize under stringent condition.

Be used for regulating (preferred increasing) this type of hybridization sequences the preferred method of the expression of plant be by in plant, introduce and express can with the nucleic acid of the nucleic acid array hybridizing that provides among the table H, or be included in introduce in the plant and express can with arbitrary aminoacid sequence of providing among the table H directly to the nucleotide sequence of the nucleic acid sequence encoding hybridization of homologue, collateral line homologue or homologue.Most preferably hybridization sequences can with by the represented nucleic acid hybridization of SEQ ID NO:229.Hybridization sequences preferably can be under the stringent condition that reduces, preferably under medium stringent condition, more preferably hybridize under stringent condition.

The hybridization sequences that is used for the inventive method comprises that length is enough to encode and falls into the nucleic acid of the polypeptide in the definition of II class HD-Zip transcription factor, promptly means to comprise following polypeptide: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.In addition, this type of hybridization sequences and II class HD-Zip transcription factor have essentially identical biologic activity.

The hybridization sequences normal length is at least 500 continuous nucleotides, preferred length is at least 750 continuous nucleotides, more preferably length is at least 1250 continuous nucleotides, described continuous nucleotide be arbitrary aminoacid sequence of providing among any nucleic acid of the nucleic acid array hybridizing that can provide among the H with table or the table H directly to any coding nucleic acid of homologue, collateral line homologue or homologue.Most preferably continuous nucleotide be have can with the nucleic acid by the represented nucleic acid hybridization of SEQ ID NO:229, or the nucleic acid of its part hybridization.

Another nucleic acid that is used for the inventive method is the splice variant of SEQ ID NO:229, or the splice variant of any nucleotide sequence of providing among the H of table, or arbitrary aminoacid sequence of providing of table H directly to the splice variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprises splice variant the expression in plant of adjusting by the represented nucleic acid of SEQ ID NO:229, or comprise the expression of splice variant in plant of the nucleotide sequence that provides among the reconciliation statement H, or comprise arbitrary aminoacid sequence of providing among the reconciliation statement H directly to the expression of splice variant in plant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

Be used for regulating (preferred increasing) this type of splice variant the preferred method of the expression of plant be by in plant, introduce and expression table H in the splice variant of the nucleotide sequence that provides, or be included in introduce in the plant and expression table H in arbitrary aminoacid sequence of providing directly to the splice variant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

The splice variant that is used for the inventive method comprises that length is enough to encode and falls into the nucleic acid of the polypeptide in the definition of II class HD-Zip transcription factor, promptly means to comprise following polypeptide: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.In addition, this type of splice variant and II class HD-Zip transcription factor have essentially identical biologic activity.

Another nucleic acid that is used to implement the inventive method is the allelic variant of SEQ ID NO:229, or the allelic variant of any nucleotide sequence of providing among the H of table, or arbitrary aminoacid sequence of providing of table H directly to the allelic variant of the coding nucleic acid of homologue, collateral line homologue or homologue.The natural existence of allelic variant, and to be included in the inventive method scope be these natural allelic purposes.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprises allelic variant the expression in plant of adjusting by the represented nucleic acid of SEQ ID NO:229, or comprise the expression of allelic variant in plant of the nucleotide sequence that provides among the reconciliation statement H, or comprise arbitrary aminoacid sequence of providing among the reconciliation statement H directly to the expression of allelic variant in plant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

Be used for regulating (preferred increasing) this type of allelic variant the preferred method of the expression of plant be by in plant, introduce and expression table H in the allelic variant of the nucleotide sequence that provides, or be included in introduce in the plant and expression table H in arbitrary aminoacid sequence of providing directly to the allelic variant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue.

The allelic variant that is used for the inventive method comprises that length is enough to encode and falls into the nucleic acid of the polypeptide in the definition of II class HD-Zip transcription factor, promptly means to comprise following polypeptide: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.In addition, this type of allelic variant and II class HD-Zip transcription factor have essentially identical biologic activity.

Another nucleic acid that is used for the inventive method is the nucleic acid that obtains by gene reorganization.Gene reorganization or orthogenesis can also be used for the variant of arbitrary nucleic acid that generation table H provides, or arbitrary aminoacid sequence of providing of table H directly to the variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

The invention provides method with respect to the content of stock in the control plant improved seed, described method comprises variant the expression in plant of adjusting by the represented nucleic acid of SEQ ID NO:229, or comprise the expression of variant in plant of the arbitrary nucleotide sequence that provides among the reconciliation statement H, or comprise arbitrary aminoacid sequence of providing among the reconciliation statement H directly to the expression of variant in plant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue, described variant nucleic acid obtains by gene reorganization.

Be used for regulating (preferred increasing) this type of variant the preferred method of the expression of plant be by in plant, introduce and expression table H in the variant of arbitrary nucleotide sequence of providing, or be included in the plant introduce and expression table H in the arbitrary amino acid sequence that provides directly to the variant of the nucleic acid sequence encoding of homologue, collateral line homologue or homologue, described variant nucleic acid obtains by gene reorganization.

This type of variant that is used for the inventive method that obtains by gene reorganization comprises that length is enough to encode and falls into the nucleic acid of the polypeptide in the definition of II class HD-Zip transcription factor, promptly means to comprise following polypeptide: (i) homeodomain box; (ii) leucine zipper; Motif I (iii) as herein described, II and III.

In addition, can also be by the site-directed mutagenic obtained nucleic acid that is used for the inventive method.Several method can be used for realizing site-directed mutagenesis, the most frequently used method that is based on PCR (CurrentProtocols in Molecular Biology.Wiley writes).

II class HD-Zip transcription factor is showed the general biologic activity (at least with its natural form) of transcription factor and is had dna binding activity and activation structure territory usually.Those skilled in the art can use conventional instrument and technology to determine that easily activation structure territory and DNA-are in conjunction with active existence.Sessa etc. 1997 (J Mol Biol 274 (3): 303-309) studied ATHB-1 and ATHB-2 (=HAT4) DNA of HD-Zip (HD-Zip-1 and-2) structural domain is in conjunction with attribute, and find that they and DNA interact as homodimer and discern two false palindrome sequences of different 9bp, be respectively CAAT (A/T) ATTG (BS-1) and CAAT (G/C) ATTG (BS-2).From the mutation analysis of HD-Zip-2 structural domain, they determine the conservative amino acid residues of spiral 3:, Val47 and Asn51 and Arg55 are crucial to the dna binding activity of HD-Zip-2 structural domain.They also report by replacing Arg55 with Methionin or eliminating by the preferred identification of HD-Zip-2 structural domain in the G/C in site, center base pairing by corresponding residue (being respectively L-Ala and tryptophane) replacement Glu46 and Thr56 with HD-Zip-1.

According to preferred feature of the present invention, the expression of being regulated is the expression that increases.The method that increases the expression of nucleic acid, gene or gene product is fully put down in writing in this area.

The coding nucleic acid that is used for regulating II class HD-Zip transcription factor comprises in the other method of the expression of plant and reducing or the basic expression of native gene in plant of removing coding II class HD-Zip transcription factor." reducing or basic the removal " of mentioning herein means native gene expression and/or polypeptide level and/or the polypeptide active reduction with respect to control plant.Compare with control plant, reducing or removing to increase progressively preferred sequence substantially is at least 10%, 20%, 30%, 40% or 50%, 60%, 70%, 80%, 85%, 90%, or 95%, 96%, 97%, 98%, 99% or more reduction.The method that reduces expression is in above " definition " part description.

In order to reduce or to remove the expression of native gene in plant substantially, need the Nucleotide of successive basically of the sufficient length of nucleotide sequence.In order to carry out gene silencing, this length can be few to 20,19,18,17,16,15,14,13,12,11,10 or still less Nucleotide, and perhaps this length can the whole gene of as many as (comprising 5 ' and/or 3 ' UTR, in part or total length).Basically successive nucleotide fragments any nucleotide sequence that can provide among the H from SEQ ID NO:229 or from table or from the arbitrary aminoacid sequence that provides in can coding schedule H directly to any nucleic acid of homologue, collateral line homologue or homologue.The nucleotide sequence of coding (functional) polypeptide is not that discussed herein to be used to reduce or to remove substantially the several different methods that native gene expresses required.

The nucleic acid that is suitable for the inventive method can be from any natural or artificial source.Can on composition and/or genome environment, modify the natural form of described nucleic acid by careful manual operation.Preferred nucleic acid is from plant, and preferred source is from dicotyledons again, and preferred source is from Cruciferae again, and more preferably nucleic acid is from Arabidopis thaliana.

The preferred method of expression that is used for regulating the coding nucleic acid of (preferred increasing) II class HD-Zip transcription factor is the coding nucleic acid of introducing and expressing II class HD-Zip transcription factor plant, yet the seed storage thing content that the effect of enforcement present method promptly improves can also use other known technologies to realize.A kind of this type of technology is that T-DNA activates mark.Can also use the TILLING technology to reproduce effect of the present invention.Can also use homologue to reproduce regeneration effect of the present invention, the selected location that described homologous recombination allows selected nucleic acid to determine in genome is introduced.

Implement the inventive method the seed that has the seed storage thing content of improvement with respect to the seed of control plant is provided.The seed storage thing content of improvement refers to one or more content with respect to the improvement of control plant respective items in lipid, oil, fat acid, starch, sugar and the protein.Preferably, the expression of coding nucleic acid in plant of regulating II class HD-Zip transcription factor produces the seed with respect to control plant, has the plant of seed of the oil-contg of increase.In the case, the expression of being regulated is the overexpression of coding nucleic acid in plant of II class HD-Zip transcription factor normally.Preferably, the expression of coding nucleic acid in plant of regulating II class HD-Zip transcription factor produces the seed with respect to control plant, has the plant of seed of the protein content of increase.In the case, the expression of being regulated normally reduces or removes substantially the expression of native gene II class HD-Zip transcription factor encoding gene.

The present invention also comprises by the obtainable plant of the inventive method or its part (especially seed).Described plant or its part comprise nucleic acid transgenosis (comprising mentioned above as being used for arbitrary nucleotide sequence of the inventive method).

The present invention also provides genetic constructs and carrier to promote to introduce in plant and/or to express mentioned above as be used for the nucleotide sequence of the inventive method.

More specifically, provide gene construct, it comprises:

(i) as the coding nucleic acid of top defined II class HD-Zip transcription factor;

(ii) one or more regulating and controlling sequences that effectively are connected with the nucleic acid of (i).

The construct that is used for the inventive method can use the well-known recombinant DNA technology of those skilled in the art to make up.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for expressing goal gene in cell transformed.The present invention also provides as defined gene construct purposes in the methods of the invention in the literary composition.

Plant transforms with the carrier that comprises aim sequence.The technician is appreciated that successfully and transforms, selects and breed the host cell that contains aim sequence and the genetic elements that must exist very much on carrier.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.Term " regulatory element ", " regulating and controlling sequence " and " promotor " are in all commutative use of this paper and in above definition.

Advantageously, can use the promotor of any kind, no matter nature or synthetic, drive the expression of nucleotide sequence.See of the definition of this paper " definition " part to various promotor types.

According to preferred feature of the present invention, purpose nucleic acid effectively is connected with constitutive promoter.Constitutive promoter is at the great majority of g and D but must not be all stages transcriptional activations all, and is generally to express basically.Preferred constitutive promoter is the GOS2 promotor, and more preferably constitutive promoter is a rice GOS2 promotor, and preferred again constitutive promoter is to be represented by the nucleotide sequence that is substantially similar to SEQ ID NO:56 or SEQ ID NO:282.Most preferably constitutive promoter such as SEQID NO:56 or SEQ ID NO:282 are represented.The example that also can be used for implementing other constitutive promoters of the inventive method above shows.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.The additional adjustment element can comprise transcribes and translates reinforcement.One skilled in the art will recognize that and go for implementing terminator of the present invention and strengthen subsequence.Intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also comprise need be used for the replication sequence starting point of keeping and/or duplicating in particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to f1-ori and colE1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Thereby genetic constructs can randomly comprise the selected marker.This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top " definition " part.

The present invention also provides the method for the transgenic plant of stock content in generation has improvement with respect to control plant the seed, it is included in the plant introduces and expresses by the represented nucleotide sequence of SEQ ID NO:229, or be included in the plant introduce and expression table H in the nucleotide sequence that provides, or be included in the plant introduce and expression table H in the arbitrary amino acid sequence that provides directly to the nucleic acid sequence encoding of homologue, collateral line homologue or homologue, or be included in introduce in the plant and express this paper definition as being used for any nucleic acid of the inventive method.

More specifically, the invention provides and be used for producing the method for transgenic plant that has the seed stock content of improvement with respect to control plant, described method comprises:

(i) in plant, plant part or vegetable cell, introduce and express by the represented nucleotide sequence of SEQ ID NO:229, or be included in the plant introduce and expression table H in the nucleotide sequence that provides, or be included in introduce in the plant and expression table H in the arbitrary amino acid sequence that provides directly to the nucleic acid sequence encoding of homologue, collateral line homologue or homologue; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth; With

(iii) from plant (ii), gather in the crops seed; Randomly

(iv) from seed (iii), extract in lipid, oil, lipid acid, starch, sugar and the protein any one or more.

The seed that can handle results is to extract specific seed storage thing, as lipid, oil, lipid acid, starch, sugar and protein.Especially, seed is used for extract oil.Can from seed that handled or undressed, extract described oil.

Nucleotide sequence is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleotide sequence is preferably by transforming introduced plant.

Term as referred to herein " conversion " as defined herein.Can follow-up clone's property propagation the plant tissue of (no matter take place or the embryo is taken place) by organ can transform and the whole strain plant that can therefrom regenerate with genetic constructs of the present invention.The concrete tissue of selecting will depend on clone's property proliferating system of the concrete species that can be used for and be suitable for just transforming most.The example organization target comprises leaf dish, pollen, embryo, cotyledon, hypocotyl, megagametophyte, callus, existing meristematic tissue (for example apical meristem, axillalry bud and root meristematic tissue) and inductive meristematic tissue (for example cotyledon meristematic tissue and hypocotyl meristematic tissue).Polynucleotide can instantaneous or stably be introduced host cell and can keep to nonconformity, for example as plasmid.Alternatively, polynucleotide can be integrated in the host genome.The transformed plant cells that produces can be used for regenerating in the manner known to persons skilled in the art the conversion plant subsequently.

The conversion of plant species is quite conventional technology now.Advantageously, the arbitrary method in several method for transformation can be used for goal gene is introduced suitable ancester cell.Method for transformation comprises that the chemical, the dna direct that use liposome, electroporation, increase dissociative DNA to take in are injected to the conversion method and the micro-projective method (microprojection) of plant, particle gun blast technique, use virus or pollen.Method for transformation can be selected from calcium/polyoxyethylene glycol method (Krens, F.A. etc., (1982) Nature 296, the 72-74 that is used for protoplastis; (1987) Plant Mol Biol 8:363-373 such as Negrutiu I); The electroporation of protoplastis ((1985) Bio/Technol 3 such as Shillito R.D., 1099-1102); Micro-injection (Crossway A etc., (1986) Mol.Gen Genet 202:179-185) to vegetable material; The particle bombardment method (Klein TM etc., (1987) Nature 327:70) of DNA or RNA coating, (nonconformity) virus infection method etc.Transgenosis rice plant preferably produces by agriculture bacillus mediated conversion method, use and be used for the known method that rice transforms arbitrarily, as in arbitrary following document, describing: disclosed European patent application EP 1198985 A1, Aldemita and Hodges (Planta 199:612-617,1996); Chan etc. (Plant Mol Biol 22 (3): 491-506,1993), Hiei etc. (Plant J 6 (2): 271-282,1994), its disclosure is incorporated herein by reference in this article, as providing fully.Under the situation that corn transforms, (Nat.Biotechnol 14 (6): 745-50 for preferable methods such as Ishida etc., 1996) or Frame etc. (Plant Physiol 129 (1): 13-22,2002) describe, its disclosure is incorporated herein by reference as fully in this article.

Can be by all methods that the technician was familiar with vegetable cell that genetic modification crosses of regenerating.S.D.Kung that can mention in the above and R.Wu, Potrykus or With find suitable method in the publication of Willmitzer.

Usually after conversion, vegetable cell or the cell colony of selecting one or more marks to exist, wherein said mark become whole strain plant with the material regeneration that transforms subsequently by the effable genes encoding of the plant that moves with the goal gene corotation.In order to select plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example, can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying.Another kind of possibility is included in the seed of growing on the agar plate that uses suitable selective agent (as required after sterilization), makes the seed that only transforms can grow into plant.Perhaps, plant transformed is by the existence screening of above-mentioned those selective markers.

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition can for example use Southern to analyze goal gene were estimated.Alternative or extraly, the expression level of newly introducing DNA can use Northern and/or Western to analyze or quantitative PCR is monitored, and all technology are that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clonal expansion method or classical breeding technique.For example, the first-generation (or T1) transforms plant can carry out selfing, and so that the s-generation (or T2) of isozygotying transformant to be provided, and the T2 plant can further breed by classical breeding technique.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's transformant (for example being transformed) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the root stock of the conversion of unconverted grafting of tender branch.

The present invention extends to any vegetable cell or the plant that produces by any means described in the literary composition clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by any preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical one or more yielding characteristicses and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.

The present invention also comprises host cell, and it contains the arbitrary nucleic acid that is used for the inventive method mentioned above.The preferred host cell of the present invention is a vegetable cell.

The inventive method advantageously is applicable to any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.

According to another preferred embodiment of the present invention, plant is to cultivate the crop plants that is used to produce oil usually.This type of example that is used to produce the crop plants of oil comprises Semen Brassicae campestris, canola oil dish, flax (linseed), soybean, Sunflower Receptacle, corn, oat, rye, barley, wheat, pepper (pepper), Flower of Aztec Marigold, cotton, oil palm, coconut (coconut palm), flax (flax), castor-oil plant (castor), peanut, olive, avocado, sesame, jatropha.

The part gathered in the crops that the present invention also extends to plant is seed particularly, also has leaf, fruit, flower, stem, root stock, stem tuber, root and bulb.The invention further relates to directly from preferred from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, oil, fat and lipid acid, starch or protein.The specific purpose product that can gather in the crops part from plant is an oil.

The present invention also comprises the purposes of any nucleic acid that is used for the inventive method that this paper mentions, is used for respect to control plant improved seed stock content.Useful especially be any peptide sequence of providing among the H of any nucleotide sequence that provides by represented nucleic acid of SEQ ID NO:229 and Table A and giving directly to any nucleic acid sequence encoding of homologue, collateral line homologue or homologue.The present invention also comprises by the purposes of the peptide sequence that provides among the purposes of the represented peptide sequence of SEQ ID NO:230 and the table H, is used for respect to control plant improved seed stock content.

These nucleic acid or encoded polypeptides can be used for the procedure of breeding, wherein identify the dna marker that can be connected with II class HD-Zip transcription factor encoding gene hereditarily.Described nucleic acid/gene or polypeptide itself can be used for defining molecule marker.This DNA or protein labeling can be used for selecting having the plant of the seed storage thing content of improvement subsequently in the procedure of breeding.

Allelic variant also can be used for the auxiliary procedure of breeding of mark.This procedure of breeding needs to introduce allelic variation by for example using the EMS mutagenesis that plant is carried out mutagenic treatment sometimes; Alternatively, this program can be from one group of allelic variant of the non-artificial what is called that causes " nature " origin.Carry out the evaluation of allelic variant subsequently, for example by the PCR method.After this be used to select discuss and cause increasing the step of excellent allelic variant of the sequence of output.Generally the growth performance that contains the plant of the different allelic variants that sequence is discussed to some extent by monitoring is implemented to select.Can be in the greenhouse or field monitoring growth performance.Other optional step comprise and will identify the plant and the another kind of plant hybridization of excellent allelic variant.This can be used for for example producing target phenotype combination of features.

Nucleic acid also can be as probe so that carry out genetic mapping or physical mapping to gene, and described probe reaches the mark of the proterties related with these genes as the part of described gene.This type of information can be used for plant breeding, so that exploitation has the strain system that wants phenotype.This purposes only needs to have the nucleotide sequence of at least 15 length of nucleotides.Nucleic acid can be used as restriction fragment length polymorphism (RFLP) mark.The Southern trace of the plant genome DNA of restrictive diges-tion (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) can be used nuclei acid probe.What produce carries out genetic analysis to make up genetic map in conjunction with graphic can use a computer subsequently program such as MapMaker people (1987) Genomics 1:174-181 such as () Lander.In addition, this nucleic acid can be used for surveying the Southern trace of the genomic dna that contains one group of individuality handling through restriction endonuclease, and wherein said one group of individual representative has the parental generation and the offspring of definite genetic cross.The separation of dna polymorphism is marked and is used for calculating the position of nucleic acid in using the previous genetic map that obtains of this colony people (1980) Am.J.Hum.Genet.32:314-331 such as () Botstein.

Generation and its purposes in genetic mapping of plant gene deutero-probe have been described in Bernatzky and Tanksley (1986) Plant Mol.Biol.Reporter 4:37-41.Numerous publications have been described the genetic mapping that uses methodology mentioned above or its modification method that specific cDNA is cloned.For example, to hand over group, the group that backcrosses, panmictic population, contiguous isozygotying mutually be can be used for mapping with other population of individuals to F2.This type of methodology is that those skilled in the art are well-known.

It (is the arrangement of sequence on physical map that described nucleic acid probe also can be used for physical mapping; See that Hoheisel etc. exists: Non-mammalian Genomic Analyasis:A Practical Guide, Academic press 1996, the 319-346 pages or leaves and the reference of wherein quoting).

In another embodiment, nucleic acid probe can directly use in fluorescence in situ hybridization (FISH) graphing method (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although large-scale clone is used in current FISH graphing method support; See people such as Laan (1995) Genome Res.5:13-20), however the improvement of sensitivity can allow to use shorter probe to carry out the FISH mapping.

The multiple method based on nucleic acid amplification that is used for genetic mapping and physical mapping can be used described nucleic acid and implement.Example is found in " definition " part in the literary composition.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med 11:95-96), pcr amplified fragment; Sheffield etc., (1993) Genomics 16:325-332), allele-specific connects (Landegren etc., (1988) Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (people such as Walter Science 241:1077-1080),, Nat.Genet.7:22-28) and Happy mapping (Dear and Cook, (1989) Nucleic Acid Res.17:6795-6807) (1997).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The inventive method produces the plant of the seed storage thing content that has improvement as previously described.These proterties also can make up with other favourable economically proterties, as other output increase proterties, to other abiotic and biological tolerances of coercing, the proterties of regulating various constructivity features and/or biochemical characteristics and/or physiologic character.

The detailed description of SYB1 polypeptide

Now be surprised to find the expression of coding nucleic acid in plant of regulating the SYB1 polypeptide and produced the plant that has the enhanced yield correlated character with respect to control plant.When under no stress conditions (non-stress conditions) when cultivating plants, being surprised to find this output increases.The specific SYB1 polypeptide classification that is suitable for strengthening output correlated character in the plant hereinafter is described in detail in detail.

The invention provides the method that is used for strengthening with respect to control plant plant output correlated character, it comprises the expression of coding nucleic acid in plant of regulating the SYB1 polypeptide.Term " adjusting " with regard to expression or genetic expression, mean such process, wherein expression level is compared with control plant because of described expression of gene changes, and preferably, increases expression level.Any kind that original expression of being regulated can be structure RNA (rRNA, tRNA) or mRNA is expressed, and is translation subsequently.

Any SYB1 polypeptide that means as defined herein of quoting of " protein that is used for the inventive method " after this.Any nucleic acid that means this type of SYB1 polypeptide of to encode of quoting of " nucleic acid that is used for the inventive method " after this.Term " polypeptide " and " protein " are used interchangeably in this article and refer to be in amino acid in the random length polymerized form.Term " polynucleotide ", " nucleotide sequence ", " nucleotide sequence " are used interchangeably in this article and refer to be in Nucleotide in the non-branch of the random length polymerized form, i.e. ribonucleotide or deoxyribonucleotide or these two combination.

Being used for regulating the preferred method of expression that (preferred increasing) be used for the proteinic coding nucleic acid of the inventive method is by introduce and be expressed as follows the proteinic coding nucleic acid that is used for the inventive method of literary composition definition plant.

The nucleic acid for the treatment of introduced plant (and thereby be used to implement the inventive method) is the proteinic any nucleic acid of the present described type of coding, hereinafter is called " SYB1 nucleic acid " or " SYB1 gene " again." SYB1 " polypeptide refers to any nucleotide sequence as defined herein, described nucleotide sequence comprises 3 RanBP type Zinc finger domain (SMART accession number: SM00547, the Interpro accession number: IPR001876) and optional one or more low-complexity structural domains, but when at the SMART database analysis, lack not with Zinc finger domain or low-complexity structural domain (if) any other structural domain of eclipsed.ZnF RBZ type Zinc finger domain appears in the Ran-conjugated protein and in other protein.In RanBPs, this structural domain is in conjunction with RanGDP.Can use the SEG algorithm of Wootton and Federhen (MethodsEnzymol.266 (1996), 554-571 page or leaf) to identify the low-complexity structural domain.The SYB1 polypeptide (that is, existing naturally as them) that preferably is in natural form did not grow 350,300,290,280 with the preferred sequence that increases, 270,260,250,240,230,220,210,190 or 180 amino acid, more preferably the length range of SYB1 polypeptide is 180 to 130 amino acid.

Known Zinc finger domain is in conjunction with zine ion and participate in protein-DNA or protein-protein interaction usually.Preferably, Zinc finger domain in the SYB1 protein with following motif at the beginning: (G/R/D/N) DW (motif 1, SEQ ID NO:344) or GSW (motif 2, SEQ ID NO:345), and at the 5th have the first conservative aminothiopropionic acid residue (wherein motif 1 or motif 2 have the 1st to the 3rd), have the second conservative aminothiopropionic acid residue between the 7th to the 10th, having the 3rd conservative aminothiopropionic acid residue between the 18th to the 21st and between the 21st to the 24th, having the 4th conservative aminothiopropionic acid residue.In addition, the Zinc finger domain in the SYB1 protein preferably includes (R/K) (A/S/P) (N/S/F) R (motif 4, SEQ ID NO 347) of R (motif 3, SEQ ID NO:346) or N (F/Y) of conservative motif NF (Q/C/S).

Randomly, the sequence that is positioned between the Zinc finger domain (start from after the 4th the conservative aminothiopropionic acid residue and motif 1 or 2 beginning finally before) is rich in glycine residue, glycine content up to 35% or even higher (about 6.93% (the SWISS PROTnotes of glycine content in the usual protein simultaneously, release in July, 44,2004)).In addition or alternatively, compare with serine content (6.89%) in the usual protein that serine content also increases in this sequence.

Preferably, when described peptide sequence is used for constructing system tree (for example shown in Fig. 2 b) when it, clusters in the SYB1 polypeptide group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

Be used for the inventive method polypeptide and the coding its nucleic acid example hereinafter the table of embodiment 40 provide.

Also be used for the inventive method be the derivative of arbitrary polypeptide of providing in the table of the homologue of arbitrary aminoacid sequence of providing of the table of embodiment 40 and embodiment 40 or any aforementioned SEQ ID NO directly to homologue or collateral line homologue.

The present invention is illustrated by the Arabidopis thaliana nucleotide sequence plant transformed of using the coded polypeptide sequence SEQ IDNO:286 that is represented by SEQ ID NO:285, yet enforcement of the present invention is not limited to these sequences.The inventive method can advantageously use coding to carry out as the defined proteinic any nucleic acid that is used for the inventive method in the literary composition, and described protein comprises directly to homologue and collateral line homologue, for example any nucleotide sequence that provides in the table of embodiment 40.Can think the aminoacid sequence that provides in the table of embodiment 40 be the SYB1 polypeptide represented by SEQ ID NO:286 directly to homologue and collateral line homologue.

Directly can easily find by carrying out so-called interactivity blast search to homologue and collateral line homologue.It generally comprises a BLAST, and described BLAST participation submission search sequence (for example using any nucleotide sequence of listing in the table of embodiment 40) is used for the blast search at arbitrary sequence library (as public's available ncbi database).When nucleotide sequence begins, use BLASTN or TBLASTX (using the standard default value) usually, and, can use BLASTP or TBLASTN (use standard default value) when when protein sequence begins.Randomly can screen BLAST result.The full length sequence of submitting The selection result and non-The selection result subsequently to is with at carry out reverse BLAST (the 2nd BLAST) from the sequence of biology, wherein search sequence is from described biology (be under the situation of SEQ ID NO:285 or SEQ ID NO:286 in search sequence wherein, the 2nd BLAST thereby will be at arabidopsis thaliana sequence).The result who compares first and second blast searches subsequently.If from the high-order position of a BLAST hit be derived from search sequence from identical species of deutero-species wherein, oppositely BLAST then identifies the collateral line homologue subsequently ideally to produce in the highest search sequence of hitting; If the high-order position among the BLAST hit be not derived from search sequence from identical species of deutero-species wherein, and preferably when reverse BLAST, produce the search sequence that belongs to the highest row that hit, then identify directly to homologue.

It is that with low E-value those hit that high-order position is hit.The E-value is low more, mark remarkable more (or in other words, this hit because of the chance odds low more).The calculating of E-value is well-known in the art.Except the E-value, comparative result is also kept the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.Under the situation of large-scale family, can use ClustalW, use subsequently in abutting connection with the tree method to show the cluster of genes involved and so that identify directly to homologue and collateral line homologue so that help.

The table of embodiment 40 provides the proteinic straight example to homologue and collateral line homologue of the SYB1 that is represented by SEQ ID NO:286.Use above-mentioned BLAST process can easily identify other directly to homologue and collateral line homologue.

Appearance by three conservative ZnF_RBZ type Zinc finger domain structural domains (shown in Figure 22) can identify protein of the present invention.Term " structural domain " and " motif " or " mark " are in the definition of " definition " part.Also there is the specialized database that is used to identify structural domain, as SMART (Schultz etc. (1998) the Proc.Natl.Acad.Sci. U.S. 95,5857-5864; Letunic etc. (2002) Nucleic Acids Res30,242-244), InterPro (Mulder etc., (2003) Nucl.Acids.Res.31,315-318), Prosite (Bucher and Bairoch (1994), be used for the broad sense collection of illustrative plates grammer of biomolecular sequence motif and in the function of automatization sequence interpretation, () ISMB-94; Second molecular biology intelligence system international conference collected works .Altman R., Brutlag D., Karp P., Lathrop R., Searls D. writes, 53-61 page or leaf, AAAIPress, Menlo Park; Hulo etc., Nucl.Acids.Res.32:D134-D137, (2004)) or Pfam (Bateman etc., Nucleic Acids Research 30 (1): 276-280 (2002)) evaluation.The one group of instrument that is used for analysing protein sequence on the computer chip can obtain (resident (Gasteiger etc. on Swiss Institute of Bioinformatics on the ExPASY proteomics server, ExPASy: be used for the protein science server of deep understanding and analysing protein, Nucleic Acids Res.31:3784-3788 (2003)).

Also use routine techniques well known in the art can easily identify structural domain, for example identify by sequence alignment.It is well-known in the art being used for the method that aligned sequences is used for comparison, and these class methods comprise GAP, BESTFIT, BLAST, FASTA and TFASTA.GAP uses Needleman and Wunsch algorithm ((1970) J Mol Biol 48:443-453) to find the overall comparison that makes the maximization of coupling number and make minimized two complete sequence of room number (the covering whole sequence).BLAST algorithm (Altschul etc. (1990) J Mol Biol 215:403-10) sequence of calculation identity percentage ratio and execution are to the statistical study of similarity between two sequences.Being used to carry out software that BLAST analyzes and being the public, to pass through NCBI (NCBI) obtainable.Homologue can use for example ClustalW multiple sequence alignment algorithm (1.83 version) evaluation easily to give tacit consent to pairing comparison parameter and methods of marking (with percentage ratio).The overall percentage ratio of similarity and identity also can use one of obtainable method in the MatGAT software package and determine (Campanella etc., BMC Bioinformatics.2003Jul 10; 4:29.MatGAT:an application that generates similarity/identitymatrices using protein or DNA sequences.).Can carry out trickle craft and write to optimize the comparison between the conservative motif, apparent as those skilled in the art.Except using full length sequence, also can use ad hoc structure territory (as one of ZnF_RBZ type Zinc finger domain or motif defined above) in addition to identify homologue.The sequence identity value of describing with per-cent among the embodiment 42 is used said procedure below, utilizes default parameters, at complete nucleic acid or aminoacid sequence/or at determining on structural domain of selecting or the conservative motif.

In addition, SYB1 protein (at least with its natural form) can and have the effect that increases seed production with protein or nucleic acid interaction when expressing according to the inventive method.Provide among the embodiment 45 and be described in further detail.

The nucleic acid that is used for the coded protein of the inventive method needs not be total length nucleic acid, because the enforcement of the inventive method does not rely on the use of total length nucleotide sequence.The nucleic acid example that is suitable for implementing the inventive method comprises the nucleotide sequence that provides in the table of embodiment 40, but is not limited to these sequences.The nucleic acid variant can be used for implementing method of the present invention.Such nucleic acid variant example comprise the proteinic coding nucleic acid that is used for the inventive method part, with the nucleic acid of the proteinic coding nucleic acid hybridization that is used for the inventive method, the splice variant that is used for the proteinic coding nucleic acid of the inventive method, the allelic variant of proteinic coding nucleic acid that is used for the inventive method and the variant of the proteinic coding nucleic acid that is used for the inventive method that obtains by gene reorganization.Term part, hybridization sequences, splice variant, allelic variant and gene reorganization now will be described.

The invention provides the method that strengthens output correlated character in the plant, described method be included in the arbitrary amino acid sequence that provides in the table of introducing and expressing the part of the arbitrary nucleotide sequence that provides in the table of embodiment 40 or embodiment 40 in the plant directly to the part of the coding nucleic acid of homologue, collateral line homologue or homologue.

The part coded polypeptide that is used for the inventive method, described polypeptide fall in the definition of the proteinic coding nucleic acid that is used for the inventive method as defined herein and have substantially the same biologic activity with aminoacid sequence that the table of embodiment 40 provides.Preferably, part is the part of arbitrary nucleic acid of providing in the table of embodiment 40.The part normal length is at least 200 continuous nucleotides, preferred length is at least 300 continuous nucleotides, more preferably length is at least 400 continuous nucleotides, and most preferably length is at least 500 continuous nucleotides, and described continuous nucleotide is the arbitrary nucleotide sequence that provides in the table of embodiment 40.Most preferred part is the part of the nucleic acid of SEQ ID NO:285.Preferably, part encoding amino acid sequence, described aminoacid sequence comprise ZnF_RBZ type Zinc finger domain as defined herein.Preferably, the part encoding amino acid sequence, when described aminoacid sequence was used to make up SYB1 genealogical tree (for example shown in Figure 23 b) when it, trend clustered in the SYB1 protein group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

The part of the proteinic coding nucleic acid of SYB1 can prepare by nucleic acid is carried out one or more disappearances as herein defined.Part can be used with isolating form, perhaps itself and other coding (or non-coding) sequence can be merged, so that for example, produce and made up some active protein.When merging with other encoding sequences, the polypeptide that is produced after translating may be bigger than the SYB1 protein portion of prediction.

Be used for another nucleic acid variant of the present invention and be under the stringent condition that reduces, preferably under stringent condition, can with the proteinic coding nucleic acid of SYB1 defined herein or with the nucleic acid of part hybridization defined herein.

The hybridization sequences coded polypeptide that is used for the inventive method, described polypeptide has three ZnF_RBZ type Zinc finger domains (seeing the comparison of Figure 23 a), and with the table of embodiment 40 in the represented SYB1 protein of the arbitrary amino acid sequence that provides have substantially the same biologic activity.The hybridization sequences normal length is at least 200 continuous nucleotides, preferred length is at least 300 continuous nucleotides, more preferably length is at least 400 continuous nucleotides, most preferably length is at least 500 continuous nucleotides, and described continuous nucleotide is the arbitrary nucleotide sequence that provides in the table of embodiment 40.Preferably, hybridization sequences be can with the sequence of any nucleic acid hybridization of providing in the table of embodiment 40 or with the sequence of the part hybridization of these sequences arbitrarily, part is definition as mentioned.Most preferred hybridization sequences be can with represented nucleic acid of SEQID NO:285 or the hybridization of its part.Preferably, hybridization sequences encoding amino acid sequence, described aminoacid sequence comprise any or a plurality of as defined herein motif or structural domain.Preferably, the hybridization sequences encoding amino acid sequence, when described aminoacid sequence was used to make up SYB1 genealogical tree (for example shown in Figure 23 b) when it, trend clustered in the SYB1 protein group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

The invention provides the method that strengthens output correlated character in the plant, described method be included in introduce in the plant and express can be with the nucleic acid of arbitrary nucleic acid hybridization of providing in the table of embodiment 40 or be included in introduce in the plant and express can with any nucleotide sequence of providing in the table of embodiment 40 directly to the nucleic acid of the coding nucleic acid hybridization of homologue, collateral line homologue or homologue.

Another nucleic acid variant that is used for the inventive method is the proteinic splice variant of SYB1 of definition as mentioned of encoding, and term " splice variant " defines as mentioned.

The invention provides the method that strengthens output correlated character in the plant, its be included in the arbitrary amino acid sequence that provides in the table of introducing and expressing the splice variant of the arbitrary nucleotide sequence that provides in the table of embodiment 40 or embodiment 40 in the plant directly to the splice variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Preferred splice variant be by the splice variant of the represented nucleic acid of SEQ ID NO:285 or SEQ ID NO:286 directly to the splice variant of the coding nucleic acid of homologue or collateral line homologue.Preferably, comprise any or a plurality of as defined herein motif or structural domain by the splice variant amino acid sequence coded.Preferably, by the splice variant amino acid sequence coded, when it was used to make up SYB1 genealogical tree (for example shown in Figure 23 b), trend clustered in the SYB1 protein group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

Another nucleic acid variant that is used for the inventive method is the splice variant of the proteinic nucleic acid of SYB1 of definition as mentioned of encoding.The natural existence of allelic variant, and to be included in the inventive method scope be the purposes of these natural equipotential volumes.The described allelic variant that is used for the inventive method has substantially the same biologic activity with the SYB1 protein of SEQ ID NO:286.

The invention provides the method that strengthens output correlated character in the plant, its be included in introduce and express the allelic variant of the arbitrary nucleic acid that provides in the table of embodiment 40 in the plant or be included in introduce in the plant and express the arbitrary amino acid sequence that provides in the table of embodiment 40 directly to the allelic variant of the coding nucleic acid of homologue, collateral line homologue or homologue.

Preferred allelic variant be the allelic variant of SEQ ID NO:285 or SEQ ID NO:286 directly to the allelic variant of the coding nucleic acid of homologue or collateral line homologue.Preferably, comprise any or a plurality of as defined herein motif or structural domain by the allelic variant amino acid sequence coded.Preferably, by the allelic variant amino acid sequence coded, when it was used to make up SYB1 genealogical tree (for example shown in Figure 23 b), trend clustered in the SYB1 protein group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

Another nucleic acid variant that is used for the inventive method is the nucleic acid variant that obtains by gene reorganization, and term " gene reorganization " is as above-mentioned.

The invention provides the method that strengthens output correlated character in the plant, its be included in the plant introduce and express the variant of the arbitrary nucleotide sequence that provides in the table of embodiment 40 or be included in introduce in the plant and express the arbitrary amino acid sequence that provides in the table of embodiment 40 directly to the variant of the coding nucleic acid of homologue, collateral line homologue or homologue, described variant nucleic acid obtains by gene reorganization.

Preferably, obtain variant nucleic acid encoding aminoacid sequence by gene reorganization, described aminoacid sequence comprises any or a plurality of as defined herein motif or structural domain.Preferably, by the aminoacid sequence that obtains the variant nucleic acid encoding by gene reorganization, when it was used to make up SYB1 genealogical tree (for example shown in Figure 23 b), trend clustered in the SYB1 protein group that comprises the aminoacid sequence shown in the SEQ ID NO:286, rather than cluster in any other group.

In addition, can pass through site-directed mutagenic obtained nucleic acid variant.Several method can be used for realizing site-directed mutagenesis; The most frequently used method that is based on PCR (Current Protocols in MolecularBiology.Wiley writes).

The proteinic nucleic acid of coding SYB1 can be from any natural or artificial source.Can on composition and/or genome environment, modify the natural form of described nucleic acid by careful manual operation.Preferred SYB1 coding nucleic acid is from plant, and preferred source more preferably is derived from Cruciferae from dicotyledons again, and most preferably nucleic acid is from Arabidopis thaliana.

Therefore proteinic the quoting of any SYB1 of this paper means the SYB1 protein of definition as mentioned.The proteinic any nucleic acid of this type of SYB1 of encoding is suitable for implementing the inventive method.

The present invention also comprises can be by plant or its part (comprising seed) of the inventive method acquisition.Described plant or its part comprise the proteinic nucleic acid transgenosis of coding SYB1 of definition as mentioned.

The present invention also provides genetic constructs and carrier to promote to introduce and/or express the nucleotide sequence that is used for the inventive method in plant.The construct that is used for the inventive method can use the well-known recombinant DNA technology of those skilled in the art to make up.This gene construct can insert the commercially available carrier that is suitable for being converted in the plant and is suitable for expressing goal gene in cell transformed.The present invention also provides as defined gene construct purposes in the methods of the invention in the literary composition.

More specifically, the invention provides construct, it comprises:

(i) as top defined SYB1 nucleic acid or its variant;

(ii) one or more regulating and controlling sequences that effectively are connected with the nucleotide sequence of (i); Randomly

(iii) transcription termination sequence.

Preferably, the nucleic acid of coding SYB1 polypeptide defines as mentioned.Term " regulating and controlling sequence " and " terminator sequence " are as defined herein.

Plant transforms (i.e. the nucleic acid of the coding SYB1 polypeptide that defines as mentioned) with the carrier that comprises aim sequence.The technician is appreciated that successfully and transforms, selects and breed the host cell that contains aim sequence and the genetic elements that must exist very much on carrier.Aim sequence is connected with one or more regulating and controlling sequences (at least with promotor) effectively.Term " regulatory element ", " regulating and controlling sequence " and " promotor " are in all commutative use of this paper and in above definition.

Advantageously, can use the expression of the promoters driven nucleotide sequence of any kind.Preferably, SYB1 nucleic acid or its variant effectively are connected with constitutive promoter.Preferred constitutive promoter also is the constitutive promoter of generally expressing basically.Also preferred constitutive promoter is from plant, more preferably monocotyledons.Most preferably be to use GOS2 promotor (from rice) (SEQ ID NO:56 or SEQ ID NO:343).Should be understood that practicality of the present invention is not limited to the SYB1 nucleic acid shown in the SEQ ID NO:285, practicality of the present invention also is not limited to the SYB1 expression of nucleic acids that is subjected to the GOS2 promoters driven.The example that has above shown other constitutive promoters that can be used for driving the SYB1 expression of nucleic acid equally.

Randomly, one or more terminator sequences can be used in the construct of introduced plant.The additional adjustment element can comprise transcribes and translates reinforcement.One skilled in the art will recognize that and go for implementing terminator of the present invention and strengthen subsequence.Intron sequences also can be added in 5 ' non-translational region (UTR) or the encoding sequence, to be increased in the amount of the ripe information that accumulates in the endochylema.Other regulating and controlling sequences (except that promotor, enhanser, silencer, intron sequences, 3 ' UTR and/or 5 ' UTR district) can be protein and/or RNA stabilization element.One skilled in the art will recognize that or can obtain this type of sequence easily.

Genetic constructs of the present invention can also comprise need be used for the replication sequence starting point of keeping and/or duplicating in particular cell types.An example is when needs are maintained additive type genetic elements (for example plasmid or clay molecule) with genetic constructs in bacterial cell.Preferred replication orgin includes, but are not limited to f1-ori and colE1.

For detecting as successful transfer of used nucleotide sequence in the methods of the invention and/or the transgenic plant that selection comprises these nucleic acid, applying marking gene (or reporter gene) is favourable.Described in more detail at definitional part " selected marker ".This marker gene can remove or excise when it no longer needs from transgenic cell.The technology that removes mark is known in the art, and useful technology is described in top " definition " part.

The present invention also provides generation to have the method for the transgenic plant of enhanced yield correlated character with respect to control plant, and described method is included in introduces and express the proteinic any coding nucleic acid of SYB1 of definition as mentioned in the plant.

(i) more specifically, the invention provides the method for the transgenic plant that are used to produce the output correlated character with increase, described method comprises: introduce and express SYB1 nucleic acid or its variant in plant or vegetable cell; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

(i) nucleic acid can be any nucleic acid of the SYB1 polypeptide of can encoding as defined herein.

Nucleic acid is introduced plant cell or introduced plant itself (comprising any other part of introducing tissue, organ or plant) directly.According to preferred feature of the present invention, nucleic acid is preferably by transforming introduced plant.The more detailed term " conversion " of having described of " definition " part.

The vegetable cell of genetic modification can be regenerated by all methods that those skilled in the art are familiar with.Suitable method be found in S.D.Kung and R Wu, Potrykus or

Above-mentioned publication with Willmitzer.

Usually after conversion, vegetable cell or the cell colony of selecting one or more marks to exist, wherein said mark become whole strain plant with the material regeneration that transforms subsequently by the effable genes encoding of the plant that moves with the goal gene corotation.In order to select plant transformed, the vegetable material that obtains in conversion is accepted selection condition in principle and is handled, to such an extent as to plant transformed can be distinguished with unconverted plant.For example, can sow with the seed that mode mentioned above obtains, after date when initial the cultivation carries out suitable selection by spraying.Another kind of possibility is included in the seed of growing on the agar plate that uses suitable selective agent (as required after sterilization), makes the seed that only transforms can grow into plant.Perhaps, plant transformed is by the existence screening of above-mentioned those selective markers.

After DNA shifted and regenerates, existence, copy number and/or genome structure that the conversion plant of supposition also can for example use Southern to analyze goal gene were estimated.Alternative or extraly, the expression level of newly introducing DNA can use Northern and/or Western to analyze and monitor, and these two kinds of technology are that those skilled in the art are well-known.

The conversion plant that produces can be bred by several different methods, as passing through clonal expansion method or classical breeding technique.For example, the first-generation (or T1) transforms plant can carry out selfing, the s-generation (or T2) transformant that selection is isozygotied, and the T2 plant can further breed by classical breeding technique subsequently.

The inverting biological that produces can be taked various ways.For example, they can be the mosaics of transformant and non-transformed cell; Clone's transformant (for example being transformed) to contain whole cells of expression cassette; The transplant of transforming tissue and unconverted tissue (for example in plant) with the root stock of the conversion of unconverted grafting of tender branch.

The present invention extends to any vegetable cell or the plant that produces by any means described in the literary composition clearly, and extends to whole plant parts and propagulum thereof.The present invention extends further to and comprises the former generation conversion that produces by any preceding method or the offspring of transfectional cell, tissue, organ or whole strain plant, unique requirement be the offspring show with by identical one or more yielding characteristicses and/or the phenotypic characteristic of those offsprings that parental generation produced in the inventive method.

The present invention also comprises host cell, and it contains the proteinic coding nucleic acid of SYB1 of definition as mentioned.The preferred host cell of the present invention is a vegetable cell.

Host plant advantageously can synthesize whole plants of used polypeptide in the inventive method in principle for used nucleic acid or carrier, expression cassette or construct or carrier in the inventive method.

The inventive method advantageously is applicable to any plant.The plant that is used in particular in the inventive method comprises the whole plants that belong to vegitabilia's superfamily, and especially monocotyledons and dicotyledons comprise being selected from following feeding or feed beans, ornamental plant, food crop, tree or shrub.According to the preferred embodiment of the invention, plant is a crop plants.The example of crop plants comprises soybean, Sunflower Receptacle, canola oil dish, clover, rape, cotton, tomato, potato and tobacco.Also preferably, plant is a monocotyledons.Monocotyledonous example comprises sugarcane.More preferably, plant is a cereal.The example of cereal comprises rice, corn, wheat, barley, grain, rye, triticale, Chinese sorghum and oat.The present invention also extend to plant the part gathered in the crops as, but be not limited to seed, leaf, fruit, flower, stem, root stock, stem tuber and bulb.The invention further relates to from, preferred directly from the product in the part gathered in the crops of this kind of plant, as dried particles or powder, oil, fat and lipid acid, starch or protein.

According to preferred feature of the present invention, the expression of being regulated is the expression that increases.As indicated above, the preferred method that is used for the expression of adjusting (preferred increasing) proteinic coding nucleic acid of SYB1 is to introduce and express the proteinic coding nucleic acid of SYB1 plant; Yet the effect of realization present method promptly strengthens the output correlated character and also can use other known technologies to realize.A kind of this type of technology is that T-DNA activates mark.Can also use TILLING (the local sudden change of the genome of directional induction) technology, or reproduce effect of the present invention with the homologue reorganization, the nucleic acid that described homologous recombination allows to select is introduced in the genome on the selected location of determining.

Implement the inventive method the plant with enhanced yield correlated character is provided.Implementing the inventive method especially provides with respect to control plant and has the output of increase, especially increases the plant of seed production.Term " output " and " seed production " are described in more detail in this paper " definition " part.

This paper means increase at the biomass (weight) of a part of plant or a plurality of parts to quoting of enhanced yield correlated character, and it comprises (can gather in the crops) part and/or (can gather in the crops) underground part on the ground.

Especially, this type of can gather in the crops part is seed, and the generation of enforcement the inventive method has the plant that has the seed production of increase with respect to the seed production of suitable control plant.

With the corn is example, and the output increase can show as following one or more indexs: the increase of the increase of the increase of every square metre of plant number of being cultivated, every strain plant spike number, the increase of line number, every row grain number, grain weight, thousand seed weight, fringe length/diameter, the full rate of seed (wherein the full rate of seed is that the full seed number is total and multiply by 100 divided by seed) and other.With the rice is example, and itself can show as the increase of following one or more indexs the output increase: the increase of every square metre of plant number, every strain plant panicle number, every panicle spikelet number, every panicle flower (Xiao Hua) number (it is expressed as the ratio of full seed number to former panicle number), the full rate of seed (wherein the full rate of seed be the full seed number divided by the seed sum and multiply by 100), the increase of thousand seed weight and other.

Because transgenic plant of the present invention have the output of increase, thereby with respect to the growth velocity of control plant, these plants might show on the corresponding stage in its life cycle increase growth velocity (its life cycle during the small part).The growth velocity that increases can be specific for one or more parts (comprising seed) of plant, or can spread all over whole strain plant basically.Plant with growth velocity of increase can possess short life cycle.The life cycle of plant can be considered as meaning from dry mature seed and grow to the needed time in stage that plant has produced the dry mature seed similar to parent material.This life cycle can be influenced by following factors, as early stage vigor, growth velocity, green degree index, flowering time and seed maturity speed.The increase of growth velocity can take place during life cycle on the one or more stages in life cycle or in whole plants basically plant.The growth velocity that increases during plant early stage in life cycle can reflect the enhanced vigor.The increase of growth velocity can change the harvest cycle of plant, allows the later sowing of plant and/or than early harvest, otherwise this can not (similar effect can obtain with flowering time early).If growth velocity increases fully, can allow further to sow the seed (for example sow and gather in the crops rice plant, sow and gather in the crops other rice plants subsequently, all rice plant is all in a conventional growth period) of identical plant species.Similarly, if growth velocity sufficiently increases, can allow further to sow the seed (for example sowing and harvesting corn plant are for example sowed and optional results soybean, potato or any other suitable plant subsequently) of different plant species.The results additional times also is possible in the situation of some crop plants from identical rhizome.The harvest cycle that changes plant can cause the increase of every acre year biomass yield (number of times (as in a year) that can grow and gather in the crops because of any specified plant increases).The increase of growth velocity also can allow cultivating transgenic plant in the geographic area widely than its wild type counterparts, because the region limits of cultivating crop is often determined by the plantation time (season early) or in the adverse environment condition of results period (season in evening).If shorten harvest cycle, then can avoid this class unfavourable condition.Growth velocity can determine that this type of parameter can be by obtain multiple parameter from growth curve: T-Mid (plant reaches the time that its 50% overall dimension is spent) and T-90 (plant reaches the time that its 90% overall dimension is spent), or the like.

According to preferred feature of the present invention, the plant of the growth velocity with increase is compared in the enforcement generation of the inventive method with control plant.Thereby, providing the method that increases plant growth rate according to the present invention, described method comprises adjusting, the preferred expression of the proteinic nucleic acid of coding SYB1 in plant that increases as defined herein.

Compare with control plant, no matter plant is under the non-stress conditions still is that plant is exposed to multiple coercing down, and the increase of output and/or growth velocity all takes place.Plant is generally replied being exposed to coerce to make by growing slowlyer.Under the condition of serious stress of soil condition, plant even can stop growing fully.On the other hand, slightly coerce and be defined as plant in this article any of its exposure coerced, the wherein said ability that does not cause plant to stop growing fully and recover growth of coercing.Compare with the control plant under the non-stress conditions, slightly coerce and in meaning of the present invention, cause being coerced the plant-growth reduction less than 40%, 35% or 30%, preferably, be more preferably less than 14%, 13%, 12%, 11% or 10% or lower less than 25%, 20% or 15%.Because the progress on the agricultural practice (irrigation, fertilising, pesticide treatments) does not often run into condition of serious stress of soil in the raise crop plant.Therefore, by the agriculture often undesirable characteristic that goes up of the impaired growth of slight stress-inducing.Slightly coerce is that the common biological and/or inanimate (environment) that plant exposes is coerced.Abiotic stress can because of arid or waterlogging, anaerobism are coerced, due to salt stress, chemical toxicity, oxidative stress and heat, cold or the freezing temperature.Abiotic stress can be to coerce (especially because arid), salt stress, oxidative stress or ion by water to coerce the osmotic stress that causes.It generally is that those that caused by pathogenic agent such as bacterium, virus, fungi and insect are coerced that biology is coerced.

Especially, method of the present invention can implemented the plant that has the output of increase with respect to control plant to produce under the non-stress conditions or under slight drought condition.As report in (Planta (2003) 218:1-14) such as Wang, abiotic stress causes influencing unfriendly a series of morphological change of plant-growth and productivity, physiology to change, biological chemistry changes and molecule changes.Known arid, salinity, extreme temperature and oxidative stress are also can damaging and primary cellular defect by induced growth by similar mechanism of connecting each other.Rabbani etc. (Plant Physiol (2003) 133:1755-1767) have described " cross-talk " that drought stress and high salinity are coerced a very high degree.For example, arid and/or salinification mainly show as osmotic stress, cause the destruction of cell homeostasis and ion distribution.Often follow the oxidative stress of high temperature or low temperature, salinity or drought stress can cause functional protein and structural protein sex change.Therefore, these various environment-stress usually activate similar cell signal approach and cell response, as producing stress protein matter, raising antioxidant, accumulation compatible solute and growth-inhibiting.Term as used in this article " non-coercing " condition is those envrionment conditionss that allow the plant optimum growhs.Those skilled in the art know that normal edaphic condition and weather condition for given place.

The enforcement of the inventive method is with respect to the suitable control plant of cultivating under comparable conditions, gives under the non-stress conditions or the output that increases of the plant of cultivating under slight drought condition.Thereby according to the present invention, be provided under the non-stress conditions or increase the method for output in the plant of under slight drought condition, cultivating, described method comprises that the nucleic acid that increases coding SYB1 polypeptide expresses in plant.

The enforcement of the inventive method produces with respect to the appropriate control plant of cultivating under comparable conditions, the plant of under the nutritive deficiency condition, especially cultivating under nitrogen shortage condition that increases output that has.Thereby, increasing the method for output in the plant that is provided under the nutritive deficiency condition, cultivating according to the present invention, described method comprises the expression of nucleic acid in plant that increases coding SYB1 polypeptide.Nutritive deficiency can be because of due to the nutraceutical shortage, and described nutrition for example is nitrogen, phosphoric acid salt and other P contained compounds, potassium, calcium, cadmium, magnesium, manganese, iron and boron etc.

In the preferred embodiment of the invention, the inventive method produces the increase of output and/or growth velocity under non-stress conditions.

The present invention also comprises the purposes and the proteinic purposes of these SYB1 of the proteinic coding nucleic acid of SYB1 described herein, is used for strengthening plant output correlated character.The present invention and then comprise the purposes of plant.Can in the procedure of breeding, use proteinic coding nucleic acid of SYB1 described herein or SYB1 protein self, wherein identify the dna marker that can be connected in the SYB1 encoding gene hereditarily.Can use nucleic acid/gene or SYB1 protein self to define molecule marker.Then this DNA or protein labeling can be used in the procedure of breeding, to select to have the plant of the enhanced yield correlated character that defines in the inventive method as mentioned.

The allelic variant of SYB1 protein coding nucleic acid/gene also can be used for the auxiliary procedure of breeding of mark.This class procedure of breeding needs to use sometimes, and for example EMS mutagenesis is introduced allele variant by the plant mutagenic treatment; Alternative, this program can begin with the allele variant of collecting what is called " natural " origin that is not intended to generation.Identify allele variant by for example PCR then.Be to select step subsequently, in order to select the better allele variant of the sequence of discussing, described allele variant is given the output that plant increases.Generally the growth behavior that contains the different allele variant plants of studying sequence to some extent by monitoring is selected.Can in greenhouse or field, monitor growth behavior.More optional step comprise, will contain plant and another plant hybridization of better allele variant through evaluation.For example, can make the combination that produces phenotypic characteristic interested in this way.

SYB1 protein coding nucleic acid can also be as probe, is used for for the part of those gene linkage proterties and carry out the mapping of heredity and physics as the gene of its mark.Such information can be used in plant breeding, to obtain having the strain of desired phenotype.This class of SYB1 protein coding nucleic acid is used the nucleotide sequence that only needs to grow to few 15 Nucleotide.SYB1 protein coding nucleic acid can be used as restriction fragment length polymorphism (RFLP) mark.The Southern trace of the plant genome DNA of available SYB1 protein coding nuclei acid probe restriction digest (Sambrook J, Fritsch EF and Maniatis T (1989) " molecular cloning: laboratory manual ").The program that uses a computer subsequently such as MapMaker (Lander etc. (1987) Genomics 1:174-181) carry out genetic analysis to the banding pattern that produces, to make up genetic map.In addition, can use nucleic acid to survey the Southern trace in the genomic dna that the restriction enzyme that contains one group of individuality is handled, described one group of individuality is the parent of the clear and definite genetic cross of representative and one group of individuality of filial generation.The separation of record dna polymorphism also is used for calculating formerly position (Botstein etc. (1980) Am.J.Hum.Genet.32:314-331) with the genetic map SYB1 protein coding nucleic acid of this colony's acquisition.

The derive generation and the purposes of probe of the plant gene that uses in genetic mapping is described among Bematzky and Tanksley (1986) the Plant Mol.Biol.Reporter 4:37-41.Described in numerous publications with aforesaid method or its flexible form specific cDNA clone was carried out genetic mapping.For example, can use F2 hybridization colony, backcross population, panmictic population, the homogenic strain of close relative and the mapping of other group of individuals.These class methods are that those skilled in the art are well-known.

Nucleic acid probe also can be used for physical mapping and (promptly settle sequence on physical map; See In:Non-mammalian Genomic Analysis:A Practical Guide such as Hoheisel, Academicpress 1996, the 319-346 pages or leaves, and the reference of wherein quoting).

In another embodiment, nucleic acid probe can be used for direct fluorescence in situ hybridization (FISH) mapping (Trask (1991) Trends Genet.7:149-154).(several kb are to a hundreds of kb although the method inclination of FISH mapping at present is used for big clone; See (1995) Genome Res.5:13-20 such as Laan), but the raising of susceptibility allows to use short probe in the FISH mapping.

The multiple method based on nucleic acid amplification that is used for heredity and physical mapping can use described nucleotide sequence to carry out.Example comprises the polymorphism (CAPS of allele specific amplification (Kazazian (1989) J.Lab.Clin.Med11:95-96), pcr amplified fragment; Sheffield etc. (1993) Genomics16:325-332), allele-specific connects (Landegren etc. (1988) Science 241:1077-1080), Nucleotide extension (Sokolov (1990) Nucleic Acid Res.18:3671), radiation hybridization mapping (Walter etc. (1997) Nat.Genet.7:22-28) and Happy mapping (Dear and Cook (1989) Nucleic Acid Res.17:6795-6807).For implementing these methods, it is right to use the nucleotide sequence design and produce the primer that is used for amplified reaction or primer extension reaction.This class primer design is that those skilled in the art are well-known.Use the method for the genetic mapping of PCR-based, may need to identify the difference of leap corresponding to dna sequence dna between the parent of nucleotide sequence of the present invention zone mapping.Yet this is dispensable usually to drawing method.

The method according to this invention is had the plant of enhanced yield correlated character as previously mentioned.These proterties can also make up other favourable economically proterties, as other improve the proterties of output, to the proterties of other abiotic or biological tolerances of coercing, the multiple structural attitude of improvement and/or biochemistry and/or physiologic character.

Accompanying drawing is described

The present invention is described referring now to following accompanying drawing, wherein:

Fig. 1 has shown the example of the structural domain structure of AZ polypeptide.SEQ ID NO:2 encoded protein matter comprises that two ankyrins repeat (boldface letter, underscore) and two C3H1 structural domains (italics, underscore).

Fig. 2 has shown according to ankyrin of SMART database (ANK) and C3H1 zinc and has referred to (C3H1) consensus sequence that the symbol that is used for each seed amino acid group provides in legend.

Fig. 3 has represented the sequence identity/similarity matrix with MATGAT (point penalty 1 is extended in the room for BLOSUM62, the open point penalty 11 in room) preparation.Sequence identity is presented on the diagonal lines with runic, provides under diagonal lines at A) full length protein sequence, B) comprises the sequence similarity of the partial protein of the Zinc finger domain that most of C-terminal is inferred.

Fig. 4 has shown binary vector p056, and it is used for Arabidopis thaliana AZ encoding sequence under WS118 protein promoter (confidential reference items PRO0151) control in the expression of rice.

Fig. 5 has shown plant and the typical structural domain structure of Mammals SYT polypeptide.Conservative SNH structural domain is positioned at the N-terminal of polypeptide.The terminal remainder of peptide C is formed by being rich in the QG structural domain in the plant SYT polypeptide, and the terminal remainder of Mammals SYT peptide C is formed by being rich in the QPGY structural domain.Be rich in the Met structural domain be generally comprised within plant be rich in QG structural domain or Mammals QPGY be rich in structural domain first half within (direction from the N-terminal to the C-terminal).Having second is rich in before the SNH structural domain that the Met structural domain is positioned at plant SYT polypeptide.

Fig. 6 has shown that the multiple ratio of several SYT polypeptide N-terminal is right, that uses is based on improved ClustalW algorithm (InforMax, Bethesda, MD, http://www.informaxinc.com) VNTI AlignX multiple ratio is to program, adopt default setting, the open point penalty in room is 10, and it is 0.05 that point penalty is extended in the room.The SNH structural domain of the intersection of plant and people SYT polypeptide shows to add frame table.Last column comprises the consensus sequence that derives from institute's aligned sequences in the comparison.

Fig. 7 has shown that the multiple ratio of several plant SYT polypeptide N-terminal is right, that uses is based on improved ClustalW algorithm (InforMax, Bethesda, MD, http://www.informaxinc.com) VNTI AlignX multiple ratio is to program, adopt default setting, the open point penalty in room is 10, and it is 0.05 that point penalty is extended in the room.Two primary structure territories from N-terminal to the C-terminal direction show to add frame table, and are accredited as the SNH structural domain and are rich in Met/ and be rich in the QG structural domain.In addition, N-terminal is rich in the Met structural domain and also shows to add frame table, and the position of SEQ ID NO:90 and SEQ ID NO 91 is represented with the runic underscore.

Fig. 8 has shown the contiguous threaded tree that utilizes ClustalW 1.83 (http://align.genome.jp/sit-bin/clustalw) to obtain by multiple SYT polypeptide comparison.SYT1 and SYT2/SYT3 clade identify with parantheses.

Fig. 9 has shown binary vector p0523, and it is used for expressing the Arabidopis thaliana AtSYT1 that is under GOS2 promotor (confidential reference items PRO0129) control rice.

Figure 10 is the general introduction of Calvin cycle.Show 13 enzymatic reactions, and implemented the title of the enzyme of these reactions.Black arrow shows the position of cpFBPase in circulation.

Figure 11 is for showing by Triphosphopyridine nucleotide photoreductase-thioredoxin system the illumination activated synoptic diagram of cpFBPase.

Figure 12 is the comparison of cyFBPase polypeptide and cpFBPase polypeptide.This peptide sequence uses Vector NTI external member, and (AlignX program MD) is compared for InforMax, Bethesda.The use room is begun point penalty 10 to multiple ratio and room extension point penalty 0.01 is carried out.The prediction chloroplast transit peptides of cpFBPase polypeptide shows to add frame table.Redox regulation and control are shown insert the halfcystine that fragment and disulfide linkage relate in forming.At last, the avtive spot district also shows to add frame table, and illustrates in conjunction with amino-acid residue Asn237, Tyr269, Tyr289 and the Arg268 of the 6-phosphoric acid of fructose-1,6-diphosphate with in conjunction with the amino-acid residue Lys299 of described fructose.

Figure 13 has shown binary vector p1597, and it is used for expressing Chlamydomonas reinhardtii (Chlamydomonasreindhardtii) nucleic acid that is in the coding cpFBPase polypeptide under GOS2 promotor (confidential reference items PRO0129) control rice.

Figure 14 is that the SIK that infers described in the coding schedule 2 is directly to the comparison of the SIK of homologue nucleic acid.

Figure 15 is rice SIK polypeptide (SEQ ID NO:2) and Arabidopis thaliana SIK polypeptide (SEQ ID NO:4) and the comparison with OSSIK polypeptide of NCBI accession number OS_BAD73441.

Figure 16 represents binary vector, and it is used at the reticent native gene of rice, uses the represented SIK nucleic acid of SEQ ID NO:1, uses the hair clip construct under the constitutive promoter GOS2 control.

Figure 17 has shown binary vector, and it is used for expressing the SIK coding nucleic acid from rice that is under the control of GOS2 promotor rice.

Figure 18 has shown the part of II class HD-Zip member's HD-Zip genealogical tree.

Figure 19 has shown the comparison of several II class HD-Zip peptide sequences.

The comparison of Figure 20 for taking from (2005) (Plant Phys 139:509-518) such as Henriksson.Leucine zipper motif is in same position (C-terminal is to HD) in all HD-Zip I and II protein.HD-Zip I is similar each other on sequence with the proteinic HD-Zip structural domain of II, although the number of amino acid position is different between HD-Zip I and the II.Is constant at the 46th amino acid in HD-ZipI and II, but is different between classification.Several other amino acid are constant as the 6th, the 25th, the 29th, the 30th, the 58th and the 61st 's amino acid in HD-Zip II, and different with the amino acid of HD-Zip I, and it has shown the variation in these positions.

Figure 21 has shown binary vector, and it is used for increasing rice the expression of the coding nucleic acid that is in the Arabidopis thaliana II class HD-Zip transcription factor (HAT4) under the control of GOS2 promotor.

Figure 22 has shown the structural domain of proteinic two examples of SYB1.Zinc finger domain is represented with the underscore boldface letter.

Figure 23 a has shown that (used sequence is different SYB1 protein: CDS2671 (SEQ IDNO:2), AAZ94630 (SEQ ID NO:4), Q53AV6 (SEQ ID NO:6), Q6Z6E6 (SEQ ID NO:8), Q8GWD1 (SEQ ID NO:10), Q9SW92 (SEQID NO:12), Q8RYZ5 (SEQ ID NO:14), beet (Beta vulgaris) (SEQ IDNO:16), Q8S8K1 (SEQ ID NO:18), Q8GZ43 (SEQ ID NO:20), Q7F1K4 (SEQ ID NO:22), Q7XHQ8 (SEQ ID NO:24)) multiple ratio is right; Figure 23 b has shown genealogical tree, and wherein black box is illustrated the SYB1 protein group.

Figure 24 has shown binary vector, and it is used for increasing the expression that is in the proteinic coding nucleic acid of Arabidopis thaliana SYB1 under GOS2 promotor (confidential reference items PRO0129) control rice.

Figure 25 detailed annotation is used to implement the example of the sequence of the inventive method.SEQ ID NO:1 to SEQID NO:56 relates to the AZ sequence.The protein sequence that SEQ ID NO:1 and 2 represents AZ encoding sequence CDS3104 and derives.The protein sequence that SEQ ID NO:53 and 47 represents AZ encoding sequence CDS3108 and derives.SEQ ID NO:3 and 6 representatives may appear at the conserved sequence in the AZ protein, SEQ ID NO:9 and 55 representatives are used for the sequence of the WS118 promotor of example part, SEQ ID NO:10 and 52 is other AZ protein and these nucleic acid sequences to proteins examples of coding, and SEQ ID NO:54 and 56 is sequences of rice GOS2 promotor.

SEQ ID NO:56 to 153 relates to the SYT sequence.Demonstration is from the SYT nucleotide sequence of origin-to-destination.The major part of these sequences checks order from low-quality EST.Therefore, may run into nucleic acid replaces.

SEQ ID NO:56 and SEQ ID NO:154 to 208 expression relates to the example series of the cpFBPase that is used to implement the inventive method.

SEQ ID NO:56 and SEQ ID NO:209 to 228 relate to the example series of the SIK protein/nucleic acid that is used to implement the inventive method, and it is used to implement the inventive method, or is used to separate this type of sequence.Sequence can be from the public EST set of inferior quality order-checking.As a result of, can expect that several nucleic acid replace.Also can use 5 and 3 ' UTR of natural transcription sequence to implement the inventive method, to reduce or to remove endogenous SIK genetic expression substantially.

SEQ ID NO:229 to 284 is the example series of the II class HD-Zip transcription factor (HAT4) that is used to implement the inventive method.

SEQ ID NO:56 and SEQ ID NO:285 to 347 are for relating to SYB1 protein and nucleic acid and being used to implement the example series of the inventive method.

Embodiment

The present invention is described referring now to as an illustration the following example only.The following example is not intended to the thoroughly definition or the restriction scope of the invention.

DNA operation: unless stated otherwise, recombinant DNA technology is according to (Sambrook (2001) Molecular Cloning:a laboratory manual, third edition Cold Spring HarborLaboratory Press, CSH, New York) in or Ausubel etc. (1994), CurrentProtocols in Molecular Biology, the standard method of describing in Current Protocols the 1st and 2 volumes is carried out.The standard material and the method that are used for plant molecular work are described at the Plant Molecular Biology Labfax (1993) of the R.D.D.Croy that is published by BIOS ScientificPublications Ltd (UK) and Blackwell Scientific Publications (UK).

Embodiment 1: identify with the inventive method in the relevant sequence of used nucleotide sequence

Use the database sequence research tool, as basic local comparison instrument (BLAST) (Altschul etc. (1990) J.Mol.Biol.215:403-410; With (1997) Nucleic Acids Res.25:3389-3402 such as Altschul) identify (full-length cDNA, EST or genome) sequence relevant in those sequences of in the Entrez Nucleotide database of NCBI (NCBI), being safeguarded with used nucleotide sequence in the inventive method.This program is used for relatively and by the significance,statistical that calculates coupling finding the zone that has local similar between sequence by nucleotide sequence or peptide sequence and sequence library.For example, use the TBLASTN algorithm, adopt default setting and filtration to offset to ignore the low-complexity sequence to being used for nucleic acid encoded polypeptide of the present invention.The result who analyzes relatively shows by pairing property, and according to probability scoring (E-value) ordering, wherein is somebody's turn to do the specific comparison result of scoring reflection because of the accidental probability (the E-value is low more, and the significance of hitting is high more) that takes place.Except the E-value, more also keep the score by identity percentage ratio.Identity percentage ratio refer to two compare identical Nucleotide (or amino acid) number in the length-specific scope between nucleic acid (or polypeptide) sequence.In some cases, can adjust default parameters to regulate the severity of search procedure.For example can increase the E-value to show the less coupling of severity.By this way, can identify short almost mating accurately.

Table A provides the tabulation of the nucleotide sequence relevant with nucleotide sequence in the inventive method.

The example of Table A AZ polypeptide

Plant origin	Nucleic acid SEQ ID NO:	Protein s EQ ID NO:
			Arabidopis thaliana	1	2
Big eucalyptus (Eucalyptus grandis)	10	11
			Rice	12	13
The puncture vine clover	14	15
			Rice	16	17
Arabidopis thaliana	18	19
			Rice	20	21
Arabidopis thaliana	22	23

Arabidopis thaliana	24	25
			Big eucalyptus	26	27
Big eucalyptus	28	29
			Soybean	30	31
Big eucalyptus	32	33
			Arabidopis thaliana	34	35
Rice	36	37
			Barley	38	39
Pine (Pinus radiata)	40	41
			Pine	42	43
Soybean		44
			Soybean		45
Soybean		46
			Arabidopis thaliana		47
Arabidopis thaliana		48
			Arabidopis thaliana		49
Arabidopis thaliana		50
			Arabidopis thaliana		51
Arabidopis thaliana		52
			Arabidopis thaliana	53

In some cases, correlated series is by institute such as the tentative compilation of Joint Genome Institute (TIGR) and open to the public.Eukaryotic gene directly can be used for identifying this type of correlated series to homologue (EGO) database, this can be by keyword search, or uses the BLAST algorithm to carry out with purpose nucleic acid or peptide sequence.

Embodiment 2: gene clone AZ

Arabidopis thaliana AZ encoding gene (CDS3104) is by PCR, and (Invitrogen, Paisley UK) increase as template to use Arabidopis thaliana seedling cDNA library.With extracting behind the RNA of seedling reverse transcription, cDNA is cloned into pCMV Sport 6.0.The insertion sequence mean size in storehouse is that 1.5kb and clone's initial number is 1.59 * 10 ⁷Cfu.6 * 10 ¹¹The original titre of first round amplification back mensuration of cfu/ml is 9.6 * 10 ⁵Cfu/ml.After plasmid extracts, the 200ng template is used for 50 μ lPCR mixtures.With primer prm06717 (justice is arranged, and the AttB1 site is an italic, and initiator codon is a boldface letter: 5 '-

Tqctqtqqatcaqacc-3 ') (SEQID NO 7) and prm06718 (antisense, complementation: the site is an italics: 5 '- Ggttag gtctctca attctgc-3 ') (SEQ ID NO8) is used for pcr amplification, and wherein said primer comprises the AttB site that is used for the Gateway reorganization.In standard conditions, use Hifi Taq archaeal dna polymerase to carry out PCR.To expect the PCR fragment amplification of size and also use standard method to give purifying.Implement the first step of Gateway method subsequently, i.e. BP reaction, " entering the clone " p07 that reorganization is named according to Gateway with generation in PCR fragment and the pDONR201 plasmid generation body during this period.Plasmid pDONR201 conduct

The part of technology is bought from Invitrogen.

Embodiment 3: vector construction

Entering clone p07 uses with p02417 (a kind of purpose carrier that is used for plant (rice) conversion) in the LR reaction subsequently.This carrier contains as functional element on the T-DNA border: plant selectable marker; Selection markers expression cassette and intention with cloned aim sequence in the described clone of entering the Gateway box of recombinating in the LR body taken place.The rice WSI18 promotor (SEQ ID NO:9) that is used for seed-specific expression (PRO0151) be positioned at this Gateway box the upstream (p056, Fig. 4).

Many different binary (with the super binary) carrier system that is used for Plant Transformation (An has for example been described, G.in Agrobacterium Protocols.Methods in Molecular Biology the 44th volume, pp 47-62, Gartland KMA and MR Davey eds.Humana Press, Totowa, NewJersey).The carrier pBIN19 that many carrier systems are described based on Bevan (Nucleic Acid Research.1984.12:8711-8721), this carrier comprise flank and connect from the left side of agrobacterium tumefaciens Ti-plasmids and the gene expression in plants box of right border sequence.The gene expression in plants box is by at least two genes, and---cDNA of selectable marker gene and regulation and control character gene or plant promoter of transcribing of genomic dna---formed.Can use different selectable marker genes, comprise the arabidopsis gene (United States Patent (USP) 57673666 and 6225105) of the acetohydroxy acid synthase (AHAS) of encoding mutant.Similarly, can use different promoter regulation character genes to realize composing type, developmental pattern, tissue-type or environmental form regulation and control to genetic transcription.

After the LR reconstitution steps, heat-shocked or electroporation scheme are converted into agrobacterium strains LBA4044 with the expression vector p056 (Fig. 4) that produces.The bacterium colony that transforms was grown 2 days with 28 ℃ on the YEP substratum, and selected with corresponding microbiotic.These Agrobacterium cultures are used for Plant Transformation.

Other agrobacterium tumefaciens bacterial strains can be used for Plant Transformation and be well known in the art.This type of bacterial strain example is C58C1 or EHA105

Embodiment 4: Plant Transformation

Rice transforms

The Agrobacterium that contains expression vector is used for transforming rice plant independently.Ripe dry seed shelling with the Japanese Cultivar Nipponbare of rice.By incubation in 70% ethanol one minute, in 2,%Hg,Cl2 30 minutes subsequently, subsequently with sterile distilled water washing 6 times 15 minutes and implement sterilization.The disinfectant seed is containing 2 subsequently, and the substratum of 4-D (callus inducing medium) is gone up and sprouted.Incubation is after 4 weeks in the dark, scultellum deutero-embryogenic callus is downcut and breeds on a kind of substratum.After 2 weeks, callus by breeding or breed uploading with a kind of substratum to be commissioned to train to support in other 2 weeks.The embryogenic callus sheet is uploaded to be commissioned to train at fresh culture and was supported 3, cultivates (to strengthen the cell fission activity) afterwards altogether.

The agrobacterium strains LBA4404 that contains expression vector is used for common cultivation.Agrobacterium is seeded in to contain on the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3.Subsequently bacterium is collected and is resuspended in liquid and cultivate altogether in the substratum to density (OD600) about 1.Suspension is transferred to culture dish subsequently and callus was soaked 15 minutes in this suspension.Callus is organized subsequently and to be cultivated on the substratum altogether and in the dark in 25 ℃ of incubations 3 days blotting and be transferred to solidified on the filter paper.Altogether the callus of cultivating in the dark in 28 ℃ in the presence of selective agent in containing 2,4 weeks of cultivation on the substratum of 4-D.During the section, form mushroom resistant calli island at this moment.To regeneration culture medium and behind incubation under the light, release of embryo generation potentiality and seedling are in 4-5 week growth subsequently in this material transfer.Seedling is downcut from callus and, wherein seedling is transferred to soil from described substratum containing incubation 2-3 week on the substratum of plant hormone.The hardened seedling is cultivated in the greenhouse under high humidity and short day.

A construct produces about 35 independent T0 rice transformant.With former generation transformant be transferred to the greenhouse from incubator for tissue culture.Behind the copy number of quantitative PCR analysis with checking T-DNA inset, the single copy transgenic plant that only keep performance selective agent tolerance are used to gather in the crops the T1 seed.Seed is gathered in the crops after transplanting subsequently the 3-5 month.Present method produces term single gene seat transformant (Aldemita and Hodges1996, Chan etc. 1993, Hiei etc. 1994) to surpass 50% ratio.

Corn transforms

The conversion of corn is carried out according to the modification method to (1996.Nature Biotech 14745-50) described methods such as Ishida.Conversion in corn be that genotype relies on and only the special genes type can operate and be used for transforming and regeneration.Inbred lines A188 (University of Minnesota) or be the good source of the donor material that is used to transform with A188 as parent's hybrid, but other genotype also can successfully be used.Mealie from maize plant after pollination about 11 days (DAP) results, this moment, the length of immature embryos was about 1 to 1.2mm.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.The embryo that downcuts is on the callus inducing medium, cultivate on the corn regeneration culture medium subsequently, and wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to the maize rooting substratum and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Wheat transforms

The conversion of wheat is carried out with the method that (1996) Nature Biotech 14 (6): 745-50 such as Ishida such as Ishida describe.Usually in conversion, use (can obtain) Cultivar Bobwhite from Mexico CIMMYT.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.With the Agrobacterium incubation after, embryo on the callus inducing medium, external cultivation on regeneration culture medium subsequently, wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to root media and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Soybean transforms

According to Texas A﹠amp; M United States Patent (USP) 5,164, the modification method soybean transformation of method described in 310.Several commercial soybean varieties are feasible for conversion by this method.Cultivar Jack (can be able to obtain from Illinois seed money) is generally used for transforming.Soybean seeds is sterilized so that external sowing.From 7 age in days seedling, downcut hypocotyl, radicle and a slice cotyledon.The cotyledon of further cultivating epicotyl and remainder is to grow the armpit tight knot.These armpit tight knots are downcut and with the agrobacterium tumefaciens incubation that contains expression vector.After cultivating processing altogether, explant is washed and is transferred to the selection substratum.The regenerated seedling is downcut and places on the seedling elongation medium.The seedling that length is no more than 1cm places on the root media until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant tolerance of performance selective agent and that contain single copy T-DNA inset, produce the T1 seed.

Semen Brassicae campestris/canola oil dish transforms

Cotyledon petiole of use 5-6 age in days seedling and hypocotyl are as being used for the explant of tissue culture and transforming according to (1998, Plant Cell Rep 17:183-188) such as Babic.Commercial Cultivar Westar (Agriculture Canada) is the standard variety that is used to transform, but also can use other kinds.Canola oil colza is done the surface sterilization so that external sowing.From external seedling, downcut and have the cotyledon petiole explant that adheres to cotyledon, and immerse bacterial suspension with the cut ends of (containing expression vector) Agrobacterium by petiole explant and inoculate.Explant subsequently on the MSBAP-3 substratum that contains 3mg/l BAP, 3% sucrose, 0.7% plant agar (Phytagar) at 23 ℃, illumination in 16 hours was cultivated 2 days down.After cultivating 2 altogether with Agrobacterium, petiole explant is transferred on the MSBAP-3 substratum of 3mg/l BAP, cefotaxime, Pyocianil or the Ticarcillin/Clavulanate Acid (300mg/l) that contain and continues 7, and cultivate containing on the MSBAP-3 substratum of cefotaxime, Pyocianil or Ticarcillin/Clavulanate Acid and selective agent subsequently, regenerate until seedling.When seedling has 5-10mm length, seedling is downcut and is transferred to seedling elongation medium (MSBAP-0.5 that contains 0.5mg/l BAP).The seedling of the about 2cm of length is transferred to the root media (MS0) that is used for root induction.The seedling that to take root migrates in the soil in greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Clover transforms

The reproducibility clone of clover (Medicago sativa) uses the method for (McKersie etc., 1999PlantPhysiol 119:839-847) to be transformed.Regeneration of clover and conversion are that genotype is dependent and thereby need aftergrowth.The method that obtains the reproducibility plant has been described.For example, these reproducibility plants any other commercial alfalfa variety that can be selected from Cultivar Rangelander (Agriculture Canada) or describe as Brown DCW and A Atanassov (1985.Plant Cell Tissue Culture 4:111-112).Alternatively, RA3 kind (University of Wisconsin) has been selected for (Walker etc., 1978Am J Bot 65:654-659) in the tissue culture.Petiole explant and the agrobacterium tumefaciens C58C1pMP90 (McKersie etc., 1999PlantPhysiol 119:839-847) or the overnight culture of LBA4404 that contain expression vector are cultivated altogether.Explant is containing 288mg/L Pro, 53mg/L Thioproline, 4.35g/L K in the dark ₂SO ₄With cultivated altogether 3 days on the SH inducing culture of 100 μ m Syringylethanones. explant half concentrate in the Murashige-Skoog substratum (Murashige and Skoog, 1962) washing and plating contain not containing Syringylethanone suitable selective agent and suitable microbiotic with the identical SH inducing culture of restraining the Agrobacterium growth on.After several weeks, somatic embryo is transferred to do not contain growth regulator, do not contain microbiotic and the BOi2Y that contains 50g/L sucrose grows in the substratum.Somatic embryo concentrates on the Murashige-Skoog substratum half subsequently to be sprouted.The sprigging that to take root is cultivated to flowerpot and in the greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 5: the evaluation of the expression of AZ in rice under the control of rice WS118 promotor is prepared

Produce about 15 to 20 T0 rice transformant independently.Transformant was transferred to the greenhouse from incubator for tissue culture and was used for growth and results T1 seed former generation.Stay 7 incidents, the T1 offspring of wherein said incident separates with 3: 1 ratios genetically modified existence/do not exist.For each incident in these incidents, select to contain genetically modified about 10 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 10 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.The T1 plant of selecting is transferred to the greenhouse.Each plant is accepted unique bar coded sticker clearly phenotype somatotype data are connected with corresponding plant.Be provided with down in following environment on the soil of T1 plant in the specially designed flowerpot of 10cm diameter (flowerpot has clear bottom so that root is visual) of selecting and cultivate: photoperiod=11.5 hour, day optical density(OD)=30,000lux, daylight temperature=28 ℃ or higher, nocturnal temperature=22 ℃, relative humidity=60-70%.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Take the utmost care of and make plant not be subjected to any coercing.Make plant pass through the digital imagery case for several times until the ripening stage from sowing time.On each time point, to every strain plant from least 6 different angles shooting digital pictures (2048 * 1536 pixels, 1,600 ten thousand colors).The vegetation period digital camera also by flowerpot bottom document image.

The arid screening

In flowerpot soil, cultivate plant under normal operation, up to entering heading stage from the T2 seed.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

The nitrogen use efficiency screening

Under normal condition except that nutritive medium, in flowerpot soil, cultivate rice plant from the T2 seed.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

Plant shoot divides area (or leaf biomass) to measure by counting is different from the pixel of background on the digital picture of dividing from plant shoot sum.This value changes into square physical surface value of mm expression to the averaging of picture of taking from different perspectives on identical time point and by correction.The over-ground part plant area that experiment confirm is measured by this way is relevant with the biomass of ground plant part.The over-ground part area is the time point that has reached its maximum leaf biomass plant.Use suitable software to derive the feature of root, for example total projection area (it can be relevant with total root volume), mean diameter and be higher than the length (the slightly length of root or the length of radicula) of the root of certain rugosity threshold value from the image that produces.

With sophisticated main panicle gather in the crops, pack, add bar code label and subsequently in loft drier 37 ℃ of dryings 3 days.Subsequently with the panicle threshing and collect whole seeds.Use blast apparatus to separate full grain and empty grain.After separation, use commercially available counter that two batches of seeds are all counted then.Discard empty grain.Full grain is weighed on analytical balance and is used digital picture to measure the cross-sectional area of seed.This method produces with next group seed relevant parameter set:

The full grain number that the full seed number passes through behind the counting separating step is determined.The whole full grain that seed ultimate production (seed gross weight) is gathered in the crops from plant by weighing is measured.The capsomere number that every strain plant seed sum is gathered in the crops from plant by counting is measured.Harvest index (HI) is defined as seed ultimate production and over-ground part area (mm in the present invention ²) between ratio, multiply by coefficient 10 ⁶Full seed number and the extrapolated thousand seed weight of gross weight (TKW) thereof from counting.As the full rate of the seed that defines among the present invention is the ratio (be expressed as %) of full seed number to seed (or Xiao Hua) sum.Can use image analysis software to derive these parameters from digital picture, then these parameters be carried out statistical analysis in the mode of automatization.Use and image analysis software link coupled customizing device (by 2 primary clusterings, weigh and imaging device, form), measure the parameter (comprising width, length, area, weight) of single seed.

Use is made gauged double factor ANOVA (variance analysis) is used for the plant phenotype feature as statistical model total appraisal to non-equilibrium design.Whole measuring parameters with whole plants of whole incidents of described gene transformation are implemented the F check.Implement the F check to check that gene is for the effect of whole transformation events and the mass action of checking gene (being called " effect of the gene overall situation " again).If it is significant that the value of F check shows these data, then can draw the conclusion that has " gene " effect, mean that not only described effect is just caused in the existence or the position of gene.For the F check, the threshold value that is used for the significance of true overall gene action is arranged on 5% probability level.

For checking the effect of gene in incident, i.e. strain is a specific effect, uses the data set from the transgenic plant and the invalid plant of correspondence to carry out the t check in each incident." invalid plant " or " invalid segregant " or " inefficacy zygote " are handled in the mode identical with transgenic plant, but transgenosis has therefrom taken place by isolating plant.The invalid plant negative plant that transforms that also can be described as isozygotying.The threshold value that is used for the significance of t check is arranged on 10% probability level.The result of some incidents can be higher or lower than this threshold value.This is based on hypothesis like this, and promptly gene may only have effect in some position in genome, and the appearance of this position dependence effect is not rare.This gene action is called " the strain system effect of gene " in this article again.The p-value distributes relatively by t-value and t-or alternatively distributes by F-value and F-and relatively obtains.It is correct probability that this p-value provides null hypothesis (being that genetically modified effect does not exist) subsequently.

The data that obtain at AZ in first experiment in second experiment of using the T2 plant, have been confirmed.Selection has 4 strains of correct expression pattern and further analyzes.By the expression of monitoring mark, screening is from the seed lot (Seed batch) of the sun plant among the T1 (heterozygote and homozygote).For the incident of each selection, keep the heterozygote seed lot then to carry out the T2 assessment.In every batch of seed, the positive of growth equal amts and heliophobous plant are to assess in the greenhouse.

In T2 generation, assess the AZ that adds up to 120 strains and transformed plant, promptly every incident 30 strain plants, wherein 15 strains are transgenic positive, 15 strains are the transgenosis feminine gender.

Because two kinds of experiments carrying out have overlapping events, therefore carry out combinatory analysis.This can be used for checking the consistence of effect in two kinds of experiments, and if situation if this is really true, thereby it can be used for collecting evidence from two kinds of experiments and increases the credibility of conclusion.The method of using is to consider the method with mixed model of the multilevel hierarchy of data (i.e. experiment-incident-segregant).The likelihood ratio that distributes by card side is relatively tested and is obtained the p value.

Embodiment 6:AZ transformant is estimated: measure the output correlation parameter

As mentioned above to seed analysis the time, the inventor finds to compare with lacking the genetically modified plant of AZ, has higher seed production with AZ gene construct plant transformed, and it is expressed as (higher) thousand seed weight.In addition, compare the vigor of discovery increase in carrying genetically modified plant and the green degree coefficient of increase with control plant.

For one of construct, thousand seed weight has increased by 2.7% in T1 generation.These positive findingses obtain (2.1% increase) at T2 in generation again.The result in associating T1 generation these T2 data of reappraising in combinatory analysis, the observed effect of P value demonstration of acquisition is a highly significant.

The gene clone of embodiment 7:AtSYT1

(Invitrogen, Paisley UK) pass through pcr amplification Arabidopis thaliana AtSYT1 gene as template to use Arabidopis thaliana seedling cDNA library.Behind the RNA that reverse transcription is extracted from seedling, cDNA is cloned into pCMV Sport 6.0.The average inset size in library is 1.5kb, and clone's original number is 1.59 * 10 ⁷The rank of cfu.6 * 10 ¹¹After the amplification first time of cfu/ml, initial titer is defined as 9.6 * 10 ⁵Cfu/ml.After plasmid extracts, the 200ng template is used for 50 μ l PCR mixtures.Primer prm06681 (the SEQ ID NO:148 that will comprise the AttB site that is used for the Gateway reorganization; Justice is arranged, and initiator codon is a boldface letter, and the AttB1 site is an italic: 5 '-

CTTAAACA

CAACAGCACCTGATG-3 ') and prm06682 (SEQ ID NO:149; Oppositely, complementation, the AttB2 site is an italic: 5 '-

CATCATTAAGATTCCTTGTGC-3 ') is used for pcr amplification.In standard conditions, use Hifi Taq archaeal dna polymerase to carry out PCR.Also use the PCR fragment (comprising the attB site) of standard method amplification and purifying 727bp.Carry out the first step BP reaction of Gateway method then, during this step, PCR fragment and pDONR201 plasmid carry out reorganization in the body, thereby produce, according to " clone crosses the threshold " of Gateway nomenclature, p07466.Plasmid pDONR201 conduct

The part of technology is available from Invitrogen.

Embodiment 8: the structure of carrier

The clone p07466 that will cross the threshold subsequently is used from the LR reaction with the purpose carrier p00640 one that is used for plant (rice) conversion.This carrier form with functional element in the T-DNA border comprises: plant selectable marker, the marker expression box that can screen and be intended to be used for carry out the Gateway box of recombinating the LR body in the aim sequence that has been cloned into the clone that crosses the threshold.The rice GOS2 promotor (SEQ ID NO:145) that is used for constitutive expression (PRO0129) is positioned at the upstream of this Gateway box.

Many different binary (with the super binary) carrier system that is used for Plant Transformation (An has for example been described, G.in Agrobacterium Protocols.Methods in Molecular Biology the 44th volume, pp 47-62, Gartland KMA and MR Davey eds.Humana Press, Totowa, NewJersey).The carrier pBIN19 that many carrier systems are described based on Bevan (Nucleic Acid Research.1984.12:8711-8721), this carrier comprise flank and connect from the left side of agrobacterium tumefaciens Ti-plasmids and the gene expression in plants box of right border sequence.The gene expression in plants box is by at least two genes, and---cDNA of selectable marker gene and regulation and control character gene or plant promoter of transcribing of genomic dna---formed.Can use different selectable marker genes, comprise the arabidopsis gene (United States Patent (USP) 57673666 and 6225105) of the acetohydroxy acid synthase (AHAS) of encoding mutant.Similarly, can use different promoter regulation character genes to realize composing type, developmental pattern, tissue-type or environmental form regulation and control to genetic transcription.

After the LR reconstitution steps, heat-shocked or electroporation scheme are converted into agrobacterium strains LBA4044 with the expression vector pGOS2::AtSYT1 (Fig. 9) that produces.The bacterium colony that transforms was grown 2 days with 28 ℃ on the YEP substratum, and selected with corresponding microbiotic.These Agrobacterium cultures are used for Plant Transformation.

Other agrobacterium tumefaciens bacterial strains can be used for Plant Transformation and be well known in the art.This type of bacterial strain example is C58C1 or EHA105.

Embodiment 9: Plant Transformation

Rice transforms

The Agrobacterium that contains expression vector is used for transforming rice plant independently.Ripe dry seed shelling with the Japanese Cultivar Nipponbare of rice.By incubation in 70% ethanol one minute, in 2,%Hg,Cl2 30 minutes subsequently, subsequently with sterile distilled water washing 6 times 15 minutes and implement sterilization.The disinfectant seed is containing 2 subsequently, and the substratum of 4-D (callus inducing medium) is gone up and sprouted.Incubation is after 4 weeks in the dark, scultellum deutero-embryogenic callus is downcut and breeds on a kind of substratum.After 2 weeks, callus by breeding or breed uploading with a kind of substratum to be commissioned to train to support in other 2 weeks.The embryogenic callus sheet is uploaded to be commissioned to train at fresh culture and was supported 3, cultivates (to strengthen the cell fission activity) afterwards altogether.

The agrobacterium strains LBA4404 that contains expression vector is used for common cultivation.Agrobacterium is seeded in to contain on the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3.Subsequently bacterium is collected and is resuspended in liquid and cultivate altogether in the substratum to density (OD600) about 1.Suspension is transferred to culture dish subsequently and callus was soaked 15 minutes in this suspension.Callus is organized subsequently and to be cultivated on the substratum altogether and in the dark in 25 ℃ of incubations 3 days blotting and be transferred to solidified on the filter paper.Altogether the callus of cultivating in the dark in 28 ℃ in the presence of selective agent in containing 2,4 weeks of cultivation on the substratum of 4-D.During the section, form mushroom resistant calli island at this moment.To regeneration culture medium and behind incubation under the light, release of embryo generation potentiality and seedling are in 4-5 week growth subsequently in this material transfer.Seedling is downcut from callus and, wherein seedling is transferred to soil from described substratum containing incubation 2-3 week on the substratum of plant hormone.The hardened seedling is cultivated in the greenhouse under high humidity and short day.

A construct produces about 35 independent T0 rice transformant.With former generation transformant be transferred to the greenhouse from incubator for tissue culture.Behind the copy number of quantitative PCR analysis with checking T-DNA inset, the single copy transgenic plant that only keep performance selective agent tolerance are used to gather in the crops the T1 seed.Seed is gathered in the crops after transplanting subsequently the 3-5 month.Present method produces term single gene seat transformant (Aldemita and Hodges1996, Chan etc. 1993, Hiei etc. 1994) to surpass 50% ratio.

Corn transforms

The conversion of corn is carried out according to the modification method to (1996.Nature Biotech 14745-50) described methods such as Ishida.Conversion in corn be that genotype relies on and only the special genes type can operate and be used for transforming and regeneration.Inbred lines A188 (University of Minnesota) or be the good source of the donor material that is used to transform with A188 as parent's hybrid, but other genotype also can successfully be used.Mealie from maize plant after pollination about 11 days (DAP) results, this moment, the length of immature embryos was about 1 to 1.2mm.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.The embryo that downcuts is on the callus inducing medium, cultivate on the corn regeneration culture medium subsequently, and wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to the maize rooting substratum and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Wheat transforms

The conversion of wheat is carried out with the method that (1996) Nature Biotech 14 (6): 745-50 such as Ishida such as Ishida describe.Usually in conversion, use (can obtain) Cultivar Bobwhite from Mexico CIMMYT.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.With the Agrobacterium incubation after, embryo on the callus inducing medium, external cultivation on regeneration culture medium subsequently, wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to root media and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Soybean transforms

According to Texas A﹠amp; M United States Patent (USP) 5,164, the modification method soybean transformation of method described in 310.Several commercial soybean varieties are feasible for conversion by this method.Cultivar Jack (can be able to obtain from Illinois seed money) is generally used for transforming.Soybean seeds is sterilized so that external sowing.From 7 age in days seedling, downcut hypocotyl, radicle and a slice cotyledon.The cotyledon of further cultivating epicotyl and remainder is to grow the armpit tight knot.These armpit tight knots are downcut and with the agrobacterium tumefaciens incubation that contains expression vector.After cultivating processing altogether, explant is washed and is transferred to the selection substratum.The regenerated seedling is downcut and places on the seedling elongation medium.The seedling that length is no more than 1cm places on the root media until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant tolerance of performance selective agent and that contain single copy T-DNA inset, produce the T1 seed.

Semen Brassicae campestris/canola oil dish transforms

Cotyledon petiole of use 5-6 age in days seedling and hypocotyl are as being used for the explant of tissue culture and transforming according to (1998, Plant Cell Rep 17:183-188) such as Babic.Commercial Cultivar Westar (Agriculture Canada) is the standard variety that is used to transform, but also can use other kinds.Canola oil colza is done the surface sterilization so that external sowing.From external seedling, downcut and have the cotyledon petiole explant that adheres to cotyledon, and immerse bacterial suspension with the cut ends of (containing expression vector) Agrobacterium by petiole explant and inoculate.Explant subsequently on the MSBAP-3 substratum that contains 3mg/l BAP, 3% sucrose, 0.7% plant agar (Phytagar) at 23 ℃, illumination in 16 hours was cultivated 2 days down.After cultivating 2 altogether with Agrobacterium, petiole explant is transferred on the MSBAP-3 substratum of 3mg/l BAP, cefotaxime, Pyocianil or the Ticarcillin/Clavulanate Acid (300mg/l) that contain and continues 7, and cultivate containing on the MSBAP-3 substratum of cefotaxime, Pyocianil or Ticarcillin/Clavulanate Acid and selective agent subsequently, regenerate until seedling.When seedling has 5-10mm length, seedling is downcut and is transferred to seedling elongation medium (MSBAP-0.5 that contains 0.5mg/l BAP).The seedling of the about 2cm of length is transferred to the root media (MS0) that is used for root induction.The seedling that to take root migrates in the soil in greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Clover transforms

Clover (reproducibility clone use the method for (McKersie etc., 1999Plant Physiol 119:839-847) to be transformed.Regeneration of clover and conversion are that genotype is dependent and thereby need aftergrowth.The method that obtains the reproducibility plant has been described.For example, these reproducibility plants any other commercial alfalfa variety that can be selected from Cultivar Rangelander (Agriculture Canada) or describe as Brown DCW and AAtanassov (1985.Plant Cell Tissue Culture 4:111-112).Alternatively, RA3 kind (University of Wisconsin) has been selected for (Walker etc., 1978Am J Bot 65:654-659) in the tissue culture.Petiole explant and the agrobacterium tumefaciens C58C1pMP90 (McKersie etc., 1999Plant Physiol 119:839-847) or the overnight culture of LBA4404 that contain expression vector are cultivated altogether.Explant is containing 288mg/L Pro, 53mg/L Thioproline, 4.35g/L K in the dark ₂SO ₄With cultivated altogether 3 days on the SH inducing culture of 100 μ m Syringylethanones. explant half concentrate in the Murashige-Skoog substratum (Murashige and Skoog, 1962) washing and plating contain not containing Syringylethanone suitable selective agent and suitable microbiotic with the identical SH inducing culture of restraining the Agrobacterium growth on.After several weeks, somatic embryo is transferred to do not contain growth regulator, do not contain microbiotic and the BOi2Y that contains 50g/L sucrose grows in the substratum.Somatic embryo concentrates on the Murashige-Skoog substratum half subsequently to be sprouted.The sprigging that to take root is cultivated to flowerpot and in the greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 10: the evaluation of AtSYT1 transgenic plant is prepared under abiotic stress

Stay 4 incidents, the T1 offspring of wherein said incident is to genetically modified existence/do not exist with the 3:1 ratio and separate.For each incident in these incidents, select to contain genetically modified about 15 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 15 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Greenhouse experiment is short day (illumination in 12 hours), 28 ℃ of medial temperatures and 22 ℃ and average relative humidity 70% in the dark under light.

The salt stress screening

Culturing plants on the matrix of making by coconut fiber and argex (3 to 1 ratios).Sprigging first two weeks to the greenhouse is being used normal nutritive medium.After first two weeks, in nutritive medium, add the salt (NaCl) of 25mM, till the results plant.Measure the seed correlation parameter subsequently.

The arid screening

Culturing plants in flowerpot soil under normal operation is up to entering heading stage.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Measure the seed correlation parameter subsequently.

To transgenic plant apply a alternative approach that water coerces be with comprise osmoticum for example the water of polyoxyethylene glycol (PEG) under the specific flow of water, handle transgenic plant.Because PEG may be deleterious, therefore only plant is provided the exposure of short-term, restart then normally to water.

Nutrient (nitrogen) the operability screening that reduces

Except that nutritive medium, in flowerpot soil, cultivating rice plant under the normal condition.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Measure the seed correlation parameter subsequently.

Statistical analysis: F check

With the statistical models of dual factors ANOVA (variance analysis) as net assessment plant phenotype feature.All measuring parameters with all incidents of all plant of gene transformation of the present invention are carried out the F check.Carry out the effect of F check with all transformation events of inspection gene pairs, and the population effect of check gene, also be called " overall genetic effect ".The significance threshold value of real overall genetic effect is set to 5% probability level of F check.If significance F test value points to certain genetic effect, this means to be not only that the existence of gene or position cause difference on the phenotype.

The measurement of biomass correlation parameter

From sowing time to the ripening stage, plant is repeatedly by the digital image-forming case.On each time point, every strain plant is obtained digital image (2048 * 1536 pixels, 1,000 6 hundred ten thousand looks) from least 6 different angles.

Plant area (leaf biomass in other words) is on the ground determined by the sum of all pixels that counting is different from the ground plant part digital image of background.This value is got the mean value of same time point from the photo of different angle shots, and is converted to the physical surface value of representing with square millimeter (physicalsurface value) by calibration.Experiment shows that the ground plant area of measuring by this method is relevant with the biomass that plant shoot divides.Area reaches the time point value of its maximum leaf biomass plant on the ground.Early stage vigor shows as plant (seedling) the ground area of sprouting three weeks of back.

The measurement of seed correlation parameter

Gather in the crops sophisticated main panicle, counting, packed, use bar code label, under 37 ℃ in baking oven dry 3 days then.To the panicle threshing, collect all seeds and count then.Use gas blower that full husk is separated with empty husk.Discard the sky husk, then the residue fraction is counted once more.On analytical balance, take by weighing the weight of full husk.Determine the number of full seed by the number of counting the full husk that behind separating step, stays.Measure total seed production by taking by weighing from the weight of all full husks of plant results.Measure total number seeds of every strain plant by counting from the number of the husk of plant results.Number and its gross weight according to the full seed of counting are calculated thousand seed weight (TKW).Harvest index (HI) is defined as seed ultimate production and ground area (mm in the present invention ²) between ratio multiply by the factor 10 ⁶The sum of every paniculiform flower is defined as ratio between seed sum and the paniculiform number of sophisticated master in the present invention.The full rate of seed accounts for the total per-cent of seed (or Xiao Hua) at the number that the present invention is defined as full seed.

Embodiment 11: the evaluation result of AtSYT1 transgenosis rice plant under abiotic stress (salt stress)

Provided evaluation result among the table B to the AtSYT1 transgenosis rice plant that is subjected to salt stress.Also show the difference percentage ratio between transgenic plant and the corresponding inefficacy zygote, be lower than 0.05 from the P value of F check.

Compare with control plant (in this case, the inefficacy zygote) under abiotic stress, ground biomass, early stage vigor, seed ultimate production, full seed number, the full rate of seed, TKW and coefficient of harvest significantly increase in the AtSYT1 transgenic plant.

Table B: the evaluation result of AtSYT1 transgenosis rice plant under abiotic stress.

Proterties	% difference
		Ground biomass	19
Early stage vigor	18
		The seed ultimate production	30
The full seed number	18
		Full rate	15
TKW	10
		Coefficient of harvest	16

Embodiment 12: the evaluation of the sequence relevant with SEQ ID NO:155 with SEQ ID NO:154

Use (BLAST) (people (1990) J.Mol.Biol.215:403-410 such as Altschul of for example basic local comparison instrument of database search instrument (Basic Local AlignmentTool); With people (1997) Nucleic Acids Res.25:3389-3402 such as Altschul), in the sequence of the Entrez of American National biotechnology information center (NCBI) Nucleotide database maintenance, identify the sequence (full-length cDNA, EST or genome sequence) relevant with SEQ ID NO:155 with SEQ IDNO:154.Usually blast program is by comparing nucleic acid or peptide sequence with sequence library, and by calculating the significance,statistical that mates, is used to seek the zone that has local similar between the sequence.To nucleic acid SEQ ID NO:154 encoded polypeptide utilization TBLASTN algorithm, use default setting, open strainer, to ignore the low complex degree sequence.The output form of analyzing is for comparing in twos, and sorts according to probability score (E value), and wherein score value reflects the occurrent probability of specific comparison (the E value is low more, and the significance of hit event is high more).Except the E value, also keep the score to relatively carrying out identity per-cent.Identity per-cent is meant that two compare the number of the identical Nucleotide (or amino acid) on length-specific between nucleic acid (or polypeptide) sequence.In some cases, can adjust the strict degree of default parameter with the change search.

Except the public obtainable nucleotide sequence that can on NCBI, obtain, also search for private sequence library according to method same as the above-mentioned method.

Table C provides the nucleotide sequence of representing with SEQ ID NO:154 relevant nucleotide sequence tabulation.

Table C: can be used for the inventive method with the relevant nucleotide sequence of nucleotide sequence (SEQ ID NO:154), and the polypeptide of deriving accordingly.

Title	The source is biological	Nucleic acid SEQ ID NO:	Polypeptide SEQ ID NO:	Database login number	State
						Chlre_cpFBPase	Chlamydomonas reinhardtii (Chlamydomonas reinhardtii)	154	155	From AW721488 BQ811919 CF555540	Total length
Bigna_cpFBPase	Bigelowiella natans		156	157	AY267678							Total length
						Aqufo_cpFBPase	Aquilegia formosa x Aquilegia pubescens	158	159	DR944021 DT749545	Total length
Arath_cpFBPase	Arabidopis thaliana		160	161	X58148							Total length
						Brana_cpFBPase	Colea
	162	163	AF081796	Total length
					Cyame_cpFBPase	Cyanidioschyzon merolae		164	165	CMO245C ＊	Total length
Glyma_cpFBPase	Soybean
			166	167	L34841	Total length
Lyces_cpFBPase	Tomato
			168	169	BT014319	Total length
Medtr_cpFBPase	The puncture vine clover						170	171	BI310407 BI265103 CO516140	Total length
		Nicta_cpFBPase	Tobacco
	172			173	DW000613 EB680028	Total length
		Orysa_cpFBPase	Rice
	174			175	AB007194	Total length

Ostlu_cpFBPase	Ostreococcus lucimarinus		176	177	jgi|Ost9901_3|92356 |ost_03_002_042 ＊＊	Total length
							Ostta_cpFBPase	Ostreococcus tauri		178	179	CR954203	Total length
Phatr_cpFBPase	Phaeodactylum tricornutum		180	181	scaffold_27：145102-145476 ＊＊	Total length
							Pissa_cpFBPase	Ripe pea	182	183	L34806	Total length
Pontr_cpFBPase	Trifoliate orange (Poncirus trifoliata)	184	185	CV705787 CV705786	Total length
						Poptr_cpFBPase	Populus tremuloides		186	187	CV241715 DT474034	Total length
Soltu_cpFBPase	Potato
			188	189	AF1340451	Total length
Spiol_cpFBPase	Spinach
			190	191	L76555	Total length
Triae_cpFBPase	Common wheat						192	193	X07780	Total length
		Zeama_cpFBPase	Zea mays	194	195	DR792524.1 DT645374.1					Total length
Phypa_cpFBPase	Small liwan moss						196	197	BY991118.1 BJ168552.1 +proprietary	Total length
		Galsu_cpFBPase	Galderia sulphuraria	198	199	AJ302644					Part
Marpo_cpFBPase	Marchantia (Marchantia polymorpha)						200	201	BJ862191	Part
		Tagpa_cpFBPase	Herba Tagetis Patulae (Tagetes patula)	202	203	Contig CON_01b-cs_scarletade-4-e3.b1 proprietary					Part
Linus_cpFBPase
		204	204	205	contig6298 proprietary	Part

^*Cyanidioschyzon merolae database is positioned at the biology department of Tokyo university

^*Ostreococcus lucimarinus and Phaeodactylum tricornutum e database are positioned at DOE JointGenome Institute, US Department of Energy

Embodiment 13: the comparison of related polypeptide sequence

From the AlignX of Vector NTI (Invitrogen) based on the clustering algorithm (Clustal algorithm) of the progressive comparison of generally using (people (1997) Nucleic Acids Res25:4876-4882 such as Thompson; People such as Chenna (2003) .Nucleic Acids Res 31:3497-3500).Can use in abutting connection with the clustering algorithm constructing system and set.The default value of the open point penalty in room is 10, and the default value that point penalty is extended in the room is 0.1, and the weight matrix of selection is Blosum 62 (if comparison polypeptide).

Can be used for implementing in the polypeptide of the inventive method in evaluation, the multisequencing comparison result that uses related polypeptide to obtain is shown among Figure 12 of the application.There is the transit peptides that supplies plastid (chloroplast(id)) ubcellular target (adding frame table among the figure shows) in chloroplast(id) FBPase polypeptide (cpFBPase) with the N-terminal that kytoplasm FBPase (cyFBPase) is different from chloroplast(id) FBPase polypeptide.In addition, the former comprises aminoacid insertion (also add frame table and show, and fragment is inserted in called after redox regulation and control), and described aminoacid insertion comprises that at least two disulfide linkage form (being the redox regulation and control) necessary cysteine residues.Conservative halfcystine is with them the position name in ripe pea (Pisum sativa) cpFBPase polypeptide, i.e. Cys153, Cys173 and Cys178.Cys153 and Cys173 normally participate in two mating partners that disulfide linkage forms (people such as Chiadmi. (1999) EMBO J 18 (23): 6809-6815).Avtive spot motif as definition cpFBPase in the Prosite database is PS00124 and corresponding with following aminoacid sequence: [AG]-[RK]-[LI]-X (1,2)-[LIV]-[FY]-E-X (2)-P-[LIVM]-[GSA], wherein [RK] is the bonded avtive spot, and X is an arbitrary amino acid.Prediction avtive spot motif and prediction avtive spot itself (being included in the described motif) are also shown by frame table in Figure 12.In Figure 12, also identify in conjunction with fructose-1, amino-acid residue Asn237, Tyr269, Tyr289 and the Arg268 of the 6-phosphoric acid of 6-bisphosphate and in conjunction with the amino-acid residue Lys299 of fructose (people such as Chiadmi. (1999) EMBO J 18 (23): 6809-6815).The back these also with they Position Numbers in ripe pea (Pisumsativa) cpFBPase polypeptide.

Embodiment 14: calculate the overall per-cent identity (global percentage identity) between the peptide sequence can be used for implementing the inventive method

A method MatGAT in the method that use can obtain in this area (matrix overall comparison instrument (Matrix Global Alignment Tool)) software (BMC Bioinformatics.20034:29.MatGAT:an application that generates similarity/identity matricesusing protein or DNA sequences.Campanella JJ, Bitincka L, Smalley J; Software by Ledion Bitincka trustship), determine to be used to implement overall similarity and identity per-cent between the full-length polypeptide sequence of method of the present invention.MatGAT software need not data are compared in advance, can produce the similarity/identity matrix of DNA or protein sequence.This program is utilized Myers and Miller overall comparison algorithm, and (the open point penalty in room is 12, and to extend point penalty be 2 in the room) carry out a series of comparison in twos, utilize for example Blosum 62 (for polypeptide) calculating similarity and identity, then the result is arranged in distance matrix.Sequence similarity is shown in the diagonal lines Lower Half, and sequence identity is shown in the diagonal lines first half.

More used parameter has:

Matrix: Blosum 62 keeps the score

First room: 12

Extend the room: 2

The overall similarity of peptide sequence length range (the part of polypeptide sequence is foreclosed) and the software analysis of identity the results are shown in table D.The diagonal lines top provides identity per-cent, and the diagonal lines below provides similarity per-cent.

Compare with SEQ ID NO:155, the per-cent identity that can be used for implementing between the peptide sequence of the inventive method can be for being low to moderate 40% amino acid identity.

Table D: the overall similarity of full-length polypeptide sequence scope and the MatGAT result of identity.

Embodiment 15: identify at the structural domain that is used to implement comprise in the peptide sequence of the inventive method

(InterPro) database is based on the integrated interface of common used tag database of the search of text and sequence to the integrated resource in protein families, structural domain and site for Integrated Resouce of ProteinFamilies, Domain and Site.The InterPro database has made up these databases, and described database uses different methods to learn biological information with in various degree relevant fully profiling protein matter to obtain protein tag.The cooperation database comprises SWISS-PROT, PROSITE, TrEMBL, PRINTS, ProDom and Pfam, Smart and TIGRFAMs.Interpro resides at the European bioinformation institute of Britain.

The result of the peptide sequence of InterPro scanning as SEQ ID NO:155 representative is in showing E.

Table E: as the InterPro scanning result of the peptide sequence of SEQ ID NO:155 representative

Database	Searching number	The retrieval title
			InterPro	IPR000146	Inositol monophosphate enzyme/fructose-1
ProDom	PD001491	Inositol monophosphate enzyme/fructose-1
			PRINTS	PR00115	FBPHPHTASE
The PIR superfamily	PIRSF000904	Fructose-1/sedoheptulose-1, the 7-diphosphatase

PANTHER	PTHR11556	Fructose-1-relevant
			Pfam	PF00316	FBPase
PROFILE	PS00124	The FBPASE avtive spot
			PROSITE	PS00124	The FBPASE avtive spot

Embodiment 16: be used to implement the topology prediction (Subcellular Localization is striden film ...) of the peptide sequence of the inventive method

The Subcellular Localization of TargetP 1.1 prediction eukaryotic proteins.The existence of the prediction that is based on any aminoterminal presequence is distributed in the position: chloroplast transit peptides (cTP), Mitochondrially targeted peptide (mTP) or Secretory Pathway signal peptide (SP).Unactual as keeping the score of final fundamentals of forecasting is probability, and they may not be integrated.Yet having the highest position of keeping the score is most possible according to TargetP, and the relation between keeping the score (reliability category) can be an index of the certainty of prediction.Reliability category (RC) is 1-5, the wherein the strongest prediction of 1 expression.TargetP safeguards on the server of Technical University Of Denmark.

Contain the sequence of aminoterminal presequence for prediction, also can predict the potential cleavage site.

Numerous parameters have been selected, as biology group (non-plant or plant), critical setting (do not have, the critical or user of predefine setting specify the critical of setting) with to the calculating of cleavage site prediction (be or not).

The result who analyzes as the TargetP 1.1 of the peptide sequence of SEQ ID NO:155 representative shows in table F.Select " plant " biological group, undefined threshold value and need the prediction length of transit peptides.Subcellular Localization as the peptide sequence of SEQ ID NO:155 representative can be a chloroplast(id), and the prediction length of transit peptides is 56 amino acid starting from the N-end (not as Subcellular Localization self reliable, can change on several amino acid whose length).It is 53 amino acid whose transit peptides that ripe Peas cpFBPase has length.When comparison Peas cpFBPase and cpFBPase SEQ lD NO:155, the length of the latter's transit peptides of may deriving also is 53 amino acid.

Table F: the TargetP 1.1 as the peptide sequence of SEQ ID NO:44 representative analyzes

Length (AA)	415
		Chloroplast transit peptides	0.832
The mitochondrial transport peptide	0.106

The Secretory Pathway signal peptide	0.003
		Other ubcellular targets	0.065
The position of prediction	Chloroplast(id)
		Reliability category	2
The transit peptides length of prediction	56

Numerous other algorithms can be used for carrying out this alanysis, comprising:

Resident ChloroP 1.1 on Technical University Of Denmark (Technical University of Denmar) server;

At (the Institute forMolecular Bioscience of molecular biosciences institute of Brisbane ,Australia University of Queensland, University of Queensland, Brisbane, resident protein Prowler Subcellular Localization predictor (Protein ProwlerSubcellular Localisation Predictor) is the 1.2nd edition on server Australia);

At Canadian Alberta province Edmonton city University of Alberta (University of Alberta, Edmonton, Alberta, resident PENCE Proteme AnalystPA-GOSUB 2.5. on server Canada)

Embodiment 17: with the relevant analytical method of peptide sequence that is used to implement the inventive method

Peptide sequence shown in SEQ ID NO:155 is to have (the EC of EC; Enzyme classification based on enzymatic reaction) numbering EC 3.1.3.11 is the enzyme of fructose-diphosphatase (being also referred to as D-fructose-1,6-diphosphate 1-phosphohydrolase).This functional assays can be based on the mensuration of the cpFBPase determination of activity of Pi colorimetric, as Huppe and Buchanan (1989), described in Naturforsch.44c:487-494.Other methods of measuring enzymic activity are described in PlantPhysiol 70:728-734 by Alscher-Herman (1982).

Embodiment 18: the clone is as the nucleotide sequence of SEQ ID NO:154 representative

Unless stated otherwise, recombinant DNA technology is according to (Sambrook (2001) MolecularCloning:a laboratory manual, third edition Cold Spring Harbor LaboratoryPress, CSH, New York) in or Ausubel etc. (1994), Current Protocols inMolecular Biology, the standard method of describing in Current Protocols the 1st and 2 volumes is carried out.The standard material and the method that are used for plant molecular work are described at the Plant Molecular Biology Labfax (1993) of the R.D.D.Croy that is published by BIOS Scientific PublicationsLtd (UK) and Blackwell Scientific Publications (UK).

Nucleotide sequence used in the inventive method passes through PCR, use is available from the Chlamy center (chlamydomonas gene center in the past) of Duke University, North Carolina, the Chlamydomonas reinhardtii CC-1690cDNA library of the U.S. (" Core library ") (in the λ ZAP II carrier from Stratagene) increases as template.Under standard conditions, use Hifi Taq archaeal dna polymerase, utilize the 200ng template in 50 μ l PCR mixtures to carry out PCR.The primer is

Prm08448SEQ ID NO:206; Justice is arranged, and the AttB1 site is a small letters: 5 '-ggggacaagtttgtacaaaaaagcaggcttaaacaatggccgccaccatg-3 '; And prm08449 (SEQ ID NO:207; Antisense, complementary AttB2 site is a small letters: 5 '-ggggaccactttgtacaagaaagctgggtagctgcttagtgcttcttggt-3 ',

It comprises the AttB site that is used for the Gateway reorganization.The amplification PCR fragment also uses standard method to give purifying.Implement the first step of Gateway method subsequently, i.e. BP reaction is recombinated in PCR fragment and the pDONR201 plasmid generation body during this period to produce " the entering the clone " according to the Gateway name, p15972.Plasmid pDONR201 conduct

The component part of technology is bought from Invitrogen.

Embodiment 19: use as SEQ ID NO:154 representative nucleotide sequence construction of expression vector

Entering clone p15972 uses with p05050 (a kind of purpose carrier that is used for the rice conversion) in the LR reaction subsequently.This carrier contains as functional element on the T-DNA border: plant selectable marker; The selection markers expression cassette is with intention and cloned the Gateway box of recombinating in the described purpose nucleotide sequence generation LR body that enters in the clone.The rice GOS2 promotor (SEQ ID NO:208) that is used for constitutive expression (PRO0129) is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector pGOS2::FBPase (Figure 13) of generation is converted among the agrobacterium strains LBA4044 according to method well-known in the art.

Embodiment 20: Plant Transformation

Rice transforms

The Agrobacterium that contains expression vector is used for transforming rice plant.Ripe dry seed shelling with the Japanese Cultivar Nipponbare of rice.By incubation in 70% ethanol one minute, in 2,%Hg,Cl2 30 minutes subsequently, subsequently with sterile distilled water washing 6 times 15 minutes and implement sterilization.The disinfectant seed is containing 2 subsequently, and the substratum of 4-D (callus inducing medium) is gone up and sprouted.Incubation is after 4 weeks in the dark, scultellum deutero-embryogenic callus is downcut and breeds on a kind of substratum.After 2 weeks, callus by breeding or breed uploading with a kind of substratum to be commissioned to train to support in other 2 weeks.The embryogenic callus sheet is uploaded to be commissioned to train at fresh culture and was supported 3, cultivates (to strengthen the cell fission activity) afterwards altogether.

The agrobacterium strains LBA4404 that contains expression vector is used for common cultivation.Agrobacterium is seeded in to contain on the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3.Subsequently bacterium is collected and is resuspended in liquid and cultivate altogether in the substratum to density (OD600) about 1.Suspension is transferred to culture dish subsequently and callus was soaked 15 minutes in this suspension.Callus is organized subsequently and to be cultivated on the substratum altogether and in the dark in 25 ℃ of incubations 3 days blotting and be transferred to solidified on the filter paper.Altogether the callus of cultivating in the dark in 28 ℃ in the presence of selective agent in containing 2,4 weeks of cultivation on the substratum of 4-D.During the section, form mushroom resistant calli island at this moment.To regeneration culture medium and behind incubation under the light, release of embryo generation potentiality and seedling are in 4-5 week growth subsequently in this material transfer.Seedling is downcut from callus and, wherein seedling is transferred to soil from described substratum containing incubation 2-3 week on the substratum of plant hormone.The hardened seedling is cultivated in the greenhouse under high humidity and short day.

A construct produces about 35 independent T0 rice transformant.With former generation transformant be transferred to the greenhouse from incubator for tissue culture.Behind the copy number of quantitative PCR analysis with checking T-DNA inset, the single copy transgenic plant that only keep performance selective agent tolerance are used to gather in the crops the T1 seed.Seed is gathered in the crops after transplanting subsequently the 3-5 month.Present method produces term single gene seat transformant (Aldemita and Hodges1996, Chan etc. 1993, Hiei etc. 1994) to surpass 50% ratio.

Corn transforms

The conversion of corn is carried out according to the modification method to (1996.Nature Biotech 14745-50) described methods such as Ishida.Conversion in corn be that genotype relies on and only the special genes type can operate and be used for transforming and regeneration.Inbred lines A188 (University of Minnesota) or be the good source of the donor material that is used to transform with A188 as parent's hybrid, but other genotype also can successfully be used.Mealie from maize plant after pollination about 11 days (DAP) results, this moment, the length of immature embryos was about 1 to 1.2mm.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.The embryo that downcuts is on the callus inducing medium, cultivate on the corn regeneration culture medium subsequently, and wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to the maize rooting substratum and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Wheat transforms

The conversion of wheat is carried out with the method that (1996) Nature Biotech 14 (6): 745-50 such as Ishida such as Ishida describe.Usually in conversion, use (can obtain) Cultivar Bobwhite from Mexico CIMMYT.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.With the Agrobacterium incubation after, embryo on the callus inducing medium, external cultivation on regeneration culture medium subsequently, wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to root media and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Soybean transforms

According to Texas A﹠amp; M United States Patent (USP) 5,164, the modification method soybean transformation of method described in 310.Several commercial soybean varieties are feasible for conversion by this method.Cultivar Jack (can be able to obtain from Illinois seed money) is generally used for transforming.Soybean seeds is sterilized so that external sowing.From 7 age in days seedling, downcut hypocotyl, radicle and a slice cotyledon.The cotyledon of further cultivating epicotyl and remainder is to grow the armpit tight knot.These armpit tight knots are downcut and with the agrobacterium tumefaciens incubation that contains expression vector.After cultivating processing altogether, explant is washed and is transferred to the selection substratum.The regenerated seedling is downcut and places on the seedling elongation medium.The seedling that length is no more than 1cm places on the root media until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant tolerance of performance selective agent and that contain single copy T-DNA inset, produce the T1 seed.

Semen Brassicae campestris/canola oil dish transforms

Cotyledon petiole of use 5-6 age in days seedling and hypocotyl are as being used for the explant of tissue culture and transforming according to (1998, Plant Cell Rep 17:183-188) such as Babic.Commercial Cultivar Westar (Agriculture Canada) is the standard variety that is used to transform, but also can use other kinds.Canola oil colza is done the surface sterilization so that external sowing.From external seedling, downcut and have the cotyledon petiole explant that adheres to cotyledon, and immerse bacterial suspension with the cut ends of (containing expression vector) Agrobacterium by petiole explant and inoculate.Explant subsequently on the MSBAP-3 substratum that contains 3mg/lBAP, 3% sucrose, 0.7% plant agar (Phytagar) at 23 ℃, illumination in 16 hours was cultivated 2 days down.After cultivating 2 altogether with Agrobacterium, petiole explant is transferred on the MSBAP-3 substratum of 3mg/l BAP, cefotaxime, Pyocianil or the Ticarcillin/Clavulanate Acid (300mg/l) that contain and continues 7, and cultivate containing on the MSBAP-3 substratum of cefotaxime, Pyocianil or Ticarcillin/Clavulanate Acid and selective agent subsequently, regenerate until seedling.When seedling has 5-10mm length, seedling is downcut and is transferred to seedling elongation medium (MSBAP-0.5 that contains 0.5mg/l BAP).The seedling of the about 2cm of length is transferred to the root media (MS0) that is used for root induction.The seedling that to take root migrates in the soil in greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Clover transforms

The reproducibility clone of clover uses the method for (McKersie etc., 1999Plant Physiol 119:839-847) to be transformed.Regeneration of clover and conversion are that genotype is dependent and thereby need aftergrowth.The method that obtains the reproducibility plant has been described.For example, these reproducibility plants any other commercial alfalfa variety that can be selected from Cultivar Rangelander (Agriculture Canada) or describe as Brown DCW and AAtanassov (1985.Plant Cell Tissue Culture 4:111-112).Alternatively, RA3 kind (University of Wisconsin) has been selected for (Walker etc., 1978Am J Bot 65:654-659) in the tissue culture.Petiole explant and the agrobacterium tumefaciens C58C1 pMP90 (McKersie etc., 1999Plant Physiol 119:839-847) or the overnight culture of LBA4404 that contain expression vector are cultivated altogether.Explant is containing 288mg/L Pro, 53mg/L Thioproline, 4.35g/L K in the dark ₂SO ₄With cultivated altogether 3 days on the SH inducing culture of 100 μ m Syringylethanones. explant half concentrate in the Murashige-Skoog substratum (Murashige and Skoog, 1962) washing and plating contain not containing Syringylethanone suitable selective agent and suitable microbiotic with the identical SH inducing culture of restraining the Agrobacterium growth on.After several weeks, somatic embryo is transferred to do not contain growth regulator, do not contain microbiotic and the BOi2Y that contains 50g/L sucrose grows in the substratum.Somatic embryo concentrates on the Murashige-Skoog substratum half subsequently to be sprouted.The sprigging that to take root is cultivated to flowerpot and in the greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 21: the phenotype evaluation method

Prepare 21.1 estimate

Produce about 35 T0 rice transformant independently.Transformant was transferred to the greenhouse from incubator for tissue culture and was used for growth and results T1 seed former generation.Stay 8 incidents, the T1 offspring of wherein said incident separates with 3: 1 ratios genetically modified existence/do not exist.For each incident in these incidents, select to contain genetically modified about 10 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 10 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Greenhouse experiment is short day (illumination in 12 hours), 28 ℃ and 22 ℃ and relative humidity 70% in the dark under light.

The arid screening

In flowerpot soil, cultivate plant under normal operation, up to entering heading stage from the T2 seed.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

The nitrogen use efficiency screening

Under normal condition except that nutritive medium, in flowerpot soil, cultivate rice plant from the T2 seed.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

4 T1 incidents T2 from generation to generation in according to as be used for T1 identical evaluation method from generation to generation and do further assessment, but that each incident adopts is more individual.Make plant pass through the digital imagery case for several times until the ripening stage from sowing time.On each time point, to every strain plant from least 6 different angles shooting digital pictures (2048x1536 pixel, 1,600 ten thousand colors).

21.2 statistical study: F-check

Use double factor ANOVA (variance analysis) to be used for the overall evaluation of plant phenotype feature as statistical model.All measuring parameters with whole plants of whole incidents of gene transformation of the present invention are implemented F check.Implement the F check to check that gene is for the effect of whole transformation events and the mass action (being called overall gene action again) of checking gene.The threshold value that is used for the significance of true overall gene action is arranged on 5% probability level for F check.Significance F test value indicates gene action, means that the existence of gene not only or position just cause the difference on the phenotype.

21.3 parameter measurement

The measurement of biomass correlation parameter

From sowing time to the ripening stage, plant is repeatedly by the digital image-forming case.On each time point, every strain plant is obtained digital image (2048 * 1536 pixels, 1,000 6 hundred ten thousand looks) from least 6 different angles.

Plant area (leaf biomass in other words) is on the ground determined by the sum of all pixels that counting is different from the ground plant part digital image of background.This value is got the mean value of same time point from the photo of different angle shots, and is converted to the physical surface value of representing with square millimeter (physicalsurface value) by calibration.Experiment shows that the ground plant area of measuring by this method is relevant with the biomass that plant shoot divides.Area reaches the time point value of its maximum leaf biomass plant on the ground.Early stage vigor shows as plant (seedling) the ground area of sprouting three weeks of back.

The measurement of seed correlation parameter

Gather in the crops sophisticated main panicle, counting, packed, use bar code label, under 37 ℃ in baking oven dry 3 days then.To the panicle threshing, collect all seeds and count then.Use gas blower that full husk is separated with empty husk.Discard the sky husk, then the residue fraction is counted once more.On analytical balance, take by weighing the weight of full husk.Determine the number of full seed by the number of counting the full husk that behind separating step, stays.Measure total seed production by taking by weighing from the weight of all full husks of plant results.Measure total number seeds of every strain plant by counting from the number of the husk of plant results.Number and its gross weight according to the full seed of counting are calculated thousand seed weight (TKW).Harvest index (HI) is defined as seed ultimate production and ground area (mm in the present invention ²) between ratio multiply by the factor 10 ⁶The sum of every paniculiform flower is defined as ratio between seed sum and the paniculiform number of sophisticated master in the present invention.The full rate of seed accounts for the total per-cent of seed (or Xiao Hua) at the number that the present invention is defined as full seed.

Embodiment 22: the phenotype evaluation result of transgenic plant

Provided the evaluation result of expressing the transgenosis rice plant of the nucleotide sequence that is used for the inventive method among the table G.Also show the difference percentage ratio between transgenic plant and the corresponding inefficacy zygote, be lower than 0.05 from the P value of F check.

Compare with control plant (in this case, the inefficacy zygote) under abiotic stress, seed ultimate production, full seed number, the full rate of seed and coefficient of harvest are used for the nucleotide sequence of the inventive method in expression transgenic plant significantly increase.

Table G: the evaluation result of expressing the transgenosis rice plant of the nucleotide sequence that is used for the inventive method.

Proterties	The increase % of T1 in the generation	The increase % of T2 in the generation
			The seed ultimate production	7	28
The full seed number	5	28
			Full rate	6	19
Coefficient of harvest	4	21

Embodiment 23: gene clone

By pcr amplification rice gene, use primer (SEQ ID NO:227; 5 '-ggggacaagtttgtacaaaaaagcaggcttaaacaatgatgggttgcttcactgtc-3 ') and (SEQ ID NO:228 justice is arranged, and initiator codon is a boldface letter, and the AttB1 site is an italic:; Antisense, complementary AttB2 site is an italic: 5 '-ggggaccactttgtacaagaaagctgggtatggacaatcaaaaaccctca-3 '), it comprises the AttB site that is used for the Gateway reorganization, is used for pcr amplification.Under standard conditions, use Hifi Taq archaeal dna polymerase to carry out PCR.The amplification PCR fragment also uses standard method to give purifying.Implement the first step of Gateway method subsequently, i.e. BP reaction, " entering the clone " that reorganization is named according to Gateway with generation in PCR fragment and the pDONR201 plasmid generation body during this period.Plasmid pDONR201 conduct

The component part of technology is bought from Invitrogen.

Embodiment 24: decrement is regulated vector construction

Entering the clone uses with the purpose carrier that is used for the conversion of decrement adjusting construct rice in the LR reaction subsequently.This carrier contains on the T-DNA border: the plant selected marker; The Gateway box of reorganization in the LR body (therefore integrating so that justice and antisense orientation to be arranged from the aim sequence that enters the clone) takes place in selection markers expression cassette and intention.The rice GOS2 promotor (SEQ IDNO:226) that is used for constitutive expression is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector (Figure 16) that produces is converted into agrobacterium strains LBA4044 and subsequent transformation to rice plant.The rice plant's growth that allows to transform is also tested to parameter described in the embodiment 26 subsequently.

Embodiment 25: the overexpression vector construction

Entering the clone uses with the purpose carrier that is used for plant (rice) conversion in the LR reaction subsequently.This carrier contains on the T-DNA border: the plant selected marker; Selection markers expression cassette and intention with cloned aim sequence in the described clone of entering the Gateway box of recombinating in the LR body taken place.The rice GOS2 promotor that is used for constitutive expression is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector (Figure 17) that produces is converted into agrobacterium strains LBA4044 and subsequent transformation to rice plant.The bacterium colony that transforms was grown 2 days with 28 ℃ on the YEP substratum, and selected with corresponding microbiotic.These Agrobacterium cultures are used for Plant Transformation.The rice plant's growth that allows to transform is also tested to parameter described in the embodiment 26 subsequently.

Embodiment 26: with the evaluation of antisense orientation SIK plant transformed

Produce about 15 to 20 T0 rice transformant independently.Transformant was transferred to the greenhouse from incubator for tissue culture and was used for growth and results T1 seed former generation.Stay 6 incidents, the T1 offspring of wherein said incident separates with 3: 1 ratios genetically modified existence/do not exist.For each incident in these incidents, select to contain genetically modified about 10 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 10 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.The T1 plant of selecting is transferred to the greenhouse.Each plant is accepted unique bar coded sticker clearly phenotype somatotype data are connected with corresponding plant.Be provided with down in following environment on the soil of T1 plant in 10cm diameter flowerpot of selecting and cultivate: photoperiod=11.5 hour, day optical density(OD)=30,000lux, daylight temperature=28 ℃ or higher, nocturnal temperature=22 ℃, relative humidity=60-70%.Culturing plants under best irrigation conditions up to entering heading stage, stops to irrigate simultaneously.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than 20%, continue moisturizing from the trend plant, reach best irrigation conditions once more.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Make plant pass through the digital imagery case for several times until the ripening stage from sowing time.On each time point, to every strain plant from least 6 different angles shooting digital pictures (2048x1536 pixel, 1,600 ten thousand colors).

The arid screening

In flowerpot soil, cultivate plant under normal operation, up to entering heading stage from the T2 seed.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

The nitrogen use efficiency screening

Under normal condition except that nutritive medium, in flowerpot soil, cultivate rice plant from the T2 seed.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

Plant shoot divides area (or leaf biomass) to measure by counting is different from the pixel of background on the digital picture of dividing from plant shoot sum.This value changes into square physical surface value of mm expression to the averaging of picture of taking from different perspectives on identical time point and by correction.The over-ground part plant area that experiment confirm is measured by this way is relevant with the biomass of ground plant part.Maximum area is the over-ground part area that has reached the time point of its maximum leaf biomass plant.

With sophisticated main panicle gather in the crops, pack, add bar code label and subsequently in loft drier 37 ℃ of dryings 3 days.Subsequently with the panicle threshing and collect whole seeds.Use blast apparatus to separate full grain and empty grain.After separation, use commercially available counter that two batches of seeds are all counted then.Discard empty grain.Full grain is weighed on analytical balance and is used digital picture to measure the cross-sectional area of seed.This method produces with next group seed relevant parameter set:

Each is paniculiform, and to spend number be the parameter of estimating the average number of each panicle Xiao Hua on the plant, this parameter from the seed sum divided by first panicle number.Be regarded as first panicle and count with the whole panicles of the highest panicle eclipsed when the highest panicle and arranged vertical with manual mode.The full grain number that the full seed number passes through behind the counting separating step is determined.The whole full grain that seed ultimate production (total seed is heavy) is gathered in the crops from plant by weighing is measured.The capsomere number that every strain plant seed sum is gathered in the crops from plant by counting is measured and corresponding every strain plant Xiao Hua number.Can use image analysis software to derive these parameters from digital picture, then these parameters be carried out statistical analysis in the mode of automatization.Use and image analysis software link coupled customizing device (by 2 primary clusterings, weigh and imaging device, form), measure the parameter (comprising width, length, area, weight) of single seed.Harvest index (HI) is defined as seed ultimate production and over-ground part area (mm in the present invention ²) between ratio, multiply by coefficient 10 ⁶

Use is made gauged double factor ANOVA (variance analysis) is used for the plant phenotype feature as statistical model total appraisal to non-equilibrium design.Whole measuring parameters with whole plants of whole incidents of described gene transformation are implemented the F check.Implement the F check to check that gene is for the effect of whole transformation events and the mass action of checking gene (being called " effect of the gene overall situation " again).If it is significant that the value of F check shows these data, then can draw the conclusion that has " gene " effect, mean that not only described effect is just caused in the existence or the position of gene.For the F check, the threshold value that is used for the significance of true overall gene action is arranged on 5% probability level.

For checking the effect of gene in incident, i.e. strain is a specific effect, uses the data set from the transgenic plant and the invalid plant of correspondence to carry out the t check in each incident." invalid plant " or " invalid segregant " or " inefficacy zygote " are handled in the mode identical with transgenic plant, but transgenosis has therefrom taken place by isolating plant.The invalid plant negative plant that transforms that also can be described as isozygotying.The threshold value that is used for the significance of t check is arranged on 10% probability level.The result of some incidents can be higher or lower than this threshold value.This is based on hypothesis like this, and promptly gene may only have effect in some position in genome, and the appearance of this position dependence effect is not rare.This gene action is called " the strain system effect of gene " in this article again.The p-value distributes relatively by t-value and t-or alternatively distributes by F-value and F-and relatively obtains.It is correct probability that this p-value provides null hypothesis (being that genetically modified effect does not exist) subsequently.

Embodiment 27: the result

1. thousand seed weight

The whole per-cent of comparing generation 3% with the transgenic strain of decrement adjusting construct conversion with control plant increases, and best strain is compared the increase of generation 10% with control plant.

2. harvest index

The whole per-cent of comparing generation 16% with the transgenic strain of decrement adjusting construct conversion with control plant increases, and best strain is compared the increase of generation 61% with control plant.

3. full rate

The whole per-cent of comparing generation 14% with the transgenic strain of decrement adjusting construct conversion with control plant increases, and best strain is compared the increase of generation 41% with control plant.

4. each paniculiform spending is counted (every strain plant flowers number)

The whole per-cent of comparing generation-3% with the transgenic strain of decrement adjusting construct conversion with control plant reduces, and best strain is compared the reduction of generation-11% with control plant.Spending several reductions is important (for example ought be used for the lawn) to grass.

The whole per-cent of comparing generation 6% with the transgenic strain of overexpression construct conversion with control plant increases, and best strain is compared the increase of generation 14% with control plant.

Embodiment 28: the evaluation of the sequence relevant with SEQ ID NO:230 with SEQ ID NO:229

Use (BLAST) (people (1990) J.Mol.Biol.215:403-410 such as Altschul of for example basic local comparison instrument of database search instrument (Basic Local AlignmentTool); With people (1997) Nucleic Acids Res.25:3389-3402 such as Altschul), in the sequence of the Entrez of American National biotechnology information center (NCBI) Nucleotide database maintenance, identify sequence (full-length cDNA, EST or genome sequence) relevant and/or the protein sequence relevant with SEQ ID NO:230 with SEQ IDNO:229.Usually blast program is by comparing nucleic acid or peptide sequence with sequence library, and by calculating the significance,statistical that mates, is used to seek the zone that has local similar between the sequence.To nucleic acid SEQ ID NO:229 encoded polypeptide utilization TBLASTN algorithm, use default setting, open strainer, to ignore the low complex degree sequence.The output form of analyzing is for comparing in twos, and sorts according to probability score (E value), and wherein score value reflects the occurrent probability of specific comparison (the E value is low more, and the significance of hit event is high more).Except the E value, also keep the score to relatively carrying out identity per-cent.Identity per-cent is meant that two compare the number of the identical Nucleotide (or amino acid) on length-specific between nucleic acid (or polypeptide) sequence.

Table H provides the nucleotide sequence of representing with SEQ ID NO:229 relevant nucleotide sequence.

Table H: be used to implement nucleic acid and peptide sequence the inventive method, that relate to nucleic acid sequence SEQ ID NO:229 and SEQID NO:230.

Title	The source is biological	Nucleic acid SEQ ID NO:	Polypeptide SEQ ID NO:	Database login number	State
						HAT4	Arabidopis thaliana		229	230	AT4G16780	Total length
LD	Arabidopis thaliana		231	232	NP 192165.1								Total length
							Rice
	233	234	Os03g47022	Total length
						Rice	235	236	Os03g12860	Total length
HAT22	Rice
			237	238	Os10g01470	Total length

HAT14	Rice		239	240	Os08g36220	Total length
								Rice	241	242	Os09g27450	Total length
HAT3	Rice
			243	244	Os06g04850	Total length
S1HDL2	Rice
			245	246	Os06g04870	Total length
HD-ZIP	Rice							247	248	Os10g41230	Total length
			Arabidopis thaliana
	249			250	NP_174025.3	Total length
		HAT14	Arabidopis thaliana					251	252	NP_974743.1	Total length
HAT4	Arabidopis thaliana				253	254						NP_193411.1	Total length
		HAT1	Arabidopis thaliana				255	256	NP_193476.1	Total length
HAT2	Arabidopis thaliana				257	258					NP_199548.1	Total length
		HB-4	Arabidopis thaliana				259	260	NP_182018.1	Total length
HAT3	Arabidopis thaliana				261	262					NP_191598.1	Total length
			Rice
	263			264	Os04g46350	Total length
			Rice
	265			266	Os10g23090	Total length
			Rice
	267			268	Os10g23090	Total length
			Rice
	269			270	Os03g08960	Total length
			Rice				271	272	Os08g37580	Total length
	Rice			273	274	Os09g29460					Total length
			Rice				275	276	Os10g39720	Total length
	Rice
			277	278	Os05g09630	Total length

Embodiment 29: the comparison of related polypeptide sequence

From the AlignX of Vector NTI (Invitrogen) based on the clustering algorithm (Clustal algorithm) of the progressive comparison of generally using (people (1997) Nucleic Acids Res25:4876-4882 such as Thompson; People such as Chenna (2003) .Nucleic Acids Res 31:3497-3500).Use is in abutting connection with clustering algorithm constructing system tree (seeing Figure 18).The default value of the open point penalty in room is 10, and the default value that point penalty is extended in the room is 0.1, and the weight matrix of selection is Blosum 62 (if comparison polypeptide).The multisequencing comparison result is shown among the Figure 19 that should use.

Embodiment 30: identify the domain protein white matter family that peptide sequence comprised, the integrated resource of structural domain and site (the Integrated Resouce of ProteinFamilies that are used for implementing the inventive method, Domain and Site, InterPro) database is based on the integrated interface of common used tag database of the search of text and sequence.The InterPro database has made up these databases, and described database uses different methods to learn biological information with in various degree relevant fully profiling protein matter to obtain protein tag.The cooperation database comprises SWISS-PROT, PROSITE, TrEMBL, PRINTS, ProDom and Pfam, Smart and TIGRFAMs.Interpro resides at the European bioinformation institute of Britain.

InterPro scanning as the result of the peptide sequence of SEQ ID NO:230 representative in Table I.

Table I: as the InterPro scanning result of the peptide sequence of SEQ ID NO:230 representative

Database	Searching number	The retrieval title
			InterPro	IPR006712	HDZIP
InterPro	IPR001356	The homology frame
			InterPro	IPR003106	HALZ
ProDom	PDA0K5J
		1	Homology frame PPHB4DNA combination
ProDom	PD728753			The DNA combination
		ProDom	PD064887		The DNA combination
The PIR superfamily	PIRPS00001			Asn glycosylation motif
		The PIR superfamily	PS00004		The cAMP phosphorylation site
The PIR superfamily	PS00005			The PKC phosphorylation site
		The PIR superfamily	PS00006		The CK2 phosphorylation site
The PIR superfamily	PS00008			Myristoylation
		The PIR superfamily	PS00009		Amidation
The PIR superfamily	PS00027			The homology frame
		The PIR superfamily	PS00029		Leucine zipper
Pfam	PF04618			HDZIP
		Pfam	PF00046		The homology frame
Pfam	PF02183			HALZ

Embodiment 31: the nucleotide sequence of clone SEQ ID NO:229 representative

Arabidopis thaliana HAT4 encoding gene is by PCR, and (Invitrogen, Paisley UK) increase as template to use Arabidopis thaliana cDNA library.With extracting behind the RNA of seedling reverse transcription, cDNA is cloned into pCMV Sport 6.0.The insertion sequence mean size in storehouse is that 1.6kb and clone's initial number is 1.67 * 10 ⁷Cfu.6 * 10 ¹⁰The original titre of first round amplification back mensuration of cfu/ml is 3.34 * 10 ⁶Cfu/ml.After plasmid extracts, the 200ng template is used for 50 μ lPCR mixtures.

Used primer is:

Forward (SEQ ID NO:283)

ggggacaagtttgtacaaaaaagcaggcttcacaatgatgttcgagaaagacgatctg

Oppositely (SEQ ID NO:284)

ggggaccactttgtacaagaaagctgggtttaggacctaggacgaagagcgt

It comprises the AttB site that is used for the Gateway reorganization.The amplification PCR fragment also uses standard method to give purifying.Implement the first step of Gateway method subsequently, i.e. BP reaction, " entering the clone " that reorganization is named according to Gateway with generation in PCR fragment and the pDONR201 plasmid generation body during this period.Plasmid pDONR201 conduct

The component part of technology is bought from Invitrogen.

Embodiment 32: use the expression vector establishment as SEQ ID NO:229 representative nucleotide sequence

Entering the clone uses with a kind of purpose carrier that is used for the rice conversion in the LR reaction subsequently.This carrier contains as functional element on the T-DNA border: plant selectable marker; The selection markers expression cassette is with intention and cloned the Gateway box of recombinating in the described purpose nucleotide sequence generation LR body that enters in the clone.The rice GOS2 promotor (SEQ IDNO:282) that is used for constitutive expression is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector of generation (Figure 21) is converted among the agrobacterium strains LBA4044 according to method well-known in the art.

Embodiment 33: Plant Transformation

Rice transforms

The Agrobacterium that contains expression vector is used for transforming rice plant.Ripe dry seed shelling with the Japanese Cultivar Nipponbare of rice.By incubation in 70% ethanol one minute, in 2,%Hg,Cl2 30 minutes subsequently, subsequently with sterile distilled water washing 6 times 15 minutes and implement sterilization.The disinfectant seed is containing 2 subsequently, and the substratum of 4-D (callus inducing medium) is gone up and sprouted.Incubation is after 4 weeks in the dark, scultellum deutero-embryogenic callus is downcut and breeds on a kind of substratum.After 2 weeks, callus by breeding or breed uploading with a kind of substratum to be commissioned to train to support in other 2 weeks.The embryogenic callus sheet is uploaded to be commissioned to train at fresh culture and was supported 3, cultivates (to strengthen the cell fission activity) afterwards altogether.

The agrobacterium strains LBA4404 that contains expression vector is used for common cultivation.Agrobacterium is seeded in to contain on the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3.Subsequently bacterium is collected and is resuspended in liquid and cultivate altogether in the substratum to density (OD600) about 1.Suspension is transferred to culture dish subsequently and callus was soaked 15 minutes in this suspension.Callus is organized subsequently and to be cultivated on the substratum altogether and in the dark in 25 ℃ of incubations 3 days blotting and be transferred to solidified on the filter paper.Altogether the callus of cultivating in the dark in 28 ℃ in the presence of selective agent in containing 2,4 weeks of cultivation on the substratum of 4-D.During the section, form mushroom resistant calli island at this moment.To regeneration culture medium and behind incubation under the light, release of embryo generation potentiality and seedling are in 4-5 week growth subsequently in this material transfer.Seedling is downcut from callus and, wherein seedling is transferred to soil from described substratum containing incubation 2-3 week on the substratum of plant hormone.The hardened seedling is cultivated in the greenhouse under high humidity and short day.

A construct produces about 35 independent T0 rice transformant.With former generation transformant be transferred to the greenhouse from incubator for tissue culture.Behind the copy number of quantitative PCR analysis with checking T-DNA inset, the single copy transgenic plant that only keep performance selective agent tolerance are used to gather in the crops the T1 seed.Seed is gathered in the crops after transplanting subsequently the 3-5 month.Present method produces term single gene seat transformant (Aldemita and Hodges1996, Chan etc. 1993, Hiei etc. 1994) to surpass 50% ratio.

Embodiment 34: the phenotype evaluation method

Prepare 34.1 estimate

Produce about 35 T0 rice transformant independently.Transformant was transferred to the greenhouse from incubator for tissue culture and was used for growth and results T1 seed former generation.Stay 8 incidents, the T1 offspring of wherein said incident separates with 3: 1 ratios genetically modified existence/do not exist.For each incident in these incidents, select to contain genetically modified about 10 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 10 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Greenhouse experiment is short day (illumination in 12 hours), 28 ℃ and 22 ℃ and relative humidity 70% in the dark under light.

4 T1 incidents T2 from generation to generation in according to as be used for T1 identical evaluation method from generation to generation and do further assessment, but that each incident adopts is more individual.

The arid screening

In flowerpot soil, cultivate plant under normal operation, up to entering heading stage from the T2 seed.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

The nitrogen use efficiency screening

Under normal condition except that nutritive medium, in flowerpot soil, cultivate rice plant from the T2 seed.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

Using Oxford QP20+ pulse (pulsed) NMR that rice is carried out nondestructive oil measures.Use unhulled whole grain seed.34.2 statistical study: F-check

Use double factor ANOVA (variance analysis) to be used for the overall evaluation of plant phenotype feature as statistical model.All measuring parameters with whole plants of whole incidents of gene transformation of the present invention are implemented F check.Implement the F check to check that gene is for the effect of whole transformation events and the mass action (being called overall gene action again) of checking gene.The threshold value that is used for the significance of true overall gene action is arranged on 5% probability level for F check.Significance F test value indicates gene action, means that the existence of gene not only or position just cause the difference on the phenotype.

34.3 result

Best product tie up on the oil-contg to compare with control plant and show 11% increase, all the oil-contg of strains on average increase by 6% and from the P value of F-check less than 0.0001.

Embodiment 35: corn transforms

The conversion of corn is carried out according to the modification method to (1996.Nature Biotech 14745-50) described methods such as Ishida.Conversion in corn be that genotype relies on and only the special genes type can operate and be used for transforming and regeneration.Inbred lines A188 (University of Minnesota) or be the good source of the donor material that is used to transform with A188 as parent's hybrid, but other genotype also can successfully be used.Mealie from maize plant after pollination about 11 days (DAP) results, this moment, the length of immature embryos was about 1 to 1.2mm.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.The embryo that downcuts is on the callus inducing medium, cultivate on the corn regeneration culture medium subsequently, and wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to the maize rooting substratum and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Embodiment 36: wheat transforms

The conversion of wheat is carried out with the method that (1996) Nature Biotech 14 (6): 745-50 such as Ishida such as Ishida describe.Usually in conversion, use (can obtain) Cultivar Bobwhite from Mexico CIMMYT.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.With the Agrobacterium incubation after, embryo on the callus inducing medium, external cultivation on regeneration culture medium subsequently, wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to root media and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Embodiment 37: soybean transforms

According to Texas A﹠amp; M United States Patent (USP) 5,164, the modification method soybean transformation of method described in 310.Several commercial soybean varieties are feasible for conversion by this method.Cultivar Jack (can be able to obtain from Illinois seed money) is generally used for transforming.Soybean seeds is sterilized so that external sowing.From 7 age in days seedling, downcut hypocotyl, radicle and a slice cotyledon.The cotyledon of further cultivating epicotyl and remainder is to grow the armpit tight knot.These armpit tight knots are downcut and with the agrobacterium tumefaciens incubation that contains expression vector.After cultivating processing altogether, explant is washed and is transferred to the selection substratum.The regenerated seedling is downcut and places on the seedling elongation medium.The seedling that length is no more than 1cm places on the root media until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant tolerance of performance selective agent and that contain single copy T-DNA inset, produce the T1 seed.

Embodiment 38: Semen Brassicae campestris/canola oil dish transforms

Cotyledon petiole of use 5-6 age in days seedling and hypocotyl are as being used for the explant of tissue culture and transforming according to (1998, Plant Cell Rep 17:183-188) such as Babic.Commercial Cultivar Westar (Agriculture Canada) is the standard variety that is used to transform, but also can use other kinds.Canola oil colza is done the surface sterilization so that external sowing.From external seedling, downcut and have the cotyledon petiole explant that adheres to cotyledon, and immerse bacterial suspension with the cut ends of (containing expression vector) Agrobacterium by petiole explant and inoculate.Explant subsequently on the MSBAP-3 substratum that contains 3mg/l BAP, 3% sucrose, 0.7% plant agar (Phytagar) at 23 ℃, illumination in 16 hours was cultivated 2 days down.After cultivating 2 altogether with Agrobacterium, petiole explant is transferred on the MSBAP-3 substratum of 3mg/l BAP, cefotaxime, Pyocianil or the Ticarcillin/Clavulanate Acid (300mg/l) that contain and continues 7, and cultivate containing on the MSBAP-3 substratum of cefotaxime, Pyocianil or Ticarcillin/Clavulanate Acid and selective agent subsequently, regenerate until seedling.When seedling has 5-10mm length, seedling is downcut and is transferred to seedling elongation medium (MSBAP-0.5 that contains 0.5mg/lBAP).The seedling of the about 2cm of length is transferred to the root media (MS0) that is used for root induction.The seedling that to take root migrates in the soil in greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 39: clover transforms

The reproducibility clone of clover uses the method for (McKersie etc., 1999 Plant Physiol 119:839-847) to be transformed.Regeneration of clover and conversion are that genotype is dependent and thereby need aftergrowth.The method that obtains the reproducibility plant has been described.For example, these reproducibility plants any other commercial alfalfa variety that can be selected from Cultivar Rangelander (Agriculture Canada) or describe as Brown DCW and AAtanassov (1985.Plant Cell Tissue Culture 4:111-112).Alternatively, RA3 kind (University of Wisconsin) has been selected for (Walker etc., 1978 Am J Bot 65:654-659) in the tissue culture.Petiole explant and the agrobacterium tumefaciens C58C1 pMP90 (McKersie etc., 1999 Plant Physiol 119:839-847) or the overnight culture of LBA4404 that contain expression vector are cultivated altogether.Explant is containing 288mg/L Pro, 53mg/L Thioproline, 4.35g/L K in the dark ₂SO ₄With cultivated altogether 3 days on the SH inducing culture of 100 μ m Syringylethanones. explant half concentrate in the Murashige-Skoog substratum (Murashige and Skoog, 1962) washing and plating contain not containing Syringylethanone suitable selective agent and suitable microbiotic with the identical SH inducing culture of restraining the Agrobacterium growth on.After several weeks, somatic embryo is transferred to do not contain growth regulator, do not contain microbiotic and the BOi2Y that contains 50g/L sucrose grows in the substratum.Somatic embryo concentrates on the Murashige-Skoog substratum half subsequently to be sprouted.The sprigging that to take root is cultivated to flowerpot and in the greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 40: the evaluation of the sequence relevant with SEQ ID NO:286 with SEQ ID NO:285

Use (BLAST) (people (1990) J.Mol.Biol.215:403-410 such as Altschul of for example basic local comparison instrument of database search instrument (Basic Local AlignmentTool); With people (1997) Nucleic Acids Res.25:3389-3402 such as Altschul), in the sequence of the Entrez of American National biotechnology information center (NCBI) Nucleotide database maintenance, identify the protein sequence that relates to the sequence (full-length cDNA, EST or genome sequence) of SEQ IDNO:285 and/or relate to SEQ ID NO:286.Usually blast program is by comparing nucleic acid or peptide sequence with sequence library, and by calculating the significance,statistical that mates, is used to seek the zone that has local similar between the sequence.To nucleic acid SEQ ID NO:285 encoded polypeptide utilization TBLASTN algorithm, use default setting, open strainer, to ignore the low complex degree sequence.The output form of analyzing is for comparing in twos, and sorts according to probability score (E value), and wherein score value reflects the occurrent probability of specific comparison (the E value is low more, and the significance of hit event is high more).Except the E value, also keep the score to relatively carrying out identity per-cent.Identity per-cent is meant that two compare the number of the identical Nucleotide (or amino acid) on length-specific between nucleic acid (or polypeptide) sequence.In some cases, can adjust the strict degree of default parameter with the change search.

Except the public obtainable nucleotide sequence that can on NCBI, obtain, according to method search proprietary sequence library same as the above-mentioned method.

Nucleic acid and protein sequence that the protein sequence that table J provides the nucleotide sequence represented with SEQ ID NO:285 and SEQ ID NO:286 to represent is relevant are tabulated.

The table J: can be used for the inventive method, with the relevant nucleotide sequence of nucleotide sequence (SEQ ID NO:1), and the derivation corresponding polypeptide.

Title	The source is biological	Nucleic acid SEQ ID NO:	Polypeptide SEQ ID NO:	Database login number	State
						SYB1	Arabidopis thaliana		285	286	NM112438	Total length
Zinc finger protein sample protein	Upland cotton		287	288	AAZ94630								Total length
						The zinc finger protein ZF2 that infers	Zea mays	289	290	Q53AV6	Total length
Zinc finger transcription factor ZFP30	Rice		291	292	Q6Z6E6							Total length
						False albuminoid At5g25490	Arabidopis thaliana		293	294	Q8GWD1		Total length
Zinc finger protein	Rice		295	296	Q9SW92							Total length
						Infer zinc finger transcription factor	Rice		297	298	Q8RYZ5		Total length
GlimmerM protein
	152	Beet	299	300	ABD83289	Total length
The a-protein t2g17975 that infers							Arabidopis thaliana	301	302	Q8S8K1	Total length
	Imagination RanBP2-type zinc finger protein At1g67325	Arabidopis thaliana		303	304	Q8GZ43						Total length
The conjugated protein sample of P53							Rice		305	306	Q7F1K4		Total length

The p53 that infers is conjugated protein	Rice		307	308	Q7XHQ8	Total length
							False albuminoid	Overgrown with weeds blue or green	309	310	CX272690	Total length
False albuminoid	Euphorbia esula L	311	312	DV143669	Total length
						False albuminoid	Upland cotton	313	314	DR459119	Total length
False albuminoid	Grape		315	316	DV221228							Total length
						False albuminoid	Comospore poplar (Populus trichocarpa)	317	318	DT496999	Total length
False albuminoid	Upland cotton		319	320	DT457997							Total length
						False albuminoid	Fireweed		321	322	DW105125		Total length
False albuminoid	Soybean	323	324	BG238374	Total length
						False albuminoid	The comospore poplar	325	226	DT494117	Total length
False albuminoid	Populus certain	327	328	DV465465	Total length
						Zinc finger transcription factor	Rice	329	330	AY219846	Total length
False albuminoid	Switchgrass (Panicum virgatum)	331	332	DN152082	Total length
						Zinc refers to, the RanBP2 type	The puncture vine clover	333	334	Q1RWK5	Total length
Zinc refers to, the RanBP2 type	The puncture vine clover	335	336	Q1S406	Total length
						False albuminoid	Yucca (Yucca filamentosa)	337	338	DT581158	Total length
False albuminoid	Barley	339	340	BQ471337	Total length

Embodiment 41: the comparison of related polypeptide sequence

From the AlignX of Vector NTI (Invitrogen) based on the clustering algorithm (Clustal algorithm) of the progressive comparison of generally using (people (1997) Nucleic Acids Res25:4876-4882 such as Thompson; People such as Chenna (2003) .Nucleic Acids Res 31:3497-3500).Can use in abutting connection with the clustering algorithm constructing system and set.The default value of the open point penalty in room is 10, and the default value that point penalty is extended in the room is 0.1, and the weight matrix of selection is Blosum 62 (if comparison polypeptide).

Can be used for implementing in the polypeptide of the inventive method in evaluation, the multisequencing comparison result that uses related polypeptide to obtain is shown among Figure 23 a that should use.Can be easily to three Zinc finger domains.

Embodiment 42: calculate the overall per-cent identity between the peptide sequence can be used for implementing the inventive method

A method MatGAT in the method that use can obtain in this area (matrix overall comparison instrument (Matrix Global Alignment Tool)) software (BMC Bioinformatics.20034:29.MatGAT:an application that generates similarity/identity matricesusing protein or DNA sequences.Campanella JJ, Bitincka L, Smalley J; Software by Ledion Bitincka trustship), determine to be used to implement overall similarity and identity per-cent between the full-length polypeptide sequence of method of the present invention.MatGAT software need not data are compared in advance, can produce the similarity/identity matrix of DNA or protein sequence.This program is utilized Myers and Miller overall comparison algorithm, and (the open point penalty in room is 12, and to extend point penalty be 2 in the room) carry out a series of comparison in twos, utilize for example Blosum 62 (for polypeptide) calculating similarity and identity, then the result is arranged in distance matrix.Sequence similarity is shown in the diagonal lines Lower Half, and sequence identity is shown in the diagonal lines first half.

More used parameter has:

Matrix: Blosum 62 keeps the score

First room: 12

Extend the room: 2

The overall similarity of peptide sequence length range (the part of polypeptide sequence is foreclosed) and the software analysis of identity the results are shown in table K.The diagonal lines top provides identity per-cent, and the diagonal lines below provides similarity per-cent.

Compare with SEQ ID NO:286, the per-cent identity that can be used for implementing between the peptide sequence of the inventive method can be for being low to moderate 16.9% amino acid identity.

Table K: the overall similarity of peptide sequence length range and the MatGAT result of identity.

Embodiment 43: identify the structural domain be included in the peptide sequence that can be used for implementing the inventive method

(InterPro) database is based on the integrated interface of common used tag database of the search of text and sequence to the integrated resource in protein families, structural domain and site for Integrated Resouce of ProteinFamilies, Domain and Site.The InterPro database has made up these databases, and described database uses different methods to learn biological information with in various degree relevant fully profiling protein matter to obtain protein tag.The cooperation database comprises SWISS-PROT, PROSITE, TrEMBL, PRINTS, ProDom and Pfam, Smart and TIGRFAMs.Interpro resides at the European bioinformation institute of Britain.

The result of the peptide sequence of InterPro scanning as SEQ ID NO:286 representative is in showing L.

Table L: as the InterPro scanning result of the peptide sequence of SEQ ID NO:286 representative

Database	Searching number	The retrieval title
			InterPro	IPR001878	Zinc refers to, the RanBP2-type
PROFILE	PS50199	ZF RANBP22
			PROSITE	PS01358	ZF RANBP21
PFAM	PF00641	zf-RanBP
			SMART	SM00547	ZnF RBZ

Embodiment 44: be used to implement the topology prediction (Subcellular Localization is striden film ...) of the peptide sequence of the inventive method

The Subcellular Localization of TargetP 1.1 prediction eukaryotic proteins.The existence of the prediction that is based on any aminoterminal presequence is distributed in the position: chloroplast transit peptides (cTP), Mitochondrially targeted peptide (mTP) or Secretory Pathway signal peptide (SP).Unactual as keeping the score of final fundamentals of forecasting is probability, and they may not be integrated.Yet having the highest position of keeping the score is most possible according to TargetP, and the relation between keeping the score (reliability category) can be an index of the certainty of prediction.Reliability category (RC) is 1-5, the wherein the strongest prediction of 1 expression.TargetP safeguards on the server of Technical University Of Denmark.

Contain the sequence of aminoterminal presequence for prediction, also can predict the potential cleavage site.

Numerous parameters have been selected, as biology group (non-plant or plant), critical setting (do not have, critical predefine setting or critical user specify settings) with to the calculating of cleavage site prediction (be or deny).

The result who analyzes as the TargetP 1.1 of the peptide sequence of SEQ ID NO:286 representative shows in table M.Select " plant " biological group, undefined threshold value and need the prediction length of transit peptides.Not clearly Subcellular Localization prediction has only the localized weak prediction of plastosome (5 grades of reliable ranks, minimum reliability).Therefore SYB1 protein also can be positioned in the kytoplasm.

Table M: the TargetP 1.1 as the peptide sequence of SEQ ID NO:286 representative analyzes

Length (AA)	164
		Chloroplast transit peptides	0.417
The mitochondrial transport peptide	0.505
		The Secretory Pathway signal peptide	0.014
Other ubcellular targets	0.184
		The position of prediction	Plastosome
Reliability category
		5
The transit peptides length of prediction			12

Numerous other algorithms can be used for carrying out this alanysis, comprising:

Resident ChloroP 1.1 on Technical University Of Denmark's server;

Resident protein Prowler Subcellular Localization predictor is the 1.2nd edition on the server of molecular biosciences institute of Brisbane ,Australia University of Queensland;

Resident PENCE Proteme Analyst PA-GOSUB 2.5 on the server of Canadian Alberta province Edmonton city University of Alberta;

Embodiment 45: the analytical method that relates to the peptide sequence that is used to implement the inventive method

The existence of peptide sequence shown in the SEQ ID NO:286 by Zinc finger domain can with nucleic acid and protein interaction.DNA is well-known in the art in conjunction with test, it comprises as the auxiliary DNA binding site of the PCR-of general reference selects and dna gel displacement combination test, referring to Current Protocols in Molecular Biology, the 1st and 2 volumes, Ausubel etc. (1994), Current Protocols.

Several protein interactions (structure is from the SMART database) shown in protein prediction shown in the SEQ ID NO:286 and the table N.The proteinic functional so usefulness candidate ligand protein test in yeast two-hybrid screening of SYB1.

Table N: the SEQ ID NO:286 of prediction interacts

Describe	Identifier
		The kytoplasm P450 that infers	At2g46960
Unknown	At5g03670
		With kytoplasm b558/566, the weak similarity of subunit B	F18B3.90
Super quick inducibility response protein matter	MQB2.6″
		Glycosylation hydrolase family 9	T21L14.7
Unknown	F26O13.50
		The protein that comprises the brix structural domain	At5g61770
peterpan	At5g61770
		The GTPase of unknown/prediction	Q9SJF1/At1g08410
Nucleotide binding protein matter	Q9SHS8/At2g27200

In addition, produce the seed production of increase as described below according to the SYB1 protein expression of the inventive method.

Embodiment 46: the nucleotide sequence of clone SEQ ID NO:285 representative

Unless stated otherwise, recombinant DNA technology is according to (Sambrook (2001) MolecularCloning:a laboratory manual, third edition Cold Spring Harbor LaboratoryPress, CSH, New York) in or Ausubel etc. (1994), Current Protocols inMolecular Biology, the standard method of describing in Current Protocols the 1st and 2 volumes is carried out.The standard material and the method that are used for plant molecular work are described at the Plant Molecular Biology Labfax (1993) of the R.D.D.Croy that is published by BIOS Scientific PublicationsLtd (UK) and Blackwell Scientific Publications (UK).

(Invitrogen, Paisley UK) pass through pcr amplification Arabidopis thaliana SYB1 gene as template to use Arabidopis thaliana seedling cDNA library.Behind the RNA that reverse transcription is extracted from seedling, cDNA is cloned into pCMV Sport 6.0.The average inset size in library is 1.5kb, and clone's original number is 1.59x10 ⁷The rank of cfu.At 6x10 ¹¹After the amplification first time of cfu/ml, initial titer is defined as 9.6x10 ⁵Cfu/ml.After plasmid extracts, the 200ng template is used for 50 μ l PCR mixtures.Primer prm5539 (the SEQ ID NO:341 that will comprise the AttB site that is used for the Gateway reorganization; 5 '-ggggacaagtttgtacaaaaaagcaggcttaaacaatgagcagacccggagatt-3 ') and prm5540 (SEQ ID NO:342 justice is arranged, and initiator codon is a boldface letter, and the AttB1 site is an italic:; Oppositely, complementation, the AttB2 site is an italic: 5 '-ggggaccactttgtacaagaaagctgggtagacaaggctacttcaaaagca-3 ') be used for pcr amplification.In standard conditions, use Hifi Taq archaeal dna polymerase to carry out PCR.Also use the PCR fragment (comprising the attB site) of standard method amplification and purifying 559bp.Carry out the first step BP reaction of Gateway method then, during this step, PCR fragment and pDONR201 plasmid carry out reorganization in the body, thereby produce, according to the Gateway nomenclature, and " clone crosses the threshold ", p58a.Plasmid pDONR201 conduct

The part of technology is available from Invitrogen.

Embodiment 47: use as SEQ ID NO:285 representative nucleotide sequence construction of expression vector

Entering clone p58a uses with p0640 (a kind of purpose carrier that is used for the rice conversion) in the LR reaction subsequently.This carrier contains as functional element on the T-DNA border: plant selectable marker; The selection markers expression cassette is with intention and cloned the Gateway box of recombinating in the described purpose nucleotide sequence generation LR body that enters in the clone.The rice GOS2 promotor (SEQ ID NO:343) that is used for constitutive expression (pGOS2) is positioned at the upstream of this Gateway box.

After the LR reconstitution steps, the expression vector pGOS2::SYB1 (Figure 24) of generation is converted among the agrobacterium strains LBA4044 according to method well-known in the art.

Embodiment 48: Plant Transformation

Rice transforms

The Agrobacterium that contains expression vector is used for transforming rice plant.Ripe dry seed shelling with the Japanese Cultivar Nipponbare of rice.By incubation in 70% ethanol one minute, in 2,%Hg,Cl2 30 minutes subsequently, subsequently with sterile distilled water washing 6 times 15 minutes and implement sterilization.The disinfectant seed is containing 2 subsequently, and the substratum of 4-D (callus inducing medium) is gone up and sprouted.Incubation is after 4 weeks in the dark, scultellum deutero-embryogenic callus is downcut and breeds on a kind of substratum.After 2 weeks, callus by breeding or breed uploading with a kind of substratum to be commissioned to train to support in other 2 weeks.The embryogenic callus sheet is uploaded to be commissioned to train at fresh culture and was supported 3, cultivates (to strengthen the cell fission activity) afterwards altogether.

The agrobacterium strains LBA4404 that contains expression vector is used for common cultivation.Agrobacterium is seeded in to contain on the suitable antibiotic AB substratum and at 28 ℃ and cultivated 3.Subsequently bacterium is collected and is resuspended in liquid and cultivate altogether in the substratum to density (OD600) about 1.Suspension is transferred to culture dish subsequently and callus was soaked 15 minutes in this suspension.Callus is organized subsequently and to be cultivated on the substratum altogether and in the dark in 25 ℃ of incubations 3 days blotting and be transferred to solidified on the filter paper.Altogether the callus of cultivating in the dark in 28 ℃ in the presence of selective agent in containing 2,4 weeks of cultivation on the substratum of 4-D.During the section, form mushroom resistant calli island at this moment.To regeneration culture medium and behind incubation under the light, release of embryo generation potentiality and seedling are in 4-5 week growth subsequently in this material transfer.Seedling is downcut from callus and, wherein seedling is transferred to soil from described substratum containing incubation 2-3 week on the substratum of plant hormone.The hardened seedling is cultivated in the greenhouse under high humidity and short day.

A construct produces about 35 independent T0 rice transformant.With former generation transformant be transferred to the greenhouse from incubator for tissue culture.Behind the copy number of quantitative PCR analysis with checking T-DNA inset, the single copy transgenic plant that only keep performance selective agent tolerance are used to gather in the crops the T1 seed.Seed is gathered in the crops after transplanting subsequently the 3-5 month.Present method produces term single gene seat transformant (Aldemita and Hodges1996, Chan etc. 1993, Hiei etc. 1994) to surpass 50% ratio.

Corn transforms

The conversion of corn is carried out according to the modification method to (1996.Nature Biotech 14745-50) described methods such as Ishida.Conversion in corn be that genotype relies on and only the special genes type can operate and be used for transforming and regeneration.Inbred lines A188 (University of Minnesota) or be the good source of the donor material that is used to transform with A188 as parent's hybrid, but other genotype also can successfully be used.Mealie from maize plant after pollination about 11 days (DAP) results, this moment, the length of immature embryos was about 1 to 1.2mm.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.The embryo that downcuts is on the callus inducing medium, cultivate on the corn regeneration culture medium subsequently, and wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to the maize rooting substratum and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Wheat transforms

The conversion of wheat is carried out with the method that (1996) Nature Biotech 14 (6): 745-50 such as Ishida such as Ishida describe.Usually in conversion, use (can obtain) Cultivar Bobwhite from Mexico CIMMYT.Immature embryos is cultivated altogether with the agrobacterium tumefaciens that contains expression vector and transgenic plant take place to recover by organ.With the Agrobacterium incubation after, embryo on the callus inducing medium, external cultivation on regeneration culture medium subsequently, wherein said regeneration culture medium contains selective agent (for example imidazolone, but can use the multiple choices mark).Culture plate is cultivated 2-3 week under illumination at 25 ℃, or grows until seedling.Green seedling is transferred to root media and cultivates 2-3 week at 25 ℃ from each embryo, until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant of performance selective agent T-DNA inset tolerance and that contain single copy, produce the T1 seed.

Soybean transforms

According to Texas A﹠amp; M United States Patent (USP) 5,164, the modification method soybean transformation of method described in 310.Several commercial soybean varieties are feasible for conversion by this method.Cultivar Jack (can be able to obtain from Illinois seed money) is generally used for transforming.Soybean seeds is sterilized so that external sowing.From 7 age in days seedling, downcut hypocotyl, radicle and a slice cotyledon.The cotyledon of further cultivating epicotyl and remainder is to grow the armpit tight knot.These armpit tight knots are downcut and with the agrobacterium tumefaciens incubation that contains expression vector.After cultivating processing altogether, explant is washed and is transferred to the selection substratum.The regenerated seedling is downcut and places on the seedling elongation medium.The seedling that length is no more than 1cm places on the root media until root development.The seedling that to take root migrates in the soil in greenhouse.From the plant tolerance of performance selective agent and that contain single copy T-DNA inset, produce the T1 seed.

Semen Brassicae campestris/canola oil dish transforms

Cotyledon petiole of use 5-6 age in days seedling and hypocotyl are as being used for the explant of tissue culture and transforming according to (1998, Plant Cell Rep 17:183-188) such as Babic.Commercial Cultivar Westar (Agriculture Canada) is the standard variety that is used to transform, but also can use other kinds.Canola oil colza is done the surface sterilization so that external sowing.From external seedling, downcut and have the cotyledon petiole explant that adheres to cotyledon, and immerse bacterial suspension with the cut ends of (containing expression vector) Agrobacterium by petiole explant and inoculate.Explant subsequently on the MSBAP-3 substratum that contains 3mg/l BAP, 3% sucrose, 0.7% plant agar (Phytagar) at 23 ℃, illumination in 16 hours was cultivated 2 days down.After cultivating 2 altogether with Agrobacterium, petiole explant is transferred on the MSBAP-3 substratum of 3mg/l BAP, cefotaxime, Pyocianil or the Ticarcillin/Clavulanate Acid (300mg/l) that contain and continues 7, and cultivate containing on the MSBAP-3 substratum of cefotaxime, Pyocianil or Ticarcillin/Clavulanate Acid and selective agent subsequently, regenerate until seedling.When seedling has 5-10mm length, seedling is downcut and is transferred to seedling elongation medium (MSBAP-0.5 that contains 0.5mg/l BAP).The seedling of the about 2cm of length is transferred to the root media (MS0) that is used for root induction.The seedling that to take root migrates in the soil in greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Clover transforms

The reproducibility clone of clover uses the method for (McKersie etc., 1999 Plant Physiol 119:839-847) to be transformed.Regeneration of clover and conversion are that genotype is dependent and thereby need aftergrowth.The method that obtains the reproducibility plant has been described.For example, these reproducibility plants any other commercial alfalfa variety that can be selected from Cultivar Rangelander (Agriculture Canada) or describe as Brown DCW and AAtanassov (1985.Plant Cell Tissue Culture 4:111-112).Alternatively, RA3 kind (University of Wisconsin) has been selected for (Walker etc., 1978 Am J Bot 65:654-659) in the tissue culture.Petiole explant and the agrobacterium tumefaciens C58C1 pMP90 (McKersie etc., 1999 Plant Physiol 119:839-847) or the overnight culture of LBA4404 that contain expression vector are cultivated altogether.Explant is containing 288mg/L Pro, 53mg/L Thioproline, 4.35g/L K in the dark ₂SO ₄With cultivated altogether 3 days on the SH inducing culture of 100 μ m Syringylethanones. explant half concentrate in the Murashige-Skoog substratum (Murashige and Skoog, 1962) washing and plating contain not containing Syringylethanone suitable selective agent and suitable microbiotic with the identical SH inducing culture of restraining the Agrobacterium growth on.After several weeks, somatic embryo is transferred to do not contain growth regulator, do not contain microbiotic and the BOi2Y that contains 50g/L sucrose grows in the substratum.Somatic embryo concentrates on the Murashige-Skoog substratum half subsequently to be sprouted.The sprigging that to take root is cultivated to flowerpot and in the greenhouse.Produce the T1 seed the plant that singly copies the T-DNA inset from showing the selective agent tolerance and containing.

Embodiment 49: the phenotype evaluation method

Prepare 49.1 estimate

Produce about 35 T0 rice transformant independently.Transformant was transferred to the greenhouse from incubator for tissue culture and was used for growth and results T1 seed former generation.Stay 6 incidents, the T1 offspring of wherein said incident separates with 3: 1 ratios genetically modified existence/do not exist.For each incident in these incidents, select to contain genetically modified about 10 strain T1 seedling (heterozygote and homozygote) and lack genetically modified about 10 strain T1 seedling (inefficacy zygote) by monitoring visual marker expression.Transgenic plant and corresponding inefficacy zygote are cultivated on random site side by side.Greenhouse experiment is short day (illumination in 12 hours), 28 ℃ and 22 ℃ and relative humidity 70% in the dark under light.

The arid screening

In flowerpot soil, cultivate plant under normal operation, up to entering heading stage from the T2 seed.Then it is transferred to " drying " zone, stop to irrigate.In the flowerpot of selecting at random, insert the humidity detection instrument, with monitoring soil water content (SWC).When SWC is lower than certain threshold value, continue moisturizing from the trend plant, up to reaching normal level once more.Then plant is transferred under the normal condition once more again.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

The nitrogen use efficiency screening

Under normal condition except that nutritive medium, in flowerpot soil, cultivate rice plant from the T2 seed.With specific nutritive medium flowerpot is watered from the plant transplanting to the maturation, the nitrogen of described nutritive medium (N) content reduces always, low 7 to 8 times usually.Remaining cultivation (plant maturation, seed results) is identical with the plant of not cultivating under the abiotic stress condition.Record growth and output are set forth parameter, as growth details under normal operation.

4 T1 incidents T2 from generation to generation in according to as be used for T1 identical evaluation method from generation to generation and do further assessment, but that each incident adopts is more individual.Make plant pass through the digital imagery case for several times until the ripening stage from sowing time.On each time point, to every strain plant from least 6 different angles shooting digital pictures (2048x1536 pixel, 1,600 ten thousand colors).

49.2 statistical study: F-check

Use double factor ANOVA (variance analysis) to be used for the overall evaluation of plant phenotype feature as statistical model.All measuring parameters with whole plants of whole incidents of gene transformation of the present invention are implemented F check.Implement the F check to check that gene is for the effect of whole transformation events and the mass action (being called overall gene action again) of checking gene.The threshold value that is used for the significance of true overall gene action is arranged on 5% probability level for F check.Significance F test value indicates gene action, means that the existence of gene not only or position just cause the difference on the phenotype.

49.3 parameter measurement

The measurement of biomass correlation parameter

From sowing time to the ripening stage, plant is repeatedly by the digital image-forming case.On each time point, every strain plant is obtained digital image (2048 * 1536 pixels, 1,000 6 hundred ten thousand looks) from least 6 different angles.

Plant area (leaf biomass in other words) is on the ground determined by the sum of all pixels that counting is different from the ground plant part digital image of background.This value is got the mean value of same time point from the photo of different angle shots, and is converted to the physical surface value of representing with square millimeter (physicalsurface value) by calibration.Experiment shows that the ground plant area of measuring by this method is relevant with the biomass that plant shoot divides.Area reaches the time point value of its maximum leaf biomass plant on the ground.Early stage vigor shows as plant (seedling) the ground area of sprouting three weeks of back.The increase of root biomass shows as total root biomass increases (be measured as plant life during observed maximum biomass); Perhaps show as root/branch exponential and increase (being measured as interim biomass of active growth of root and branch and the ratio between the branch biomass).

The measurement of seed correlation parameter

Gather in the crops sophisticated main panicle, counting, packed, use bar code label, under 37 ℃ in baking oven dry 3 days then.To the panicle threshing, collect all seeds and count then.Use gas blower that full husk is separated with empty husk.Discard the sky husk, then the residue fraction is counted once more.On analytical balance, take by weighing the weight of full husk.Determine the number of full seed by the number of counting the full husk that behind separating step, stays.Measure total seed production by taking by weighing from the weight of all full husks of plant results.Measure total number seeds of every strain plant by counting from the number of the husk of plant results.Number and its gross weight according to the full seed of counting are calculated thousand seed weight (TKW).Harvest index (HI) is defined as seed ultimate production and ground area (mm in the present invention ²) between ratio multiply by the factor 10 ⁶The sum of every paniculiform flower is defined as ratio between seed sum and the paniculiform number of sophisticated master in the present invention.The full rate of seed accounts for the total per-cent of seed (or Xiao Hua) at the number that the present invention is defined as full seed.

Embodiment 50: the phenotype evaluation result of transgenic plant

Provided the evaluation result of expressing the transgenosis rice plant of the nucleotide sequence that is used for the inventive method among the table O.Also show the difference percentage ratio between transgenic plant and the corresponding inefficacy zygote, be lower than 0.05 from the P value of F check.

Compare with control plant (in this case, the inefficacy zygote) under abiotic stress, seed ultimate production, full seed number, the full rate of seed, coefficient of harvest and thousand seed weight are used for the nucleotide sequence of the inventive method in expression transgenic plant significantly increase.

Table O: the evaluation result of expressing the transgenosis rice plant of the nucleotide sequence be used to implement the inventive method.

Proterties	T1 increases % from generation to generation	T2 increases % from generation to generation
			The seed ultimate production	20	23
The full seed number	17	19
			Full rate	12	20
Coefficient of harvest	15	20
			Thousand seed weight	2	4

Sequence table

＜110〉Cropdesign NV

＜120〉has the plant and preparation method thereof of improved characteristics

<130>PF58401-prio

<160>347

＜170〉PatentIn version 3 .3

<210>1

<211>2151

<212>DNA

＜213〉Arabidopis thaliana (Arabidopsis thaliana)

<400>1

atgtgctgtg gatcagaccg attaaaccag atcgtgtcat caagatcttc gttgccaatt 60

tctttcgagg aagataacaa tcttgttacc aacacagaca tgaatcactt aacagtcgaa 120

acagaggata cgtttgcgag cttgcttgag cttgcagcta acaacgatgt tgaaggtgta 180

aggctatcta tcgagagaga cccttcttgt gtagacgaag ctggtctctg gtacggtcgt 240

caaaaaggtt ctaaagctat ggtcaacgat tacaggactc cgttgatggt tgctgctact 300

tacggaagca ttgatgtgat caagcttatt gtttctttga ctgatgctga cgtgaaccgt 360

gcttgcggga atgatcagac cactgcgtta cactgcgctg cttctggagg agctgtgaat 420

gctatccaag ttgttaagct gcttcttgca gctggagctg atttgaatct gttggatgct 480

gaaggtcaac gagctggtga tgttattgtt gttcctccta agcttgaagg cgtgaagctg 540

atgcttcagg agcttctttc tgctgatgga tcatctactg cggagcggaa tctacgggtt 600

gtgacaaatg ttccgaatag aagctcatct ccgtgtcatt ctcctactgg agagaatggt 660

ggatcagggt ctggttcacc gctcggctct ccttttaagc tgaaatctac tgaattcaag 720

aaagagtatc cggttgatcc gtctttgcca gatatcaaga acagtatcta cgcgactgat 780

gagtttagaa tgtattcctt caaggtccgg ccttgctctc gtgcttattc acatgattgg 840

actgagtgtc cttttgttca cccgggtgaa aacgcgagga ggagagaccc gaggaagttc 900

cattacagct gcgttccttg cccggatttt aggaaaggag cttgtaggag aggagatatg 960

tgtgagtatg cgcacggtgt gtttgaatgc tggcttcatc cggctcagta caggacccgt 1020

ctttgcaaag atggaacagg ctgtgctcgg cgggtttgtt tctttgcgca tacacccgag 1080

gagcttcgac ctttgtacgc atcaactggt tcagcggttc cttcgcctag atcgaatgct 1140

gattatgcag ctgctttgag tctccttcct ggttctccat caggagtctc tgtcatgtcc 1200

ccgctttccc catcagcagc ggggaacgga atgtctcatt cgaatatggc ttggccacaa 1260

ccaaatgtcc ctgcgttgca cttaccagga agcaatctac agtcaagcag gctaaggtct 1320

tctctcaatg caagggatat cccgacggat gagttcaata tgttagcgga ttacgagcag 1380

cagcaactcc tcaacgagta ttccaatgct ctgagccgtt ctggtcggat gaaatcaatg 1440

cctccttcga atcttgaaga tcttttctca gcagaaggct cttcatctcc ccggttcact 1500

gattccgctt tagcttccgc ggtgttctcg cctacacaca agtcagctgt cttcaaccag 1560

ttccaacaac agcaacagca gcagcagagc atgttgtctc caatcaacac aagcttttct 1620

tcaccaaaga gcgttgacca ctcattgttt tcaggtggag gaagaatgtc tcctcggaat 1680

gttgttgaac caatatcacc catgagtgct cgggtttcca tgttggctca gtgcgtgaag 1740

caacaacaac agcaacagca gcagcagcag cagcaacatc agttccgtag ccttagctcc 1800

agagagctca gaacaaactc tagcccaatc gttggttcac cggtaaacaa caacacatgg 1860

tcatcaaaat ggggatcttc aaatggtcaa ccggattggg gaatgagctc agaagcactt 1920

ggtaagttga gatcttcgtc atcgtttgat ggtgatgagc ctgatgtgtc atgggtccag 1980

tcactggtga aggagactcc agcagaagcc aaagagaaag cagcaacatc ttcctcaggg 2040

gaacacgtga tgaagcagcc aaatccggtt gaaccggtaa tggatcatgc tgggctagaa 2100

gcttggattg agcaaatgca gctcgatcag cttgtggctc agcagaattg a 2151

<210>2

<211>716

<212>PRT

＜213〉Arabidopis thaliana

<400>2

Met Cys Cys Gly Ser Asp Arg Leu Asn Gln Ile Val Ser Ser Arg Ser

1 5 10 15

Ser Leu Pro Ile Ser Phe Glu Glu Asp Asn Asn Leu Val Thr Asn Thr

20 25 30

Asp Met Asn His Leu Thr Val Glu Thr Glu Asp Thr Phe Ala Ser Leu

35 40 45

Leu Glu Leu Ala Ala Asn Asn Asp Val Glu Gly Val Arg Leu Ser Ile

50 55 60

Glu Arg Asp Pro Ser Cys Val Asp Glu Ala Gly Leu Trp Tyr Gly Arg

65 70 75 80

Gln Lys Gly Ser Lys Ala Met Val Asn Asp Tyr Arg Thr Pro Leu Met

85 90 95

Val Ala Ala Thr Tyr Gly Ser Ile Asp Val Ile Lys Leu Ile Val Ser

100 105 110

Leu Thr Asp Ala Asp Val Asn Arg Ala Cys Gly Asn Asp Gln Thr Thr

115 120 125

Ala Leu His Cys Ala Ala Ser Gly Gly Ala Val Asn Ala Ile Gln Val

130 135 140

Val Lys Leu Leu Leu Ala Ala Gly Ala Asp Leu Asn Leu Leu Asp Ala

145 150 155 160

Glu Gly Gln Arg Ala Gly Asp Val Ile Val Val Pro Pro Lys Leu Glu

165 170 175

Gly Val Lys Leu Met Leu Gln Glu Leu Leu Ser Ala Asp Gly Ser Ser

180 185 190

Thr Ala Glu Arg Asn Leu Arg Val Val Thr Asn Val Pro Asn Arg Ser

195 200 205

Ser Ser Pro Cys His Ser Pro Thr Gly Glu Asn Gly Gly Ser Gly Ser

210 215 220

Gly Ser Pro Leu Gly Ser Pro Phe Lys Leu Lys Ser Thr Glu Phe Lys

225 230 235 240

Lys Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp Ile Lys Asn Ser Ile

245 250 255

Tyr Ala Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Arg Pro Cys

260 265 270

Ser Arg Ala Tyr Ser Hi s Asp Trp Thr Glu Cys Pro Phe Val His Pro

275 280 285

Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Ser Cys

290 295 300

Val Pro Cys Pro Asp Phe Arg Lys Gly Ala Cys Arg Arg Gly Asp Met

305 310 315 320

Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln

325 330 335

Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr Gly Cys Ala Arg Arg Val

340 345 350

Cys Phe Phe Ala His Thr Pro Glu Glu Leu Arg Pro Leu Tyr Ala Ser

355 360 365

Thr Gly Ser Ala Val Pro Ser Pro Arg Ser Asn Ala Asp Tyr Ala Ala

370 375 380

Ala Leu Ser Leu Leu Pro Gly Ser Pro Ser Gly Val Ser Val Met Ser

385 390 395 400

Pro Leu Ser Pro Ser Ala Ala Gly Asn Gly Met Ser His Ser Asn Met

405 410 415

Ala Trp Pro Gln Pro Asn Val Pro Ala Leu Hi s Leu Pro Gly Ser Asn

420 425 430

Leu Gln Ser Ser Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp Ile Pro

435 440 445

Thr Asp Glu Phe Asn Met Leu Ala Asp Tyr Glu Gln Gln Gln Leu Leu

450 455 460

Asn Glu Tyr Ser Asn Ala Leu Ser Arg Ser Gly Arg Met Lys Ser Met

465 470 475 480

Pro Pro Ser Asn Leu Glu Asp Leu Phe Ser Ala Glu Gly Ser Ser Ser

485 490 495

Pro Arg Phe Thr Asp Ser Ala Leu Ala Ser Ala Val Phe Ser Pro Thr

500 505 510

His Lys Ser Ala Val Phe Asn Gln Phe Gln Gln Gln Gln Gln Gln Gln

515 520 525

Gln Ser Met Leu Ser Pro Ile Asn Thr Ser Phe Ser Ser Pro Lys Ser

530 535 540

Val Asp His Ser Leu Phe Ser Gly Gly Gly Arg Met Ser Pro Arg Asn

545 550 555 560

Val Val Glu Pro Ile Ser Pro Met Ser Ala Arg Val Ser Met Leu Ala

565 570 575

Gln Cys Val Lys Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln

580 585 590

His Gln Phe Arg Ser Leu Ser Ser Arg Glu Leu Arg Thr Asn Ser Ser

595 600 605

Pro Ile Val Gly Ser Pro Val Asn Asn Asn Thr Trp Ser Ser Lys Trp

610 615 620

Gly Ser Ser Asn Gly Gln Pro Asp Trp Gly Met Ser Ser Glu Ala Leu

625 630 635 640

Gly Lys Leu Arg Ser Ser Ser Ser Phe Asp Gly Asp Glu Pro Asp Val

645 650 655

Ser Trp Val Gln Ser Leu Val Lys Glu Thr Pro Ala Glu Ala Lys Glu

660 665 670

Lys Ala Ala Thr Ser Ser Ser Gly Glu Hi s Val Met Lys Gln Pro Asn

675 680 685

Pro Val Glu Pro Val Met Asp Hi s Ala Gly Leu Glu Ala Trp Ile Glu

690 695 700

Gln Met Gln Leu Asp Gln Leu Val Ala Gln Gln Asn

705 710 715

<210>3

<211>13

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif 1

<220>

＜221〉variant

<222>(1)..(1)

＜223〉replace=" Ala "

<220>

＜221〉variant

<222>(7)..(7)

＜223〉replace=" Thr "

<400>3

Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys

1 5 10

<210>4

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif 2

<400>4

His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg

1 5 10

<210>5

<211>19

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif 3

<220>

＜221〉variant

<222>(3)..(3)

＜223〉replace=" Ile "

<220>

＜221〉variant

<222>(6)..(6)

＜223〉replace=" Ser "

<220>

＜221〉variant

<222>(11)..(11)

＜223〉replace=" Ser "

<220>

＜221〉variant

<222>(14)..(14)

＜223〉replace=" Lys "

<400>5

His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg

1 5 10 15

Leu Cys Lys

<210>6

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif 4

<400>6

Cys Phe Phe Ala His

1 5

<210>7

<211>54

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm06717

<400>7

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgtg ctgtggatca gaec 54

<210>8

<211>50

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm06718

<400>8

ggggaccact ttgtacaaga aagctgggtg gttaggtctc tcaattctgc 50

<210>9

<211>981

<212>DNA

＜213〉rice (Oryza sativa)

<400>9

aagaggcaag agcatccgta ttaaccagcc ttttgagact tgagagtgtg tgtgactcga 60

tccagcgtag tttcagttcg tgtgttggtg agtgattcca gccaagtttg cgatggcttc 120

tcagcaggaa cgggctagct accacgccgg cgagaccaag gcccgcgccg aggagaagac 180

ggggcgcatg atgggcacgg cgcaggagaa ggcgcgggag gccaaggaca cggcgtccga 240

cgccgcgggg cgcgcgatgg gcaggggaca cggcgccaag gaggcgacca aggagaaggc 300

gtacgagacc aaggacgcga ccaaggagaa ggcgtacgag gcaaaggacg cggcctccga 360

cgccaccggc cgcgccatgg acaagggccg cggcgccgcg ggcgccacga gggacaaggc 420

gtacgatgcc aaggacaggg cggctgacac ggcgcagtcc gccgccgacc gcgcccgcga 480

cggcgccggg cagaccggga gctacattgg acagaccgcc gaggccgcca agcagaaagc 540

ggccggcgcc gcgcagtacg ccaaggagac cgcgatcgcc ggcaaggaca agaccggcgc 600

cgtgctccag caggcagggg agcaggtgaa gagcgtggcg gtgggggcga aggacgcggt 660

gatgtacacg ctcgggatgt caggcgataa caagaacaac gccgctgccg gcaaggacac 720

cagcacctac aagcctggaa ctgggagtga ctaccagtaa tacggtagaa gaagcatgtg 780

tcgtctttgg cactgatgcc aaagtgtacg tgttgtatcc tcttttttaa gtttcagctc 840

gacttcgacg tgttcggtgt cacactttgg tttttcagtt gtgctcaact gttcatgttt 900

ctggttccat ggagggccag tgtggaggtc aatgtttaag ctttcgtttt aaaatctgat 960

aataaagttg gttaagacct g 981

<210>10

<211>3372

<212>DNA

＜213〉big eucalyptus (Eucalyptus grandis)

<400>10

ggaagacgaa gagcaacaaa ataggtctca ctccctcctc tctcctctct cctctctctt 60

ctttctctct cttctgcttc taacaaagtc tcttccttga gagacagggc tgcgtcgtcg 120

tctttctctc tcctcgctgc gagcttctga gaaagttcaa ttctttttct cttgttctct 180

ctctctaccc ttctgggtac cactgtgaag ctccggtctt ttctattttt tttttttttg 240

ggctatctgg gtctgggcaa atccatcgcg cgctctgctc tggactgaga ggccgtcagt 300

ggctttagat ctgcgacgcc tcttgcttgc tcagtgagct gggctagttc aaatcgacga 360

agaaagcatg cgctagtgat tggtgtgggt aatacactgc attcgatctc tactaagtat 420

ccccaagtat actaagatcc cgttctcagc catgagtcaa ctgaccattc agactgagga 480

cacttttgcc agcttgcttg agcttgctgc taacaacgac acagaatctt tcggacggtg 540

tgtggaacgt gatccttcga gcatagatga aattggatat tggtatggtc gccaaaaggg 600

ttcgaagcag gtggtcaata tgcaaagaac tcctcttatg gtggctgcta catatggtag 660

tgttgatgta atgagactca ttctttgcct atctgatgct gatgtgaatc gaacctgcag 720

cacagacaag agcacagccc ttcactgtgc tgcctctggt ggtgctgtga atgctgtaga 780

tgctgtgagg ctactcctgt cagctggtgc tgacccaagt ttagcagatg ctaacggtca 840

gcggcctgtg gatgttattg ttgttcctcc aaagctcctt tcaataaagt ttgctcttga 900

agagctcttg tcgaccgaag gatctgtaaa tgaacacaat ctgagagtgt ccgtagccac 960

ttccaattca acctctcccc cactttcatc ttccccggat aatggttccc cagcatctgc 1020

taattgttct tcccccaaga actcaaagtt aagtgatgcc cctgttcttt atgcatcaga 1080

aaagaaggaa tacccggtgg atccatctct tccagatatc aagaatagca tttactcaac 1140

agatgaattc cgaatgtatt cttttaaagt gcggccttgt tcacgagcgt actcgcatga 1200

ttggacggag tgcccttttg ttcatccagg ggagaatgcc cgtagaaggg atccaaggaa 1260

gttccactac agctgtgtcc cttgccctga tttccggaag ggtgcttgta gacgtggaga 1320

tatgtgtgaa tatgctcatg gtgtttttga gtgctggctc catcctgctc agtatcggac 1380

tcgattatgc aaggatggta caagttgtgc tcggagagtg tgcttctttg cccacacgga 1440

gcaagagctg cgtccattgt acgtctccac tggttctgct gttccgtctc ctcgctcgag 1500

tacctctgga gctgctgcca tggattttgc tgcagccatg agcctcttac ctggttcccc 1560

atcatcagta tccatcatgt ccccttcacc cttcactcct cccatgtctc catctgctaa 1620

tggtatttct cacccatctg ttgcctggcc ccagcaaaat gtaccaactt tgcatcttcc 1680

cggaagcaat cttcagtcca gccgcttgag atcttctctt aatgcaagag atattcctca 1740

ggaggatttt gacttgctgt cagattatga tgtgcaacag cagcagctcc taaatgagtt 1800

ttccatcctt tcacaacaat cgatgggtgc taattccttg aaccgttctg gtcggctgaa 1860

aactttgacc ccctcaaacc ttgatgatct cttctctgct gagagctcat cccctcgcta 1920

cgctgatcaa gccctggctt ctgctgtttt ttcaccaacg cacaaatctg cagtaatcaa 1980

tcaatttcag cagcagcagc agagcatgtt atcacccatc aacacaacct tctctcctaa 2040

gagtgtcgac caccctttgt tgcaagcgtc tttcggtgtt caatctgggc gaatgtcccc 2100

tcgtaacatg gatcccatct ctcctataag ttctcgtgtg tcgatgttgg cccaacgaga 2160

gaaacagcaa cagcaattac gcagcctaag ctctcgtgaa ctcggttcca attcagccgc 2220

cattgtgggt tcccccgtgg gttcttggtc gaaatgggga gctacaaatg ggaaaccaga 2280

ctgggctgtt agtgcagatg aactaggtaa gcttcgcagg tctaattcat ttgagcttgg 2340

gaacaatggt gaggagccag atctttcatg ggttcaatcc ctcgttaaag aatctcctac 2400

cgagatgaaa gaaaagcttt cgtcaactct ctctggtgtt ccagcccccg ctacatccag 2460

tgaggttccg agtatcagct cgcagatgga atcggttgat cacgaagtgc taggagcatg 2520

gctccagcag atgcagctcg atccgctcgt ggctcagcaa aactaggttg ttttttttcc 2580

tacatggcct tgaggaagta gacagcggaa agtttttttt ggtaaatact atgttttttc 2640

tggaaatttt tgatgctggg ggtggggtct ggaagaagat aacaaggcag gaaaggggtc 2700

agtgaagtca ctggagaaaa ggaattcatt tttaaccatt ttatcattct attacaacag 2760

aaagtaggga aaaaaaagga agaccctctg ggttatgaag agaaattaaa cccaggctag 2820

gcgttctcct ttctaatatt tccaatttta ggtccatatt actgtcattt cctttttgcc 2880

gtcttatcat atttcatcaa aatggaactg gggactaatg tttgttccat tctttcgctc 2940

ttctgattta tttgcaccct tggggtaaga tcaaaagaga aattatgatc attttctttt 3000

gaggatattt ttttttccca atatttgtga gaatgaaagt taagagggga tatgatgtgt 3060

ctggtgttgt agtatgaaaa accaataacc gagttcacct gttgctgctg gtggtagaag 3120

aagtggagaa gaagctatga tcctttgatg taacagtcaa tcaaacattt taataccttt 3180

attttttgtt tcctcatgta atccatcctt tgtgattgtc ctctctctct ctctctctct 3240

ctctctctcc ctccccgtgt tctttcttca taagcgtctt gcttgtcgat ctgtaaatta 3300

ttgaaaaggg tcatggaaag ccgtgccggt gtggattctc atttttgcaa aaaaaaaaaa 3360

aaaaaaaaaa aa 3372

<210>11

<211>704

<212>PRT

＜213〉big eucalyptus

<400>11

Met Ser Gln Leu Thr Ile Gln Thr Glu Asp Thr Phe Ala Ser Leu Leu

1 5 10 15

Glu Leu Ala Ala Asn Asn Asp Thr Glu Ser Phe Gly Arg Cys Val Glu

20 25 30

Arg Asp Pro Ser Ser Ile Asp Glu Ile Gly Tyr Trp Tyr Gly Arg Gln

35 40 45

Lys Gly Ser Lys Gln Val Val Asn Met Gln Arg Thr Pro Leu Met Val

50 55 60

Ala Ala Thr Tyr Gly Ser Val Asp Val Met Arg Leu Ile Leu Cys Leu

65 70 75 80

Ser Asp Ala Asp Val Asn Arg Thr Cys Ser Thr Asp Lys Ser Thr Ala

85 90 95

Leu His Cys Ala Ala Ser Gly Gly Ala Val Asn Ala Val Asp Ala Val

100 105 110

Arg Leu Leu Leu Ser Ala Gly Ala Asp Pro Ser Leu Ala Asp Ala Asn

115 120 125

Gly Gln Arg Pro Val Asp Val Ile Val Val Pro Pro Lys Leu Leu Ser

130 135 140

Ile Lys Phe Ala Leu Glu Glu Leu Leu Ser Thr Glu Gly Ser Val Asn

145 150 155 160

Glu His Asn Leu Arg Val Ser Val Ala Thr Ser Asn Ser Thr Ser Pro

165 170 175

Pro Leu Ser Ser Ser Pro Asp Asn Gly Ser Pro Ala Ser Ala Asn Cys

180 185 190

Ser Ser Pro Lys Asn Ser Lys Leu Ser Asp Ala Pro Val Leu Tyr Ala

195 200 205

Ser Glu Lys Lys Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp Ile Lys

210 215 220

Asn Ser Ile Tyr Ser Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val

225 230 235 240

Arg Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe

245 250 255

Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His

260 265 270

Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg Lys Gly Ala Cys Arg Arg

275 280 285

Gly Asp Met Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His

290 295 300

Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr Ser Cys Ala

305 310 315 320

Arg Arg Val Cys Phe Phe Ala His Thr Glu Gln Glu Leu Arg Pro Leu

325 330 335

Tyr Val Ser Thr Gly Ser Ala Val Pro Ser Pro Arg Ser Ser Thr Ser

340 345 350

Gly Ala Ala Ala Met Asp Phe Ala Ala Ala Met Ser Leu Leu Pro Gly

355 360 365

Ser Pro Ser Ser Val Ser Ile Met Ser Pro Ser Pro Phe Thr Pro Pro

370 375 380

Met Ser Pro Ser Ala Asn Gly Ile Ser His Pro Ser Val Ala Trp Pro

385 390 395 400

Gln Gln Asn Val Pro Thr Leu His Leu Pro Gly Ser Asn Leu Gln Ser

405 410 415

Ser Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp Ile Pro Gln Glu Asp

420 425 430

Phe Asp Leu Leu Ser Asp Tyr Asp Val Gln Gln Gln Gln Leu Leu Asn

435 440 445

Glu Phe Ser Ile Leu Ser Gln Gln Ser Met Gly Ala Asn Ser Leu Asn

450 455 460

Arg Ser Gly Arg Leu Lys Thr Leu Thr Pro Ser Asn Leu Asp Asp Leu

465 470 475 480

Phe Ser Ala Glu Ser Ser Ser Pro Arg Tyr Ala Asp Gln Ala Leu Ala

485 490 495

Ser Ala Val Phe Ser Pro Thr His Lys Ser Ala Val Ile Asn Gln Phe

500 505 510

Gln Gln Gln Gln Gln Ser Met Leu Ser Pro Ile Asn Thr Thr Phe Ser

515 520 525

Pro Lys Ser Val Asp His Pro Leu Leu Gln Ala Ser Phe Gly Val Gln

530 535 540

Ser Gly Arg Met Ser Pro Arg Asn Met Asp Pro Ile Ser Pro Ile Ser

545 550 555 560

Ser Arg Val Ser Met Leu Ala Gln Arg Glu Lys Gln Gln Gln Gln Leu

565 570 575

Arg Ser Leu Ser Ser Arg Glu Leu Gly Ser Asn Ser Ala Ala Ile Val

580 585 590

Gly Ser Pro Val Gly Ser Trp Ser Lys Trp Gly Ala Thr Asn Gly Lys

595 600 605

Pro Asp Trp Ala Val Ser Ala Asp Glu Leu Gly Lys Leu Arg Arg Ser

610 615 620

Asn Ser Phe Glu Leu Gly Asn Asn Gly Glu Glu Pro Asp Leu Ser Trp

625 630 635 640

Val Gln Ser Leu Val Lys Glu Ser Pro Thr Glu Met Lys Glu Lys Leu

645 650 655

Ser Ser Thr Leu Ser Gly Val Pro Ala Pro Ala Thr Ser Ser Glu Val

660 665 670

Pro Ser Ile Ser Ser Gln Met Glu Ser Val Asp His Glu Val Leu Gly

675 680 685

Ala Trp Leu Gln Gln Met Gln Leu Asp Pro Leu Val Ala Gln Gln Asn

690 695 700

<210>12

<211>1860

<212>DNA

＜213〉rice

<400>12

atgggggagc ctgggggcgc cgaggcggcc gtctccgcga ggctgctcga gctggcggcc 60

gacgacaacg cggcggggct cggggagctc ctcgcggcgt ggccctccct cgccgacgag 120

cccgcgccgt ggtacacccc ggcgcggggc gcggagccgc tgaccccgct catggtcgcc 180

gccgtgtacg gctcggtggg ctgcctcgac gcgctcctct cgccgcccta cctcgtggac 240

cccaaccgcg cctcggcgtc gtcgctctcc accccgctcc acctcgccgc cgcgggcggg 300

tccgcctccg cccccgcggc ggtctcccgc ctcctcgccg ccggcgccga cccggccctc 360

ctcgaccacc tccagcgccg ggcgtccgac ctcgtcgcgc tcccgcccaa ctcgctcccg 420

ctcaagaacc acctcctctc cctcctcggc gcccgcaagg agtggcctcc cgacccctcc 480

ctccccgaca tcaagaacgg cgcctacgcc tccgacgact tcaggatgta ctcgttcaag 540

gtgcgcgcgt gctcgcgggc ctactcccat gactggacgg agtgcccctt cgtccacccc 600

ggcgagaacg cgcggcggcg cgacccgagg aagtaccact acagctgcgt gccgtgcccg 660

gagttcaaga agggggccgg gtgcaggaga ggggacatgt gcgagtacgc gcacggggtg 720

ttcgagagct ggctccaccc ggcgcagtac cggacgcgcc tctgcaagga cggcgtcggc 780

tgcgcccgcc gcgtctgctt cttcgcccac acgcccgacg agctccgccc gctctacgtc 840

tccacgggct ccgccgtgcc gtcgccgcgc ggggcgttgg agatggcggc ggcggcggcg 900

gcgatgggga tggggctgtc gtcgccgggg tcgtcgtcgt tcacgccgcc gctatcgccg 960

tcggccggcg ggggcggggg cgggggcggg ggcagcggcg gcggcggcgc gtggccgcag 1020

cagccgagcg tgccggcgct ctgcctgccc gggagcgccg ggaacctcca cctgagccgg 1080

ctgcgcacgt cgctgagcgc gcgcgacatg gccgtcgacg agctgctcgc cgcggcggcg 1140

gcggcggcgg actacgacgg cctcgtcgcc tcccccgcct ccatccggtc cgcgaggggg 1200

aaggcgcttg tgccgtcaaa tctcgacgag ctcttctccg ctgagctcgc cgccgccgcg 1260

gcgtcgcgct cgccgcgcta cgccgaccaa ggcggcgccg cgttctcccc gacccgcaag 1320

gccaccgtgc tcaaccaatt ccagctgcag cagcagcata gcttgctctc gccgcgggcg 1380

gccgcggtga caccagagcc ggtctcccca atgagctccc gcctcctcgc cgcgctggcg 1440

cagcgggaga agatgcagca gcagacgctg cggagcatga gctcacggga cctcggcaac 1500

gccgcgtcgc tgctggtcgg ctcgccggtg agctcgagca tgtccaaatg ggggttcccc 1560

tccggcaacc cggactgggg cgccgacgac gaggagctcg gccgcctcaa gcgttgctcc 1620

tcgttcgagc tccggtccgg agccgccaat ggcaaccatg agcctgacct ctcatgggtc 1680

aacaccctag tgaaggagcc gacaccggag aagatgatga cgacgacatc ggcaatggat 1740

tccattggca tcttgggaca gaacacaagc cgtgatcaca tcgtcggagg cgaggatgac 1800

actgccggag tcatcagcag ctggcttgaa cagctccagc tcgatgagat ggttgtctag 1860

<210>13

<211>619

<212>PRT

＜213〉rice

<400>13

Met Gly Glu Pro Gly Gly Ala Glu Ala Ala Val Ser Ala Arg Leu Leu

1 5 10 15

Glu Leu Ala Ala Asp Asp Asn Ala Ala Gly Leu Gly Glu Leu Leu Ala

20 25 30

Ala Trp Pro Ser Leu Ala Asp Glu Pro Ala Pro Trp Tyr Thr Pro Ala

35 40 45

Arg Gly Ala Glu Pro Leu Thr Pro Leu Met Val Ala Ala Val Tyr Gly

50 55 60

Ser Val Gly Cys Leu Asp Ala Leu Leu Ser Pro Pro Tyr Leu Val Asp

65 70 75 80

Pro Asn Arg Ala Ser Ala Ser Ser Leu Ser Thr Pro Leu His Leu Ala

85 90 95

Ala Ala Gly Gly Ser Ala Ser Ala Pro Ala Ala Val Ser Arg Leu Leu

100 105 110

Ala Ala Gly Ala Asp Pro Ala Leu Leu Asp His Leu Gln Arg Arg Ala

115 120 125

Ser Asp Leu Val Ala Leu Pro Pro Asn Ser Leu Pro Leu Lys Asn His

130 135 140

Leu Leu Ser Leu Leu Gly Ala Arg Lys Glu Trp Pro Pro Asp Pro Ser

145 150 155 160

Leu Pro Asp Ile Lys Asn Gly Ala Tyr Ala Ser Asp Asp Phe Arg Met

165 170 175

Tyr Ser Phe Lys Val Arg Ala Cys Ser Arg Ala Tyr Ser His Asp Trp

180 185 190

Thr Glu Cys Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp

195 200 205

Pro Arg Lys Tyr His Tyr Ser Cys Val Pro Cys Pro Glu Phe Lys Lys

210 215 220

Gly Ala Gly Cys Arg Arg Gly Asp Met Cys Glu Tyr Ala His Gly Val

225 230 235 240

Phe Glu Ser Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys

245 250 255

Asp Gly Val Gly Cys Ala Arg Arg Val Cys Phe Phe Ala His Thr Pro

260 265 270

Asp Glu Leu Arg Pro Leu Tyr Val Ser Thr Gly Ser Ala Val Pro Ser

275 280 285

Pro Arg Gly Ala Leu Glu Met Ala Ala Ala Ala Ala Ala Met Gly Met

290 295 300

Gly Leu Ser Ser Pro Gly Ser Ser Ser Phe Thr Pro Pro Leu Ser Pro

305 310 315 320

Ser Ala Gly Gly Gly Gly Gly Gly Gly Gly Gly Ser Gly Gly Gly Gly

325 330 335

Ala Trp Pro Gln Gln Pro Ser Val Pro Ala Leu Cys Leu Pro Gly Ser

340 345 350

Ala Gly Asn Leu His Leu Ser Arg Leu Arg Thr Ser Leu Ser Ala Arg

355 360 365

Asp Met Ala Val Asp Glu Leu Leu Ala Ala Ala Ala Ala Ala Ala Asp

370 375 380

Tyr Asp Gly Leu Val Ala Ser Pro Ala Ser Ile Arg Ser Ala Arg Gly

385 390 395 400

Lys Ala Leu Val Pro Ser Asn Leu Asp Glu Leu Phe Ser Ala Glu Leu

405 410 415

Ala Ala Ala Ala Ala Ser Arg Ser Pro Arg Tyr Ala Asp Gln Gly Gly

420 425 430

Ala Ala Phe Ser Pro Thr Arg Lys Ala Thr Val Leu Asn Gln Phe Gln

435 440 445

Leu Gln Gln Gln His Ser Leu Leu Ser Pro Arg Ala Ala Ala Val Thr

450 455 460

Pro Glu Pro ValSer Pro Met Ser Ser Arg Leu Leu Ala Ala Leu Ala

465 470 475 480

Gln Arg Glu Lys Met Gln Gln Gln Thr Leu Arg Ser Met Ser Ser Arg

485 490 495

Asp Leu Gly Asn Ala Ala Ser Leu Leu Val Gly Ser Pro Val Ser Ser

500 505 510

Ser Met Ser Lys Trp Gly Phe Pro Ser Gly Asn Pro Asp Trp Gly Ala

515 520 525

Asp Asp Glu Glu Leu Gly Arg Leu Lys Arg Cys Ser Ser Phe Glu Leu

530 535 540

Arg Ser Gly Ala Ala Asn Gly Asn His Glu Pro Asp Leu Ser Trp Val

545 550 555 560

Asn Thr Leu Val Lys Glu Pro Thr Pro Glu Lys Met Met Thr Thr Thr

565 570 575

Ser Ala Met Asp Ser Ile Gly Ile Leu Gly Gln Asn Thr Ser Arg Asp

580 585 590

His Ile Val Gly Gly Glu Asp Asp Thr Ala Gly Val Ile Ser Ser Trp

595 600 605

Leu Glu Gln Leu Gln Leu Asp Glu Met Val Val

610 615

<210>14

<211>2106

<212>DNA

＜213〉puncture vine clover (Medicago truncatula)

<400>14

atgaaaaatc taactgttcg tactgatgat tctttttcca gcttacttga acatgcttct 60

aacaatgatt ttgaagattt caaggtagct ctagatagtg atgcttcact tattaatgaa 120

gttggcttct ggtatgtccg tcaaaaggga tctaaccaaa ttgttcttga gcaccgaacc 180

cctttaatgg tggctgcttc ctatgggagt attgatattc taaagcttat actctcatat 240

cccgaggctg atgttaattt ctcctgtgga actgataaaa gcactgctct tcactgtgct 300

gcctcaagtg gttcagttaa tgctgttgat gctataaaat tgcttttatc agctggtgct 360

gatatcaatt ctgtggatgc taatgggaaa cgccctgtgg atgttatcgt tgttcctatt 420

gttgttcctc ataagctcga aggtgttaaa acaattcttg aagaacttct ctcagacagt 480

gcttctgaag gatctgtgga tgattgctct cttcccctgt ctcttatttc atcgagtcct 540

ggttcatctg cccctttatc atctgctgaa aatggatctc catcctctcc tgtggctccc 600

aagtttacag atacagctgt taattctaca tcagaaaaga aagagtatcc agttgaccca 660

tctcttcctg acataaaaaa cagcatgtat gccacagatg aattccgcat gtattcattc 720

aaggttcgtc cttgttctcg tgcatactct catgattgga ctgagtgtcc ttttgtgcat 780

cctggagaga atgctcgaag gagagaccct agaaagtttc actacagctg tgtgccatgc 840

cctgatttta ggaaaggggc ttgccgacgt tcggatatgt gtgaatatgc tcatggagta 900

ttcgagtgct ggctacaccc agctcagtat cggacaaggc tgtgcaaaga cggtatgggt 960

tgtaaccgaa gggtgtgctt cttcgctcac tcacctgaag agctgcgtcc gctgtatgtg 1020

tccactggtt ctgctgttcc ttcaccccga tcagctgctt ctactgctaa tgtcatggac 1080

atggctgctg ctatgagcct tttccctggt tcaccatcat caatctcttt gatgtctcaa 1140

tcaccctttg cacagcctcc tctatctcca tctgcaaatg gcaataatgc ttggccacag 1200

cccaatgtgc cagctcttca tttaccagga agcattaatc aaactagtcg tttgagatct 1260

tctcttagtg cccgtgatat gccacacgac gacttcaaca atatgttgca agactttgat 1320

gggcagcagc agatactaaa tgacttgagc tgtttctcac agccccgtcc tggtgctatt 1380

tcagttggtc gatctggccg ccctaaaaca ctaactccct caaatctgga tgatcttttt 1440

tgtgctgaga ttgcttcatc tcctaggtat tccgaccccg ctgcggcttc tgtattttcc 1500

ccaacacaca aatctgctgt cttcaaccag tttcaacagc ttcaaagctc cttatcaccc 1560

atcaacacaa atgtcatgtc tcctacaaac gtagagcatc ccctgttcca ccaggcttca 1620

tatggtctct cttctcctgg aaggatgtca ccaagaagta tggaagccct atctccaatg 1680

agttctcggc tgtcagcttt tgctcagcgt gagaaacaac agcagcagca gcaacagctg 1740

cgtagcctca gctcaagaga actcggtgct aacaatcctc tctcagctgt tgggtcccct 1800

gttaactcct ggtccaagtg gggatcatcc cctattggaa aagctgattg gtcggtaaat 1860

ccaaatgact tcggtcaaac acagagatca acttcttttg agcatggaaa caatggagaa 1920

gagcctgatg taggttgggt ccattccctt gtcaaggatc ccacacctga gaagaaagag 1980

aagcttgcag gttccggccc aattccatcc gttgaaaaga atcccaatcc tcaagcggac 2040

ggcattgatc actctgtttt gggagcttgg ctcgagcaac tgcagctgga tcaacttgta 2100

gtctag 2106

<210>15

<211>701

<212>PRT

＜213〉puncture vine clover

<400>15

Met Lys Asn Leu Thr Val Arg Thr Asp Asp Ser Phe Ser Ser Leu Leu

1 5 10 15

Glu His Ala Ser Asn Asn Asp Phe Glu Asp Phe Lys Val Ala Leu Asp

20 25 30

Ser Asp Ala Ser Leu Ile Asn Glu Val Gly Phe Trp Tyr Val Arg Gln

35 40 45

Lys Gly Ser Asn Gln Ile Val Leu Glu His Arg Thr Pro Leu Met Val

50 55 60

Ala Ala Ser Tyr Gly Ser Ile Asp Ile Leu Lys Leu Ile Leu Ser Tyr

65 70 75 80

Pro Glu Ala Asp Val Asn Phe Ser Cys Gly Thr Asp Lys Ser Thr Ala

85 90 95

Leu His Cys Ala Ala Ser Ser Gly Ser Val Asn Ala Val Asp Ala Ile

100 105 110

Lys Leu Leu Leu Ser Ala Gly Ala Asp Ile Asn Ser Val Asp Ala Asn

115 120 125

Gly Lys Arg Pro Val Asp Val Ile Val Val Pro Ile Val Val Pro His

130 135 140

Lys Leu Glu Gly Val Lys Thr Ile Leu Glu Glu Leu Leu Ser Asp Ser

145 150 155 160

Ala Ser Glu Gly Ser Val Asp Asp Cys Ser Leu Pro Leu Ser Leu Ile

165 170 175

Ser Ser Ser Pro Gly Ser Ser Ala Pro Leu Ser Ser Ala Glu Asn Gly

180 185 190

Ser Pro Ser Ser Pro Val Ala Pro Lys Phe Thr Asp Thr Ala Val Asn

195 200 205

Ser Thr Ser Glu Lys Lys Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp

210 215 220

Ile Lys Asn Ser Met Tyr Ala Thr Asp Glu Phe Arg Met Tyr Ser Phe

225 230 235 240

Lys Val Arg Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys

245 250 255

Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys

260 265 270

Phe His Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg Lys Gly Ala Cys

275 280 285

Arg Arg Ser Asp Met Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp

290 295 300

Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Gly Met Gly

305 310 315 320

Cys Asn Arg Arg Val Cys Phe Phe Ala His Ser Pro Glu Glu Leu Arg

325 330 335

Pro Leu Tyr Val Ser Thr Gly Ser Ala Val Pro Ser Pro Arg Ser Ala

340 345 350

Ala Ser Thr Ala Asn Val Met Asp Met Ala Ala Ala Met Ser Leu Phe

355 360 365

Pro Gly Ser Pro Ser Ser Ile Ser Leu Met Ser Gln Ser Pro Phe Ala

370 375 380

Gln Pro Pro Leu Ser Pro Ser Ala Asn Gly Asn Asn Ala Trp Pro Gln

385 390 395 400

Pro Asn Val Pro Ala Leu His Leu Pro Gly Ser Ile Asn Gln Thr Ser

405 410 415

Arg Leu Arg Ser Ser Leu Ser Ala Arg Asp Met Pro His Asp Asp Phe

420 425 430

Asn Asn Met Leu Gln Asp Phe Asp Gly Gln Gln Gln Ile Leu Asn Asp

435 440 445

Leu Ser Cys Phe Ser Gln Pro Arg Pro Gly Ala Ile Ser Val Gly Arg

450 455 460

Ser Gly Arg Pro Lys Thr Leu Thr Pro Ser Asn Leu Asp Asp Leu Phe

465 470 475 480

Cys Ala Glu Ile Ala Ser Ser Pro Arg Tyr Ser Asp Pro Ala Ala Ala

485 490 495

Ser Val Phe Ser Pro Thr His Lys Ser Ala Val Phe Asn Gln Phe Gln

500 505 510

Gln Leu Gln Ser Ser Leu Ser Pro Ile Asn Thr Asn Val Met Ser Pro

515 520 525

Thr Asn Val Glu His Pro Leu Phe His Gln Ala Ser Tyr Gly Leu Ser

530 535 540

Ser Pro Gly Arg Met Ser Pro Arg Ser Met Glu Ala Leu Ser Pro Met

545 550 555 560

Ser Ser Arg Leu Ser Ala Phe Ala Gln Arg Glu Lys Gln Gln Gln Gln

565 570 575

Gln Gln Gln Leu Arg Ser Leu Ser Ser Arg Glu Leu Gly Ala Asn Asn

580 585 590

Pro Leu Ser Ala Val Gly Ser Pro Val Asn Ser Trp Ser Lys Trp Gly

595 600 605

Ser Ser Pro Ile Gly Lys Ala Asp Trp Ser Val Asn Pro Asn Asp Phe

610 615 620

Gly Gln Thr Gln Arg Ser Thr Ser Phe Glu His Gly Asn Asn Gly Glu

625 630 635 640

Glu Pro Asp Val Gly Trp Val His Ser Leu Val Lys Asp Pro Thr Pro

645 650 655

Glu Lys Lys Glu Lys Leu Ala Gly Ser Gly Pro Ile Pro Ser Val Glu

660 665 670

Lys Asn Pro Asn Pro Gln Ala Asp Gly Ile Asp His Ser Val Leu Gly

675 680 685

Ala Trp Leu Glu Gln Leu Gln Leu Asp Gln Leu Val Val

690 695 700

<210>16

<211>2841

<212>DNA

＜213〉rice

<400>16

atgaacggca cgccgatctc cgcgtccgcc gcggccggcg tcgacggagt cggcgcggcg 60

gtggcgctgg cggccgcgac caagaagagt gccgccgcgg cggccgccgt cgccgagatg 120

gcgaaaaccc tcaccgtcga cacggacgac gccttcgcgg ggctcctcga gctcgccgcg 180

gacgacgacg cggagggcct gcgccgcgcg ctggagcgcg ccccgcccgc cgccgcggac 240

gaggcgggcc tctggtacgg ccgccgcaag gtcctcgagc accgcacgcc gctgatggtc 300

gcggccacct atggcagcct cgcggtgctt cgcctgctgc tgtccctccc gtccgtcgat 360

gtcaatcgcc gctgtggctc cgacggcacc accgccctcc actgtgcggc gtctggtggc 420

tcgccgtctt gtgtggaggc cgtcaagctg ctgcttgctg ctggggctga tgctgatgcc 480

acggatgctt ccggatatcg tccagctgat gtgatctctg ttcctccaaa gatgtttgac 540

gccaagattg ccctccaaga tcttcttgga tgcccaaagg ctgggcatgg cgttctccgg 600

gtggtgacaa gggccgcaaa ctctatgttg tcacctgtat catcccctac agcagaagat 660

gcacgatctc catcagctgc tgtgatgatg acgacaaagt ttgcagatct tccaagggtt 720

gtgacatcgg aaaagaaaga atatccagtg gatccgtccc ttcccgatat caagaacagc 780

atctatgctt ccgatgagtt ccgcatgtac tcatttaaga tcaggccatg ctcgcgggcg 840

tactcacatg attggactga gtgcccgttt gttcacccag gggagaacgc acggcgtcgg 900

gaccctcgca agtatcacta cagctgtgtg ccatgccccg actttagaaa gggagtttgc 960

cggcgtggtg acatgtgtga atatgctcat ggcgtgttcg agtgttggct ccatccagca 1020

cagtaccgta ctcgcctttg caaggatggc acaagctgta atcgccgtgtctgtttcttt 1080

gcgcatacaa ctgatgagct ccgaccacta tatgtttcca ctggatctgc agtaccatcc 1140

ccaagagcct cggcaacagc tacaatggag atggctgcag caatgggctt gatgcctggt 1200

tctccatcat cagtttcagc agtcatgtcc ccatttacac caccaatgtc cccttcaggc 1260

aatgggatgc ccccttcatt gggctggcag cagccaaatg ttccgacact acaccttcca 1320

ggcagcagcc ttcagtcgag ccggctccgt acctcactta gtgcaaggga tatgcctgct 1380

gatgattact ccctgatgca ggatattgat tcacagctta taaatgattt gtgctattca 1440

cgtattggtt catcaacagg aaaccacacg tctcggacca agtccctaaa tccgtcaaac 1500

ttggatgatc tcttctctgc tgagatggtc tcttccccga ggtatagtaa tgctgatcag 1560

ggtggtatgt tttcaccatc tcacaaggct gctttcctta atcagttcca gcaacagcag 1620

caggcacttc tttcaccaat caacacagtc ttctccccga agtctgtgga caaccagcag 1680

ttgccttcac actcatctct gttgcaagca tcacttggta tatcctcccc tggccgcatg 1740

tctcctcgat gtgttgaatc tgggtcccct atgaactctc atcttgctgc tgctcttgct 1800

cagcgtgaga agcaacagca gacaatgaga agtctcagtt ctcgtgatct tgggccgagt 1860

gctgcaagag catcaggtgt tgttggctcc cctctaagct catcatggtc aaagtgggga 1920

tcaccttcag ggacacctga ctggggtgtt aatggtgaag aattgggcaa gcttcgccgg 1980

tcatcatcgt ttgagctgag atctggtggt gatgatccag atctctcttg ggtacacaca 2040

ctggttaagg aatctccacc agagaagcaa gtcactactg ctgaatccat aaactctgtt 2100

ggaccttcac cactgatgcc tcccagtgta agcaacggtg aaggtcctag tctgaatgcc 2160

ccgctggatg ggcatgacca agctgctgtt attggagcat tgcttgaaca gatgcagctt 2220

gatcagcata ttggtagtct agcaacataa gcgctgaatg agcctggaaa gtgcaaggag 2280

ttattattct tagttaatga atttggagta atttttttcc tgttcattaa gatggtcagc 2340

aagcaaaagg atggatagct gatggtggtg attcagagat tggttttctt tactttattg 2400

aggtaaatca tatacattat tgaggttcca gtaggttgaa agattgaagt accttgattg 2460

gggtcgtttc aagaccgacc caggtagaat cgcaccccgg cagcttcaat tcatcggtca 2520

aaaatatttc cctgttttgt taattaaccc cgttaaaaaa gaagactcgt ttggtgtttc 2580

ggaattcttt tctttacctt agcggtgttt attttgttta ttatgatatt gatacttgat 2640

gtactgatgg gtataaggtt ggttaccagg catgctatag tggtatatca agtcccaaag 2700

tattcttttt ctccctttca ccatttgtcg aggatcatac tatggccttg ttttggtcag 2760

atcttgaggc ctgtataatc cttggatttg taataatgta atattgtcat tgaacttaca 2820

ttgctattgt tttgcaatcg c 2841

<210>17

<211>749

<212>PRT

＜213〉rice

<400>17

Met Asn Gly Thr Pro Ile Ser Ala Ser Ala Ala Ala Gly Val Asp Gly

1 5 10 15

Val Gly Ala Ala Val Ala Leu Ala Ala Ala Thr Lys Lys Ser Ala Ala

20 25 30

Ala Ala Ala Ala Val Ala Glu Met Ala Lys Thr Leu Thr Val Asp Thr

35 40 45

Asp Asp Ala Phe Ala Gly Leu Leu Glu Leu Ala Ala Asp Asp Asp Ala

50 55 60

Glu Gly Leu Arg Arg Ala Leu Glu Arg Ala Pro Pro Ala Ala Ala Asp

65 70 75 80

Glu Ala Gly Leu Trp Tyr Gly Arg Arg Lys Val Leu Glu His Arg Thr

85 90 95

Pro Leu Met Val Ala Ala Thr Tyr Gly Ser Leu Ala Val Leu Arg Leu

100 105 110

Leu Leu Ser Leu Pro Ser Val Asp Val Asn Arg Arg Cys Gly Ser Asp

115 120 125

Gly Thr Thr Ala Leu His Cys Ala Ala Ser Gly Gly Ser Pro Ser Cys

130 135 140

Val Glu Ala Val Lys Leu Leu Leu Ala Ala Gly Ala Asp Ala Asp Ala

145 150 155 160

Thr Asp Ala Ser Gly Tyr Arg Pro Ala Asp Val Ile Ser Val Pro Pro

165 170 175

Lys Met Phe Asp Ala Lys Ile Ala Leu Gln Asp Leu Leu Gly Cys Pro

180 185 190

Lys Ala Gly His Gly Val Leu Arg Val Val Thr Arg Ala Ala Asn Ser

195 200 205

Met Leu Ser Pro Val Ser Ser Pro Thr Ala Glu Asp Ala Arg Ser Pro

210 215 220

Ser Ala Ala Val Met Met Thr Thr Lys Phe Ala Asp Leu Pro Arg Val

225 230 235 240

Val Thr Ser Glu Lys Lys Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp

245 250 255

Ile Lys Asn Ser Ile Tyr Ala Ser Asp Glu Phe Arg Met Tyr Ser Phe

260 265 270

Lys Ile Arg Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys

275 280 285

Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys

290 295 300

Tyr His Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg Lys Gly Val Cys

305 310 315 320

Arg Arg Gly Asp Met Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp

325 330 335

Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr Ser

340 345 350

Cys Asn Arg Arg Val Cys Phe Phe Ala His Thr Thr Asp Glu Leu Arg

355 360 365

Pro Leu Tyr Val Ser Thr Gly Ser Ala Val Pro Ser Pro Arg Ala Ser

370 375 380

Ala Thr Ala Thr Met Glu Met Ala Ala Ala Met Gly Leu Met Pro Gly

385 390 395 400

Ser Pro Ser Ser Val Ser Ala Val Met Ser Pro Phe Thr Pro Pro Met

405 410 415

Ser Pro Ser Gly Asn Gly Met Pro Pro Ser Leu Gly Trp Gln Gln Pro

420 425 430

Asn Val Pro Thr Leu His Leu Pro Gly Ser Ser Leu Gln Ser Ser Arg

435 440 445

Leu Arg Thr Ser Leu Ser Ala Arg Asp Met Pro Ala Asp Asp Tyr Ser

450 455 460

Leu Met Gln Asp Ile Asp Ser Gln Leu Ile Asn Asp Leu Cys Tyr Ser

465 470 475 480

Arg Ile Gly Ser Ser Thr Gly Asn His Thr Ser Arg Thr Lys Ser Leu

485 490 495

Asn Pro Ser Asn Leu Asp Asp Leu Phe Ser Ala Glu Met Val Ser Ser

500 505 510

Pro Arg Tyr Ser Asn Ala Asp Gln Gly Gly Met Phe Ser Pro Ser His

515 520 525

Lys Ala Ala Phe Leu Asn Gln Phe Gln Gln Gln Gln Gln Ala Leu Leu

530 535 540

Ser Pro Ile Asn Thr Val Phe Ser Pro Lys Ser Val Asp Asn Gln Gln

545 550 555 560

Leu Pro Ser His Ser Ser Leu Leu Gln Ala Ser Leu Gly Ile Ser Ser

565 570 575

Pro Gly Arg Met Ser Pro Arg Cys Val Glu Ser Gly Ser Pro Met Asn

580 585 590

Ser His Leu Ala Ala Ala Leu Ala Gln Arg Glu Lys Gln Gln Gln Thr

595 600 605

Met Arg Ser Leu Ser Ser Arg Asp Leu Gly Pro Ser Ala Ala Arg Ala

610 615 620

Ser Gly Val Val Gly Ser Pro Leu Ser Ser Ser Trp Ser Lys Trp Gly

625 630 635 640

Ser Pro Ser Gly Thr Pro Asp Trp Gly Val Asn Gly Glu Glu Leu Gly

645 650 655

Lys Leu Arg Arg Ser Ser Ser Phe Glu Leu Arg Ser Gly Gly Asp Asp

660 665 670

Pro Asp Leu Ser Trp Val His Thr Leu Val Lys Glu Ser Pro Pro Glu

675 680 685

Lys Gln Val Thr Thr Ala Glu Ser Ile Asn Ser Val Gly Pro Ser Pro

690 695 700

Leu Met Pro Pro Ser Val Ser Asn Gly Glu Gly Pro Ser Leu Asn Ala

705 710 715 720

Pro Leu Asp Gly His Asp Gln Ala Ala Val Ile Gly Ala Leu Leu Glu

725 730 735

Gln Met Gln Leu Asp Gln His Ile Gly Ser Leu Ala Thr

740 745

<210>18

<211>2769

<212>DNA

＜213〉Arabidopis thaliana

<400>18

acaccagtta ccctctcatc cgttttcgtt ttttttttct ctctttcaaa aatctctcag 60

ctgaggttga tcgatcttct tcttcttctt cctcactctt tagatttgtt ccttttgcat 120

tttacacttt tggatctgaa aatgtggttc tctgtttcgc ccgttaccgt ttagattcag 180

ttctgttttt ttcttacccg atcgcttgat tcggactgtg atctttgatc ttttttcttc 240

tccagtgccg tgaaggatgt gtggtcttgc taagaagctg gatatagagg atactttgac 300

atcactgtca gaccaagaga atgaatcttt ggccaaaccc atgaatgatg ctgctgaatg 360

ggaacattcg ttttctgcct tgcttgagtt tgctgcagac aacgatgtgg aggggtttag 420

gcggcaactc tctgatgtgt cttgtatcaa ccagatgggt ctttggtaca gacggcagag 480

gtttgttaga agaatggttc ttgagcaaag aaccccgctg atggttgctt cgttatatgg 540

gagtttagat gttgtgaagt ttattctttc tttcccggaa gcggagttga atctgtcttg 600

tggtcctgat aaaagtactg ctcttcattg cgctgcttct ggtgcttctg tgaattcctt 660

ggatgttgtc aagttgcttt tgagtgtagg agcagatcct aatatccctg atgctcatgg 720

aaatcgtcct gttgatgttc ttgttgtgtc tccacacgct cctggtttga gaaccatcct 780

tgaagagatc ttgaagaaag acgagattat atctgaagat ctgcatgcct cgtcatctag 840

cttgggatca agtttccggt ctctctcatc atcccctgat aatggttcct cgttactctc 900

cttagattca gtatcctctc cgactaagcc acacggtact gatgtaactt tcgcatcaga 960

gaagaaagag tacccaattg atccatcatt gcctgatatc aaaagcggga tttattcaac 1020

cgatgagttt cgtatgttct cgttcaagat ccgcccatgt tctcgagcat attcccatga 1080

ctggactgaa tgtccatttg cacacccagg tgagaatgca aggagaagag acccgaggaa 1140

gtttcactat acgtgtgttc catgcccgga ttttaagaaa ggatcctgta agcaaggtga 1200

tatgtgtgaa tatgctcatg gggtttttga atgctggcta caccctgctc agtacagaac 1260

acgattgtgc aaggacggaa tgggttgcaa ccgaagggtt tgcttctttg ctcacgcaaa 1320

tgaggagttg cgtcccttgt acccttccac aggatctgga ttgccatctc ctcgggcttc 1380

gtctgctgtt tccgcctcta ctatggacat ggcgtcagtt ttgaacatgt taccaggctc 1440

accatctgct gctcaacatt cgttcacccc accaatatct ccttctggaa atggtagtat 1500

gccccattca tcgatgggtt ggcctcagca gaacataccg gcgttgaatc ttcctggaag 1560

caatatccag ttgagtcgtc tgagatcttc tcttaacgct agagatattc cttctgagca 1620

gcttagcatg ctgcatgagt ttgaaatgca acgtcagctt gctggcgata tgcacagtcc 1680

acgctttatg aatcattccg ctcgtcctaa gacactgaac ccttcaaatc tggaggaact 1740

cttctcagct gaggttgcat ctcctcgttt ctctgatcaa cttgctgttt catctgttct 1800

atcgccttcc cacaagtccg cgcttcttaa tcagctgcag aataataagc agagcatgct 1860

ttctcctatc aagacaaatc taatgtcttc tccaaagaat gtggagcaac attctcttct 1920

gcagcaagcc tcgtcacccc gaggcggaga gcctatttcc ccaatgaatg ctcgaatgaa 1980

acagcagcta cattcacgca gcctaagctc ccgtgatttt ggatctagtc tgccccgtga 2040

tttaatgccg actgattctg gttcgccatt aagtccatgg tcaagttggg accagaccca 2100

tggaagcaag gtggattggt cagtccaatc agatgagtta ggtcggttga gaaaatctca 2160

ttccttggct aataacccaa acagggaagc agatgtttca tgggctcagc agatgttaaa 2220

agactcttca tcacctagga acggaaaccg tgttgtgaac atgaatggtg caaggccatt 2280

gactcaaggt ggttcgagtg tgaatcctca caacagtgac actcgtgaga gcgacattct 2340

tgatgcgtgg cttgaacagc tgcacctaga tcgctgagcc tcagctgcga gagagaggtt 2400

cacatttctg tgaagctgtg aaactgatga ttcgtttatt tattattcaa gaaagcaaac 2460

ggaaacaaaa gcaaactccg ggtaagcttt tttcgattct aataacccta aaaggctcag 2520

ttttttcagg cttctttctg aaatttcttt actttcttat ttttatcacc tcattaaatt 2580

aattattgta tcatctctgt tgtaacaatg gccaaagtgc gcctctatta cttcccggat 2640

ttctgattta cattttttgt atcctctcag tttgtcaatt gtttctaata tctccttcat 2700

atttgtcaaa gaacactgta tgagaaataa taacatattg tttcagctaa taagattcat 2760

tcatttcct 2769

<210>19

<211>706

<212>PRT

＜213〉Arabidopis thaliana

<400>19

Met Cys Gly Leu Ala Lys Lys Leu Asp Ile Glu Asp Thr Leu Thr Ser

1 5 10 15

Leu Ser Asp Gln Glu Asn Glu Ser Leu Ala Lys Pro Met Asn Asp Ala

20 25 30

Ala Glu Trp Glu Hi s Ser Phe Ser Ala Leu Leu Glu Phe Ala Ala Asp

35 40 45

Asn Asp Val Glu Gly Phe Arg Arg Gln Leu Ser Asp Val Ser Cys Ile

50 55 60

Asn Gln Met Gly Leu Trp Tyr Arg Arg Gln Arg Phe Val Arg Arg Met

65 70 75 80

Val Leu Glu Gln Arg Thr Pro Leu Met Val Ala Ser Leu Tyr Gly Ser

85 90 95

Leu Asp Val Val Lys Phe Ile Leu Ser Phc Pro Glu Ala Glu Leu Asn

100 105 110

Leu Ser Cys Gly Pro Asp Lys Ser Thr Ala Leu His Cys Ala Ala Ser

115 120 125

Gly Ala Ser Val Asn Ser Leu Asp Val Val Lys Leu Leu Leu Ser Val

130 135 140

Gly Ala Asp Pro Asn Ile Pro Asp Ala His Gly Asn Arg Pro Val Asp

145 150 155 160

Val Leu Val Val Ser Pro His Ala Pro Gly Leu Arg Thr Ile Leu Glu

165 170 175

Glu Ile Leu Lys Lys Asp Glu Ile Ile Ser Glu Asp Leu His Ala Ser

180 185 190

Ser Ser Ser Leu Gly Ser Ser Phe Arg Ser Leu Ser Ser Ser Pro Asp

195 200 205

Asn Gly Ser Ser Leu Leu Ser Leu Asp Ser Val Ser Ser Pro Thr Lys

210 215 220

Pro His Gly Thr Asp Val Thr Phe Ala Ser Glu Lys Lys Glu Tyr Pro

225 230 235 240

Ile Asp Pro Ser Leu Pro Asp Ile Lys Ser Gly Ile Tyr Ser Thr Asp

245 250 255

Glu Phe Arg Met Phe Ser Phe Lys Ile Arg Pro Cys Ser Arg Ala Tyr

260 265 270

Ser His Asp Trp Thr Glu Cys Pro Phe Ala His Pro Gly Glu Asn Ala

275 280 285

Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Thr Cys Val Pro Cys Pro

290 295 300

Asp Phe Lys Lys Gly Ser Cys Lys Gln Gly Asp Met Cys Glu Tyr Ala

305 310 315 320

His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg

325 330 335

Leu Cys Lys Asp Gly Met Gly Cys Asn Arg Arg Val Cys Phe Phe Ala

340 34 5350

His Ala Asn Glu Glu Leu Arg Pro Leu Tyr Pro Ser Thr Gly Ser Gly

355 360 365

Leu Pro Ser Pro Arg Ala Ser Ser Ala Val Ser Ala Ser Thr Met Asp

370 375 380

Met Ala Ser Val Leu Asn Met Leu Pro Gly Ser Pro Ser Ala Ala Gln

385 390 395 400

His Ser Phe Thr Pro Pro Ile Ser Pro Ser Gly Asn Gly Ser Met Pro

405 410 415

His Ser Ser Met Gly Trp Pro Gln Gln Asn Ile Pro Ala Leu Asn Leu

420 425 430

Pro Gly Ser Asn Ile Gln Leu Ser Arg Leu Arg Ser Ser Leu Asn Ala

435 440 445

Arg Asp Ile Pro Ser Glu Gln Leu Ser Met Leu His Glu Phe Glu Met

450 455 460

Gln Arg Gln Leu Ala Gly Asp Met His Ser Pro Arg Phe Met Asn His

465 470 475 480

Ser Ala Arg Pro Lys Thr Leu Asn Pro Ser Asn Leu Glu Glu Leu Phe

485 490 495

Ser Ala Glu Val Ala Ser Pro Arg Phe Ser Asp Gln Leu Ala Val Ser

500 505 510

Ser Val Leu Ser Pro Ser His Lys Ser Ala Leu Leu Asn Gln Leu Gln

515 520 525

Asn Asn Lys Gln Ser Met Leu Ser Pro Ile Lys Thr Asn Leu Met Ser

530 535 540

Ser Pro Lys Asn Val Glu Gln His Ser Leu Leu Gln Gln Ala Ser Ser

545 550 555 560

Pro Arg Gly Gly Glu Pro Ile Ser Pro Met Asn Ala Arg Met Lys Gln

565 570 575

Gln Leu His Ser Arg Ser Leu Ser Ser Arg Asp Phe Gly Ser Ser Leu

580 585 590

Pro Arg Asp Leu Met Pro Thr Asp Ser Gly Ser Pro Leu Ser Pro Trp

595 600 605

Ser Ser Trp Asp Gln Thr His Gly Ser Lys Val Asp Trp Ser Val Gln

610 615 620

Ser Asp Glu Leu Gly Arg Leu Arg Lys Ser His Ser Leu Ala Asn Asn

625 630 635 640

Pro Asn Arg Glu Ala Asp Val Ser Trp Ala Gln Gln Met Leu Lys Asp

645 650 655

Ser Ser Ser Pro Arg Asn Gly Asn Arg Val Val Asn Met Asn Gly Ala

660 665 670

Arg Pro Leu Thr Gln Gly Gly Ser Ser Val Asn Pro His Asn Ser Asp

675 680 685

Thr Arg Glu Ser Asp Ile Leu Asp Ala Trp Leu Glu Gln Leu His Leu

690 695 700

Asp Arg

705

<210>20

<211>2674

<212>DNA

＜213〉rice

<400>20

tgcgagtcct cctcctcttc tcgtcgccgt gctactctcg ctttctctct ctctctctct 60

cacttgttcc ccaaggcgag aagcagccgc cgccggcgag cgtcgcgggg gaggggaggg 120

aagggaggga ggagcggtgg atccgggctt gattggattg ggtcggattc gattttggat 180

caaccccgga gggcgggagc ggttgctaca gatgcgttga gctttggtta atctatccgg 240

cgagagataa tgggcgagct tgctgatctc gttgtcgtgc cgtcgcagcc gccgctcgcc 300

ggcggccggc gggacagget ggcggcgctg ctggagctcg cggcggcgga tgatgttgat 360

gggctcaggg gggcgctcgc ggagggaggc gaggaggcgg cggagttggc tgatggggtc 420

gggctgtggt atggtcggag caaggcgtac gaggcgcgca cgccgctgat ggtggcggcg 480

acgtacggca gcgccggggt ggtctcgctg ctggtgggcc tcggcggttg cgtcgacgtc 540

aaccgtcgcc ctggagccga cggcgccacc gcgctccact gcgccgcctc cggtggctcg 600

cgcaacgccg tcgctgttgt caagctgctt ttggccgctg gcgccgatcc ggccaccccc 660

gattccgccg gccgcttccc cgccgacgtc atcctagctc ctccggcttc gccagatgcc 720

cttggcgatc tcgaggtgct cctcggccgc cgccgagcac tcgccgtggc gacctcggtg 780

gcttcaggtt cgtcatcccc tccgctctcg tcctcaccag atgagggcaa caggtcgccc 840

tcgtcgcgtt cgtcgtcgct gtctcccatc actgtggatc gtgggaagaa ggagtatccg 900

gtggatccaa ctctgccgga catcaagagc agcgtgtatg cttcggatga gttccgcatg 960

tttgcgttca aggtccggcc ctgctcccgt gcctactcac acgactggac tgagtgcccg 1020

tttgtgcacc ccggcgagaa cgcccgccgc cgtgatcccc gcaagcaccc atacactgct 1080

gtgccttgcc ccaactttcg ccggcctggt ggctgcccta gcggcgatag ctgtgagttc 1140

tcgcatggcg tgtttgagag ctggctacac ccatcacagt atcgcacaag gctctgcaag 1200

gagggagcag cttgcgcccg tcgcatttgc ttctttgccc atgatgagga tgagctccgc 1260

catgtgcctc acaacagtgg tgccggcctg ctgtctcccc gcgcttcttc atccattgat 1320

atgactgctg cagctgcgct cgggcttctt ccaggttctc ctaccagaca ctttgcaccg 1380

ccgcctgtgt caccatctgc tgggagcaat ggaggagctg ctgctgcgca ttggctccaa 1440

ggcagtaggc tgcgttcttc tttcaatgca agggatgctg ctgttgatga ccttggcatg 1500

ctcctcgaat gggaatcaca ataccttggg gcactctgcc tgccacccag cagccgcccc 1560

caaccacgcc tttcagctgg tctgagtatc aggccaacaa ttgctccatc caatcttgaa 1620

gacatgtatg cttcagacat ggcaatgtct ccgaggttcc ctaatgacca aggtcactca 1680

gtctactcac cagcccacaa atcagccctc ctcaacaagc ttcatcaaca gaagggcctc 1740

ttatcacctg ttaacaccaa cagaatgtac tccccaaggg ctcttgatcc gtcatctttg 1800

gcacattctc catttggtgg catgtctccc cggtcccccc gtaccatgga acctacatca 1860

cccctaagtg ctcgtgtagg agcccctgcc acacagcggc cttctgttgg ttcaccacgg 1920

aattccagtg cttggggcac cgtggggtcc ccgatgggta aggttgactg gggtgtcgat 1980

agcgaggagc tagtccgctt gagacgccct gcacaaccag ggtttggaga agatgagaca 2040

gatgtatcat gggtgcagtc actggtaagc aatgctgagc ttaatggcaa gaggggcgaa 2100

gtacaaggca tgcctggtac ttctgcattg atgaacaggc ctgacctgaa caatcagggt 2160

gacttgttgg accagacggt gatcggtgct tggcttgagc agatgcacct ggatcagaag 2220

tgatttccaa gggaagccat gaagtcccaa agtggatgaa gcctttattt tgccaaggtt 2280

atttaccaaa gaatagttgt tggtcctagt aaataataat ttattctttt taattcttga 2340

aatttttggt gggcaaagtc agagatggtg gtcaagttca acaaaacatt tggtcacaga 2400

ttggtagctg aaatcagttc cagagattgg taaacaacct cattacttgg ggtcctaact 2460

agtattcttt tgattagctc agatgagtct ttattttagt gggttaaaat tcatatgttc 2520

cccatggtta ttatgtccat gatctcttcc taacaaaaga gagattataa ttgtccattt 2580

ttcatttatc aatgaatgat tttgttaaaa caatgtaagt tacattctta attttttctc 2640

tgttcaatgg aattaccttc cttggttagt cctc 2674

<210>21

<211>657

<212>PRT

＜213〉rice

<400>21

Met Gly Glu Leu Ala Asp Leu Val Val Val Pro Ser Gln Pro Pro Leu

1 5 10 15

Ala Gly Gly Arg Arg Asp Arg Leu Ala Ala Leu Leu Glu Leu Ala Ala

20 25 30

Ala Asp Asp Val Asp Gly Leu Arg Gly Ala Leu Ala Glu Gly Gly Glu

35 40 45

Glu Ala Ala Glu Leu Ala Asp Gly Val Gly Leu Trp Tyr Gly Arg Ser

50 55 60

Lys Ala Tyr Glu Ala Arg Thr Pro Leu Met Val Ala Ala Thr Tyr Gly

65 70 75 80

Ser Ala Gly Val Val Ser Leu Leu Val Gly Leu Gly Gly Cys Val Asp

85 90 95

Val Asn Arg Arg Pro Gly Ala Asp Gly Ala Thr Ala Leu His Cys Ala

100 105 110

Ala Ser Gly Gly Ser Arg Asn Ala Val Ala Val Val Lys Leu Leu Leu

115 120 125

Ala Ala Gly Ala Asp Pro Ala Thr Pro Asp Ser Ala Gly Arg Phe Pro

130 135 140

Ala Asp Val Ile Leu Ala Pro Pro Ala Ser Pro Asp Ala Leu Gly Asp

145 150 155 160

Leu Glu Val Leu Leu Gly Arg Arg Arg Ala Leu Ala Val Ala Thr Ser

165 170 175

Val Ala Ser Gly Ser Ser Ser Pro Pro Leu Ser Ser Ser Pro Asp Glu

180 185 190

Gly Asn Arg Ser Pro Ser Ser Arg Ser Ser Ser Leu Ser Pro Ile Thr

195 200 205

Val Asp Arg Gly Lys Lys Glu Tyr Pro Val Asp Pro Thr Leu Pro Asp

210 215 220

Ile Lys Ser Ser Val Tyr Ala Ser Asp Glu Phe Arg Met Phe Ala Phe

225 230 235 240

Lys Val Arg Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys

245 250 255

Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys

260 265 270

His Pro Tyr Thr Ala Val Pro Cys Pro Asn Phe Arg Arg Pro Gly Gly

275 280 285

Cys Pro Ser Gly Asp Ser Cys Glu Phe Ser His Gly Val Phe Glu Ser

290 295 300

Trp Leu His Pro Ser Gln Tyr Arg Thr Arg Leu Cys Lys Glu Gly Ala

305 310 315 320

Ala Cys Ala Arg Arg Ile Cys Phe Phe Ala His Asp Glu Asp Glu Leu

325 330 335

Arg His Val Pro His Asn Ser Gly Ala Gly Leu Leu Ser Pro Arg Ala

340 345 350

Ser Ser Ser Ile Asp Met Thr Ala Ala Ala Ala Leu Gly Leu Leu Pro

355 360 365

Gly Ser Pro Thr Arg His Phe Ala Pro Pro Pro Val Ser Pro Ser Ala

370 375 380

Gly Ser Asn Gly Gly Ala Ala Ala Ala His Trp Leu Gln Gly Ser Arg

385 390 395 400

Leu Arg Ser Ser Phe Asn Ala Arg Asp Ala Ala Val Asp Asp Leu Gly

405 410 415

Met Leu Leu Glu Trp Glu Ser Gln Tyr Leu Gly Ala Leu Cys Leu Pro

420 425 430

Pro Ser Ser Arg Pro Gln Pro Arg Leu Ser Ala Gly Leu Ser Ile Arg

435 440 445

Pro Thr Ile Ala Pro Ser Asn Leu Glu Asp Met Tyr Ala Ser Asp Met

450 455 460

Ala Met Ser Pro Arg Phe Pro Asn Asp Gln Gly His Ser Val Tyr Ser

465 470 475 480

Pro Ala His Lys Ser Ala Leu Leu Asn Lys Leu His Gln Gln Lys Gly

485 490 495

Leu Leu Ser Pro Val Asn Thr Asn Arg Met Tyr Ser Pro Arg Ala Leu

500 505 510

Asp Pro Ser Ser Leu Ala His Ser Pro Phe Gly Gly Met Ser Pro Arg

515 520 525

Ser Pro Arg Thr Met Glu Pro Thr Ser Pro Leu Ser Ala Arg Val Gly

530 535 540

Ala Pro Ala Thr Gln Arg Pro Ser Val Gly Ser Pro Arg Asn Ser Ser

545 550 555 560

Ala Trp Gly Thr Val Gly Ser Pro Met Gly Lys Val Asp Trp Gly Val

565 570 575

Asp Ser Glu Glu Leu Val Arg Leu Arg Arg Pro Ala Gln Pro Gly Phe

580 585 590

Gly Glu Asp Glu Thr Asp Val Ser Trp Val Gln Ser Leu Val Ser Asn

595 600 605

Ala Glu Leu Asn Gly Lys Arg Gly Glu Val Gln Gly Met Pro Gly Thr

610 6l5 620

Ser Ala Leu Met Asn Arg Pro Asp Leu Asn Asn Gln Gly Asp Leu Leu

625 630 635 640

Asp Gln Thr Val Ile Gly Ala Trp Leu Glu Gln Met His Leu Asp Gln

645 650 655

Lys

<210>22

<211>2223

<212>DNA

＜213〉Arabidopis thaliana

<400>22

ttcttcaaaa accccaacca cttcttctcc ccaaaaacct ccaaagtttc aatctttact 60

tctctctttt tctccaagtt atcttctttt ctaggaagag atatgtgcgg tgcaaagagc 120

aacctttgct catctaaaac cctaacagaa gtcgaattca tgaggcagaa atcagaagac 180

ggagcttccg ccacgtgtct cctcgaattc gccgcctgtg atgatctttc atcgtttaag 240

agagagatcg aagagaatcc atcggtggag attgatgagt cagggttttg gtattgcaga 300

cgggtcgggt ctaagaagat gggttttgaa gaaagaacac cacttatggt tgctgctatg 360

tatggaagca tggaagtgtt gaattacata attgccacag gaagatccga tgtgaacaga 420

gtttgcagtg acgagaaagt cactgctctt cactgtgcag tttctggctg ttctgtttct 480

atcgttgaga tcatcaagat cttgcttgat gcttctgctt cacctaattg tgttgacgct 540

aatgggaaca aaccggttga tttgttggct aaagattctc ggtttgttcc taaccagagt 600

agaaaggcgg ttgaggtttt actgaccggg attcatggtt cggttatgga agaagaggag 660

gaggaactga agagtgttgt gactaagtat ccagctgatg catcacttcc tgatattaac 720

gaaggtgttt atggaactga tgattttagg atgtttagct ttaaggttaa gccatgttct 780

agggcttatt cacatgattg gactgaatgt ccttttgttc atcctggtga gaatgcaagg 840

aggagagatc ctaggaagta tccttacact tgtgtgcctt gtcccgagtt tcgtaaaggg 900

tcttgtccta aaggagattc gtgtgagtac gcgcacggtg ttttcgagtc ttggcttcac 960

ccggcgcagt ataggacacg gctttgcaaa gatgagactg gttgtgctag gagagtttgt 1020

ttctttgctc atagacggga tgagttaaga ccggttaatg cttctactgg ttctgcaatg 1080

gtttcaccaa ggtcgtctaa tcagtctcct gagatgtctg ttatgtctcc tttgacgctg 1140

ggatcatcgc caatgaactc tcctatggct aatggtgttc ctttgtctcc aagaaatggt 1200

ggtttatggc agaacagagt taatagcctt acaccaccac cgttgcagct taatggtagc 1260

agattgaagt cgactttgag tgctagagat atggatatgg agatggaact taggtttcgc 1320

ggtttggata accggagact tggtgatctc aagccatcca acctcgaaga gactttcgga 1380

tcatatgact cagcttctgt gatgcaactt caatcaccaa gcaggcattc tcagatgaac 1440

cactatccgt cttcacctgt gaggcagcct cctcctcatg gattcgaatc ttcagcagcc 1500

atggcagctg cagtgatgaa tgcaagatcc tcagcgtttg cgaaacgcag cttgagtttc 1560

aaaccagctc cagtagcttc taatgtctcc gattggggat caccaaatgg gaagcttgag 1620

tggggaatgc aaagagatga gctgaacaag ttgaggagaa gtgcctcctt cggcattcat 1680

ggaaacaaca acaacagtgt gtcacgccct gctagagact acagtgacga gccagatgtg 1740

tcgtgggtga actcactggt gaaagagaat gcaccagaga gagtgaatga gagggttggg 1800

aatacggtga atggtgcagc gagtagagac aagtttaagc tgccgtcgtg ggcagagcaa 1860

atgtatatag accatgagca gcagattgtg gcataagaag cagaaagaaa gatgtgggat 1920

ttatattgct tttgtcttct gggcctctct acacagaatc taacaaatct ggcaataatt 1980

ctttgatttg tgtttgaccc atagtttggt tactagtata tgttttttta tgttcttttt 2040

ttctttgtca ttctcttgtc cttcgtgaca ctatgtaatg attaaaagca aataattgat 2100

gcatgagttc aaatgttctt tgaaggatcc atcttattag ctttgtaatt gttgtgatat 2160

cttaatctta ttggttacgt atttcaagtg ctttagaaaa aatgggccta agagattttg 2220

ggg 2223

<210>23

<211>597

<212>PRT

＜213〉Arabidopis thaliana

<400>23

Met Cys Gly Ala Lys Ser Asn Leu Cys Ser Ser Lys Thr Leu Thr Glu

1 5 10 15

Val Glu Phe Met Arg Gln Lys Ser Glu Asp Gly Ala Ser Ala Thr Cys

20 25 30

Leu Leu Glu Phe Ala Ala Cys Asp Asp Leu Ser Ser Phe Lys Arg Glu

35 40 45

Ile Glu Glu Asn Pro Ser Val Glu Ile Asp Glu Ser Gly Phe Trp Tyr

50 55 60

Cys Arg Arg Val Gly Ser Lys Lys Met Gly Phe Glu Glu Arg Thr Pro

65 70 75 80

Leu Met Val Ala Ala Met Tyr Gly Ser Met Glu Val Leu Asn Tyr Ile

85 90 95

Ile Ala Thr Gly Arg Ser Asp Val Asn Arg Val Cys Ser Asp Glu Lys

100 105 110

Val Thr Ala Leu His Cys Ala Val Ser Gly Cys Ser Val Ser Ile Val

115 120 125

Glu Ile Ile Lys Ile Leu Leu Asp Ala Ser Ala Ser Pro Asn Cys Val

130 135 140

Asp Ala Asn Gly Asn Lys Pro Val Asp Leu Leu Ala Lys Asp Ser Arg

145 150 155 160

Phe Val Pro Asn Gln Ser Arg Lys Ala Val Glu Val Leu Leu Thr Gly

165 170 175

Ile His Gly Ser Val Met Glu Glu Glu Glu Glu Glu Leu Lys Ser Val

180 185 190

Val Thr Lys Tyr Pro Ala Asp Ala Ser Leu Pro Asp Ile Asn Glu Gly

195 200 205

Val Tyr Gly Thr Asp Asp Phe Arg Met Phe Ser Phe Lys Val Lys Pro

210 215 220

Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His

225 230 235 240

Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr Pro Tyr Thr

245 250 255

Cys Val Pro Cys Pro Glu Phe Arg Lys Gly Ser Cys Pro Lys Gly Asp

260 265 270

Ser Cys Glu Tyr Ala His Gly Val Phe Glu Ser Trp Leu His Pro Ala

275 280 285

Gln Tyr Arg Thr Arg Leu Cys Lys Asp Glu Thr Gly Cys Ala Arg Arg

290 295 300

Val Cys Phe Phe Ala His Arg Arg Asp Glu Leu Arg Pro Va1 Asn Ala

305 310 315 320

Ser Thr Gly Ser Ala Met Val Ser Pro Arg Ser Ser Asn Gln Ser Pro

325 330 335

Glu Met Ser Val Met Ser Pro Leu Thr Leu Gly Ser Ser Pro Met Asn

340 345 350

Ser Pro Met Ala Asn Gly Val Pro Leu Ser Pro Arg Asn Gly Gly Leu

355 360 365

Trp Gln Asn Arg Val Asn Ser Leu Thr Pro Pro Pro Leu Gln Leu Asn

370 375 380

Gly Ser Arg Leu Lys Ser Thr Leu Ser Ala Arg Asp Met Asp Met Glu

385 390 395 400

Met Glu Leu Arg Phe Arg Gly Leu Asp Asn Arg Arg Leu Gly Asp Leu

405 410 415

Lys Pro Ser Asn Leu Glu Glu Thr Phe Gly Ser Tyr Asp Ser Ala Ser

420 425 430

Val Met Gln Leu Gln Ser Pro Ser Arg His Ser Gln Met Asn His Tyr

435 440 445

Pro Ser Ser Pro Val Arg Gln Pro Pro Pro His Gly Phe Glu Ser Ser

450 455 460

Ala Ala Met Ala Ala Ala Val Met Asn Ala Arg Ser Ser Ala Phe Ala

465 470 475 480

Lys Arg Ser Leu Ser Phe Lys Pro Ala Pro Val Ala Ser Asn Val Ser

485 490 495

Asp Trp Gly Ser Pro Asn Gly Lys Leu Glu Trp Gly Met Gln Arg Asp

500 505 510

Glu Leu Asn Lys Leu Arg Arg Ser Ala Ser Phe Gly Ile His Gly Asn

515 520 525

Asn Asn Asn Ser Val Ser Arg Pro Ala Arg Asp Tyr Ser Asp Glu Pro

530 535 540

Asp Val Ser Trp Val Asn Ser Leu Val Lys Glu Asn Ala Pro Glu Arg

545 550 555 560

Val Asn Glu Arg Val Gly Asn Thr Val Asn Gly Ala Ala Ser Arg Asp

565 570 575

Lys Phe Lys Leu Pro Ser Trp Ala Glu Gln Met Tyr Ile Asp His Glu

580 585 590

Gln Gln Ile Val Ala

595

<210>24

<211>1761

<212>DNA

＜213〉Arabidopis thaliana

<400>24

atggaaaaag atagtattat gtgcagtgga ccaaagagca atctctgctc ttcaagaacc 60

ttaacagaaa tcgaatcaag gcaaaaggaa gaagaaacaa tgcttctcct cgaattcgct 120

gcttgtgatg atcttgactc gttcaagaga gaggttgaag agaaagggct tgatttggat 180

gagtcagggt tatggtattg cagacgtgtc ggttctaaga agatgggtct tgaagaaaga 240

acacctttaa tggttgcagc tatgtatgga agcataaagg ttttgacttt catcgtttcc 300

actggaaaat ctgatgtgaa cagagcttgt ggtgaagaga gagttactcc gcttcactgt 360

gctgttgctg gctgttctgt gaatatgatt gaagtcatca atgtcttgct tgatgcttct 420

gctttggtta actctgttga tgctaatggg aatcaacctt tggatgtgtt tgttcgagtt 480

tcgaggtttg tggctagtcc gaggaggaaa gcggttgagt tgttgctgag aggaggaggt 540

gttggaggat tgatcgatga ggcggttgaa gaagagatca agattgtctc taagtatcca 600

gctgatgctt ctttaccgga tataaacgaa ggggtttatg gaagtgatga gtttaggatg 660

tatagcttta aggttaagcc atgttctagg gcttattctc atgattggac cgagtgtgct 720

tttgttcatc cgggagaaaa tgcgaggagg agagatccga ggaagtatcc ttacacttgt 780

gtcccctgtc ccgagttccg taaaggatca tgcccgaaag gagattcttg cgagtatgct 840

cacggggttt tcgagtcgtg gcttcacccc gcgcagtata aaacccggct ttgtaaagat 900

gaaacgggtt gtgcaaggaa agtttgtttc tttgctcata aacgcgaaga gatgagacct 960

gttaatgctt caactggctc tgccgtggct cagtctccgt ttagcagctt ggagatgatg 1020

ccagggttgt ctcctcttgc ttattcttca ggagtttcga ctcctccggt ttctccaatg 1080

gctaatggtg ttccttcctc tccaagaaac ggcggatcat ggcagaacag agtcaatacc 1140

cttactccac cggctttgca gctcaatggt ggaagcagat tgaagtccac actgagcgct 1200

agagatatcg atatggagat ggagatggaa ttgagactcc gcggttttgg caacaatgtg 1260

gaagagacgt tcgggtctta tgtttcctct ccaagtagga attctcaaat gggtcaaaac 1320

atgaaccaac attatccatc ttccccggtg agacaaccgc catctcaaca cgggttcgaa 1380

tcttcagcag ctgcagcggt tgcagtgatg aaagcgagat caaccgcctt tgcgaaacgt 1440

agcttgagct tcaaaccagc tactcaagca gcaccacagt cgaatctctc ggattgggga 1500

tctccaaacg ggaagctgga atggggaatg aaaggagaag agctgaataa gatgagaaga 1560

agtgtttcct ttggaatcca tggaaacaac aacaataacg cagctagaga ctacagggac 1620

gagccagatg tgtcatgggt taactcttta gttaaagaca gtactgtggt gtctgagaga 1680

agctttggaa tgaatgagag ggttcggata atgtcgtggg ctgagcaaat gtacagagag 1740

aaggagcaga ctgtggtgta a 1761

<210>25

<211>586

<212>PRT

＜213〉Arabidopis thaliana

<400>25

Met Glu Lys Asp Ser Ile Met Cys Ser Gly Pro Lys Ser Asn Leu Cys

1 5 10 15

Ser Ser Arg Thr Leu Thr Glu Ile Glu Ser Arg Gln Lys Glu Glu Glu

20 25 30

Thr Met Leu Leu Leu Glu Phe Ala Ala Cys Asp Asp Leu Asp Ser Phe

35 40 45

Lys Arg Glu Val Glu Glu Lys Gly Leu Asp Leu Asp Glu Ser Gly Leu

50 55 60

Trp Tyr Cys Arg Arg Val Gly Ser Lys Lys Met Gly Leu Glu Glu Arg

65 70 75 80

Thr Pro Leu Met Val Ala Ala Met Tyr Gly Ser Ile Lys Val Leu Thr

85 90 95

Phe Ile Val Ser Thr Gly Lys Ser Asp Val Asn Arg Ala Cys Gly Glu

100 105 110

Glu Arg Val Thr Pro Leu His Cys Ala Val Ala Gly Cys Ser Val Asn

115 120 125

Met Ile Glu Val Ile Asn Val Leu Leu Asp Ala Ser Ala Leu Val Asn

130 135 140

Ser Val Asp Ala Asn Gly Asn Gln Pro Leu Asp Val Phe Val Arg Val

145 150 155 160

Ser Arg Phe Val Ala Ser Pro Arg Arg Lys Ala Val Glu Leu Leu Leu

165 170 175

Arg Gly Gly Gly Val Gly Gly Leu Ile Asp Glu Ala Val Glu Glu Glu

180 185 190

Ile Lys Ile Val Ser Lys Tyr Pro Ala Asp Ala Ser Leu Pro Asp Ile

195 200 205

Asn Glu Gly Val Tyr Gly Ser Asp Glu Phe Arg Met Tyr Ser Phe Lys

210 215 220

Val Lys Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Ala

225 230 235 240

Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr

245 250 255

Pro Tyr Thr Cys Val Pro Cys Pro Glu Phe Arg Lys Gly Ser Cys Pro

260 265 270

Lys Gly Asp Ser Cys Glu Tyr Ala His Gly Val Phe Glu Ser Trp Leu

275 280 285

His Pro Ala Gln Tyr Lys Thr Arg Leu Cys Lys Asp Glu Thr Gly Cys

290 295 300

Ala Arg Lys Val Cys Phe Phe Ala His Lys Arg Glu Glu Met Arg Pro

305 310 315 320

Val Asn Ala Ser Thr Gly Ser Ala Val Ala Gln Ser Pro Phe Ser Ser

325 330 335

Leu Glu Met Met Pro Gly Leu Ser Pro Leu Ala Tyr Ser Ser Gly Val

340 345 350

SerThr Pro Pro Val Ser Pro Met Ala Asn Gly Val Pro Ser Ser Pro

355 360 365

Arg Asn Gly Gly Ser Trp Gln Asn Arg Val Asn Thr Leu Thr Pro Pro

370 375 380

Ala Leu Gln Leu Asn Gly Gly Ser Arg Leu Lys Ser Thr Leu Ser Ala

385 390 395 400

Arg Asp Ile Asp Met Glu Met Glu Met Glu Leu Arg Leu Arg Gly Phe

405 410 415

Gly Asn Asn Val Glu Glu Thr Phe Gly Ser Tyr Val Ser Ser Pro Ser

420 425 430

Arg Asn Ser Gln Met Gly Gln Asn Met Asn Gln His Tyr Pro Ser Ser

435 440 445

Pro Val Arg Gln Pro Pro Ser Gln His Gly Phe Glu Ser Ser Ala Ala

450 455 460

Ala Ala Val Ala Val Met Lys Ala Arg Ser Thr Ala Phe Ala Lys Arg

465 470 475 480

Ser Leu Ser Phe Lys Pro Ala Thr Gln Ala Ala Pro Gln Ser Asn Leu

485 490 495

Ser Asp Trp Gly Ser Pro Asn Gly Lys Leu Glu Trp Gly Met Lys Gly

500 505 510

Glu Glu Leu Asn Lys Met Arg Arg Ser Val Ser Phe Gly Ile His Gly

515 520 525

Asn Asn Asn Asn Asn Ala Ala Arg Asp Tyr Arg Asp Glu Pro Asp Val

530 535 540

Ser Trp Val Asn Ser Leu Val Lys Asp Ser Thr Val Val Ser Glu Arg

545 550 555 560

Ser Phe Gly Met Asn Glu Arg Val Arg Ile Met Ser Trp Ala Glu Gln

565 570 575

Met Tyr Arg Glu Lys Glu Gln Thr Val Val

580 585

<210>26

<211>2709

<212>DNA

＜213〉big eucalyptus

<400>26

cttctgaaag ctttttgact taagacgaga gagaaggaga gaaggtcccc ctcctcgtcc 60

tcgtcccccc gtggattttg aagaagaaaa gtcgcacctt ccttctcctt tcccactcct 120

ccctctgctc gaagcttttc tcttccgcag aattacataa aaacctcgac tttgcgcatc 180

attccgattc acctcacacc ttcactttcc cactcgaggt ctcccccctc ttttcctagc 240

tctttccctt tccctccctc tctctcgaga atcgccgcat ttggaggagc tccaatctgc 300

tttgctttgc tttgctctct tcttgctcgg ttccccctca taaggagtcg attatgtgca 360

acggttcttc gaagggtaaa cttttcccct cgagtatggg catggagggc gaattccaca 420

acaaggatgg cgaagcaccc cgtaaatgct ctgccttgct tgaattggca gcctcggacg 480

atctctcgtc gttcaaaagt gaagtggaag agaagggctg cgacgttgat gaggccagct 540

tttggtatgg taggagaatc gggtcgaaga agatgggttt tgaagagagg actccattga 600

tgatctctgc tttgtttgga agcaccaagg tcttgaaata cataatcgag accgccagag 660

ctgatgtcaa caggtcttgt gggtccgaca aggtggccgc cctccattgc gcagccgcgg 720

gtgggtccag ttcttcactt gaaattgtga agctcttgat tgaggcctca gcggatatta 780

attctgtaga tggcaatgga aataggccca tcgacgtgct tgccccggca gggaagtctc 840

gctgcaattc cagaaataag tttgttagat cgttgctgaa aggtgaaaac tatgtcgtgg 900

aaggtgacca atcctttgac atagaaggag aggagaagct agtcgctctt ccaaaggagg 960

gaggcgagaa gaaagagtat cctgttgatg tctctctacc tgacataaac aatgggttct 1020

acagtaccga tgagttccgg atgtatgctt tcaaggtgaa gccttgctcg agggcttact 1080

cccacgactg gaccgagtgc ccgtttgtgc accctgggga gaacgcgagg aggagggacc 1140

cacgcaagta cccttacagc tgtgtccctt gtcctgagtt tcgcaagggt tcgtgcgtaa 1200

ggggggatgc ttgtgagtat gctcatggag tctttgagtc gtggcttcac ccagcgcaat 1260

accgaacccg gctgtgcaag gatgaaactg gttgtactcg caaagtttgc ttctttgctc 1320

acaagtccga agaattgcgt cccgtgtatg cttccacagg ttctgctatg ccctcaccca 1380

agtccttttc agctaatgcc ctagacatga caaccctgag ccccttatcc cttaattcac 1440

catctctgcc tttgcctgct acttccacgc cccccatgtc acctttggct gcctcatctt 1500

cacccaaggg catgaacttg tggcataaca aaattaacct gaccccacca agcctgcagc 1560

ttcctggcag ccggctgaag acggctatga gtgcgcggga cttcgatttt gagttggaat 1620

ttcttgggct ggaaaagcaa gcttctcagc ggcagcaact gatagaagag atttctcgtc 1680

tctcatcgcc ctctcatatg tggaactcgg aatttggcag aaccgcagag ctgaagccca 1740

ctaaccttga tgatgcgttt ggatctcttg acacttctct tttgtctccg ttgcaggggt 1800

cgtcgatgaa aacatcgact cctacccagt tgcaatcccc cacagggctt aaaatttcga 1860

atttgaacca actccgtgcg agctacccgt ctagcagctt gtcgtcctct cctgtgagga 1920

agacctcttc ttttgggttc gactcatcca gtgcagttgc tgcagcagtc atgaactcac 1980

ggtctgctgc tatgacgaag cggagccaga gcttcattga ccgtggagca gtgggtcaac 2040

ggtctggact cattggacct gctaattctg ctcctaggat gtccaacctt tcggactggg 2100

gctcgcctga tgggaagttg gattggggtg ttcaagggga cgagctcaac aagcttagga 2160

agtccgcttc cttcggcttt agaaacaaca gtatggcgaa cccaaacaac gtggcgtctc 2220

ccagtgctga tgagccggac gtgtcgtggg ttggttcatt ggtgaaggat gtggctccgc 2280

ccgaagggta tccacagtat ctgtacatag aacaggagca gatggtggca taactaaagc 2340

gaagagcacc acacgaactc tctcctgatg gcttaagatg acttgtttga cattctttat 2400

attcttacaa acagcgcgtt cttaggagtt agctggagga aagaaggaaa cggtattgag 2460

tttgagattc aggctcttag ctggacagcg aaaatttggg gaaggaagag aatttggttt 2520

cttgcccaac ttagataatg atgcttttga aggcttaaaa gaaagatgaa ggcaaacatt 2580

cttttgttag tattgtatta ttgttttaat ttttcatccc ctctgtcggg gtgtggtggg 2640

tgtcgatgtt tctttcatca gtaaaatata taatgaggtt tactcatcta ttttctacta 2700

aaaaaaaaa 2709

<210>27

<211>659

<212>PRT

＜213〉big eucalyptus

<400>27

Met Cys Asn Gly Ser Ser Lys Gly Lys Leu Phe Pro Ser Ser Met Gly

1 5 10 15

Met Glu Gly Glu Phe His Asn Lys Asp Gly Glu Ala Pro Arg Lys Cys

20 25 30

Ser Ala Leu Leu Glu Leu Ala Ala Ser Asp Asp Leu Ser Ser Phe Lys

35 40 45

Ser Glu Val Glu Glu Lys Gly Cys Asp Val Asp Glu Ala Ser Phe Trp

50 55 60

Tyr Gly Arg Arg Ile Gly Ser Lys Lys Met Gly Phe Glu Glu Arg Thr

65 70 75 80

Pro Leu Met Ile Ser Ala Leu Phe Gly Ser Thr Lys Val Leu Lys Tyr

85 90 95

Ile Ile Glu Thr Ala Arg Ala Asp Val Asn Arg Ser Cys Gly Ser Asp

100 105 110

Lys Val Ala Ala Leu His Cys Ala Ala Ala Gly Gly Ser Ser Ser Ser

115 120 125

Leu Glu Ile Val Lys Leu Leu Ile Glu Ala Ser Ala Asp Ile Asn Ser

130 135 140

Val Asp Gly Asn Gly Asn Arg Pro Ile Asp Val Leu Ala Pro Ala Gly

145 150 155 160

Lys Ser Arg Cys Asn Ser Arg Asn Lys Phe Val Arg Ser Leu Leu Lys

165 170 175

Gly Glu Asn Tyr Val Val Glu Gly Asp Gln Ser Phe Asp Ile Glu Gly

180 185 190

Glu Glu Lys Leu Val Ala Leu Pro Lys Glu Gly Gly Glu Lys Lys Glu

195 200 205

Tyr Pro Val Asp Val Ser Leu Pro Asp Ile Asn Asn Gly Phe Tyr Ser

210 215 220

Thr Asp Glu Phe Arg Met Tyr Ala Phe Lys Val Lys Pro Cys Ser Arg

225 230 235 240

Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu

245 250 255

Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr Pro Tyr Ser Cys Val Pro

260 265 270

Cys Pro Glu Phe Arg Lys Gly Ser Cys Val Arg Gly Asp Ala Cys Glu

275 280 285

Tyr Ala His Gly Val Phe Glu Ser Trp Leu His Pro Ala Gln Tyr Arg

290 295 300

Thr Arg Leu Cys Lys Asp Glu Thr Gly Cys Thr Arg Lys Val Cys Phe

305 310 315 320

Phe Ala His Lys Ser Glu Glu Leu Arg Pro Val Tyr Ala Ser Thr Gly

325 330 335

Ser Ala Met Pro Ser Pro Lys Ser Phe Ser Ala Asn Ala Leu Asp Met

340 345 350

Thr Thr Leu Ser Pro Leu Ser Leu Asn Ser Pro Ser Leu Pro Leu Pro

355 360 365

Ala Thr Ser Thr Pro Pro Met Ser Pro Leu Ala Ala Ser Ser Ser Pro

370 375 380

Lys Gly Met Asn Leu Trp His Asn Lys Ile Asn Leu Thr Pro Pro Ser

385 390 395 400

Leu Gln Leu Pro Gly Ser Arg Leu Lys Thr Ala Met Ser Ala Arg Asp

405 410 415

Phe Asp Phe Glu Leu Glu Phe Leu Gly Leu Glu Lys Gln Ala Ser Gln

420 425 430

Arg Gln Gln Leu Ile Glu Glu Ile Ser Arg Leu Ser Ser Pro Ser His

435 440 445

Met Trp Asn Ser Glu Phe Gly Arg Thr Ala Glu Leu Lys Pro Thr Asn

450 455 460

Leu Asp Asp Ala Phe Gly Ser Leu Asp Thr Ser Leu Leu Ser Pro Leu

465 470 475 480

Gln Gly Ser Ser Met Lys Thr Ser Thr Pro Thr Gln Leu Gln Ser Pro

485 490 495

Thr Gly Leu Lys Ile Ser Asn Leu Asn Gln Leu Arg Ala Ser Tyr Pro

500 505 510

Ser Ser Ser Leu Ser Ser Ser Pro Val Arg Lys Thr Ser Ser Phe Gly

515 520 525

Phe Asp Ser Ser Ser Ala Val Ala Ala Ala Val Met Asn Ser Arg Ser

530 535 540

Ala Ala Met Thr Lys Arg Ser Gln Ser Phe Ile Asp Arg Gly Ala Val

545 550 555 560

Gly Gln Arg Ser Gly Leu Ile Gly Pro Ala Asn Ser Ala Pro Arg Met

565 570 575

Ser Asn Leu Ser Asp Trp Gly Ser Pro Asp Gly Lys Leu Asp Trp Gly

580 585 590

Val Gln Gly Asp Glu Leu Asn Lys Leu Arg Lys Ser Ala Ser Phe Gly

595 600 605

Phe Arg Asn Asn Ser Met Ala Asn Pro Asn Asn Val Ala Ser Pro Ser

610 615 620

Ala Asp Glu Pro Asp Val Ser Trp Val Gly Ser Leu Val Lys Asp Val

625 630 635 640

Ala Pro Pro Glu Gly Tyr Pro Gln Tyr Leu Tyr Ile Glu Gln Glu Gln

645 650 655

Met Val Ala

<210>28

<211>2518

<212>DNA

＜213〉big eucalyptus

<400>28

tctccttcga gtttctttct tcactagaat tcgctcccga gtctgttgtt gctcgtgtag 60

ttttgcttac tccgtccttc gtttagctcg ctgaccagcg cggagctagg agcggtcgct 120

aaaggattac tcgtacaaaa cgtaaactca gctctgccaa ttttcccatg gagggggaat 180

cttacttcga gaaagatgaa aaatattcta attgctcaat cttgctcgaa ttatctgctt 240

cggacgatct cccagctttt gaaaggaaag cgaaagagaa gggctgtaac attgatggtg 300

ctagcttctg gtacggtaga agaattggct caaggaagat gggtcttgaa gagaggactc 360

ctctcatggt ggcttccttg tttggaagct ctagggttgt gaagtacatt ctcgaatctg 420

gcaaagtcga tgtaaatagg gcttgtggtt cggacaaggt cactgccctt cactgtgctg 480

ttgccagtgg ctctgcttct gcggtggagg ttgtcaagct cttgcttcac gcatctgccg 540

atgctaattg cattgatggc aatggaaaga agccaattga tgtgatagcc cttccattaa 600

agtcacgcgg cgattcaagg aggaagctga tggagctgtt gctgaaaggc gataattctg 660

atggggaatt tgaatcccac gaggagaagc cgattgccgc accgcaagca tccaaagagg 720

gaagcgaaaa gaaagagtat caatttcctg ttgatatctc tctgcctgac ataaatgttg 780

ggatttacag tactgatgag ttcagaatgt atgctttcaa agtaaagcct tgctcgcggg 840

catactccca tgactggaca gagtgcccat ttgttcatcc tggcgagaat gcgaggaggc 900

gggaccctcg caagtacccc tacagctgcg tcccttgccc tgaatttcgg aagggatctt 960

gecaaaaggg tgactcctgt gagtacgcgc acggcgtatt tgagtcgtgg cttcatcctg 1020

cacagtatag aacaagactg tgcaaggatg agactggatg tgctcgcaaa gtttgtttct 1080

ttgctcacaa gcccgaagaa ttaaggcctg tctatgcttc gacgggatca gctatgcctt 1140

ccccaaaatc ctactcatca agtgggctgg acatgtccac attgagtcct ctctcaatca 1200

gttctccgtc agcatcgttg cctgttactt caacagcacc catgtctcct cttgcagcct 1260

cgtcatctcc gatgtctgtg aacatgtggc agagcaaggc taacaagctc tccccgccaa 1320

tgctgcagct ctcaggtagt aggctgaaga ctgctttgag tgctagggac ttggacctgg 1380

agatggaatt gcgtggtcta gagagtcaga tggccactca acagcatcag ttgatggaag 1440

agatatctcg tctctcctca ccatcatcct gctttagtag taggattggg gaagtgaaac 1500

ccactaacct cgatgacgtt tttgggtctc cggatcctgc tttgctgcct caattgcagg 1560

ggctgtcaag accttcaaca ccaagccagt tgcaatctcc aactgggctt cagatgcgcc 1620

agaatgcaac ccagtttcgt ggggcgtacc agagcaatgc aaatgcattg tcatctccag 1680

caatgaagca ggcaccttct tatgggtttg actcatctag tgcagttgca gcagcggtga 1740

tgaattcgag gtcagccgct tttgcgaagc ggagtcagag ttttatcgac aggggaatgg 1800

cgtgccctgg aattgccaat tcttccccta tgatgtcttc agctatgtcg agctggagct 1860

cacctcatgg gaaattggat tggggcgtcc aaggagatga gttgaatagg ctgaggaaag 1920

ctgcttcctt taagatgaga agcagcaccg gagcaggtgc taatactgtc tcggcagcag 1980

ccatggctga tgagccagat atttcttggg tcagttcatt ggttaaggac gtgccttctg 2040

cggaggacgc gatgttcgct gcagagaaag gacagcgcac ttatgggaaa gacatccgcg 2100

aaaggattac cccatgggtg gagcagctgt acagagaagt gccacggatg gcgatgtaag 2160

attgccactg caagtcggat gccttagtat gctgactaat tgatattctt tgcatttgtt 2220

ttgaggcatt tggtagccat tagatacgag aaaaggccaa gcagcaggtg gtgtcttggc 2280

aaggaatagg atgcacatag tctgttatcg agtagaatag acttgggaac aatggttata 2340

gccaaatgtt aaaagttatg atattctttt ccaattcttt ctcttcctca tagtaggttt 2400

ctcaccaagt cttttagtga gagcctgcgg gatgtactat atgtttccct tatgtaacgt 2460

ctcttcgttg aaagaaatgg ctttataata taaagcatca agttttttaa aaaaaaaa 2518

<210>29

<211>663

<212>PRT

＜213〉big eucalyptus

<400>29

Met Glu Gly Glu Ser Tyr Phe Glu Lys Asp Glu Lys Tyr Ser Asn Cys

1 5 10 15

Ser Ile Leu Leu Glu Leu Ser Ala Ser Asp Asp Leu Pro Ala Phe Glu

20 25 30

Arg Lys Ala Lys Glu Lys Gly Cys Asn Ile Asp Gly Ala Ser Phe Trp

35 40 45

Tyr Gly Arg Arg Ile Gly Ser Arg Lys Met Gly Leu Glu Glu Arg Thr

50 55 60

Pro Leu Met Val Ala Ser Leu Phe Gly Ser Ser Arg Val Val Lys Tyr

65 70 75 80

Ile Leu Glu Ser Gly Lys Val Asp Val Asn Arg Ala Cys Gly Ser Asp

85 90 95

Lys Val Thr Ala Leu His Cys Ala Val Ala Ser Gly Ser Ala Ser Ala

100 105 110

Val Glu Val Val Lys Leu Leu Leu His Ala Ser Ala Asp Ala Asn Cys

115 120 125

Ile Asp Gly Asn Gly Lys Lys Pro Ile Asp Val Ile Ala Leu Pro Leu

130 135 140

Lys Ser Arg Gly Asp Ser Arg Arg Lys Leu Met Glu Leu Leu Leu Lys

145 150 155 160

Gly Asp Asn Ser Asp Gly Glu Phe Glu Ser His Glu Glu Lys Pro Ile

165 170 175

Ala Ala Pro Gln Ala Ser Lys Glu Gly Ser Glu Lys Lys Glu Tyr Gln

180 185 190

Phe Pro Val Asp Ile Ser Leu Pro Asp Ile Asn Val Gly Ile Tyr Ser

195 200 205

Thr Asp Glu Phe Arg Met Tyr Ala Phe Lys Val Lys Pro Cys Ser Arg

210 215 220

Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu

225 230 235 240

Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr Pro Tyr Ser Cys Val Pro

245 250 255

Cys Pro Glu Phe Arg Lys Gly Ser Cys Gln Lys Gly Asp Ser Cys Glu

260 265 270

Tyr Ala His Gly Val Phe Glu Ser Trp Leu His Pro Ala Gln Tyr Arg

275 280 285

Thr Arg Leu Cys Lys Asp Glu Thr Gly Cys Ala Arg Lys Val Cys Phe

290 295 300

Phe Ala His Lys Pro Glu Glu Leu Arg Pro Val Tyr Ala Ser Thr Gly

305 310 315 320

Ser Ala Met Pro Ser Pro Lys Ser Tyr Ser Ser Ser Gly Leu Asp Met

325 330 335

Ser Thr Lcu Ser Pro Leu Ser Ile Ser Ser Pro Ser Ala Ser Leu Pro

340 345 350

Val Thr Ser Thr Ala Pro Met Ser Pro Leu Ala Ala Ser Ser Ser Pro

355 360 365

Met Ser Val Asn Met Trp Gln Ser Lys Ala Asn Lys Leu Ser Pro Pro

370 375 380

Met Leu Gln Leu Ser Gly Ser Arg Leu Lys Thr Ala Leu Ser Ala Arg

385 390 395 400

Asp Leu Asp Leu Glu Met Glu Leu Arg Gly Leu Glu Ser Gln Met Ala

405 410 415

Thr Gln Gln His Gln Leu Met Glu Glu Ile Ser Arg Leu Ser Ser Pro

420 425 430

Ser Ser Cys Phe Ser Ser Arg Ile Gly Glu Val Lys Pro Thr Asn Leu

435 440 445

Asp Asp Val Phe Gly Ser Pro Asp Pro Ala Leu Leu Pro Gln Leu Gln

450 455 460

Gly Leu Ser Arg Pro Ser Thr Pro Ser Gln Leu Gln Ser Pro Thr Gly

465 470 475 480

Leu Gln Met Arg Gln Asn Ala Thr Gln Phe Arg Gly Ala Tyr Gln Ser

485 490 495

Asn Ala Asn Ala Leu Ser Ser Pro Ala Met Lys Gln Ala Pro Ser Tyr

500 505 510

Gly Phe Asp Ser Ser Ser Ala Val Ala Ala Ala Val Met Asn Ser Arg

515 520 525

Ser Ala Ala Phe Ala Lys Arg Ser Gln Ser Phe Ile Asp Arg Gly Met

530 535 540

Ala Cys Pro Gly Ile Ala Asn Ser Ser Pro Met Met Ser Ser Ala Met

545 550 555 560

Ser Ser Trp Scr Ser Pro His Gly Lys Leu Asp Trp Gly Val Gln Gly

565 570 575

Asp Glu Leu Asn Arg Leu Arg Lys Ala Ala Ser Phe Lys Met Arg Ser

580 585 590

Ser Thr Gly Ala Gly Ala Asn Thr Val Ser Ala Ala Ala Met Ala Asp

595 600 605

Glu Pro Asp Ile Ser Trp Val Ser Ser Leu Val Lys Asp Val Pro Ser

610 615 620

Ala Glu Asp Ala Met Phe Ala Ala Glu Lys Gly Gln Arg Thr Tyr Gly

625 630 635 640

Lys Asp Ile Arg Glu Arg Ile Thr Pro Trp Val Glu Gln Leu Tyr Arg

645 650 655

Glu Val Pro Arg Met Ala Met

660

<210>30

<211>2001

<212>DNA

＜213〉common wheat (Triticum aestivum)

<220>

<221>misc_feature

<222>(481)..(530)

＜223〉n is a, c, g or t

<400>30

cgaattccgg tcgacgattt ctcgatttcc ttctctataa cacaacgctc tcttctcttg 60

caaccaaagt acttgttcca gtgtctactc tactcaaaaa ggatttggga catcatgtgc 120

agtgattcga aaagtaaact ttcttcccca accctcgtcg tcatggagaa tagtaacatt 180

cagaagcaga atctggatgg tctctacaac tcggttttgc ttgaattgtc tgcatctgat 240

gattatgaag ctttcaaaag agaggtggag gaaaaaggct tagatgtgaa cgaggcaggc 300

ttttggtacg gtagaagaat tgggtcaaag aagatgggat ctgaaacgag gacccctctg 360

atgattgctt ctttgtttgg aagcgccaag gtgctcaatt atattcttct tcagaaagga 420

ggaggtgttg atgtgaacag ggtctgtggt tctgataggg ccactgctct ccattgtgct 480

nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn tcccgggtcg 540

cgatttcgta ggagattcac agagagaaaa gaagcaatta gtgataataa gaaagaatac 600

cctgttgata tatcactgcc agacataaac aacggtgtat atggaacaga tgattttagg 660

atgtacaact tcaaggtgaa gccttgctca agggcttact cccatgactg gaccgagtgt 720

ccattcgttc acccagggga gaacgctagg aggagagacc cacggaaata cccttacagc 780

tgtgttcctt gccctgagtt ccgcaaaggg acctgccaga agggtgattc ctgtgagtat 840

gctcatggtg tttttgagtc ctggetgcat cctgcccaat accggacaag gctttgcaag 900

gatgagactg gctgcgctag aaaagtctgc ttctttgccc acaaacctga agagctacgc 960

cctgtgtatg cttccactgg gtcggctatg ccatcaccaa aatcatattc agctagtgga 1020

cttgacatga cagcgatgag tccattggct ctaagttcca catctttgcc taatgccccc 1080

ccgtttccag cctcacccta tcgtgcgccc tcgttcttct ctcagagtga agctgtgcag 1140

aacaaaataa accttactcc accatcgttg cagctccctg gtagccgact gaaggctgct 1200

ttgagtgcca gggatctgga gatggagatg gaactgctcg gtctagaaag ccctgctcgc 1260

caacaacagc agcagcagca acaattgatc gaagagattg ccaggatctc ttccccatct 1320

ttccggagca aggaattcaa taggattgtt gatttgaatc ctactaacct tgatgacctg 1380

ttagcatctg ctgacccttc tgtattttct caactacatg gactttctgt gcaaccttca 1440

acacccacac aaagtgggct tcagatgcgc caaaacatga accacctccg tgcgagttat 1500

ccatccaaca tcccttcctc tcctgtgagg aagccctcag cttttgggtt tgactcatca 1560

gctgctgtgg caactgcagt gatgaattct aggtctgctg ccttcgcaaa gcgaagccaa 1620

agtttcattg atcgtggagc tgcaacccac catcttgggc tgtcttcagc ttccaactct 1680

tcttgcaggg tatcctctac cctttcagat tggagttccc ctaccgggaa actggattgg 1740

ggtgtaaacg gagacaagct gaacaagctg aggaaatcta cttcctttgg attcagaaac 1800

agtggggtaa ctgcatcccc catagcacag cctgaatttg gtgctgagcc ggatgtctca 1860

tgggttcatt cattggttaa agatgttccc tccgagaggt ctgagatatt tggtgctgag 1920

aagcaacaat atgatctcag taaagagatg cttccaccat ggatggagca gctgtatata 1980

gagcaggagc agatggtagc a 2001

<210>31

<211>667

<212>PRT

＜213〉common wheat

<220>

＜221〉uncertain

<222>(161)..(177)

＜223〉Xaa can make the amino acid of any natural generation

<400>31

Arg Ile Pro Val Asp Asp Phe Ser Ile Ser Phe Ser Ile Thr Gln Arg

1 5 10 15

Ser Leu Leu Leu Gln Pro Lys Tyr Leu Phe Gln Cys Leu Leu Tyr Ser

20 25 30

Lys Arg Ile Trp Asp Ile Met Cys Ser Asp Ser Lys Ser Lys Leu Ser

35 40 45

Ser Pro Thr Leu Val Val Met Glu Asn Ser Asn Ile Gln Lys Gln Asn

50 55 60

Leu Asp Gly Leu Tyr Asn Ser Val Leu Leu Glu Leu Ser Ala Ser Asp

65 70 75 80

Asp Tyr Glu Ala Phe Lys Arg Glu Val Glu Glu Lys Gly Leu Asp Val

85 90 95

Asn Glu Ala Gly Phe Trp Tyr Gly Arg Arg Ile Gly Ser Lys Lys Met

100 105 110

Gly Ser Glu Thr Arg Thr Pro Leu Met Ile Ala Ser Leu Phe Gly Ser

115 120 125

Ala Lys Val Leu Asn Tyr Ile Leu Leu Gln Lys Gly Gly Gly Val Asp

130 135 140

Val Asn Arg Val Cys Gly Ser Asp Arg Ala Thr Ala Leu His Cys Ala

145 150 155 160

Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa Xaa

165 170 175

Xaa Pro Gly Ser Arg Phe Arg Arg Arg Phe Thr Glu Arg Lys Glu Ala

180 185 190

Ile Ser Asp Asn Lys Lys Glu Tyr Pro Val Asp Ile Ser Leu Pro Asp

195 200 205

Ile Asn Asn Gly Val Tyr Gly Thr Asp Asp Phe Arg Met Tyr Asn Phe

210 215 220

Lys Val Lys Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys

225 230 235 240

Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys

245 250 255

Tyr Pro Tyr Ser Cys Val Pro Cys Pro Glu Phe Arg Lys Gly Thr Cys

260 265 270

Gln Lys Gly Asp Ser Cys Glu Tyr Ala His Gly Val Phe Glu Ser Trp

275 280 285

Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Glu Thr Gly

290 295 300

Cys Ala Arg Lys Val Cys Phe Phe Ala His Lys Pro Glu Glu Leu Arg

305 310 315 320

Pro Val Tyr Ala Ser Thr Gly Ser Ala Met Pro Ser Pro Lys Ser Tyr

325 330 335

Ser Ala Ser Gly Leu Asp Met Thr Ala Met Ser Pro Leu Ala Leu Ser

340 345 350

Ser Thr Ser Leu Pro Asn Ala Pro Pro Phe Pro Ala Ser Pro Tyr Arg

355 360 365

Ala Pro Ser Phe Phe Ser Gln Ser Glu Ala Val Gln Asn Lys Ile Asn

370 375 380

Leu Thr Pro Pro Ser Leu Gln Leu Pro Gly Ser Arg Leu Lys Ala Ala

385 390 395 400

Leu Ser Ala Arg Asp Leu Glu Met Glu Met Glu Leu Leu Gly Leu Glu

405 410 415

Ser Pro Ala Arg Gln Gln Gln Gln Gln Gln Gln Gln Leu Ile Glu Glu

420 425 430

Ile Ala Arg Ile Ser Ser Pro Ser Phe Arg Ser Lys Glu Phe Asn Arg

435 440 445

Ile Val Asp Leu Asn Pro Thr Asn Leu Asp Asp Leu Leu Ala Ser Ala

450 455 460

Asp Pro Ser Val Phe Ser Gln Leu His Gly Leu Ser Val Gln Pro Ser

465 470 475 480

Thr Pro Thr Gln Ser Gly Leu Gln Met Arg Gln Asn Met Asn His Leu

485 490 495

Arg Ala Ser Tyr Pro Ser Asn Ile Pro Ser Ser Pro Val Arg Lys Pro

500 505 510

Ser Ala Phe Gly Phe Asp Ser Ser Ala Ala Val Ala Thr Ala Val Met

515 520 525

Asn Ser Arg Ser Ala Ala Phe Ala Lys Arg Ser Gln Ser Phe Ile Asp

530 535 540

Arg Gly Ala Ala Thr His His Leu Gly Leu Ser Ser Ala Ser Asn Ser

545 550 555 560

Ser Cys Arg Val Ser Ser Thr Leu Ser Asp Trp Ser Ser Pro Thr Gly

565 570 575

Lys Leu Asp Trp Gly Val Asn Gly Asp Lys Leu Asn Lys Leu Arg Lys

580 585 590

Ser Thr Ser Phe Gly Phe Arg Asn Ser Gly Val Thr Ala Ser Pro Ile

595 600 605

Ala Gln Pro Glu Phe Gly Ala Glu Pro Asp Val Ser Trp Val His Ser

610 615 620

Leu Val Lys Asp Val Pro Ser Glu Arg Ser Glu Ile Phe Gly Ala Glu

625 630 635 640

Lys Gln Gln Tyr Asp Leu Ser Lys Glu Met Leu Pro Pro Trp Met Glu

645 650 655

Gln Leu Tyr Ile Glu Gln Glu Gln Met Val Ala

660 665

<210>32

<211>2683

<212>DNA

＜213〉big eucalyptus

<400>32

gcaaaggtcg atcacttcct ccctagaaag cgagtgtgga gttgaagctt gataaccaga 60

ggccgcctct cgtctcgtct cgcccgcctg cgcttgctct gctctccgcg tgccaaggga 120

gtgttcctag gtgctgaatc tttccatgtg tagcggttca aaagggaagg gagagtgaat 180

tcgagaagca gaggatgtcg gcccgtcagt tctcgatcct gctcgagtta tctgctgcgg 240

atgatctgac gaactttaag aaagcagttg aggaagacgg ctacgatatt gatgagtcga 300

gcttgtggta tggtaggagg atcgggtcga agaagattgg gcttgaagag agaactcccc 360

tcatgattgc cgcgatgttc ggcagtatgt ccgtgctgga ttatattatc aagtctggcc 420

gggccaatgt aaacaaggcg tgtggttcag atggtgctac cgcgcttcac tgtgctgcgg 480

ctggtggctc ggtacaatct cctgaggtgg tcaagctgtt gcttgattct tcagcgaatg 540

ctaactccat tgatgcgaat gggaaacgag cgggagactt gatttctgag gtctctggtt 600

cgcccttcaa ttcgagaagg aagactttgg atgtcatgtt gactggaggt gggactgttg 660

agtttgttga ggaaacttac aatctgcctg agaatctggg tagtcaaatt gaaggaaacg 720

aacaaagaga gagtccaacg gcccgcgctt ccaaggatgg ttctgaaaag aaagagtatc 780

ctgtcgacct ttctcttccg gacatcaaca atggaatata tagcacagat gagtttagga 840

tgtattcttt caaagtgaag ccttgctcga gagcttactc tcatgactgg actgagtgtc 900

catttgttea ccctggggag aatgcaagac ggcgtgaccc acggaaatat caetacagct 960

gtgtgccttg ccctgagttc cgcaaggggt catgcaggca aggggatggc tgcgagtatg 1020

ctcatggtat atttgagtgc tggcttcacc cagctcaata tcgcacccgt ctctgtaagg 1080

atgagattgg atgcaccaga aaagtctgtt tctttgccca caaacatgaa gagcttcgtc 1140

cattgtatgc atcaactggt tcggcgcttc cttctccaag atcattttcg cccgttgctg 1200

cttctctaga catgggatca ctgagccctc tctctctcgg ttcttcttca gtccggatac 1260

cgccaacttc aacaccacct atgactccat caggggcctc ttctcccctt ggtgggtcga 1320

tgtggaaaag ccaaattaat agcactccgc ctggcttgca gcttccaggt agcaggttga 1380

gaagcgcatt gagtgctaga gacatggatt tagatgttga cttgatcgat ctagaaaata 1440

attatcgttt gcagaagcag ttgctcgaac actttcctga tctgtcctct cctcgtggtt 1500

ggaacaactc ttcatccacc acgtcggctt tccctgagta ttcaggtgac atgactggag 1560

aaataagtag gttaggagta aaaccaaata atctcgagga tagtttcagg tcattggacc 1620

tgaccctctt gtctcagtta caagggctgt cacttgatgg tgcaatatcc cagctgcaat 1680

ctcctactgg aatgaagatt cggcagaaca tgacccagca gctctactca aactatactg 1740

acaagctttc ctcgtcacct agggcaatgc catcatttgg aaccgatcct tccagagctt 1800

cagcagcagc cactctgagt tccaggtcat tggcatttgc aaaaaggagc cacagcttca 1860

ttgagcggag tacagtgaac agtcagtctg gatattcagc aggtgctgct tctccaactg 1920

caaggatgtc ttcccagaat gactggggct cgcccgatgg caaactagac tggggcattc 1980

aaggggagga gctgaacaag ctgaggaaat ctgcatcatt cgggctcagg agcagcagca 2040

accgcttcca tgcgtctgca gattctgcga cagcaactgt aggggaccca gacatgccct 2100

ggattcagtc cttggcaaag gaagccccgt cacaaaaccc tggcaatttt ggagcagagc 2160

atcagcagca gcagcagcag cagcagcagc agcagtatca tcttaattct ggaggtactg 2220

agctgcttcc agcttgggtg gagcagttgt acgcggatca ggagcagatg gtcgcctgag 2280

atcaacattg gcttcttatc taaccactat tagtcatttc gttattgctt taattttttt 2340

tcttctgagt ctagtattaa tgtctaggat tcgaacgaac tggaaaatta aatctagagg 2400

gaagatggga agaaaagagc aggatggaag gtttctgctc ggtccgagat ttctcatagt 2460

ctattataga ctatcgtatt tctcgttctt ttccgtccca atgttcttga tttggttctc 2520

agcatgtttt ctggatgagg cttacaaact atgtaatctt gtcttgctaa aagaatcaga 2580

gctgcacctg caccaaaggt tgtgatacta ccgcttattg atgatgatga taataataat 2640

aattcggaca tttagtacca agtccgatgt ctcaaaaaaa aaa 2683

<210>33

<211>694

<212>PRT

＜213〉big eucalyptus

<400>33

Met Ser Ala Arg Gln Phe Ser Ile Leu Leu Glu Leu Ser Ala Ala Asp

1 5 10 15

Asp Leu Thr Asn Phe Lys Lys Ala Val Glu Glu Asp Gly Tyr Asp Ile

20 25 30

Asp Glu Ser Ser Leu Trp Tyr Gly Arg Arg Ile Gly Ser Lys Lys Ile

35 40 45

Gly Leu Glu Glu Arg Thr Pro Leu Met Ile Ala Ala Met Phe Gly Ser

50 55 60

Met Ser Val Leu Asp Tyr Ile Ile Lys Ser Gly Arg Ala Asn Val Asn

65 70 75 80

Lys Ala Cys Gly Ser Asp Gly Ala Thr Ala Leu His Cys Ala Ala Ala

85 90 95

Gly Gly Ser Val Gln Ser Pro Glu Val Val Lys Leu Leu Leu Asp Ser

100 105 110

Ser Ala Asn Ala Asn Ser Il e Asp Ala Asn Gly Lys Arg Ala Gly Asp

115 120 125

Leu Ile Ser Glu Val Ser Gly Ser Pro Phe Asn Ser Arg Arg Lys Thr

130 135 140

Leu Asp Val Met Leu Thr Gly Gly Gly Thr Val Glu Phe Val Glu Glu

145 150 155 160

Thr Tyr Asn Leu Pro Glu Asn Leu Gly Ser Gln Ile Glu Gly Asn Glu

165 170 175

Gln Arg Glu Ser Pro Thr Ala Arg Ala Ser Lys Asp Gly Ser Glu Lys

180 185 190

Lys Glu Tyr Pro Val Asp Leu Ser Leu Pro Asp Ile Asn Asn Gly Ile

195 200 205

Tyr Ser Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Lys Pro Cys

210 215 220

Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro

225 230 235 240

Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr His Tyr Ser Cys

245 250 255

Val Pro Cys Pro Glu Phe Arg Lys Gly Ser Cys Arg Gln Gly Asp Gly

260 265 270

Cys Glu Tyr Ala His Gly Ile Phe Glu Cys Trp Leu His Pro Ala Gln

275 280 285

Tyr Arg Thr Arg Leu Cys Lys Asp Glu Ile Gly Cys Thr Arg Lys Val

290 295 300

Cys Phe Phe Ala His Lys His Glu Glu Leu Arg Pro Leu Tyr Ala Ser

305 310 315 320

Thr Gly Ser Ala Leu Pro Ser Pro Arg Ser Phe Ser Pro Val Ala Ala

325 330 335

Ser Leu Asp Met Gly Ser Leu Ser Pro Leu Ser Leu Gly Ser Ser Ser

340 345 350

Val Arg Ile Pro Pro Thr Ser Thr Pro Pro Met Thr Pro Ser Gly Ala

355 360 365

Ser Ser Pro Leu Gly Gly Ser Met Trp Lys Ser Gln Ile Asn Ser Thr

370 375 380

Pro Pro Gly Leu Gln Leu Pro Gly Ser Arg Leu Arg Ser Ala Leu Ser

385 390 395 400

Ala Arg Asp Met Asp Leu Asp Val Asp Leu Ile Asp Leu Glu Asn Asn

405 410 415

Tyr Arg Leu Gln Lys Gln Leu Leu Glu His Phe Pro Asp Leu Ser Ser

420 425 430

Pro Arg Gly Trp Asn Asn Ser Ser Ser Thr Thr Ser Ala Phe Pro Glu

435 440 445

Tyr Ser Gly Asp Met Thr Gly Glu Ile Ser Arg Leu Gly Val Lys Pro

450 455 460

Asn Asn Leu Glu Asp Ser Phe Arg Ser Leu Asp Leu Thr Leu Leu Ser

465 470 475 480

Gln Leu Gln Gly Leu Ser Leu Asp Gly Ala Ile Ser Gln Leu Gln Ser

485 490 495

Pro Thr Gly Met Lys Ile Arg Gln Asn Met Thr Gln Gln Leu Tyr Ser

500 505 510

Asn Tyr Thr Asp Lys Leu Ser Ser Ser Pro Arg Ala Met Pro Ser Phe

515 520 525

Gly Thr Asp Pro Ser Arg Ala Ser Ala Ala Ala Thr Leu Ser Ser Arg

530 535 540

Ser Leu Ala Phe Ala Lys Arg Ser His Ser Phe Ile Glu Arg Ser Thr

545 550 555 560

Val Asn Ser Gln Ser Gly Tyr Ser Ala Gly Ala Ala Ser Pro Thr Ala

565 570 575

Arg Met Ser Ser Gln Asn Asp Trp Gly Ser Pro Asp Gly Lys Leu Asp

580 585 590

Trp Gly Ile Gln Gly Glu Glu Leu Asn Lys Leu Arg Lys Ser Ala Ser

595 600 605

Phe Gly Leu Arg Ser Ser Ser Asn Arg Phe His Ala Ser Ala Asp Ser

610 615 620

Ala Thr Ala Thr Val Gly Asp Pro Asp Met Pro Trp Ile Gln Ser Leu

625 630 635 640

Ala Lys Glu Ala Pro Ser Gln Asn Pro Gly Asn Phe Gly Ala Glu His

645 650 655

Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Tyr His Leu Asn Ser

660 665 670

Gly Gly Thr Glu Leu Leu Pro Ala Trp Val Glu Gln Leu Tyr Ala Asp

675 680 685

Gln Glu Gln Met Val Ala

690

<210>34

<211>2499

<212>DNA

＜213〉Arabidopis thaliana

<400>34

attttgacct taagaagaaa gtgacaagga gaggaagaag aagaaaaaaa acataatttg 60

aggaagaaga aaaaaaattc ggatttgttt tttcaataaa ttgactaatt gagtactcgt 120

ttaaaggaag tgaagagcgg ttttttggta gtggtggtcg agaaaagaga gagtttgtct 180

ctgtgactca gagtgaaatc aatagagcgg gaaaagattg ttgctttttt ttgccatggg 240

agttgatgag ctgtctcacc tcaaattctc tcttctgcta gaatcatcag cctgcaatga 300

tttgtccggt tttaagtctc tagttgaaga agaaggtctt gagagcattg atggctctgg 360

tttgtggtat gggaggagat taggatcaaa gaagatgggt tttgaggaga ggacgcctct 420

tatgattgct gccttgtttg gaagcaaaga ggttgttgat tacatcatta gtactggtct 480

tgttgacgtg aaccgctctt gtggctctga tggtgccacg gctcttcact gtgcggtctc 540

tggcttgtct gccaatagcc ttgagattgt tactcttctg ctgaagggct ctgcgaatcc 600

ggattcttgt gatgcttatg gtaacaagcc tggagatgtg attttccctt gtttgagtcc 660

ggtttttagc gcgaggatga aggttttgga gcgtttgttg aaaggaaatg atgatttgaa 720

tgaagttaat gggcaagaag aaagcgagcc agaggttgag gttgaggttg aggtttcgcc 780

tcctcggggg tctgagagga aggagtatcc ggttgatcca acgcttcctg atatcaagaa 840

cggtgtatat gggacggatg agttccggat gtatgctttc aagatcaagc cgtgctctag 900

agcatactct cacgactgga cggaatgtcc ctttgttcat ccgggtgaga acgcaaggag 960

gcgtgatccg aggaagtacc attatagttg tgtcccttgt cctgaattcc ggaaggggtc 1020

ttgttccaga ggtgatactt gcgagtatgc tcatggtatc tttgagtgct ggcttcaccc 1080

ggctcagtac cggactcgtc tctgcaagga cgagacgaat tgctcgagaa gagtttgttt 1140

ctttgcccac aaacccgagg agctgcgtcc tttgtaccct tcaactggat caggtgttcc 1200

gtccccgcgg tcttccttct catcttgcaa ttcctcgacc gctttcgaca tgggaccgat 1260

tagtccgctt cctatcggag caacaaccac acctcctttg agtcctaacg gtgtatcctc 1320

tccaataggt ggaggaaaaa cgtggatgaa ctggcctaac ataacccctc ctgcattgca 1380

gcttccaggg agcagattga aatctgcatt gaatgcaaga gaaatcgatt tctctgaaga 1440

gatgcaaagt cttacttctc caactacatg gaacaacacg ccaatgtcat ctccattctc 1500

cggaaagggc atgaacaggc ttgcaggagg agcaatgagc ccggtgaata gtctcagtga 1560

tatgtttggg acagaggata atacatcggg tttgcagatc cgacgcagcg tcattaaccc 1620

gcagctgcat tccaacagtc tttcttcatc acctgtggga gccaattctc tgttttcgat 1680

ggattcctcc gcagtcttgg cttcaagagc ggctgaattt gctaaacagc gaagccaaag 1740

cttcatagaa cgcaacaacg gactgaatca ccatcccgca atctcttcca tgactacaac 1800

ttgtttaaac gattggggct cattggatgg gaagcttgac tggagcgtcc aaggagacga 1860

gctacagaag ctcagaaaat ccacttcttt ccgtctcaga gccggtggca tggaatcaag 1920

actgcctaac gaagggactg ggctcgaaga gccagatgtc tcatgggtgg agccgctggt 1980

gaaagagcca caggagacaa gactagctcc ggtttggatg gagcaatcat acatggagac 2040

agaacagacc gtggcttgaa tcaaaagttt tgaactttca ttaaccgttc cacaagaagc 2100

aaagtcagaa agattccgag aggtcgatgc taatctattt cattttattt gtttaatgct 2160

ttgttatttt tctttagaat aaaaagaaaa aattcttagg ggacaaaaga gagttcgttt 2220

gtctctctct ctgtctccaa agaaaaacag aggtgaaaaa aggtttcaaa acctaagaaa 2280

ccttgaatta cctcacctca cttccttgat tctttactat tcacaatgag taatcgattt 2340

ttttttttct tggtaacact ctcacgctga atatatatgt tttttagtaa taatataatt 2400

ggaatacaga aatgtattta cacttgtgaa gttagggaaa gtgttgtaat tgtttcttct 2460

aagagttgat ctaagatgtt tgagactata tcttcgctt 2499

<210>35

<211>607

<212>PRT

＜213〉Arabidopis thaliana

<400>35

Met Gly Val Asp Glu Leu Ser His Leu Lys Phe Ser Leu Leu Leu Glu

1 5 10 15

Ser Ser Ala Cys Asn Asp Leu Ser Gly Phe Lys Ser Leu Val Glu Glu

20 25 30

Glu Gly Leu Glu Ser Ile Asp Gly Ser Gly Leu Trp Tyr Gly Arg Arg

35 40 45

Leu Gly Ser Lys Lys Met Gly Phe Glu Glu Arg Thr Pro Leu Met Ile

50 55 60

Ala Ala Leu Phe Gly Ser Lys Glu Val Val Asp Tyr Ile Ile Ser Thr

65 70 75 80

Gly Leu Val Asp Val Asn Arg Ser Cys Gly Ser Asp Gly Ala Thr Ala

85 90 95

Leu His Cys Ala Val Ser Gly Leu Ser Ala Asn Ser Leu Glu Ile Val

100 105 110

Thr Leu Leu Leu Lys Gly Ser Ala Asn Pro Asp Ser Cys Asp Ala Tyr

115 120 125

Gly Asn Lys Pro Gly Asp Val Ile Phe Pro Cys Leu Ser Pro Val Phe

130 135 140

Ser Ala Arg Met Lys Val Leu Glu Arg Leu Leu Lys Gly Asn Asp Asp

145 150 155 160

Leu Asn Glu Val Asn Gly Gln Glu Glu Ser Glu Pro Gl u Val Glu Val

165 170 175

Glu Val Glu Val Ser Pro Pro Arg Gly Ser Glu Arg Lys Glu Tyr Pro

180 185 190

Val Asp Pro Thr Leu Pro Asp Ile Lys Asn Gly Val Tyr Gly Thr Asp

195 200 205

Glu Phe Arg Met Tyr Ala Phe Lys Ile Lys Pro Cys Ser Arg Ala Tyr

210 215 220

Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu Asn Ala

225 230 235 240

Arg Arg Arg Asp Pro Arg Lys Tyr His Tyr Ser Cys Val Pro Cys Pro

245 250 255

Glu Phe Arg Lys Gly Ser Cys Ser Arg Gly Asp Thr Cys Glu Tyr Ala

260 265 270

His Gly Ile Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg

275 280 285

LeL Cys Lys Asp Glu Thr Asn Cys Ser Arg Arg Val Cys Phe Phe Ala

290 295 300

His Lys Pro Glu Glu Leu Arg Pro Leu Tyr Pro Ser Thr Gly Ser Gly

305 310 315 320

Val Pro Ser Pro Arg Ser Ser Phe Ser Ser Cys Asn Ser Ser Thr Ala

325 330 335

Phe Asp Met Gly Pro Ile Ser Pro Leu Pro Ile Gly Ala Thr Thr Thr

340 345 350

Pro Pro Leu Ser Pro Asn Gly Val Ser Ser Pro Ile Gly Gly Gly Lys

355 360 365

Thr Trp Met Asn Trp Pro Asn Ile Thr Pro Pro Ala Leu Gln Leu Pro

370 375 380

Gly Ser Arg Leu Lys Ser Ala Leu Asn Ala Arg Glu Ile Asp Phe Ser

385 390 395 400

Glu Glu Met Gln Ser Leu Thr Ser Pro Thr Thr Trp Asn Asn Thr Pro

405 410 415

Met Ser Ser Pro Phe Ser Gly Lys Gly Met Asn Arg Leu Ala Gly Gly

420 425 430

Ala Met Ser Pro Val Asn Ser Leu Ser Asp Met Phe Gly Thr Glu Asp

435 440 445

Asn Thr Ser Gly Leu Gln Ile Arg Arg Ser Val Ile Asn Pro Gln Leu

450 455 460

His Ser Asn Ser Leu Ser Ser Ser Pro Val Gly Ala Asn Ser Leu Phe

465 470 475 480

Ser Met Asp Ser Ser Ala Val Leu Ala Ser Arg Ala Ala Glu Phe Ala

485 490 495

Lys Gln Arg Ser Gln Ser Phe Ile Glu Arg Asn Asn Gly Leu Asn His

500 505 510

His Pro Ala Ile Ser Ser Met Thr Thr Thr Cys Leu Asn Asp Trp Gly

515 520 525

Ser Leu Asp Gly Lys Leu Asp Trp Ser Val Gln Gly Asp Glu Leu Gln

530 535 540

Lys Leu Arg Lys Ser Thr Ser Phe Arg Leu Arg Ala Gly Gly Met Glu

545 550 555 560

Ser Arg Leu Pro Asn Glu Gly Thr Gly Leu Glu Glu Pro Asp Val Ser

565 570 575

Trp Val Glu Pro Leu Val Lys Glu Pro Gln Glu Thr Arg Leu Ala Pro

580 585 590

Val Trp Met Glu Gln Ser Tyr Met Glu Thr Glu Gln Thr Val Ala

595 600 605

<210>36

<211>1806

<212>DNA

＜213〉rice

<400>36

atgtgctctg ggccgcgcaa gccgtccaca ccgccgctgc cgcagcagca gaaggaggcg 60

acggtgatgg cggcgtcctt gcttcttgag ctggcggcag cggacgacgt ggcggcggtg 120

aggagggtcg tggaggagga gaaggtgtct cttggcgtgg ctgggttgtg gtatgggcct 180

tcggcgagcg gcgtggcgag gctcgggatg gagcggagga cggcggcgat ggtggcggcg 240

ctgtacggga gcacgggggt gcttgggtat gtcgtggcgg cagcgccggc ggaggccgcg 300

cgcgcgtcgg agacggatgg ggccacgccg ctgcacatgg cggctgccgg tggcgcggcg 360

aacgcggtcg cggccacgcg cctgttgctc gccgcggggg cgtcggtcga cgcgctctcg 420

gcttcggggc tccgcgccgg tgacctcctc ccgcgcgcca ccgcggcgga gaaggccatc 480

cggctgctgc tcaagtcgcc ggccgtgtcg ccgtcgtcgt cgccgaagaa gtcggcctcg 540

ccgccgtcgc cgccgccgcc gcaggaggcg aagaaggagt acccgcctga cctgacgctg 600

cccgacctca agagcggact gttcagcacc gacgagttcc gcatgtacag cttcaaggtg 660

aagccgtgct cccgcgccta ctcccatgac tggaccgagt gccccttcgt ccaccccggc 720

gagaacgcgc gccgccgcga ccctcgccgc tactcctaca gctgcgtgcc ttgcccggag 780

ttccgcaagg gcggctcgtg ccgcaagggc gacgcgtgcg agtacgccca tggcgtgttc 840

gagtgctggc tccacccggc gcagtacagg acgcgcctct gcaaggacga ggtcggctgc 900

gcgcgccgca tctgcttctt cgcccacaag cccgacgagc tccgcgccgt caacccctcc 960

gccgtgtccg tcggcatgca gcccaccgta tcgtcgccgc gctcctcgcc gcccaacggg 1020

ctcgacatgg cggcggcggc ggcggcgatg atgagccccg cctggccgtc gtccccagcg 1080

agccgcctca agacggcgct cggcgcgcgg gagctcgact tcgacctcga gatgctcgcg 1140

ctggaccagt accagcagaa gctgttcgac aaggtgtccg gcgcgccgtc gccgagggcg 1200

agctggggcg ccgcggcgaa cggcctcgcc accgcgtcgc cggcgagggc cgtgccggac 1260

tacaccgacc tgctcggctc cgtcgacccg gccatgctgt cccagctcca cgcgctgtcc 1320

ctcaagcagg ccggcgacat gcccgcgtac agctccatgg cggacaccac gcagatgcac 1380

atgccgacct cgccgatggt gggcggcgcg aacaccgcgt tcgggctgga ccactccatg 1440

gcgaaggcga tcatgagctc ccgcgcctcg gcgttcgcca agcgcagcca gagcttcatc 1500

gaccgcggag gccgcgcccc ggcggcgcgt tcgctcatgt cgccggcgac gaccggcgcg 1560

ccgtccattc tctcggactg gggctcgccg gacggcaagc tggactgggg cgtccagggc 1620

gacgagctgc acaagctccg caagtcggcg tcgttcgcgt tccgcggcca atccgccatg 1680

ccggtggcga cgcacgccgc ggcggcggag ccggacgtgt catgggtgaa ctctcttgtc 1740

aaggacggcc acgccgccgg cgacatattc gcgcagtggc cggagcagga gcagatggtg 1800

gcatga 1806

<210>37

<211>601

<212>PRT

＜213〉rice

<400>37

Met Cys Ser Gly Pro Arg Lys Pro Ser Thr Pro Pro Leu Pro Gln Gln

1 5 10 15

Gln Lys Glu Ala Thr Val Met Ala Ala Ser Leu Leu Leu Glu Leu Ala

20 25 30

Ala Ala Asp Asp Val Ala Ala Val Arg Arg Val Val Glu Glu Glu Lys

35 40 45

Val Ser Leu Gly Val Ala Gly Leu Trp Tyr Gly Pro Ser Ala Ser Gly

50 55 60

Val Ala Arg Leu Gly Met Glu Arg Arg Thr Ala Ala Met Val Ala Ala

65 70 75 80

Leu Tyr Gly Ser Thr Gly Val Leu Gly Tyr Val Val Ala Ala Ala Pro

85 90 95

Ala Glu Ala Ala Arg Ala Ser Glu Thr Asp Gly Ala Thr Pro Leu His

100 105 110

Met Ala Ala Ala Gly Gly Ala Ala Asn Ala Val Ala Ala Thr Arg Leu

115 120 125

Leu Leu Ala Ala Gly Ala Ser Val Asp Ala Leu Ser Ala Ser Gly Leu

130 135 140

Arg Ala Gly Asp Leu Leu Pro Arg Ala Thr Ala Ala Glu Lys Ala Ile

145 150 155 160

Arg Leu Leu Leu Lys Ser Pro Ala Val Ser Pro Ser Ser Ser Pro Lys

165 170 175

Lys Ser Ala Ser Pro Pro Ser Pro Pro Pro Pro Gl n Glu Ala Lys Lys

180 185 190

Glu Tyr Pro Pro Asp Leu Thr Leu Pro Asp Leu Lys Ser Gly Leu Phe

195 200 205

Ser Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Lys Pro Cys Ser

210 215 220

Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly

225 230 235 240

Glu Asn Ala Arg Arg Arg Asp Pro Arg Arg Tyr Ser Tyr Ser Cys Val

245 250 255

Pro Cys Pro Glu Phe Arg Lys Gly Gly Ser Cys Arg Lys Gly Asp Ala

260 265 270

Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu Hi s Pro Ala Gln

275 280 285

Tyr Arg Thr Arg Leu Cys Lys Asp Glu Val Gly Cys Ala Arg Arg Ile

290 295 300

Cys Phe Phe Ala His Lys Pro Asp Glu Leu Arg Ala Val Asn Pro Ser

305 310 315 320

Ala Val Ser Val Gly Met Gln Pro Thr Val Ser Ser Pro Arg Ser Ser

325 330 335

Pro Pro Asn Gly Leu Asp Met Ala Ala Ala Ala Ala Ala Met Met Ser

340 345 350

Pro Ala Trp Pro Ser Ser Pro Ala Ser Arg Leu Lys Thr Ala Leu Gly

355 360 365

Ala Arg Glu Leu Asp Phe Asp Leu Glu Met Leu Ala Leu Asp Gln Tyr

370 375 380

Gln Gln Lys Leu Phe Asp Lys Val Ser Gly Ala Pro Ser Pro Arg Ala

385 390 395 400

Ser Trp Gly Ala Ala Ala Asn Gly Leu Ala Thr Ala Ser Pro Ala Arg

405 410 415

Ala Val Pro Asp Tyr Thr Asp Leu Leu Gly Ser Val Asp Pro Ala Met

420 425 430

Leu Ser Gln Leu His Ala Leu Ser Leu Lys Gln Ala Gly Asp Met Pro

435 440 445

Ala Tyr Ser Ser Met Ala Asp Thr Thr Gln Met His Met Pro Thr Ser

450 455 460

Pro Met Val Gly Gly Ala Asn Thr Ala Phe Gly Leu Asp His Ser Met

465 470 475 480

Ala Lys Ala Ile Met Ser Ser Arg Ala Ser Ala Phe Ala Lys Arg Ser

485 490 495

Gln Ser Phe Ile Asp Arg Gly Gly Arg Ala Pro Ala Ala Arg Ser Leu

500 505 510

Met Ser Pro Ala Thr Thr Gly Ala Pro Ser Ile Leu Ser Asp Trp Gly

515 520 525

Ser Pro Asp Gly Lys Leu Asp Trp Gly Val Gln Gly Asp Glu Leu His

530 535 540

Lys Leu Arg Lys Ser Ala Ser Phe Ala Phe Arg Gly Gln Ser Ala Met

545 550 555 560

Pro Val Ala Thr His Ala Ala Ala Ala Glu Pro Asp Val Ser Trp Val

565 570 575

Asn Scr Leu Val Lys Asp Gly His Ala Ala Gly Asp Ile Phe Ala Gln

580 585 590

Trp Pro Glu Gln Glu Gln Met Val Ala

595 600

<210>38

<211>1692

<212>DNA

＜213〉barley (Hordeum vulgare)

<400>38

cggcacgagg cacatccatc atctaacctc acctctcctc tcctcccctc tcctcctacc 60

aaacccaaaa ccaagcagag caagagcaag agcaagagca agagcaagca agcatgtgcc 120

ctggcctgcg caacctcgcc gccgccatgc caccctccgc ccacgaccac ccctcctcct 180

acctgctcga gctcgccgcc gacgacgacc tccccgcctt ccgccgcgcc gtccaggagg 240

acaacctctc cctcgacgcc gcatccccga ggtacgagcc atcccccaaa tcagaccaac 300

aacaacaaca cgccccagct cgcgctccac ctgcgcaccc ccgccatggt cgccgcgctc 360

tacggcagca ccaccgtcct ctcctacgtc ctctccatcg ccccctccga ggccgcccgc 420

gcctccgcat ccgacggcgc caccccgctc ctcctcgccc accagggccg cgcgccatcc 480

gcgccccacg ccgcacgcct cctcctcacc gacggcgcat catcgtcctc cctactcgcg 540

ccccaagctc accctctcaa ccaccaaaac caaaaccaaa acagccccac caagaaagac 600

tcgccgccgg actccaggag gaccaccacc aagaaggact actcctccgc ctccgactcc 660

cagacggagg acatcaacgc gggcgtcttc gccaccgacg acttccggat gtacagcttc 720

aaggtgaacc cgtgctcccg cgcctacacg cacgactgga ccgagtgccc cttcgcccac 780

cccggcgaga acgcgcgccg ccgcgacccg cgccgcgtgc catactcgtg cgtcccatgc 840

ccggacttcc gccgcgaccc ggccgcatgc cgcaagggcg acgcctgcga gtacgcgcac 900

ggcgtcttcg agtcatggct ccaccccgcg cagtaccgca ccaggctctg caaggacgag 960

gtcggatgcc cgcgccgcat ctgcttcttc gcgcacggcg cccgacagct acgcgccgtc 1020

aacccctccg ccgcatccat ggactcgcca tccccaactt cctcttcgcc gccgcgaacc 1080

tccaggccgg ccgcgctcac cgcgtcgctc agctcgcggg acctcgactt ggacgccgac 1140

aaccaggccc agtacgcgcg caggatgatg atggccaggg ccaactcccc gccggactac 1200

tcgcccgacc tcgtcgccgc ctacgtacag gcgctctcct ccctgcaaca gcagcagcat 1260

cagcagaacc agcaacagca gcatcagcag cagaaccagc accagcagca acatcagcag 1320

aaccagcacc agcagcatca gcagcaacat cagcagagca tggggatggg ggggctgagc 1380

gcccgcgccg ccgccttcac caaccgcagc cagaccttcg tgcaccgctc tccgtccccg 1440

gctccggcgc ggtcgttcaa gtctccggcg ccgtcgtcca tgctcgcgga ctgggggtcg 1500

ccggacggga agctggactg gggcgtgcag gccgcggagc tgcgcaagtc cacgtctttc 1560

ggagtcagaa gcagcagcag gccgcatcat gagacgacga gggcggagga caacatgtac 1620

ccgtcgtgga tgaaggacgg cagcgatatg ctgctggcgg cgcggtggtc ggacctggag 1680

cagatggtcg cc 1692

<210>39

<211>564

<212>PRT

＜213〉barley

<400>39

Arg His Glu Ala His Pro Ser Ser Asn Leu Thr Ser Pro Leu Leu Pro

1 5 10 15

Ser Pro Pro Thr Lys Pro Lys Thr Lys Gln Ser Lys Ser Lys Ser Lys

20 25 30

Ser Lys Ser Lys Gln Ala Cys Ala Leu Ala Cys Ala Thr Ser Pro Pro

35 40 45

Pro Cys His Pro Pro Pro Thr Thr Thr Pro Pro Pro Thr Cys Ser Ser

50 55 60

Ser Pro Pro Thr Thr Thr Ser Pro Pro Ser Ala Ala Pro Ser Arg Arg

65 70 75 80

Thr Thr Ser Pro Ser Thr Pro His Pro Arg Gly Thr Ser His Pro Pro

85 90 95

Asn Gln Thr Asn Asn Asn Asn Thr Pro Gln Leu Ala Leu His Leu Arg

100 105 110

Thr Pro Ala Met Val Ala Ala Leu Tyr Gly Ser Thr Thr Val Leu Ser

115 120 125

Tyr Val Leu Ser Ile Ala Pro Ser Glu Ala Ala Arg Ala Ser Ala Ser

130 135 140

Asp Gly Ala Thr Pro Leu Leu Leu Ala His Gln Gly Arg Ala Pro Ser

145 150 155 160

Ala Pro His Ala Ala Arg Leu Leu Leu Thr Asp Gly Ala Ser Ser Ser

165 170 175

Ser Leu Leu Ala Pro Gln Ala His Pro Leu Asn His Gln Asn Gln Asn

180 185 190

Gln Asn Ser Pro Thr Lys Lys Asp Ser Pro Pro Asp Ser Arg Arg Thr

195 200 205

Thr Thr Lys Lys Asp Tyr Ser Ser Ala Ser Asp Ser Gln Thr Glu Asp

210 215 220

Ile Asn Ala Gly Val Phe Ala Thr Asp Asp Phe Arg Met Tyr Ser Phe

225 230 235 240

Lys Val Asn Pro Cys Ser Arg Ala Tyr Thr Hi s Asp Trp Thr Glu Cys

245 250 255

Pro Phe Ala His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Arg

260 265 270

Val Pro Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg Arg Asp Pro Ala

275 280 285

Ala Cys Arg Lys Gly Asp Ala Cys Glu Tyr Ala His Gly Val Phe Glu

290 295 300

Ser Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Glu

305 310 315 320

Val Gly Cys Pro Arg Arg Ile Cys Phe Phe Ala His Gly Ala Arg Gln

325 330 335

Leu Arg Ala Val Asn Pro Ser Ala Ala Ser Met Asp Ser Pro Ser Pro

340 345 350

Thr Ser Ser Ser Pro Pro Arg Thr Ser Arg Pro Ala Ala Leu Thr Ala

355 360 365

Ser Leu Ser Ser Arg Asp Leu Asp Leu Asp Ala Asp Asn Gln Ala Gln

370 375 380

Tyr Ala Arg Arg Met Met Met Ala Arg Ala Asn Ser Pro Pro Asp Tyr

385 390 395 400

Ser Pro Asp Leu Val Ala Ala Tyr Val Gln Ala Leu Ser Ser Leu Gln

405 410 415

Gln Gln Gln His Gln Gln Asn Gln Gln Gln Gln His Gln Gln Gln Asn

420 425 430

Gln His Gln Gln Gln His Gln Gln Asn Gln His Gln Gln Hi s Gln Gln

435 440 445

Gln His Gln Gln Ser Met Gly Met Gly Gly Leu Ser Ala Arg Ala Ala

450 455 460

Ala Phe Thr Asn Arg Ser Gln Thr Phe Val His Arg Ser Pro Ser Pro

465 470 475 480

Ala Pro Ala Arg Ser Phe Lys Ser Pro Ala Pro Ser Ser Met Leu Ala

485 490 495

Asp Trp Gly Ser Pro Asp Gly Lys Leu Asp Trp Gly Val Gln Ala Ala

500 505 510

Glu Leu Arg Lys Ser Thr Ser Phe Gly Val Arg Ser Ser Ser Arg Pro

515 520 525

His His Glu Thr Thr Arg Ala Glu Asp Asn Met Tyr Pro Ser Trp Met

530 535 540

Lys Asp Gly Ser Asp Met Leu Leu Ala Ala Arg Trp Ser Asp Leu Glu

545 550 555 560

Gln Met Val Ala

<210>40

<211>3610

<212>DNA

＜213〉pine (Pinus radiata)

<400>40

tgtttccagg cgggcactaa agcaagggag ggggtaggct ttactttctg ctctgcgcaa 60

agaacgttga aatcaatcgc cctggctggt ctggcgtgac tactagattc aatttcttca 120

tggccgtctt cacataccca ttctttaccg gttcagagct gtgatcttta tttttaacag 180

ccacaatcat ggtttgtgtt tccagtgtta tgatctgagt gaagttcgtt ctttttctcg 240

tgacccaggc ttgatactag gccggacctt tctgaggtgg aagagatcta tacatttgag 300

gcctattttg tgtagccatg tgtggaggcc cagaacattt gaagcctgcc agcccacacg 360

aaggagaaga taaagtcaaa atggccgaga atcagtctat caaagtgaag gaattgtctg 420

aatcttgttc aagtctacat gaactagctg ctaataatga ccttattggc tttaagaaag 480

caatggagga agaagggtca aagatagatg aggttaactt ttggtacggg aggcagaatg 540

gttctaatca gatggtcctg gagcaaagga ctccattgat ggttgctgca ctttatggca 600

gtgtagatgc gctgagttac atcttatcca tttatgtaac ttgtggagca gatgttaacc 660

aagcctgtgg gtcagataac tccactgcct tgcattgtgc ggctgtggga gggtctgcct 720

gtgcagttga aactgtaaaa ttgttacttc atgcaggcag tgatgtgaat cgcttggatg 780

cttatggcag aagaccagca gatgtgatta tggtttctcc taagctaacc gaaatcaagg 840

ccaagctaga agaaatgtta aacgcagctg gttcatgtca aacttctccg gcaaagttgc 900

ctaacatagt ttcagggcca cctgggtttg agtcaaaggg gatggagtcc atgtccccat 960

tgccattgtt gcctctttca ttgtctttag aagcatccaa taatagatca ggttgtgtga 1020

attctccaac atcttcgcca aagtccatgg aagcattaaa gggtttcggt gatgttaatg 1080

agaagaagga atatcctgtg gacccttctt ttccagacat aaagaatagc atctatacta 1140

cagatgaatt tcggatgttt tccttcaagg tgcggccatg ttcacgggca tattctcatg 1200

attggactga atgcccattt gtgcatcctg gtgaaaatgc cagaaggcgg gatccaagaa 1260

ggtatcatta tagctgtgtt ccttgcccag attttcggaa agggacttgt aggcgcagtg 1320

atgtttgtga atatgcacac ggtgtttttg agtgctggtt acatcctgct caatatagga 1380

cacggttgtg caaagatggg actaattgtt cacgtagagt ttgcttcttt gctcacacat 1440

ctgaggaact acgccctctc attgtctcta ctgggtctgc tgttccatcc ccaagggcat 1500

catcatctct ggacatgaca tctgtcatga gtcctcttgc ccctggttct ccctcttcag 1560

tttcaatgat gtcacccttc ctatcaaatc ctcagcaagg cagtgtgctt actccgccta 1620

tgtctccatc agcgtcctct gtaaatggat atggaggctg gccacagcct aatgtaccaa 1680

ccttacacct tcctggtagc aatgttcaaa ccagccgtct tagagcggaa cttaatgcca 1740

gagacatgcc tgttgaggat tctcctcgaa tttcagacta tgaagggcag caactcctga 1800

atgatttttc tccactgtcc acacaagcca ggctgaatgc tgctgctgct gttatatctg 1860

gtggcgggaa caccacaaca aggtctggaa aatacaagag tcacgggatc aatactgttg 1920

ctccaacgaa tcttgaagac ttgtttgcct ccgaggtaac atctcctaga gtagcagttc 1980

ttgaaccttc catcttttct cagatgagtc cccaaatgca agctcataag actgcccagg 2040

catatatgca gattcaaaac cagatgctgc ctcctataaa tacacaggca ttttcgcagg 2100

gaattacaca gatgcagcag gctgcaatag agcctcagag ccctggacat tctttgatgc 2160

aatcaccttt ccaatcttcc tcgtatgggt tgggatcccc tggtagaatg tcacctcgtt 2220

gtgtggatgt ggaacgtcat aatacatgtg ggtctccctt atcaccggct atggctgcaa 2280

cgataaattc aagaatggct atggctgctt ttgttcagag ggaaaaacgg agccatagtt 2340

cccgtgactt gggagctaat gtgaatccca gttcatggtc tgattggggc tcgcctacag 2400

gtaaagttga ctggggggtt caaggagaag agttgagcaa attaagaaag tcggcttcat 2460

ttggtccccg cagttatgaa gaaccggatt tgtcttgggt tcaaacactg gtaaaggaaa 2520

ctacaccaga gggtaaagat ggaggaaatg taagctgttc tggggaaact ccacacaagg 2580

ggcaaataga aaatgttgat cattcagttt tgggtgcctg gattgaacag atgcagcttg 2640

atcagattgt agcttgagat taggattatt tatttggagt ggtggtaggg ataggctcat 2700

ttaaaattca atttctcatt ttttactatt tcttttataa aaattcccca ttatagttta 2760

ggaaatagtc tggttttcta cctattatca gaattacacc tgcaggaaat tttggaggaa 2820

agcatgcaaa aagtagatag ggatgttatt cctatcagca ggttgacaag ctgaaaatca 2880

cttgggtggt agaccagaga atgacactat tttttgttga catggcaact gaagatgctg 2940

ttttctttac ttatcattaa caaccctata tatatttgtt ttgaaagaac tgagcggaga 3000

aatgttgtca gttggttact ctgcgcaagg ccttggaaga aatccaagat gtggcatctt 3060

ggtgcatttt taatttatca agtgtgaaat ccataacagg tttcagtgag tgacttctga 3120

ggttgtatat ggaaaaacct atgatgttgg ctgtctactg ctatttttct gtgcctaaac 3180

tgtcaactaa agtttgcagg tggcaatttt gtggcagcat atttgcacat tgaagcggat 3240

ggtctgcacc tgctatagaa gttttcgagt ctgtagaatt tgatggtgca agatgatttt 3300

ctagttgata tatttggaag gctttgccaa agtagtggca tgtacatttt gcaaaaattt 3360

aaaggatggc aatccattgt tttgccatgt agcttcactt tattgattag gtggaaagga 3420

attttgagac acttcaattt gtgcatactt ttgttctgaa ctgcaaaatc agtctcttgt 3480

gatgtcctca aggctattat gctcagggat ttgcctaaaa ccataagtgg ccttagataa 3540

ggtaccattg tattaccttt tattgtttgg atattttatt tatgaaagtg aatttatttt 3600

aaaaaaaaaa 3610

<210>41

<211>779

<212>PRT

＜213〉pine

<400>41

Met Cys Gly Gly Pro Glu His Leu Lys Pro Ala Ser Pro His Glu Gly

1 5 10 15

Glu Asp Lys Val Lys Met Ala Glu Asn Gln Ser Ile Lys Val Lys Glu

20 25 30

Leu Ser Glu Ser Cys Ser Ser Leu His Glu Leu Ala Ala Asn Asn Asp

35 40 45

Leu Ile Gly Phe Lys Lys Ala Met Glu Glu Glu Gly Ser Lys Ile Asp

50 55 60

Glu Val Asn Phe Trp Tyr Gly Arg Gln Asn Gly Ser Asn Gln Met Val

65 70 75 80

Leu Glu Gln Arg Thr Pro Leu Met Val Ala Ala Leu Tyr Gly Ser Val

85 90 95

Asp Ala Leu Ser Tyr Ile Leu Ser Ile Tyr Val Thr Cys Gly Ala Asp

100 105 110

Val Asn Gln Ala Cys Gly Ser Asp Asn Ser Thr Ala Leu His Cys Ala

115 120 125

Ala Val Gly Gly Ser Ala Cys Ala Val Glu Thr Val Lys Leu Leu Leu

130 135 140

His Ala Gly Ser Asp Val Asn Arg Leu Asp Ala Tyr Gly Arg Arg Pro

145 150 155 160

Ala Asp Val Ile Met Val Ser Pro Lys Leu Thr Glu Ile Lys Ala Lys

165 170 175

Leu Glu Glu Met Leu Asn Ala Ala Gly Ser Cys Gln Thr Ser Pro Ala

180 185 190

Lys Leu Pro Asn Ile Val Ser Gly Pro Pro Gly Phe Glu Ser Lys Gly

195 200 205

Met Glu Ser Met Ser Pro Leu Pro Leu Leu Pro Leu Ser Leu Ser Leu

210 215 220

Glu Ala Ser Asn Asn Arg Ser Gly Cys Val Asn Ser Pro Thr Ser Ser

225 230 235 240

Pro Lys Ser Met Glu Ala Leu Lys Gly Phe Gly Asp Val Asn Glu Lys

245 250 255

Lys Glu Tyr Pro Val Asp Pro Ser Phe Pro Asp Ile Lys Asn Ser Ile

260 265 270

Tyr Thr Thr Asp Glu Phe Arg Met Phe Ser Phe Lys Val Arg Pro Cys

275 280 285

Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro

290 295 300

Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Arg Tyr His Tyr Ser Cys

305 310 315 320

Val Pro Cys Pro Asp Phe Arg Lys Gly Thr Cys Arg Arg Ser Asp Val

325 330 335

Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln

340 345 350

Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr Asn Cys Ser Arg Arg Val

355 360 365

Cys Phe Phe Ala His Thr Ser Glu Glu Leu Arg Pro Leu Ile Val Ser

370 375 380

Thr Gly Ser Ala Val Pro Ser Pro Arg Ala Ser Ser Ser Leu Asp Met

385 390 395 400

Thr Ser Val Met Ser Pro Leu Ala Pro Gly Ser Pro Ser Ser Val Ser

405 410 415

Met Met Ser Pro Phe Leu Ser Asn Pro Gln Gln Gly Ser Val Leu Thr

420 425 430

Pro Pro Met Ser Pro Ser Ala Ser Ser Val Asn Gly Tyr Gly Gly Trp

435 440 445

Pro Gln Pro Asn Val Pro Thr Leu His Leu Pro Gly Ser Asn Val Gln

450 455 460

Thr Ser Arg Leu Arg Ala Glu Leu Asn Ala Arg Asp Met Pro Val Glu

465 470 475 480

Asp Ser Pro Arg Ile Ser Asp Tyr Glu Gly Gln Gln Leu Leu Asn Asp

485 490 495

Phe Ser Pro Leu Ser Thr Gln Ala Arg Leu Asn Ala Ala Ala Ala Val

500 505 510

Ile Ser Gly Gly Gly Asn Thr Thr Thr Arg Ser Gly Lys Tyr Lys Ser

515 520 525

His Gly Ile Asn Thr Val Ala Pro Thr Asn Leu Glu Asp Leu Phe Ala

530 535 540

Ser Glu Val Thr Ser Pro Arg Val Ala Val Leu Glu Pro Ser Ile Phe

545 550 555 560

Ser Gln Met Ser Pro Gln Met Gln Ala His Lys Thr Ala Gln Ala Tyr

565 570 575

Met Gln Ile Gln Asn Gln Met Leu Pro Pro Ile Asn Thr Gln Ala Phe

580 585 590

Ser Gln Gly Ile Thr Gln Met Gln Gln Ala Ala Ile Glu Pro Gln Ser

595 600 605

Pro Gly His Ser Leu Met Gln Ser Pro Phe Gln Ser Ser Ser Tyr Gly

610 615 620

Leu Gly Ser Pro Gly Arg Met Ser Pro Arg Cys Val Asp Val Glu Arg

625 630 635 640

His Asn Thr Cys Gly Ser Pro Leu Ser Pro Ala Met Ala Ala Thr Ile

645 650 655

Asn Ser Arg Met Ala Met Ala Ala Phe Val Gln Arg Glu Lys Arg Ser

660 665 670

His Ser Ser Arg Asp Leu Gly Ala Asn Val Asn Pro Ser Ser Trp Ser

675 680 685

Asp Trp Gly Ser Pro Thr Gly Lys Val Asp Trp Gly Val Gln Gly Glu

690 695 700

Glu Leu Ser Lys Leu Arg Lys Ser Ala Ser Phe Gly Pro Arg Ser Tyr

705 710 715 720

Glu Glu Pro Asp Leu Ser Trp Val Gln Thr Leu Val Lys Glu Thr Thr

725 730 735

Pro Glu Gly Lys Asp Gly Gly Asn Val Ser Cys Ser Gly Glu Thr Pro

740 745 750

His Lys Gly Gln Ile Glu Asn Val Asp His Ser Val Leu Gly Ala Trp

755 760 765

Ile Glu Gln Met Gln Leu Asp Gln Ile Val Ala

770 775

<210>42

<211>3610

<212>DNA

＜213〉pine

<400>42

tgtttccagg cgggcactaa agcaagggag ggggtaggct ttactttctg ctctgcgcaa 60

agaacgttga aatcaatcgc cctggctggt ctggcgtgac tactagattc aatttcttca 120

tggccgtctt cacataccca ttctttaccg gttcagagct gtgatcttta tttttaacag 180

ccacaatcat ggtttgtgtt tccagtgtta tgatctgagt gaagttcgtt ctttttctcg 240

tgacccaggc ttgatactag gccggacctt tctgaggtgg aagagatcta tacatttgag 300

gcctattttg tgtagccatg tgtggaggcc cagaacattt gaagcctgcc agcccacacg 360

aaggagaaga taaagtcaaa atggccgaga atcagtctat caaagtgaag gaattgtctg 420

aatcttgttc aagtctacat gaactagctg ctaataatga ccttattggc tttaagaaag 480

caatggagga agaagggtca aagatagatg aggttaactt ttggtacggg aggcagaatg 540

gttctaatca gatggtcctg gagcaaagga ctccattgat ggttgctgca ctttatggca 600

gtgtagatgc gctgagttac atcttatcca tttatgtaac ttgtggagca gatgttaacc 660

aagcctgtgg gtcagataac tccactgcct tgcattgtgc ggctgtggga gggtctgcct 720

gtgcagttga aactgtaaaa ttgttacttc atgcaggcag tgatgtgaat cgcttggatg 780

cttatggcag aagaccagca gatgtgatta tggtttctcc taagctaacc gaaatcaagg 840

ccaagctaga agaaatgtta aacgcagctg gttcatgtca aacttctccg gcaaagttgc 900

ctaacatagt ttcagggcca cctgggtttg agtcaaaggg gatggagtcc atgtccccat 960

tgccattgtt gcctctttca ttgtctttag aagcatccaa taatagatca ggttgtgtga 1020

attctccaac atcttcgcca aagtccatgg aagcattaaa gggtttcggt gatgttaatg 1080

agaagaagga atatcctgtg gacccttctt ttccagacat aaagaatagc atctatacta 1140

cagatgaatt tcggatgttt tccttcaagg tgcggccatg ttcacgggca tattctcatg 1200

attggactga atgcccattt gtgcatcctg gtgaaaatgc cagaaggcgg gatccaagaa 1260

ggtatcatta tagctgtgtt ccttgcccag attttcggaa agggacttgt aggcgcagtg 1320

atgtttgtga atatgcacac ggtgtttttg agtgctggtt acatcctgct caatatagga 1380

cacggttgtg caaagatggg actaattgtt cacgtagagt ttgcttcttt gctcacacat 1440

ctgaggaact acgccctctc attgtctcta ctgggtctgc tgttccatcc ccaagggcat 1500

catcatctct ggacatgaca tctgtcatga gtcctcttgc ccctggttct ccctcttcag 1560

tttcaatgat gtcacccttc ctatcaaatc ctcagcaagg cagtgtgctt actccgccta 1620

tgtctccatc agcgtcctct gtaaatggat atggaggctg gccacagcct aatgtaccaa 1680

ccttacacct tcctggtagc aatgttcaaa ccagccgtct tagagcggaa cttaatgcca 1740

gagacatgcc tgttgaggat tctcctcgaa tttcagacta tgaagggcag caactcctga 1800

atgatttttc tccactgtcc acacaagcca ggctgaatgc tgctgctgct gttatatctg 1860

gtggcgggaa caccacaaca aggtctggaa aatacaagag tcacgggatc aatactgttg 1920

ctccaacgaa tcttgaagac ttgtttgcct ccgaggtaac atctcctaga gtagcagttc 1980

ttgaaccttc catcttttct cagatgagtc cccaaatgca agctcataag actgcccagg 2040

catatatgca gattcaaaac cagatgctgc ctcctataaa tacacaggca ttttcgcagg 2100

gaattacaca gatgcagcag gctgcaatag agcctcagag ccctggacat tctttgatgc 2160

aatcaccttt ccaatcttcc tcgtatgggt tgggatcccc tggtagaatg tcacctcgtt 2220

gtgtggatgt ggaacgtcat aatacatgtg ggtctccctt atcaccggct atggctgcaa 2280

cgataaattc aagaatggct atggctgctt ttgttcagag ggaaaaacgg agccatagtt 2340

cccgtgactt gggagctaat gtgaatccca gttcatggtc tgattggggc tcgcctacag 2400

gtaaagttga ctggggggtt caaggagaag agttgagcaa attaagaaag tcggcttcat 2460

ttggtccccg cagttatgaa gaaccggatt tgtcttgggt tcaaacactg gtaaaggaaa 2520

ctacaccaga gggtaaagat ggaggaaatg taagctgttc tggggaaact ccacacaagg 2580

ggcaaataga aaatgttgat cattcagttt tgggtgcctg gattgaacag atgcagcttg 2640

atcagattgt agcttgagat taggattatt tatttggagt ggtggtaggg ataggctcat 2700

ttaaaattca atttctcatt ttttactatt tcttttataa aaattcccca ttatagttta 2760

ggaaatagtc tggttttcta cctattatca gaattacacc tgcaggaaat tttggaggaa 2820

agcatgcaaa aagtagatag ggatgttatt cctatcagca ggttgacaag ctgaaaatca 2880

cttgggtggt agaccagaga atgacactat tttttgttga catggcaact gaagatgctg 2940

ttttctttac ttatcattaa caaccctata tatatttgtt ttgaaagaac tgagcggaga 3000

aatgttgtca gttggttact ctgcgcaagg ccttggaaga aatccaagat gtggcatctt 3060

ggtgcatttt taatttatca agtgtgaaat ccataacagg tttcagtgag tgacttctga 3120

ggttgtatat ggaaaaacct atgatgttgg ctgtctactg ctatttttct gtgcctaaac 3180

tgtcaactaa agtttgcagg tggcaatttt gtggcagcat atttgcacat tgaagcggat 3240

ggtctgcacc tgctatagaa gttttcgagt ctgtagaatt tgatggtgca agatgatttt 3300

ctagttgata tatttggaag gctttgccaa agtagtggca tgtacatttt gcaaaaattt 3360

aaaggatggc aatccattgt tttgccatgt agcttcactt tattgattag gtggaaagga 3420

attttgagac acttcaattt gtgcatactt ttgttctgaa ctgcaaaatc agtctcttgt 3480

gatgtcctca aggctattat gctcagggat ttgcctaaaa ccataagtgg ccttagataa 3540

ggtaccattg tattaccttt tattgtttgg atattttatt tatgaaagtg aatttatttt 3600

aaaaaaaaaa 3610

<210>43

<211>749

<212>PRT

＜213〉pine

<400>43

Met Lys Glu Met Ala Glu Tyr Cys Ser Pro Ala Leu Leu Glu Leu Ala

1 5 10 15

Ala Asn Asn Asp Leu Ser Gly Phe Lys Gln Ala Val Glu Glu Gly Gly

20 25 30

Ser Ser Val Asn Glu Arg Gly Leu Trp Tyr Gly Arg Gln Ile Gly Ser

35 40 45

Gly Gln Lys Met Val Leu Glu Gln Arg Thr Pro Leu Met Val Ala Ala

50 55 60

Leu Tyr Gly Ser Leu Asp Val Leu Ser Tyr Met Leu Ser Gly Gly Arg

65 70 75 80

Val Asp Val Asn Gln Ser Cys Gly Ser Asp Met Ser Thr Ala Leu His

85 90 95

Cys Ala Ala Ala Gly Gly Ser Ile Leu Ala Ile Glu Thr Val Gly Met

100 105 110

Leu Ile Lys Ala Gly Ala Asp Val Asn Phe Met Asn Ala Gly Gly Arg

115 120 125

Lys Pro Ala Asp Val Ile Met Val Ser Pro Lys Leu Ala His Phe Lys

130 135 140

Asn Val Leu Glu Asp Leu Leu Ile Met Gly Ser Asn Ser Pro Met Lys

145 150 155 160

Ile Pro Cys Arg Val Ser Gly Ser Gly Phe Tyr Leu Pro Glu Gly Gly

165 170 175

Gly Cys Phe Phe Asp Glu His Gly Cys Val Val Ser Val Pro Thr Ser

180 185 190

Ser Pro Leu Phe Ser Ser Pro Asp Ala Thr Ser Pro Ala Thr Val Asn

195 200 205

Ser Pro Leu Ser Ser Pro Pro Thr Ser Leu Asp Thr Pro Lys Asn Leu

210 215 220

Cys Asp Cys Gly Gln Lys Lys Glu Phe Ala Val Asp Ser Ser Leu Pro

225 230 235 240

Asp Ile Lys Asn Ser Ile Tyr Ser Thr Asp Glu Phe Arg Met Tyr Ser

245 250 255

Phe Lys Val Arg Pro Cys Ser Arg Ala Tyr Ser His Asp Trp Thr Glu

260 265 270

Cys Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg

275 280 285

Lys Tyr His Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg Lys Gly Ala

290 295 300

Cys Arg Arg Gly Asp Val Cys Glu Tyr Ala His Gly Val Phe Glu Cys

305 310 315 320

Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr

325 330 335

Asn Cys Ser Arg Arg Val Cys Phe Phe Ala His Thr Pro Glu Glu Leu

340 345 350

Arg Pro Leu Tyr Pro Pro Ala Cys Ser Ser Met Leu Ser Gln Arg Thr

355 360 365

Thr Met Thr Ser Ser Asp Lys Met Ala Val Met His Pro Leu Ala Pro

370 375 380

Gly Ser Ala Ser Ser Val Leu Met Met Ser Ser Ser Asn Ser Ser Gln

385 390 395 400

Ser Ser Phe Pro Asn Ser Pro Val Ser Pro Leu Ser Ser Ala Asn Thr

405 410 415

Ser Ser His Ser Ser Phe Gly Gly Gly Ser Trp Ala His Pro Asn Leu

420 425 430

Pro Thr Leu His Leu Ser Asn Gly Ala Leu Gln Ala Ser Arg Leu Arg

435 440 445

Thr Ala Val Asn Ala Arg Asp Met His Pro Asp Cys Ser Ile Glu Ser

450 455 460

Gly Asp Tyr Glu Gly Gln Leu Leu Asn Glu Phe Ala Tyr Leu Ser Thr

465 470 475 480

Gln Ala Arg Gly Asn Gly Pro Met Ala Thr Val Ser Ser Ser Gly Asn

485 490 495

Thr Pro Cys Arg Pro Arg Lys Phe Arg Ala His Asn Val Ala Pro Thr

500 505 510

Asn Leu Glu Asp Leu Phe Ala Ser Glu Val Phe Ser Pro Lys Met Thr

515 520 525

Ala Ser Glu Ser Ala Phe Leu Ser Glu Ile Gln Ser His Lys Ser Ala

530 535 540

Gln Leu Ser Pro Gln Leu Gln Ser Gln Met Leu Ser Ser Phe Asn Thr

545 550 555 560

Gln Val Tyr Pro Gln Gly Ser Thr Gln Gly Gln Met His Met Gln His

565 570 575

Gly Gly Val Asp Cys Gln Ser Pro Ser Val Phe Leu Ser Pro Pro Pro

580 585 590

Val Gln Leu Ala Ser Tyr Ser Leu Ser Ser Leu Gly Pro Leu Ser Ser

595 600 605

Leu Thr Gly Glu Leu Glu Arg Gln Asn Ser Asn Gly Ser Pro Leu Ser

610 615 620

Pro Ile Met Ser Thr Ala Ala Asp Ser Arg Ala Val Ala Phe Ser Gln

625 630 635 640

Arg Asp Lys Gly Ser Ser Arg Ser Gly Asp Leu Gly Gly Ala Thr Thr

645 650 655

Trp Ser Glu Trp Gly Ser Pro Thr Gly Lys Val Asn Trp Gly Ile Arg

660 665 670

Gly Glu Glu Leu Gln Lys Phe Arg Lys Ser Ala Ser Phe Gly Ile Arg

675 680 685

Ser Ser Asp Glu Pro Asp Leu Ser Trp Val Gln Lys Leu Phe Lys Glu

690 695 700

Ala Pro Met Glu Ser Met Asp Arg Gly Thr Met Gly Arg Ser Met Asp

705 710 715 720

Ile Ala Asn Ser Val Gln Met Glu Ala Thr Asp Leu Gly Gly Trp Ile

725 730 735

Ser Gln Ile Asn Pro Asp Gln Val Ala Pro Leu Thr Leu

740 745

<210>44

<211>711

<212>PRT

＜213〉soybean (Glycine max)

<400>44

Lys Ser Ala Asn Asp Lys Glu Met Lys Ser Leu Thr Val Asn Thr Glu

1 5 10 15

Asp Ser Phe Ser Ser Leu Leu Glu Leu Ala Ser Asn Asn Asp Ile Glu

20 25 30

Gly Phe Lys Val Leu Leu Glu Lys Asp Ser Ser Ser Ile Asn Glu Val

35 40 45

Gly Leu Trp Tyr Gly Arg Gln Asn Gly Ser Lys Gln Phe Val Leu Glu

50 55 60

His Arg Thr Pro Leu Met Val Ala Ala Thr Tyr Gly Ser Ile Asp Val

65 70 75 80

Met Lys Ile Ile Leu Leu Cys Pro Glu Ala Asp Val Asn Phe Ala Cys

85 90 95

Gly Ala Asn Lys Thr Thr Ala Leu His Cys Ala Ala Ser Gly Gly Ser

100 105 110

Ala Asn Ala Val Asp Ala Val Lys Ile Leu Leu Ser Ala Gly Ala Asp

115 120 125

Val Asn Gly Val Asp Ala Asn Gly Asn Arg Pro Ile Asp Val Ile Ala

130 135 140

Val Pro Pro Lys Leu Gln Gly Ala Lys Ala Val Leu Glu Glu Leu Leu

145 150 155 160

Ser Asp Ser Ala Ser Glu Gly Ser Ile Gly Glu Phe Ser Val Pro Val

165 170 175

Ser Val Asn Thr Ser Ser Leu Gly Ser Pro Gly His Ser Ser Asn Gly

180 185 190

Met Pro Tyr Thr Pro Ser Ser Ser Pro Pro Ser Pro Val Val Ala Lys

195 200 205

Phe Thr Asp Ala Ala Val Cys Ser Leu Ser Glu Lys Lys Glu Tyr Pro

210 215 220

Ile Asp Pro Ser Leu Pro Asp Ile Lys Asn Ser Ile Tyr Ala Thr Asp

225 230 235 240

Glu Phe Arg Met Phe Ser Phe Lys Val Arg Pro Cys Ser Arg Ala Tyr

245 250 255

Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu Asn Ala

260 265 270

Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Ser Cys Val Pro Cys Pro

275 280 285

Asp Phe Arg Lys Gly Ala Cys Arg Arg Gly Asp Met Cys Glu Tyr Ala

290 295 300

His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg

305 310 315 320

Leu Cys Lys Asp Gly Thr Ser Cys Asn Arg Arg Val Cys Phe Phe Ala

325 330 335

His Thr Ala Glu Glu Leu Arg Pro Leu Tyr Val Ser Thr Gly Ser Ala

340 345 350

Val Pro Ser Pro Arg Ser Ser Ala Ser Ala Pro Asn Val Met Asp Met

355 360 365

Ala Ala Ala Met Ser Leu Leu Pro Gly Ser Pro Ser Ser Val Ser Ser

370 375 380

Met Ser Pro Ser His Phe Gly Gln Pro Met Ser Pro Ser Ala Asn Gly

385 390 395 400

Met Ser Leu Ser Ser Ala Trp Ala Gln Pro Asn Val Ser Ala Leu His

405 410 415

Leu Pro Gly Ser Asn Leu Gln Ser Ser Arg Leu Arg Ser Ser Leu Ser

420 425 430

Ala Arg Asp Met Pro Pro Asp Asp Leu Asn Met Met Ser Asp Leu Asp

435 440 445

Gly Gln Gln Gln His Pro Leu Asn Asp Leu Ser Cys Tyr Leu Gln Pro

450 455 460

Arg Pro Gly Ala Gly Ser Val Ser Arg Ser Gly Arg Ser Lys Ile Leu

465 470 475 480

Thr Pro Ser Asn Leu Glu Asp Leu Phe Ser Ala Glu Ile Ser Ser Ser

485 490 495

Pro Arg Tyr Ser Asp Pro Ala Ala Gly Ser Val Phe Ser Pro Thr His

500 505 510

Lys Ser Ala Val Leu Asn Gln Phe Gln Gln Leu Gln Ser Met Leu Ser

515 520 525

Pro Ile Asn Thr Asn Leu Leu Ser Pro Lys Asn Val Glu His Pro Leu

530 535 540

Leu Gln Ala Ser Phe Gly Val Ser Pro Ser Gly Arg Met Ser Pro Arg

545 550 555 560

Ser Val Glu Pro Ile Ser Pro Met Ser Ser Arg Ile Ser Ala Phe Ala

565 570 575

Gln Arg Glu Lys Gln Gln Gln Gln Gln Gln Gln Leu Arg Ser Leu Ser

580 585 590

Ser Arg Asp Leu Gly Ala Asn Ser Pro Ala Ser Leu Val Gly Ser Pro

595 600 605

Ala Asn Pro Trp Ser Lys Trp Gly Ser Pro Asn Gly Lys Ala Asp Trp

610 615 620

Ser Val Asn Gly Asp Thr Leu Gly Arg Gln Met Arg Arg Ser Ser Ser

625 630 635 640

Phe Glu Leu Lys Asn Asn Gly Glu Glu Pro Asp Leu Ser Trp Val Gln

645 650 655

Ser Leu Val Lys Glu Ser Pro Pro Glu Met Ile Lys Glu Lys Phe Ala

660 665 670

Ser Pro Met Pro Thr Ala Ser Ala Asp Gly Pro Asn Ser Asn Ser Gln

675 680 685

Ile Glu Ser Ile Asp His Ser Val Leu Gly Ala Trp Leu Glu Gln Met

690 695 700

Gln Leu Asp Gln Leu Val Val

705 710

<210>45

<211>643

<212>PRT

＜213〉soybean

<400>45

Arg Gly Ser Gly Phe Pro Gly Arg Pro Thr Arg Pro Arg Ser Gly Arg

1 5 10 15

Thr Arg Gly Arg Thr Arg Gly Val Asn Phe Ala Cys Gly Ala Asn Lys

20 25 30

Thr Thr Ala Leu His Cys Ala Ala Ser Gly Ala Ser Thr Lys Ala Val

35 40 45

Asp Ala Val Lys Leu Leu Leu Ser Ala Gly Ala Asp Val Asn Cys Val

50 55 60

Asp Ala Asn Gly Asn Arg Pro Ile Asp Val Ile Ala Val Pro Pro Lys

65 70 75 80

Leu Gln Gly Ala Lys Ala Val Leu Glu Glu Leu Leu Ser Asp Asn Ala

85 90 95

Ser Asp Val Ser Val Gly Glu Phe Ser Val Pro Val Ser Val Asn Ser

100 105 110

Ser Ser Pro Gly Ser Pro Ala His Ser Ser Asn Gly Met Pro Tyr Thr

115 120 125

Pro Ser Val Ser Pro Pro Ser Pro Val Ala Ala Lys Phe Thr Asp Ala

130 135 140

Ala Ile Cys Ser Leu Ser Glu Lys Ala Arg Glu Tyr Pro Ile Asp Pro

145 150 155 160

Ser Leu Pro Asp Ile Lys Asn Ser Ile Tyr Ala Thr Asp Glu Phe Arg

165 170 175

Met Phe Ser Phe Lys Val Arg Pro Cys Ser Arg Ala Tyr Ser His Asp

180 185 190

Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg Arg

195 200 205

Asp Pro Arg Lys Phe His Tyr Ser Cys Val Pro Cys Pro Asp Phe Arg

210 215 220

Lys Gly Ala Cys Arg Arg Gly Asp Met Cys Glu Tyr Ala His Gly Val

225 230 235 240

Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys Lys

245 250 255

Asp Gly Thr Ser Cys Asn Arg Arg Val Cys Phe Phe Ala His Thr Ala

260 265 270

Glu Glu Leu Arg Pro Leu Tyr Val Ser Thr Gly Ser Ala Ala Pro Ser

275 280 285

Pro Arg Ser Ser Ala Ser Gly Pro Asn Val Met Asp Met Ala Ala Ala

290 295 300

Met Ser Leu Phe Pro Gly Ser Pro Ser Ser Gly Ser Ser Ile Ser Leu

305 310 315 320

Ser Ile Ser Phe Ser Leu Asp Pro Met Ser Pro Ser Ala Asn Gly Met

325 330 335

Pro Leu Ser Ser Ala Trp Ala Gln Pro Asn Val Pro Ala Leu His Leu

340 345 350

Pro Gly Ser Asn Leu Gln Ser Ser Arg Leu Arg Ser Ser Leu Ser Ala

355 360 365

Arg Asp Ile Pro Pro Glu Asp Leu Asn Met Met Ser Asp Leu Asp Gly

370 375 380

Gln Gln Gln His His Leu Asn Asp Leu Ser Cys Tyr Ile Gln Pro Arg

385 390 395 400

Pro Gly Ala Ser Ser Val Ser Arg Ser Gly Arg Ser Lys Thr Leu Thr

405 410 415

Pro Ser Asn Leu Glu Glu Leu Phe Ser Ala Glu Ile Ser Leu Ser Pro

420 425 430

Arg Tyr Ser Asp Pro Ala Ala Gly Ser Val Phe Ser Pro Thr His Lys

435 440 445

Ser Ala Val Leu Asn Gln Phe Gln Gln Leu Gln Ser Met Leu Ser Pro

450 455 460

Ile Asn Thr Asn Leu Leu Ser Pro Lys Asn Val Glu His Pro Leu Phe

465 470 475 480

Gln Ala Ser Phe Gly Val Ser Pro Ser Gly Arg Met Ser Pro Arg Ser

485 490 495

Val Glu Pro Ile Ser Pro Met Ser Ala Arg Leu Ser Ala Phe Ala Gln

500 505 510

Arg Glu Lys Gln Gln Gln Gln Leu Arg Ser Val Ser Ser Arg Asp Leu

515 520 525

Gly Ala Asn Ser Pro Ala Ser Leu Val Gly Ser Pro Ala Asn Pro Trp

530 535 540

Ser Lys Trp Gly Ser Pro Ile Gly Lys Ala Asp Trp Ser Val Asn Gly

545 550 555 560

Asp Ser Leu Gly Arg Gln Met Arg Arg Ser Ser Ser Phe Glu Arg Lys

565 570 575

Asn Asn Gly Glu Glu Pro Asp Leu Ser Trp Val Gln Ser Leu Val Lys

580 585 590

Glu Ser Pro Pro Glu Met Ile Lys Glu Lys Phe Ala Ser Pro Met Pro

595 600 605

Thr Ala Ser Ala Asp Gly Pro Asn Ser Asn Ser Gln Ile Glu Ser Ile

610 615 620

Asp His Ser Val Leu Gly Ala Trp Leu Glu Gln Met Gln Leu Asp Gln

625 630 635 640

Leu Val Val

<210>46

<211>669

<212>PRT

＜213〉soybean

<400>46

Ser His Glu Met Asn His Leu Ser Leu Asp Thr Glu Asp Ser Leu Ala

1 5 10 15

Ser Leu Leu Leu Glu Leu Ala Ala Asn Asn Asp Val Ser Gly Phe Lys

20 25 30

Arg Leu Ile Glu Cys Glu Pro Ser Ser Ile Asp Glu Val Gly Leu Trp

35 40 45

Tyr Gly Arg His Lys Glu Ser Lys Lys Met Val Asn Glu Gln Arg Thr

50 55 60

Pro Leu Met Val Ala Ala Thr Tyr Gl y Ser Ile Asp Val Met Thr Leu

65 70 75 80

Ile Leu Ser Leu Ser Glu Ala Asp Val Asn Arg Ser Ser Gly Leu Asp

85 90 95

Lys Ser Thr Ala Leu His Cys Ala Ala Ser Gly Gly Ser Glu Asn Ala

100 105 110

Val Asp Ala Val Lys Leu Leu Leu Glu Ala Gly Ala Asp Arg Asn Ser

115 120 125

Val Asp Ala Asn Gly Arg Arg Pro Gly Asp Val Ile Val Ser Pro Pro

130 135 140

Lys Leu Asp Tyr Val Lys Lys Ser Leu Glu Glu Leu Leu Gly Ser Asp

145 150 155 160

Asp Trp Ser Leu Leu Arg Val Met Arg Ser Thr Cys Asn Gly Cys Ser

165 170 175

Ala Glu Asp Leu Lys Met Lys Thr Asn Glu Val Ser Glu Lys Lys Glu

180 185 190

Tyr Pro Val Asp Leu Ser Leu Pro Asp Ile Lys Asn Ser Ile Tyr Ser

195 200 205

Ser Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Arg Pro Cys Ser Arg

210 215 220

Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu

225 230 235 240

Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Ser Cys Val Pro

245 250 255

Cys Pro Glu Phe Arg Lys Gly Ala Cys Arg Arg Gly Asp Met Cys Glu

260 265 270

Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg

275 280 285

Thr Arg Leu Cys Lys Asp Gly Thr Asn Cys Ala Arg Arg Val Cys Phe

290 295 300

Phe Ala His Thr Asn Glu Glu Leu Arg Pro Leu Tyr Val Ser Thr Gly

305 310 315 320

Ser Ala Val Pro Ser Pro Arg Ser Ser Ala Ser Ser Ala Met Asp Phe

325 330 335

Val Ala Ala Ile Ser Pro Ser Ser Met Ser Val Met Ser Pro Ser Pro

340 345 350

Phe Thr Pro Pro Met Ser Pro Ser Ser Ala Ser Ile Ala Trp Pro Gln

355 360 365

Pro Asn Ile Pro Ala Leu His Leu Pro Gly Ser Asn Phe His Ser Ser

370 375 380

Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp Phe Ser Val Asp Asp Phe

385 390 395 400

Asp Leu Leu Leu Pro Asp Tyr Asp His His His His Gln Gln Gln Gln

405 410 415

Gln Gln Phe Leu Asn Glu Leu Ser Cys Leu Ser Pro His Ala Met Asn

420 425 430

Cys Asn Thr Met Asn Arg Ser Gly Arg Met Lys Pro Leu Thr Pro Ser

435 440 445

Asn Leu Asp Asp Leu Phe Ser Ala Glu Ser Ser Ser Pro Arg Tyr Ala

450 455 460

Asp Pro Ala Leu Ala Ser Ala Val Phe Ser Pro Thr His Lys Ser Ala

465 470 475 480

Val Phe Asn Gln Phe Gln His Gln Gln Ser Met Leu Ala Pro Leu Asn

485 490 495

Thr Asn Phe Ala Ser Lys Asn Phe Glu His Pro Leu Leu Gln Ala Ser

500 505 510

Leu Gly Met Ser Pro Arg Asn Val Glu Pro Ile Ser Pro Met Gly Ser

515 520 525

Arg Ile Ser Met Leu Ala Gln Arg Glu Lys Gln Gln Phe Arg Ser Leu

530 535 540

Ser Phe Arg Glu Leu Gly Ser Asn Ser Ala Ala Ala Ser Ala Asp Ser

545 550 555 560

Trp Ser Lys Trp Gly Ser Pro Asn Val Lys Leu Asp Trp Pro Val Gly

565 570 575

Ala Gly Glu Val Gly Lys Leu Arg Arg Ser Ser Ser Phe Glu Leu Gly

580 585 590

Asn Asn Gly Glu Glu Pro Asp Leu Ser Trp Val Gln Ser Leu Val Lys

595 600 605

Glu Ser Pro Ala Glu Val Lys Asp Lys Leu Ala Thr Thr Val Ser Tyr

610 615 620

Val Ala Ala Ala Ala Ala Gly Ser Ser Ser Glu Gly Ser Asn Ile Ser

625 630 635 640

Thr Gln Met Glu Ser Val Val Asp His Ala Val Leu Gly Ala Trp Leu

645 650 655

Glu Gln Met Gln Leu Asp His Leu Val Ala Gln Gln Asn

660 665

<210>47

<211>580

<212>PRT

＜213〉Arabidopis thaliana

<400>47

Met Cys Ser Gly Pro Lys Ser Asn Leu Cys Ser Ser Arg Thr Leu Thr

1 5 10 15

Glu Ile Glu Ser Arg Gln Lys Glu Glu Glu Thr Met Leu Leu Leu Glu

20 25 30

Phe Ala Ala Cys Asp Asp Leu Asp Ser Phe Lys Arg Glu Val Glu Glu

35 40 45

Lys Gly Leu Asp Leu Asp Glu Ser Gly Leu Trp Tyr Cys Arg Arg Val

50 55 60

Gly Ser Lys Lys Met Gly Leu Glu Glu Arg Thr Pro Leu Met Val Ala

65 70 75 80

Ala Met Tyr Gly Ser Ile Lys Val Leu Thr Phe Ile Val Ser Thr Gly

85 90 95

Lys Ser Asp Val Asn Arg Ala Cys Gly Glu Glu Arg Val Thr Pro Leu

100 105 110

His Cys Ala Val Ala Gly Cys Ser Val Asn Met Ile Glu Val Ile Asn

115 120 125

Val Leu Leu Asp Ala Ser Ala Leu Val Asn Ser Val Asp Ala Asn Gly

130 135 140

Asn Gln Pro Leu Asp Val Phe Val Arg Val Ser Arg Phe Val Ala Ser

145 150 155 160

Pro Arg Arg Lys Ala Val Glu Leu Leu Leu Arg Gly Gly Gly Val Gly

165 170 175

Gly Leu Ile Asp Glu Ala Val Glu Glu Glu Ile Lys Ile Val Ser Lys

180 185 190

Tyr Pro Ala Asp Ala Ser Leu Pro Asp Ile Asn Glu Gly Val Tyr Gly

195 200 205

Ser Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Lys Pro Cys Ser Arg

210 215 220

Ala Tyr Ser His Asp Trp Thr Glu Cys Ala Phe Val His Pro Gly Glu

225 230 235 240

Asn Ala Arg Arg Arg Asp Pro Arg Lys Tyr Pro Tyr Thr Cys Val Pro

245 250 255

Cys Pro Glu Phe Arg Lys Gly Ser Cys Pro Lys Gly Asp Ser Cys Glu

260 265 270

Tyr Ala His Gly Val Phe Glu Ser Trp Leu His Pro Ala Gln Tyr Lys

275 280 285

Thr Arg Leu Cys Lys Asp Glu Thr Gly Cys Ala Arg Lys Val Cys Phe

290 295 300

Phe Ala His Lys Arg Glu Glu Met Arg Pro Val Asn Ala Ser Thr Gly

305 310 315 320

Ser Ala Val Ala Gln Ser Pro Phe Ser Ser Leu Glu Met Met Pro Gly

325 330 335

Leu Ser Pro Leu Ala Tyr Ser Ser Gly Val Ser Thr Pro Pro Val Ser

340 345 350

Pro Met Ala Asn Gly Val Pro Ser Ser Pro Arg Asn Gly Gly Ser Trp

355 360 365

Gln Asn Arg Val Asn Thr Leu Thr Pro Pro Ala Leu Gln Leu Asn Gly

370 375 380

Gly Ser Arg Leu Lys Ser Thr Leu Ser Ala Arg Asp Ile Asp Met Glu

385 390 395 400

Met Glu Met Glu Leu Arg Leu Arg Gly Phe Gly Asn Asn Val Glu Glu

405 410 415

Thr Phe Gly Ser Tyr Val Ser Ser Pro Ser Arg Asn Ser Gln Met Gly

420 425 430

Gln Asn Met Asn Gln His Tyr Pro Ser Ser Pro Val Arg Gln Pro Pro

435 440 445

Ser Gln His Gly Phe Glu Ser Ser Ala Ala Ala Ala Val Ala Val Met

450 455 460

Lys Ala Arg Ser Thr Ala Phe Ala Lys Arg Ser Leu Ser Phe Lys Pro

465 470 475 480

Ala Thr Gln Ala Ala Pro Gln Ser Asn Leu Ser Asp Trp Gly Ser Pro

485 490 495

Asn Gly Lys Leu Glu Trp Gly Met Lys Gly Glu Glu Leu Asn Lys Met

500 505 510

Arg Arg Ser Val Ser Phe Gly Ile His Gly Asn Asn Asn Asn Asn Ala

515 520 525

Ala Arg Asp Tyr Arg Asp Glu Pro Asp Val Ser Trp Val Asn Ser Leu

530 535 540

Val Lys Asp Ser Thr Val Val Ser Glu Arg Ser Phe Gly Met Asn Glu

545 550 555 560

Arg Val Arg Ile Met Ser Trp Ala Glu Gln Met Tyr Arg Glu Lys Glu

565 570 575

Gln Thr Val Val

580

<210>48

<211>719

<212>PRT

＜213〉Arabidopis thaliana

<400>48

Met Cys Cys Gly Ser Asp Arg Leu Asn Gln Ile Val Ser Ser Arg Ser

1 5 10 15

Ser Leu Pro Ile Ser Phe Glu Glu Asp Asn Asn Leu Val Thr Asn Thr

20 25 30

Asp Met Asn His Ile Thr Val Glu Thr Glu Asp Thr Phe Ala Ser Leu

35 40 45

Leu Glu Leu Ala Ala Asn Asn Asp Val Glu Gly Val Arg Leu Ser Ile

50 55 60

Glu Arg Asp Pro Ser Cys Val Asp Glu Ala Gly Leu Trp Tyr Gly Arg

65 70 75 80

Gln Lys Gly Ser Lys Ala Met Val Asn Asp Tyr Arg Thr Pro Leu Met

85 90 95

Val Ala Ala Thr Tyr Gly Ser Ile Asp Val Ile Lys Leu Ile Val Ser

100 105 110

Leu Thr Asp Ala Asp Val Asn Arg Ala Cys Gly Asn Asp Gln Thr Thr

115 120 125

Ala Leu His Cys Ala Ala Ser Gly Gly Ala Val Asn Ala Ile Gln Val

130 135 140

Val Lys Leu Leu Leu Ala Ala Gly Ala Asp Leu Asn Leu Leu Asp Ala

145 150 155 160

Glu Gly Gln Arg Ala Gly Asp Val Ile Val Val Pro Pro Lys Leu Glu

165 170 175

Gly Val Lys Leu Met Leu Gln Glu Leu Leu Ser Ala Asp Gly Ser Ser

180 185 190

Thr Ala Glu Arg Asn Leu Arg Val Val Thr Asn ValPro Asn Arg Ser

195 200 205

Ser Ser Pro Cys His Ser Pro Thr Gly Glu Asn Gly Gly Ser Gly Ser

210 215 220

Gly Ser Pro Leu Gly Ser Pro Phe Lys Leu Lys Ser Thr Glu Phe Lys

225 230 235 240

Lys Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp Ile Lys Asn Ser Ile

245 250 255

Tyr Ala Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Arg Pro Cys

260 265 270

Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro

275 280 285

Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Ser Cys

290 295 300

Val Pro Cys Pro Asp Phe Arg Lys Gly Ala Cys Arg Arg Gly Asp Met

305 310 315 320

Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln

325 330 335

Tyr Arg Thr Arg Leu Cys Lys Asp Gly Thr Gly Cys Ala Arg Arg Val

340 345 350

Cys Phe Phe Ala His Thr Pro Glu Glu Leu Arg Pro Leu Tyr Ala Ser

355 360 365

Thr Gly Ser Ala Val Pro Ser Pro Arg Ser Asn Ala Asp Tyr Ala Ala

370 375 380

Ala Leu Ser Leu Leu Pro Gly Ser Pro Ser Gly Val Ser Val Met Ser

385 390 395 400

Pro Leu Ser Pro Ser Ala Ala Gly Asn Gly Met Ser His Ser Asn Met

405 410 415

Ala Trp Pro Gln Pro Asn Val Pro Ala Leu His Leu Pro Gly Ser Asn

420 425 430

Leu Gln Ser Ser Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp Ile Pro

435 440 445

Thr Asp Glu Phe Asn Met Leu Ala Asp Tyr Glu Gln Gln Gln Leu Leu

450 455 460

Asn Glu Tyr Ser Asn Ala Leu Ser Arg Ser Gly Arg Met Lys Ser Met

465 470 475 480

Pro Pro Ser Asn Leu Glu Asp Leu Phe Ser Ala Glu Gly Ser Ser Ser

485 490 495

Pro Arg Phe Thr Asp Ser Ala Leu Ala Ser Ala Val Phe Ser Pro Thr

500 505 510

His Lys Ser Ala Val Phe Asn Gln Phe Gln Gln Gln Gln Gln Gln Gln

515 520 525

Gln Gln Gln Ser Met Leu Ser Pro Ile Asn Thr Ser Phe Ser Ser Pro

530 535 540

Lys Ser Val Asp His Ser Leu Phe Ser Gly Gly Gly Arg Met Ser Pro

545 550 555 560

Arg Asn Val Val Glu Pro Ile Ser Pro Met Ser Ala Arg Val Ser Met

565 570 575

Leu Ala Gln Cys Val Lys Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln

580 585 590

Gln Gln Gln His Gln Phe Arg Ser Leu Ser Ser Arg Glu Leu Arg Thr

595 600 605

Asn Ser Ser Pro Ile Val Gly Ser Pro Val Asn Asn Asn Thr Trp Ser

610 615 620

Ser Lys Trp Gly Ser Ser Asn Gly Gln Pro Asp Trp Gly Met Ser Ser

625 630 635 640

Glu Ala Leu Gly Lys Leu Arg Ser Ser Ser Ser Phe Asp Gly Asp Glu

645 650 655

Pro Asp Val Ser Trp Val Gln Ser Leu Val Lys Glu Thr Pro Ala Glu

660 665 670

Ala Lys Glu Lys Ala Ala Thr Ser Ser Ser Gly Glu His Val Met Lys

675 680 685

Gln Pro Asn Pro Val Glu Pro Val Met Asp His Ala Gly Leu Glu Ala

690 695 700

Trp Ile Glu Gln Met Gln Leu Asp Gln Leu Val Ala Gln Gln Asn

705 710 715

<210>49

<211>686

<212>PRT

＜213〉Arabidopis thaliana

<400>49

Met Asn His Ile Thr Val Glu Thr Glu Asp Thr Phe Ala Ser Leu Leu

1 5 10 15

Glu Leu Ala Ala Asn Asn Asp Val Glu Gly Val Arg Leu Ser Ile Glu

20 25 30

Arg Asp Pro Ser Cys Val Asp Glu Ala Gly Leu Trp Tyr Gly Arg Gln

35 40 45

Lys Gly Ser Lys Ala Met Val Asn Asp Tyr Arg Thr Pro Leu Met Val

50 55 60

Ala Ala Thr Tyr Gly Ser Ile Asp Val Ile Lys Leu Ile Val Ser Leu

65 70 75 80

Thr Asp Ala Asp Val Asn Arg Ala Cys Gly Asn Asp Gln Thr Thr Ala

85 90 95

Leu His Cys Ala Ala Ser Gly Gly Ala Val Asn Ala Ile Gln Val Val

100 105 110

Lys Leu Leu Leu Ala Ala Gly Ala Asp Leu Asn Leu Leu Asp Ala Glu

115 120 125

Gly Gln Arg Ala Gly Asp Val Ile Val Val Pro Pro Lys Leu Glu Gly

130 135 140

Val Lys Leu Met Leu Gln Glu Leu Leu Ser Ala Asp Gly Ser Ser Thr

145 150 155 160

Ala Glu Arg Asn Leu Arg Val Val Thr Asn Val Pro Asn Arg Ser Ser

165 170 175

Ser Pro Cys His Ser Pro Thr Gly Glu Asn Gly Gly Ser Gly Ser Gly

180 185 190

Ser Pro Leu Gly Ser Pro Phe Lys Leu Lys Ser Thr Glu Phe Lys Lys

195 200 205

Glu Tyr Pro Val Asp Pro Ser Leu Pro Asp Ile Lys Asn Ser Ile Tyr

210 215 220

Ala Thr Asp Glu Phe Arg Met Tyr Ser Phe Lys Val Arg Pro Cys Ser

225 230 235 240

Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly

245 250 255

Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Ser Cys Val

260 265 270

Pro Cys Pro Asp Phe Arg Lys Gly Ala Cys Arg Arg Gly Asp Met Cys

275 280 285

Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr

290 295 300

Arg Thr Arg Leu Cys Lys Asp Gly Thr Gly Cys Ala Arg Arg Val Cys

305 310 315 320

Phe Phe Ala His Thr Pro Glu Glu Leu Arg Pro Leu Tyr Ala Ser Thr

325 330 335

Gly Ser Ala Val Pro Ser Pro Arg Ser Asn Ala Asp Tyr Ala Ala Ala

340 345 350

Leu Ser Leu Leu Pro Gly Ser Pro Ser Gly Val Ser Val Met Ser Pro

355 360 365

Leu Ser Pro Ser Ala Ala Gly Asn Gly Met Ser His Ser Asn Met Ala

370 375 380

Trp Pro Gln Pro Asn Val Pro Ala Leu His Leu Pro Gly Ser Asn Leu

385 390 395 400

Gln Ser Ser Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp Ile Pro Thr

405 410 415

Asp Glu Phe Asn Met Leu Ala Asp Tyr Glu Gln Gln Gln Leu Leu Asn

420 425 430

Glu Tyr Ser Asn Ala Leu Ser Arg Ser Gly Arg Met Lys Ser Met Pro

435 440 445

Pro Ser Asn Leu Glu Asp Leu Phe Ser Ala Glu Gly Ser Ser Ser Pro

450 455 460

Arg Phe Thr Asp Ser Ala Leu Ala Ser Ala Val Phe Ser Pro Thr His

465 470 475 480

Lys Ser Ala Val Phe Asn Gln Phe Gln Gln Gln Gln Gln Gln Gln Gln

485 490 495

Gln Gln Ser Met Leu Ser Pro Ile Asn Thr Ser Phe Ser Ser Pro Lys

500 505 510

Ser Val Asp His Ser Leu Phe Ser Gly Gly Gly Arg Met Ser Pro Arg

515 520 525

Asn Val Val Glu Pro Ile Ser Pro Met Ser Ala Arg Val Ser Met Leu

530 535 540

Ala Gln Cys Val Lys Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln

545 550 555 560

Gln Gln His Gln Phe Arg Ser Leu Ser Ser Arg Glu Leu Arg Thr Asn

565 570 575

Ser Ser Pro Ile Val Gly Ser Pro Val Asn Asn Asn Thr Trp Ser Ser

580 585 590

Lys Trp Gly Ser Ser Asn Gly Gln Pro Asp Trp Gly Met Ser Ser Glu

595 600 605

Ala Leu Gly Lys Leu Arg Ser Ser Ser Ser Phe Asp Gly Asp Glu Pro

610 615 620

Asp Val Ser Trp Val Gln Ser Leu Val Lys Glu Thr Pro Ala Glu Ala

625 630 635 640

Lys Glu Lys Ala Ala Thr Ser Ser Ser Gly Glu His Val Met Lys Gln

645 650 655

Pro Asn Pro Val Glu Pro Val Met Asp His Ala Gly Leu Glu Ala Trp

660 665 670

Ile Glu Gln Met Gln Leu Asp Gln Leu Val Ala Gln Gln Asn

675 680 685

<210>50

<211>633

<212>PRT

＜213〉Arabidopis thaliana

<400>50

Met Val Asn Asp Tyr Arg Thr Pro Leu Met Val Ala Ala Thr Tyr Gly

1 5 10 15

Ser Ile Asp Val Ile Lys Leu Ile Val Ser Leu Thr Asp Ala Asp Val

20 25 30

Asn Arg Ala Cys Gly Asn Asp Gln Thr Thr Ala Leu His Cys Ala Ala

35 40 45

Ser Gly Gly Ala Val Asn Ala Ile Gln Val Val Lys Leu Leu Leu Ala

50 55 60

Ala Gly Ala Asp Leu Asn Leu Leu Asp Ala Glu Gly Gln Arg Ala Gly

65 70 75 80

Asp Val Ile Val Val Pro Pro Lys Leu Glu Gly Val Lys Leu Met Leu

85 90 95

Gln Glu Leu Leu Ser Ala Asp Gly Ser Ser Thr Ala Glu Arg Asn Leu

100 105 110

Arg Val Val Thr Asn Val Pro Asn Arg Ser Ser Ser Pro Cys His Ser

115 120 125

Pro Thr Gly Glu Asn Gly Gly Ser Gly Ser Gly Ser Pro Leu Gly Ser

130 135 140

Pro Phe Lys Leu Lys Ser Thr Glu Phe Lys Lys Glu Tyr Pro Val Asp

145 150 155 160

Pro Ser Leu Pro Asp Ile Lys Asn Ser Ile Tyr Ala Thr Asp Glu Phe

165 170 175

Arg Met Tyr Ser Phe Lys Val Arg Pro Cys Ser Arg Ala Tyr Ser His

180 185 190

Asp Trp Thr Glu Cys Pro Phe Val His Pro Gly Glu Asn Ala Arg Arg

195 200 205

Arg Asp Pro Arg Lys Phe His Tyr Ser Cys Val Pro Cys Pro Asp Phe

210 215 220

Arg Lys Gly Ala Cys Arg Arg Gly Asp Met Cys Glu Tyr Ala His Gly

225 230 235 240

Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys

245 250 255

Lys Asp Gly Thr Gly Cys Ala Arg Arg Val Cys Phe Phe Ala His Thr

260 265 270

Pro Glu Glu Leu Arg Pro Leu Tyr Ala Ser Thr Gly Ser Ala Val Pro

275 280 285

Ser Pro Arg Ser Asn Ala Asp Tyr Ala Ala Ala Leu Ser Leu Leu Pro

290 295 300

Gly Ser Pro Ser Gly Val Ser Val Met Ser Pro Leu Ser Pro Ser Ala

305 310 315 320

Ala Gly Asn Gly Met Ser His Ser Asn Met Ala Trp Pro Gln Pro Asn

325 330 335

Val Pro Ala Leu His Leu Pro Gly Ser Asn Leu Gln Ser Ser Arg Leu

340 345 350

Arg Ser Ser Leu Asn Ala Arg Asp Ile Pro Thr Asp Glu Phe Asn Met

355 360 365

Leu Ala Asp Tyr Glu Gln Gln Gln Leu Leu Asn Glu Tyr Ser Asn Ala

370 375 380

Leu Ser Arg Ser Gly Arg Met Lys Ser Met Pro Pro Ser Asn Leu Glu

385 390 395 400

Asp Leu Phe Ser Ala Glu Gly Ser Ser Ser Pro Arg Phe Thr Asp Ser

405 410 415

Ala Leu Ala Ser Ala Val Phe Ser Pro Thr His Lys Ser Ala Val Phe

420 425 430

Asn Gln Phe Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Ser Met Leu

435 440 445

Ser Pro Ile Asn Thr Ser Phe Ser Ser Pro Lys Ser Val Asp His Ser

450 455 460

Leu Phe Ser Gly Gly Gly Arg Met Ser Pro Arg Asn Val Val Glu Pro

465 470 475 480

Ile Ser Pro Met Ser Ala Arg Val Ser Met Leu Ala Gln Cys Val Lys

485 490 495

Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln Gln His Gln Phe

500 505 510

Arg Ser Leu Ser Ser Arg Glu Leu Arg Thr Asn Ser Ser Pro Ile Val

515 520 525

Gly Ser Pro Val Asn Asn Asn Thr Trp Ser Ser Lys Trp Gly Ser Ser

530 535 540

Asn Gly Gln Pro Asp Trp Gly Met Ser Ser Glu Ala Leu Gly Lys Leu

545 550 555 560

Arg Ser Ser Ser Ser Phe Asp Gly Asp Glu Pro Asp Val Ser Trp Val

565 570 575

Gln Ser Leu Val Lys Glu Thr Pro Ala Glu Ala Lys Glu Lys Ala Ala

580 585 590

Thr Ser Ser Ser Gly Glu His Val Met Lys Gln Pro Asn Pro Val Glu

595 600 605

Pro Val Met Asp His Ala Gly Leu Glu Ala Trp Ile Glu Gln Met Gln

610 615 620

Leu Asp Gln Leu Val Ala Gln Gln Asn

625 630

<210>51

<211>678

<212>PRT

＜213〉Arabidopis thaliana

<400>51

Met Asn Asp Ala Ala Glu Trp Glu His Ser Phe Ser Ala Leu Leu Glu

1 5 10 15

Phe Ala Ala Asp Asn Asp Val Glu Gly Phe Arg Arg Gln Leu Ser Asp

20 25 30

Val Ser Cys Ile Asn Gln Met Gly Leu Trp Tyr Arg Arg Gln Arg Phe

35 40 45

Val Arg Arg Met Val Leu Glu Gln Arg Thr Pro Leu Met Val Ala Ser

50 55 60

Leu Tyr Gly Ser Leu Asp Val Val Lys Phe Ile Leu Ser Phe Pro Glu

65 70 75 80

Ala Glu Leu Asn Leu Ser Cys Gly Pro Asp Lys Ser Thr Ala Leu His

85 90 95

Cys Ala Ala Ser Gly Ala Ser Val Asn Ser Leu Asp Val Val Lys Leu

100 105 110

Leu Leu Ser Val Gly Ala Asp Pro Asn Ile Pro Asp Ala His Gly Asn

115 120 125

Arg Pro Val Asp Val Leu Val Val Ser Pro His Ala Pro Gly Leu Arg

130 135 140

Thr Ile Leu Glu Glu Ile Leu Lys Lys Asp Glu Ile Ile Ser Glu Asp

145 150 155 160

Leu His Ala Ser Ser Ser Ser Leu Gly Ser Ser Phe Arg Ser Leu Ser

165 170 175

Ser Ser Pro Asp Asn Gly Ser Ser Leu Leu Ser Leu Asp Ser Val Ser

180 185 190

Ser Pro Thr Lys Pro His Gly Thr Asp Val Thr Phe Ala Ser Glu Lys

195 200 205

Lys Glu Tyr Pro Ile Asp Pro Ser Leu Pro Asp Ile Lys Ser Gly Ile

210 215 220

Tyr Ser Thr Asp Glu Phe Arg Met Phe Ser Phe Lys Ile Arg Pro Cys

225 230 235 240

Ser Arg Ala Tyr Ser His Asp Trp Thr Glu Cys Pro Phe Ala His Pro

245 250 255

Gly Glu Asn Ala Arg Arg Arg Asp Pro Arg Lys Phe His Tyr Thr Cys

260 265 270

Val Pro Cys Pro Asp Phe Lys Lys Gly Ser Cys Lys Gln Gly Asp Met

275 280 285

Cys Glu Tyr Ala His Gly Val Phe Glu Cys Trp Leu His Pro Ala Gln

290 295 300

Tyr Arg Thr Arg Leu Cys Lys Asp Gly Met Gly Cys Asn Arg Arg Val

305 310 315 320

Cys Phe Phe Ala His Ala Asn Glu Glu Leu Arg Pro Leu Tyr Pro Ser

325 330 335

Thr Gly Ser Gly Leu Pro Ser Pro Arg Ala Ser Ser Ala Val Ser Ala

340 345 350

Ser Thr Met Asp Met Ala Ser Val Leu Asn Met Leu Pro Gly Ser Pro

355 360 365

Ser Ala Ala Gln His Ser Phe Thr Pro Pro Ile Ser Pro Ser Gly Asn

370 375 380

Gly Ser Met Pro His Ser Ser Met Gly Trp Pro Gln Gln Asn Ile Pro

385 390 395 400

Ala Leu Asn Leu Pro Gly Ser Asn Ile Gln Leu Ser Arg Leu Arg Ser

405 410 415

Ser Leu Asn Ala Arg Asp Ile Pro Ser Glu Gln Leu Ser Met Leu His

420 425 430

Glu Phe Glu Met Gln Arg Gln Leu Ala Gly Asp Met His Ser Pro Arg

435 440 445

Phe Met Asn His Ser Ala Arg Pro Lys Thr Leu Asn Pro Ser Asn Leu

450 455 460

Glu Glu Leu Phe Ser Ala Glu Val Ala Ser Pro Arg Phe Ser Asp Gln

465 470 475 480

Leu Ala Val Ser Ser Val Leu Ser Pro Ser His Lys Ser Ala Leu Leu

485 490 495

Asn Gln Leu Gln Asn Asn Lys Gln Ser Met Leu Ser Pro Ile Lys Thr

500 505 510

Asn Leu Met Ser Ser Pro Lys Asn Val Glu Gln His Ser Leu Leu Gln

515 520 525

Gln Ala Ser Ser Pro Arg Gly Gly Glu Pro Ile Ser Pro Met Asn Ala

530 535 540

Arg Met Lys Gln Gln Leu His Ser Arg Ser Leu Ser Ser Arg Asp Phe

545 550 555 560

Gly Ser Ser Leu Pro Arg Asp Leu Met Pro Thr Asp Ser Gly Ser Pro

565 570 575

Leu Ser Pro Trp Ser Ser Trp Asp Gln Thr His Gly Ser Lys Val Asp

580 585 590

Trp Ser Val Gln Ser Asp Glu Leu Gly Arg Leu Arg Lys Ser His Ser

595 600 605

Leu Ala Asn Asn Pro Asn Arg Glu Ala Asp Val Ser Trp Ala Gln Gln

610 615 620

Met Leu Lys Asp Ser Ser Ser Pro Arg Asn Gly Asn Arg Val Val Asn

625 630 635 640

Met Asn Gly Ala Arg Pro Leu Thr Gln Gly Gly Ser Ser Val Asn Pro

645 650 655

His Asn Ser Asp Thr Arg Glu Ser Asp Ile Leu Asp Ala Trp Leu Glu

660 665 670

Gln Leu His Leu Asp Arg

675

<210>52

<211>640

<212>PRT

＜213〉Arabidopis thaliana

<400>52

Met Gly Leu Trp Tyr Arg Arg Gln Arg Phe Val Arg Arg Met Val Leu

1 5 10 15

Glu Gln Arg Thr Pro Leu Met Val Ala Ser Leu Tyr Gly Ser Leu Asp

20 25 30

Val Val Lys Phe Ile Leu Ser Phe Pro Glu Ala Glu Leu Asn Leu Ser

35 40 45

Cys Gly Pro Asp Lys Ser Thr Ala Leu His Cys Ala Ala Ser Gly Ala

50 55 60

Ser Val Asn Ser Leu Asp Val Val Lys Leu Leu Leu Ser Val Gly Ala

65 70 75 80

Asp Pro Asn Ile Pro Asp Ala His Gly Asn Arg Pro Val Asp Val Leu

85 90 95

Val Val Ser Pro His Ala Pro Gly Leu Arg Thr Ile Leu Glu Glu Ile

100 105 110

Leu Lys Lys Asp Glu Ile Ile Ser Glu Asp Leu His Ala Ser Ser Ser

115 120 125

Ser Leu Gly Ser Ser Phe Arg Ser Leu Ser Ser Ser Pro Asp Asn Gly

130 135 140

Ser Ser Leu Leu Ser Leu Asp Ser Val Ser Ser Pro Thr Lys Pro His

145 150 155 160

Gly Thr Asp Val Thr Phe Ala Ser Glu Lys Lys Glu Tyr Pro Ile Asp

165 170 175

Pro Ser Leu Pro Asp Ile Lys Ser Gly Ile Tyr Ser Thr Asp Glu Phe

180 185 190

Arg Met Phe Ser Phe Lys Ile Arg Pro Cys Ser Arg Ala Tyr Ser His

195 200 205

Asp Trp Thr Glu Cys Pro Phe Ala His Pro Gly Glu Asn Ala Arg Arg

210 215 220

Arg Asp Pro Arg Lys Phe His Tyr Thr Cys Val Pro Cys Pro Asp Phe

225 230 235 240

Lys Lys Gly Ser Cys Lys Gln Gly Asp Met Cys Glu Tyr Ala His Gly

245 250 255

Val Phe Glu Cys Trp Leu His Pro Ala Gln Tyr Arg Thr Arg Leu Cys

260 265 270

Lys Asp Gly Met Gly Cys Asn Arg Arg Val Cys Phe Phe Ala His Ala

275 280 285

Asn Glu Glu Leu Arg Pro Leu Tyr Pro Ser Thr Gly Ser Gly Leu Pro

290 295 300

Ser Pro Arg Ala Ser Ser Ala Val Ser Ala Ser Thr Met Asp Met Ala

305 310 315 320

Ser Val Leu Asn Met Leu Pro Gly Ser Pro Ser Ala Ala Gln His Ser

325 330 335

Phe Thr Pro Pro Ile Ser Pro Ser Gly Asn Gly Ser Met Pro His Ser

340 345 350

Ser Met Gly Trp Pro Gln Gln Asn Ile Pro Ala Leu Asn Leu Pro Gly

355 360 365

Ser Asn Ile Gln Leu Ser Arg Leu Arg Ser Ser Leu Asn Ala Arg Asp

370 375 380

Ile Pro Ser Glu Gln Leu Ser Met Leu His Glu Phe Glu Met Gln Arg

385 390 395 400

Gln Leu Ala Gly Asp Met His Ser Pro Arg Phe Met Asn His Ser Ala

405 410 415

Arg Pro Lys Thr Leu Asn Pro Ser Asn Leu Glu Glu Leu Phe Ser Ala

420 425 430

Glu Val Ala Ser Pro Arg Phe Ser Asp Gln Leu Ala Val Ser Ser Va1

435 440 445

Leu Ser Pro Ser His Lys Ser Ala Leu Leu Asn Gln Leu Gln Asn Asn

450 455 460

Lys Gln Ser Met Leu Ser Pro Ile Lys Thr Asn Leu Met Ser Ser Pro

465 470 475 480

Lys Asn Val Glu Gln His Ser Leu Leu Gln Gln Ala Ser Ser Pro Arg

485 490 495

Gly Gly Glu Pro Ile Ser Pro Met Asn Ala Arg Met Lys Gln Gln Leu

500 505 510

His Ser Arg Ser Leu Ser Ser Arg Asp Phe Gly Ser Ser Leu Pro Arg

515 520 525

Asp Leu Met Pro Thr Asp Ser Gly Ser Pro Leu Ser Pro Trp Ser Ser

530 535 540

Trp Asp Gln Thr His Gly Ser Lys Val Asp Trp Ser Val Gln Ser Asp

545 550 555 560

Glu Leu Gly Arg Leu Arg Lys Ser His Ser Leu Ala Asn Asn Pro Asn

565 570 575

Arg Glu Ala Asp Val Ser Trp Ala Gln Gln Met Leu Lys Asp Ser Ser

580 585 590

Ser Pro Arg Asn Gly Asn Arg Val Val Asn Met Asn Gly Ala Arg Pro

595 600 605

Leu Thr Gln Gly Gly Ser Ser Val Asn Pro His Asn Ser Asp Thr Arg

610 615 620

Glu Ser Asp Ile Leu Asp Ala Trp Leu Glu Gln Leu His Leu Asp Arg

625 630 635 640

<210>53

<211>2158

<212>DNA

＜213〉rice

<400>53

tgctccttct tcattgcaaa gaaagcacca ccttttaaag aatcctcctc acactccatt 60

cttcttaaaa aacccaccac aacacaattc ccacttgttt cttcatcatc acctacttca 120

atcaaaaaac attccaactt tcttctcaat ttcattccag gatagtatta tgtgcagtgg 180

accaaagagc aatctctgct cttcaagaac cttaacagaa atcgaatcaa ggcaaaagga 240

agaagaaaca atgcttctcc tcgaattcgc tgcttgtgat gatcttgact cgttcaagag 300

agaggttgaa gagaaagggc ttgatttgga tgagtcaggg ttatggtatt gcagacgtgt 360

cggttctaag aagatgggtc ttgaagaaag aacaccttta atggttgcag ctatgtatgg 420

aagcataaag gttttgactt tcatcgtttc cactggaaaa tctgatgtga acagagcttg 480

tggtgaagag agagttactc cgcttcactg tgctgttgct ggctgttctg tgaatatgat 540

tgaagtcatc aatgtcttgc ttgatgcttc tgctttggtt aactctgttg atgctaatgg 600

gaatcaacct ttggatgtgt ttgttcgagt ttcgaggttt gtggctagtc cgaggaggaa 660

agcggttgag ttgttgctga gaggaggagg tgttggagga ttgatcgatg aggcggttga 720

agaagagatc aagattgtct ctaagtatcc agctgatgct tctttaccgg atataaacga 780

aggggtttat ggaagtgatg agtttaggat gtatagcttt aaggttaagc catgttctag 840

ggcttattct catgattgga ccgagtgtgc ttttgttcat ccgggagaaa atgcgaggag 900

gagagatccg aggaagtatc cttacacttg tgtcccctgt cccgagttcc gtaaaggatc 960

atgcccgaaa ggagattctt gcgagtatgc tcacggggtt ttcgagtcgt ggcttcaccc 1020

cgcgcagtat aaaacccggc tttgtaaaga tgaaacgggt tgtgcaagga aagtttgttt 1080

ctttgctcat aaacgcgaag agatgagacc tgttaatgct tcaactggct ctgccgtggc 1140

tcagtctccg tttagcagct tggagatgat gccagggttg tctcctcttg cttattcttc 1200

aggagtttcg actcctccgg tttctccaat ggctaatggt gttccttcct ctccaagaaa 1260

cggcggatca tggcagaaca gagtcaatac ccttactcca ccggctttgc agctcaatgg 1320

tggaagcaga ttgaagtcca cactgagcgc tagagatatc gatatggaga tggagatgga 1380

attgagactc cgcggttttg gcaacaatgt ggaagagacg ttcgggtctt atgtttcctc 1440

tccaagtagg aattctcaaa tgggtcaaaa catgaaccaa cattatccat cttccccggt 1500

gagacaaccg ccatctcaac acgggttcga atcttcagca gctgcagcgg ttgcagtgat 1560

gaaagcgaga tcaaccgcct ttgcgaaacg tagcttgagc ttcaaaccag ctactcaagc 1620

agcaccacag tcgaatctct cggattgggg atctccaaac gggaagctgg aatggggaat 1680

gaaaggagaa gagctgaata agatgagaag aagtgtttcc tttggaatcc atggaaacaa 1740

caacaataac gcagctagag actacaggga cgagccagat gtgtcatggg ttaactcttt 1800

agttaaagac agtactgtgg tgtctgagag aagctttgga atgaatgaga gggttcggat 1860

aatgtcgtgg gctgagcaaa tgtacagaga gaaggagcag actgtggtgt aaacacacac 1920

aaagatggtt tcttatatat attgcttttg ggccatctct gcaaatttga ttctttaatt 1980

tttgtgactt tctttagttg ttactgttat tagtagtata tggtttgttg tcactacgag 2040

tctacgtgat gaaaagatag aagttaattg cattagtttc tatattcgtt tctcatcctc 2100

ttgtaattta tcaaaccatg aaatggctaa gcaatccaaa ccgaaaaaaa aaaaaaaa 2158

<210>54

<211>2193

<212>DNA

＜213〉rice

<400>54

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140

cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200

tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260

gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320

ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380

gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440

aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500

ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560

atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620

acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680

cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740

ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800

gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860

tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920

attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980

tgtcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040

cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100

agctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160

tggtgtagct tgccactttc accagcaaag ttc 2193

<210>55

<211>1827

<212>DNA

＜213〉rice

<400>55

gcttgagtca tagggagaaa acaaatcgat catatttgac tcttttccct ccatctctct 60

taccggcaaa aaaagtagta ctggtttata tgtaaagtaa gattctttaa ttatgtgaga 120

tccggcttaa tgcttttctt ttgtcacata tactgcattg caacaattgc catatattca 180

cttctgccat cccattatat agcaactcaa gaatggattg atatatcccc tattactaat 240

ctagacatgt taaggctgag ttgggcagtc catcttccca acccaccacc ttcgtttttc 300

gcgcacatac ttttcaaact actaaatggt gtgtttttta aaaatatttt caatacaaaa 360

gttgctttaa aaaattatat tgatccattt ttttaaaaaa aatagctaat acttaattaa 420

tcacgtgtta aaagaccgct ccgttttgcg tgcaggaggg ataggttcac atcctgcatt 480

accgaacaca gcctaaatct tgttgtctag attcgtagta ctggatatat taaatcatgt 540

tctaagttac tatatactga gatgaataga ataagtaaaa ttagacccac cttaagtctt 600

gatgaagtta ctactagctg cgtttgggag gacttcccaa aaaaaaaagt attagccatt 660

agcacgtgat taattaagta ctagtttaaa aaacttaaaa aataaattaa tatgattctc 720

ttaagtaact ctcctataga aaacttttac aaaattacac cgtttaatag tttggaaaat 780

atgtcagtaa aaaataagag agtagaagtt atgaaagtta gaaaaagaat tgttttagta 840

gtatacagtt ataaactatt ccctctgttc taaaacataa gggattatgg atggattcga 900

catgtaccag taccatgaat cgaatccaga caagtttttt atgcatattt attctactat 960

aatatatcac atctgctcta aatatcttat atttcgaggt ggagactgtc gctatgtttt 1020

tctgcccgtt gctaagcaca cgccaccccc gatgcgggga cgcctctggc cttcttgcca 1080

cgataattga atggaacttc cacattcaga ttcgataggt gaccgtcgac tccaagtgct 1140

ttgcacaaaa caactccggc ctcccggcca ccagtcacac gactcacggc actaccaccc 1200

ctgactccct gaggcggacc tgccactgtt ctgcatgcga agctatctaa aattctgaag 1260

caaagaaagc acagcacatg ctccgggaca cgcgccaccc ggcggaaaag ggctcggtgt 1320

ggcgatctca cagccgcata tcgcatttca caagccgccc atctccaccg gcttcacgag 1380

gctcatcgcg gcacgaccgc gcacggaacg cacgcggccg acccgcgcgc ctcgatgcgc 1440

gagcccatcc gccgcgtcct ccctttgcct ttgccgctat cctctcggtc gtatcccgtt 1500

tctctgtctt ttgctccccg gcgcgcgcca gttcggagta ccagcgaaac ccggacacct 1560

ggtacacctc cgccggccac aacgcgtgtc cccctacgtg gccgcgcagc acatgcccat 1620

gcgcgacacg tgcacctcct catccaaact ctcaagtctc aacggtccta taaatgcacg 1680

gatagcctca agctgctcgt cacaaggcaa gaggcaagag gcaagagcat ccgtattaac 1740

cagccttttg agacttgaga gtgtgtgtga ctcgatccag cgtagtttca gttcgtgtgt 1800

tggtgagtga ttccagccaa gtttgcg 1827

<210>56

<211>2194

<212>DNA

＜213〉rice

<400>56

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctccttctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt ctcgatccat atcttccggt cgagttcttg gtcgatctct tccctcctcc 1140

acctcctcct cacagggtat gtgcctccct tcggttgttc ttggatttat tgttctaggt 1200

tgtgtagtac gggcgttgat gttaggaaag gggatctgta tctgtgatga ttcctgttct 1260

tggatttggg atagaggggt tcttgatgtt gcatgttatc ggttcggttt gattagtagt 1320

atggttttca atcgtctgga gagctctatg gaaatgaaat ggtttaggga tcggaatctt 1380

gcgattttgt gagtaccttt tgtttgaggt aaaatcagag caccggtgat tttgcttggt 1440

gtaataaagt acggttgttt ggtcctcgat tctggtagtg atgcttctcg atttgacgaa 1500

gctatccttt gtttattccc tattgaacaa aaataatcca actttgaaga cggtcccgtt 1560

gatgagattg aatgattgat tcttaagcct gtccaaaatt tcgcagctgg cttgtttaga 1620

tacagtagtc cccatcacga aattcatgga aacagttata atcctcagga acaggggatt 1680

ccctgttctt ccgatttgct ttagtcccag aatttttttt cccaaatatc ttaaaaagtc 1740

actttctggt tcagttcaat gaattgattg ctacaaataa tgcttttata gcgttatcct 1800

agctgtagtt cagttaatag gtaatacccc tatagtttag tcaggagaag aacttatccg 1860

atttctgatc tccattttta attatatgaa atgaactgta gcataagcag tattcatttg 1920

gattattttt tttattagct ctcacccctt cattattctg agctgaaagt ctggcatgaa 1980

ctgtcctcaa ttttgttttc aaattcacat cgattatcta tgcattatcc tcttgtatct 2040

acctgtagaa gtttcttttt ggttattcct tgactgcttg attacagaaa gaaatttatg 2100

aagctgtaat cgggatagtt atactgcttg ttcttatgat tcatttcctt tgtgcagttc 2160

ttggtgtagc ttgccacttt caccagcaaa gttc 2194

<210>57

<211>46

<212>PRT

＜213〉artificial sequence

<220>

＜223〉SNH structural domain

<220>

＜221〉variant

<222>(3)..(3)

＜223 〉/replace=" Lys "

<220>

＜221〉uncertain

<222>(4)..(4)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(6)..(6)

＜223 〉/replace=" Glu "

<220>

＜221〉variant

<222>(7)..(7)

＜223 〉/replace=" Asp "

<220>

＜221〉variant

<222>(9)..(9)

＜223 〉/replace=" Asn "

<220>

＜221〉uncertain

<222>(10)..(10)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉uncertain

<222>(13)..(13)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(14)..(14)

＜223 〉/replace=" Ala "/replace=" Lys "

<220>

＜221〉variant

<222>(16)..(16)

＜223 〉/replace=" Val "/replace=" Met "

<220>

＜221〉variant

<222>(17)..(17)

＜223 〉/replace=" Asp "/replace=" Ser "

<220>

＜221〉variant

<222>(18)..(18)

＜223 〉/replace=" Asn "

<220>

＜221〉variant

<222>(19)..(19)

＜223 〉/replace=" Leu "

<220>

＜221〉uncertain

<222>(21)..(21)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(23)..(23)

＜223 〉/replace=" Arg "

<220>

＜221〉uncertain

<222>(24)..(25)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(28)..(28)

＜223 〉/replace=" Asp "/replace=" Ser "

<220>

＜221〉uncertain

<222>(29)..(30)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(32)..(32)

＜223 〉/replace=" Ser "/replace=" Gln "

<220>

＜221〉uncertain

<222>(33)..(33)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(35)..(35)

＜223 〉/replace=" His "

<220>

＜221〉uncertain

<222>(36)..(36)

＜223〉Xaa can make the amino acid of any natural generation

<220>

＜221〉variant

<222>(39)..(39)

＜223 〉/replace=" Leu "/replace=" Val "

<220>

＜221〉variant

<222>(43)..(43)

＜223 〉/replace=" Thr "

<400>57

Ile Gln Gln Xaa Leu Asp Glu Asn Lys Xaa Leu Ile Xaa Cys Ile Leu

1 5 10 15

Glu Ser Gln Asn Xaa Gly Lys Xaa Xaa Glu Cys Ala Xaa Xaa Gln Ala

20 25 30

Xaa Leu Gln Xaa Asn Leu Met Tyr Leu Ala Ala Ile Ala Asp

35 40 45

<210>58

<211>46

<212>PRT

＜213〉Arabidopis thaliana

<400>58

Ile Gln Gln Tyr Leu Asp Glu Asn Lys Ser Leu Ile Leu Lys Ile Val

1 5 10 15

Glu Ser Gln Asn Ser Gly Lys Leu Ser Glu Cys Ala Glu Asn Gln Ala

20 25 30

Arg Leu Gln Arg Asn Leu Met Tyr Leu Ala Ala Ile Ala Asp

35 40 45

<210>59

<211>633

<212>DNA

＜213〉Arabidopis thaliana

<400>59

atgcaacagc acctgatgca gatgcagccc atgatggctg gttactaccc cagcaatgtt 60

acctctgatc atatccaaca gtacttggac gaaaacaaat cgttgattct gaagattgtt 120

gagtctcaaa actctggaaa gcttagcgaa tgcgccgaga atcaagcaag gcttcaacgc 180

aacctaatgt acctagctgc aatagcagat tctcagcctc agccaccaag tgtgcatagc 240

cagtatggat ctgctggtgg tgggatgatt cagggagaag gagggtcaca ctatttgcag 300

cggcaacaag cgactcaaca gcaacagatg actcagcagt ctctaatggc ggctcgatct 360

tcaatgttgt atgctcagca acagcagcag cagcagcctt acgcgacgct tcagcatcag 420

caattgcacc atagccagct tggaatgagc tcgagcagcg gaggaggagg aagcagtggt 480

ctccatatcc ttcagggaga ggctggtggg tttcatgatt ttggccgtgg gaagccggaa 540

atgggaagtg gtggtggcgg tgaaggcaga ggaggaagtt caggggatgg tggagaaacc 600

ctttacttga aatcatcaga tgatgggaat tga 633

<210>60

<211>210

<212>PRT

＜213〉Arabidopis thaliana

<400>60

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Gly Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Ser Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Ser Val His Ser

60 70 75 80

Gln Tyr Gly Ser Ala Gly Gly Gly Met Ile Gln Gly Glu Gly Gly Ser

85 90 95

His Tyr Leu Gln Gln Gln Gln Ala Thr Gln Gln Gln Gln Met Thr Gln

100 105 110

Gln Ser Leu Met Ala Ala Arg Ser Ser Met Leu Tyr Ala Gln Gln Gln

115 120 125

Gln Gln Gln Gln Pro Tyr Ala Thr Leu Gln His Gln Gln Leu His His

130 135 140

Ser Gln Leu Gly Met Ser Ser Ser Ser Gly Gly Gly Gly Ser Ser Gly

145 150 155 160

Leu His Ile Leu Gln Gly Glu Ala Gly Gly Phe His Asp Phe Gly Arg

165 170 175

Gly Lys Pro Glu Met Gly Ser Gly Gly Gly Gly Glu Gly Arg Gly Gly

180 185 190

Ser Ser Gly Asp Gly Gly Glu Thr Leu Tyr Leu Lys Ser Ser Asp Asp

195 200 205

Gly Asn

210

<210>61

<211>588

<212>DNA

＜213〉Arabidopis thaliana

<400>61

atgcagcagc agcagtctcc gcaaatgttt ccgatggttc cgtcgattcc ccctgctaac 60

aacatcacta ccgaacagat ccaaaagtac cttgatgaga acaagaagct gattatggcc 120

atcatggaaa accagaatct cggtaaactt gctgagtgcg cccagtacca agctcttctc 180

cagaagaact tgatgtatct tgctgcaatt gctgatgctc aacccccacc acctacgcca 240

ggaccttcac catctacagc tgtcgctgcc cagatggcaa caccgcattc tgggatgcaa 300

ccacctagct acttcatgca acacccacaa gcatcccctg cagggatttt cgctccaagg 360

ggtcctttac agtttggtag cccactccag tttcaggatc cgcaacagca gcagcagata 420

catcagcaag ctatgcaagg acacatgggg attagaccaa tgggtatgac caacaacggg 480

atgcagcatg cgatgcaaca accagaaacc ggtcttggag gaaacgtggg gcttagagga 540

ggaaagcaag atggagcaga tggacaagga aaagatgatg gcaagtga 588

<210>62

<211>195

<212>PRT

＜213〉Arabidopis thaliana

<400>62

Met Gln Gln Gln Gln Ser Pro Gln Met Phe Pro Met Val Pro Ser Ile

1 5 10 15

Pro Pro Ala Asn Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp

20 25 30

Glu Asn Lys Lys Leu Ile Met Ala Ile Met Glu Asn Gln Asn Leu Gly

35 40 45

Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Leu Leu Gln Lys Asn Leu

50 55 60

Met Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Pro Pro Pro Thr Pro

65 70 75 80

Gly Pro Ser Pro Ser Thr Ala Val Ala Ala Gln Met Ala Thr Pro His

85 90 95

Ser Gly Met Gln Pro Pro Ser Tyr Phe Met Gln His Pro Gln Ala Ser

100 105 110

Pro Ala Gly Ile Phe Ala Pro Arg Gly Pro Leu Gln Phe Gly Ser Pro

115 120 125

Leu Gln Phe Gln Asp Pro Gln Gln Gln Gln Gln Ile His Gln Gln Ala

130 135 140

Met Gln Gly His Met Gly Ile Arg Pro Met Gly Met Thr Asn Asn Gly

145 150 155 160

Met Gln His Ala Met Gln Gln Pro Glu Thr Gly Leu Gly Gly Asn Val

165 170 175

Gly Leu Arg Gly Gly Lys Gln Asp Gly Ala Asp Gly Gln Gly Lys Asp

180 185 190

Asp Gly Lys

195

<210>63

<211>672

<212>DNA

＜213〉Arabidopis thaliana

<400>63

atgcagcaat ctccacagat gattccgatg gttcttcctt catttccgcc caccaataat 60

atcaccaccg aacagatcca aaagtatctt gatgagaaca agaagctgat aatggcgatc 120

ttggaaaatc agaacctcgg taaacttgca gaatgtgctc agtatcaagc tcttctccag 180

aagaatttga tgtatctcgc tgcaattgcg gatgctcaac ctcagccacc agcagctaca 240

ctaacatcag gagccatgac tccccaagca atggctccta atccgtcatc aatgcagcca 300

ccaccaagct acttcatgca gcaacatcaa gctgtgggaa tggctcaaca aatacctcct 360

gggattttcc ctcctagagg tccattgcaa tttggtagcc cgcatcagtt tctggatccg 420

cagcaacagt tacatcaaca agctatgcaa gggcacatgg ggattagacc aatgggtttg 480

aataataaca acggactgca acatcaaatg caccaccatg aaactgctct tgccgcaaac 540

aatgcgggtc ctaacgatgc tagtggagga ggtaaaccgg atgggaccaa tatgagccag 600

agtggagctg atgggcaagg tggctcagcc gctagacatg gcggtggtga tgcaaaaact 660

gaaggaaaat ga 672

<210>64

<211>223

<212>PRT

＜213〉Arabidopis thaliana

<400>64

Met Gln Gln Ser Pro Gln Met Ile Pro Met Val Leu Pro Ser Phe Pro

1 5 10 15

Pro Thr Asn Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu

20 25 30

Asn Lys Lys Leu Ile Met Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys

35 40 45

Leu Ala Glu Cys Ala Gln Tyr Gln Ala Leu Leu Gln Lys Asn Leu Met

50 55 60

Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Pro Pro Ala Ala Thr

65 70 75 80

Leu Thr Ser Gly Ala Met Thr Pro Gln Ala Met Ala Pro Asn Pro Ser

85 90 95

Ser Met Gln Pro Pro Pro Ser Tyr Phe Met Gln Gln His Gln Ala Val

100 105 110

Gly Met Ala Gln Gln Ile Pro Pro Gly Ile Phe Pro Pro Arg Gly Pro

115 120 125

Leu Gln Phe Gly Ser Pro His Gln Phe Leu Asp Pro Gln Gln Gln Leu

130 135 140

His Gln Gln Ala Met Gln Gly His Met Gly Ile Arg Pro Met Gly Leu

145 150 155 160

Asn Asn Asn Asn Gly Leu Gln His Gln Met His His His Glu Thr Ala

165 170 175

Leu Ala Ala Asn Asn Ala Gly Pro Asn Asp Ala Ser Gly Gly Gly Lys

180 185 190

Pro Asp Gly Thr Asn Met Ser Gln Ser Gly Ala Asp Gly Gln Gly Gly

195 200 205

Ser Ala Ala Arg His Gly Gly Gly Asp Ala Lys Thr Glu Gly Lys

210 215 220

<210>65

<211>633

<212>DNA

<213>Aspergillus officinalis

<400>65

atgcagcagc acctgatgca gatgcagccc atgatggcaa cctacggttc accgaatcag 60

gtcaccaccg atatcattca gcagtatctg gacgagaaca agcagttgat tctggctatt 120

cttgaaaacc aaaattcagg aaaagctgat gaatgtgctg agaatcaggc taagcttcag 180

aggaatctga tgtatcttgc agccattgcg gatagccagc cccaagttcc taccattgct 240

cagtatcctc ccaacgctgt tgctgctatg caatcgagtg ctcgctacat gcaacaacac 300

caagcagctc aacagatgac ccctcaatct ctcatggctg ctcgctcctc aatgctctac 360

tcacagtccc caatgtctgc actccagcag caacagcagc aagcagcaat gcatagccag 420

ctcgccatga gctccggagg caacaacagc agcaccggag gattcaccat tcttcatggt 480

gaagctagca taggaggcaa tggctcaatg aattctggtg gagtctttgg agattttgga 540

cggagcagcg gtgggaagca agagactggg agcgaagggc acgggacaga gactcctatg 600

tacctgaaag gctctgaaga agaaggaaac tga 633

<210>66

<211>210

<212>PRT

<213>Aspergillus officinalis

<400>66

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Thr Tyr Gly

1 5 10 15

Ser Pro Asn Gln Val Thr Thr Asp Ile Ile Gln Gln Tyr Leu Asp Glu

20 25 30

Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Ser Gly Lys

35 40 45

Ala Asp Glu Cys Ala Glu Asn Gln Ala Lys Leu Gln Arg Asn Leu Met

50 55 60

Tyr Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Val Pro Thr Ile Ala

65 70 75 80

Gln Tyr Pro Pro Asn Ala Val Ala Ala Met Gln Ser Ser Ala Arg Tyr

85 90 95

Met Gln Gln His Gln Ala Ala Gln Gln Met Thr Pro Gln Ser Leu Met

100 105 110

Ala Ala Arg Ser Ser Met Leu Tyr Ser Gln Ser Pro Met Ser Ala Leu

115 120 125

Gln Gln Gln Gln Gln Gln Ala Ala Met His Ser Gln Leu Ala Met Ser

130 135 140

Ser Gly Gly Asn Asn Ser Ser Thr Gly Gly Phe Thr Ile Leu His Gly

145 150 155 160

Glu Ala Ser Ile Gly Gly Asn Gly Ser Met Asn Ser Gly Gly Val Phe

165 170 175

Gly Asp Phe Gly Arg Ser Ser Gly Gly Lys Gln Glu Thr Gly Ser Glu

180 185 190

Gly His Gly Thr Glu Thr Pro Met Tyr Leu Lys Gly Ser Glu Glu Glu

195 200 205

Gly Asn

210

<210>67

<211>591

<212>DNA

＜213〉colea (Brassica napus)

<400>67

atgcagccca tgatggctgg ttactacccc agcaatgtca cctctgatca tatccagcag 60

tacttggatg agaacaagtc tttgattctg aagatagttg agtctcaaaa ctcaggaaag 120

ctcagcgagt gtgccgagaa tcaggcaagg cttcaacgca acctcatgta cttggctgca 180

atagcagatt ctcagcctca acctccaagc gtgcatagcc agtatggatc tgctggtggt 240

gggttgattc agggagaagg agcgtcacac tatttgcagc agcaacaggc gactcaacag 300

cagcagatga ctcagcagtc tcttatggca gctcgttctt caatgatgta tcagcagcag 360

caacagcctt atgcaacgct tcagcatcag cagttgcacc atagccagct tgggatgagc 420

tctagcagcg gaggaggaag cagtggtctc catatccttc agggagaggc tggtgggttt 480

catgaatttg gccgtgggaa gccggagatg ggaagtggtg aaggcagggg tggaagctca 540

ggggatggtg gagaaacact ctacttgaag tcatcagatg atgggaactg a 591

<210>68

<211>203

<212>PRT

＜213〉colea

<400>68

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Gly Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Ser Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Ser Val His Ser

65 70 75 80

Gln Tyr Gly Ser Ala Gly Gly Gly Leu Ile Gln Gly Glu Gly Ala Ser

85 90 95

His Tyr Leu Gln Gln Gln Gln Ala Thr Gln Gln Gln Gln Met Thr Gln

100 105 110

Gln Ser Leu Met Ala Ala Arg Ser Ser Met Met Tyr Gln Gln Gln Gln

115 120 125

Gln Pro Tyr Ala Thr Leu Gln His Gln Gln Leu His His Ser Gln Leu

130 135 140

Gly Met Ser Ser Ser Ser Gly Gly Gly Ser Ser Gly Leu His Ile Leu

145 150 155 160

Gln Gly Glu Ala Gly Gly Phe His Glu Phe Gly Arg Gly Lys Pro Glu

165 170 175

Met Gly Ser Gly Glu Gly Arg Gly Gly Ser Ser Gly Asp Gly Gly Glu

180 185 190

Thr Leu Tyr Leu Lys Ser Ser Asp Asp Gly Asn

195 200

<210>69

<211>663

<212>DNA

＜213〉sweet orange (Citrus sinensis)

<400>69

atgcaacagc acctgatgca gatgcagccc atgatggcag cttattatcc caacaacgtc 60

actactgacc acattcaaca gtatctagat gagaacaaat cattgatttt gaagattgtt 120

gagagccaga attcagggaa actgagcgag tgtgcagaga accaggcaag attgcagcgg 180

aatctcatgt acctggctgc tattgctgat gctcaacccc aaccacctag cgttcatgcc 240

cagttctctt ctggtggcat tatgcagcca ggagctcact atatgcaaca ccagcaatct 300

cagccaatga caccacagtc acttatggct gcacgctcat ccatggtgta ctctcaacag 360

caattttcag tgcttcagca acagcaagcc ttgcatggtc agcttggcat gagctctggt 420

ggtagctcag gacttcacat gctgcaaagt gagggtagta ctgcaggagg tagtggttca 480

cttgggggtg ggggattccc tgattttggc cgtggctcat ctggtgaagg cttgcactca 540

aggggaatgg ggagcaagca tgatataggc agttctggat ctgctgaagg acgaggaggg 600

agctcaggaa gccaagatgg aggcgaaact ctctacttga aaggggctga tgatggaaat 660

taa 663

<210>70

<211>219

<212>PRT

＜213〉sweet orange

<400>70

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Pro Pro Ser Val His Ala

65 70 75 80

Gln Phe Ser Ser Gly Gly Ile Met Gln Pro Gly Ala His Tyr Met Gln

85 90 95

His Gln Gln Ser Gln Pro Met Thr Pro Gln Ser Leu Met Ala Ala Arg

100 105 110

Ser Ser Met Val Tyr Ser Gln Gln Gln Phe Ser Val Leu Gln Gln Gln

115 120 125

Gln Ala Leu His Gly Gln Leu Gly Met Ser Ser Gly Gly Ser Ser Gly

130 135 140

Leu His Met Leu Gln Ser Glu Gly Ser Thr Ala Gly Gly Ser Gly Ser

145 150 155 160

Leu Gly Gly Gly Gly Phe Pro Asp Phe Gly Arg Gly Ser Ser Gly Glu

165 170 175

Gly Leu His Ser Arg Gly Met Gly Ser Lys His Asp Ile Gly Ser Ser

180 185 190

Gly Ser Ala Glu Gly Arg Gly Gly Ser Ser Gly Ser Gln Asp Gly Gly

195 200 205

Glu Thr Leu Tyr Leu Lys Gly Ala Asp Asp Gly

210 215

<210>71

<211>660

<212>DNA

＜213〉tree cotton (Gossypium arboreum)

<220>

<221>misc_feature

<222>(309)..(309)

＜223〉n is a, c, g or t

<400>71

atgcagcagc acctgatgca gatgcagccc atgatggcag cttattatcc caacaacgtc 60

actactgatc atattcaaca gtatctcgat gagaacaagt cattgatctt aaagattgtt 120

gagagccaga attctgggaa attgagtgaa tgtgctgaga accaagcaag gctgcagcga 180

aacctcatgt acctggctgc cattgcggat tctcaacccc aaccacccac cgtgcatgca 240

cagtttccat ctggtggtat catgcagcaa ggagctgggc actacatgca gcaccaacaa 300

gctcaacana tgacacaaca gtcgcttatg gctgctcggt cctcaatgtt gtattctcag 360

caaccatttt ctgcactgca acaacaacaa caacaaggct ttgcacagtc agcttggcat 420

gagctctggc gggagcacag gcctttcata tgctgcaaac tgaatctagt actgcagggg 480

gcagtgagac accttgggcc cgagggttgt cctgatttgg acgggggtct tttggagagg 540

catccctggt ggcaggccaa tggccggggg aacaaccaaa aatccgggga ggccggctca 600

cctaagggcc gggaggagcc cttggggcag gggggggtga tggggggaac ctcttcttaa 660

<210>72

<211>219

<212>PRT

＜213〉tree cotton

<220>

＜221〉uncertain

<222>(103)..(103)

＜223〉Xaa can make the amino acid of any natural generation

<400>72

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Thr Val His Ala

65 70 75 80

Gln Phe Pro Ser Gly Gly Ile Met Gln Gln Gly Ala Gly His Tyr Met

85 90 95

Gln His Gln Gln Ala Gln Xaa Met Thr Gln Gln Ser Leu Met Ala Ala

100 105 110

Arg Ser Ser Met Leu Tyr Ser Gln Gln Pro Phe Ser Ala Leu Gln Gln

115 120 125

Gln Gln Gln Gln Gly Phe Ala Gln Ser Ala Trp His Glu Leu Trp Arg

130 135 140

Glu His Arg Pro Phe Ile Cys Cys Lys Leu Asn Leu ValLeu Gln Gly

145 150 155 160

Ala Val Arg His Leu Gly Pro Glu Gly Cys Pro Asp Leu Asp Gly Gly

165 170 175

Leu Leu Glu Arg His Pro Trp Trp Gln Ala Asn Gly Arg Gly Asn Asn

180 185 190

Gln Lys Ser Gly Glu Ala Gly Ser Pro Lys Gly Arg Glu Glu Pro Leu

195 200 205

Gly Gln Gly Gly Val Met Gly Gly Thr Ser Ser

210 215

<210>73

<211>636

<212>DNA

＜213〉puncture vine clover (Medicago trunculata)

<400>73

atgcagcagc acctgatgca gatgcagccc atgatggcag cttactatcc taacaacgtc 60

actactgatc atattcaaca gtatcttgat gagaacaagt ccttgattct caagattgtt 120

gaaagccaga acactggcaa gctcaccgag tgtgctgaga accaatcaag gcttcagaga 180

aatctcatgt acctagctgc aatagctgat tctcaacccc aaccacctac tatgcctggc 240

cagtaccctt caagtggaat gatgcagcag ggaggacact acatgcaggc tcaacaagct 300

cagcagatga cacaacaaca attaatggct gcacgttcct ctcttatgta tgctcaacag 360

cttcaacagc agcaagcctt gcaaagccaa cttggtatga attccagtgg aagtcaaggc 420

cttcacatgt tgcatagtga aggggctaat gttggaggca attcatctct aggggctggt 480

tttcctgatt ttggccgtag ctcagccggt gatggtttgc acggcagtgg taagcaagac 540

attggaagca ctgatggccg cggtggaagc tctagtggtc actctggtga tggcggcgaa 600

acactttacc tgaaatcttc tggtgatggg aattag 636

<210>74

<211>211

<212>PRT

＜213〉puncture vine clover

<400>74

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Thr Gly Lys Leu

35 40 45

Thr Glu Cys Ala Glu Asn Gln Ser Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Thr Met Pro Gly

65 70 75 80

Gln Tyr Pro Ser Ser Gly Met Met Gln Gln Gly Gly His Tyr Met Gln

85 90 95

Ala Gln Gln Ala Gln Gln Met Thr Gln Gln Gln Leu Met Ala Ala Arg

100 105 110

Ser Ser Leu Met Tyr Ala Gln Gln Leu Gln Gln Gln Gln Ala Leu Gln

115 120 125

Ser Gln Leu Gly Met Asn Ser Ser Gly Ser Gln Gly Leu His Met Leu

130 135 140

His Ser Glu Gly Ala Asn Val Gly Gly Asn Ser Ser Leu Gly Ala Gly

145 150 155 160

Phe Pro Asp Phe Gly Arg Ser Ser Ala Gly Asp Gly Leu His Gly Ser

165 170 175

Gly Lys Gln Asp Ile Gly Ser Thr Asp Gly Arg Gly Gly Ser Ser Ser

180 185 190

Gly His Ser Gly Asp Gly Gly Glu Thr Leu Tyr Leu Lys Ser Ser Gly

195 200 205

Asp Gly Asn

210

<210>75

<211>684

<212>DNA

＜213〉rice

<400>75

atgcagcagc aacacctgat gcagatgaac cagggcatga tggggggata tgcttcccct 60

accaccgtca ccactgatct cattcagcag tatctggatg agaacaagca gctgatcctg 120

gccatccttg acaaccagaa caatgggaag gtggaagagt gcgctcggaa ccaagctaag 180

ctccagcaca atctcatgta cctcgccgcc atcgccgaca gccagccgcc gcagacggcc 240

gccatgtccc agtatccgtc gaacctgatg atgcagtccg gggcgaggta catgccgcag 300

cagtcggcgc agatgatggc gccgcagtcg ctgatggcgg cgaggtcttc gatgatgtac 360

gcgcagccgg cgctgtcgcc gctccagcag cagcagcagc agcaggcggc ggcggcgcac 420

gggcagctgg gcatgggctc ggggggcacc accagcgggt tcagcatcct ccacggcgag 480

gccagcatgg gcggcggcgg cggcggcggt ggcgccggta acagcatgat gaacgccggc 540

gtgttctccg acttcggacg cggcggcggc ggcggcggca aggaggggtc cacctcgctg 600

tccgtcgacg tccggggcgc caactccggc gcccagagcg gcgacgggga gtacctcaag 660

ggcaccgagg aggaaggcagctag 684

<210>76

<211>227

<212>PRT

＜213〉rice

<400>76

Met Gln Gln Gln His Leu Met Gln Met Asn Gln Gly Met Met Gly Gly

1 5 10 15

Tyr Ala Ser Pro Thr Thr Val Thr Thr Asp Leu Ile Gln Gln Tyr Leu

20 25 30

Asp Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Asn

35 40 45

Gly Lys Val Glu Glu Cys Ala Arg Asn Gln Ala Lys Leu Gln His Asn

50 55 60

Leu Met Tyr Leu Ala Ala Ile Ala Asp Ser Gln Pro Pro Gln Thr Ala

65 70 75 80

Ala Met Ser Gln Tyr Pro Ser Asn Leu Met Met Gln Ser Gly Ala Arg

85 90 95

Tyr Met Pro Gln Gln Ser Ala Gln Met Met Ala Pro Gln Ser Leu Met

100 105 110

Ala Ala Arg Ser Ser Met Met Tyr Ala Gln Pro Ala Leu Ser Pro Leu

115 120 125

Gln Gln Gln Gln Gln Gln Gln Ala Ala Ala Ala His Gly Gln Leu Gly

130 135 140

Met Gly Ser Gly Gly Thr Thr Ser Gly Phe Ser Ile Leu Hi s Gly Glu

145 150 155 160

Ala Ser Met Gly Gly Gly Gly Gly Gly Gly Gly Ala Gly Asn Ser Met

165 170 175

Met Asn Ala Gly Val Phe Ser Asp Phe Gly Arg Gly Gly Gly Gly Gly

180 185 190

Gly Lys Glu Gly Ser Thr Ser Leu Ser Val Asp ValArg Gly Ala Asn

195 200 205

Ser Gly Ala Gln Ser Gly Asp Gly Glu Tyr Leu Lys Gly Thr Glu Glu

210 215 220

Glu Gly Ser

225

<210>77

<211>558

<212>DNA

＜213〉rice

<400>77

atgcagcagc agccgatgcc gatgcccgcg caggcgccgc cgacggccgg aatcaccacc 60

gagcagatcc aaaagtatct ggatgaaaac aagcagctta ttttggctat tttggaaaat 120

cagaatctgg gaaagttggc agaatgtgct cagtatcaag cgcagcttca gaagaatctc 180

ttgtacttgg ctgcaattgc tgatactcaa ccgcagacca ctataagccg tccccagatg 240

gtgccgcatg gtgcatcgcc ggggttaggg gggcaataca tgtcgcaggt gccaatgttc 300

ccccccagga cccctctaac gccccagcag atgcaggagc agcagctgca gcaacagcaa 360

gcccagctgc tctcgttcgg cggtcagatg gttatgaggc ctggcgttgt gaatggcatt 420

cctcagcttc tgcaaggcga aatgcaccgc ggagcagatc accagaacgc tggcggggcc 480

acctcggagc cttccgagag ccacaggagc accggcaccg aaaatgacgg tggaagcgac 540

ttcggcgatc aatcctaa 558

<210>78

<211>185

<212>PRT

＜213〉rice

<400>78

Met Gln Gln Gln Pro Met Pro Met Pro Ala Gln Ala Pro Pro Thr Ala

1 5 10 15

Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys Gln

20 25 30

Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys Leu Ala Glu

35 40 45

Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Leu Tyr Leu Ala

50 55 60

Ala Ile Ala Asp Thr Gln Pro Gln Thr Thr Ile Ser Arg Pro Gln Met

65 70 75 80

Val Pro His Gly Ala Ser Pro Gly Leu Gly Gly Gln Tyr Met Ser Gln

85 90 95

Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln Gln Met Gln

100 105 110

Glu Gln Gln Leu Gln Gln Gln Gln Ala Gln Leu Leu Ser Phe Gly Gly

115 120 125

Gln Met Val Met Arg Pro Gly Val Val Asn Gly Ile Pro Gln Leu Leu

130 135 140

Gln Gly Glu Met His Arg Gly Ala Asp His Gln Asn Ala Gly Gly Ala

145 150 155 160

Thr Ser Glu Pro Ser Glu Ser His Arg Ser Thr Gly Thr Glu Asn Asp

165 170 175

Gly Gly Ser Asp Phe Gly Asp Gln Ser

180 185

<210>79

<211>618

<212>DNA

213〉rice

<400>79

atgcagcagc agatggccat gccggcgggg gccgccgccg ccgcggtgcc gccggcggcc 60

ggcatcacca ccgagcagat ccaaaagtat ttggatgaaa ataaacagct aattttggcc 120

atcctggaaa atcaaaacct agggaagttg gctgaatgtg ctcagtacca agctcagctt 180

caaaagaatc tcttgtatct ggctgccatt gcagatgccc aaccacctca gaatccagga 240

agtcgccctc agatgatgca gcctggtgct accccaggtg ctgggcatta catgtcccaa 300

gtaccgatgt tccctccaag aactccctta accccacaac agatgcaaga gcagcagcag 360

cagcaactcc agcaacagca agctcaggct ctagccttcc ccggccagat gctaatgaga 420

ccaggtactg tcaatggcat gcaatctatc ccagttgctg accctgctcg cgcagccgat 480

cttcagacgg cagcaccggg ctcggtagat ggccgaggaa acaagcagga tgcaacctcg 540

gagccttccg ggaccgagag ccacaagagt gcgggagcag ataacgacgc aggcggtgac 600

atagcggaga agtcctga 618

<210>80

<211>205

<212>PRT

＜213〉rice

<400>80

Met Gln Gln Gln Met Ala Met Pro Ala Gly Ala Ala Ala Ala Ala Val

1 5 10 15

Pro Pro Ala Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp

20 25 30

Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly

35 40 45

Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu

50 55 60

Leu Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Pro Gln Asn Pro Gly

65 70 75 80

Ser Arg Pro Gln Met Met Gln Pro Gly Ala Thr Pro Gly Ala Gly His

85 90 95

Tyr Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro

100 105 110

Gln Gln Met Gln Glu Gln Gln Gln Gln Gln Leu Gln Gln Gln Gln Ala

115 120 125

Gln Ala Leu Ala Phe Pro Gly Gln Met Leu Met Arg Pro Gly Thr Val

130 135 140

Asn Gly Met Gln Ser Ile Pro Val Ala Asp Pro Ala Arg Ala Ala Asp

145 150 155 160

Leu Gln Thr Ala Ala Pro Gly Ser Val Asp Gly Arg Gly Asn Lys Gln

165 170 175

Asp Ala Thr Ser Glu Pro Ser Gly Thr Glu Ser His Lys Ser Ala Gly

180 185 190

Ala Asp Asn Asp Ala Gly Gly Asp Ile Ala Glu Lys Ser

195 200 205

<210>81

<211>540

<212>DNA

＜213〉potato (Solanum tuberosum)

<400>81

atgcagcagc agcacctgat gcagatgcag cccatgatgg cagcctatta tcccaacaat 60

gtcactactg atcatattca acagttcctg gatgagaaca aatcacttat tctgaagatt 120

gttgagagcc agaactctgg gaaaataagt gaatgtgcag agtcccaagc taaacttcag 180

agaaatctta tgtaccttgc agctattgct gattcacagc cccagcctcc tagtatgcat 240

tcacagttag cttctggtgg gatgatgcag ggaggggcac attatatgca gcaacaacaa 300

gctcaacaac tcacaacgca atcgcttatg gctgcagcaa gatcctcctc ctcaatgctc 360

tatggacaac aacaacaaca acaacaacaa caactatcat cattgcaaca acagcaagca 420

gcctttcata gccagcaact cggaatgagc agctctggtg gaggaagcag tagtggactt 480

cacatgctac aaagcgaaaa cactcatagt gctagcactg gtggtgggtg gtttccctga 540

<210>82

<211>179

<212>PRT

＜213〉potato

<400>82

Met Gln Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr

1 5 10 15

Tyr Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Phe Leu Asp Glu

20 25 30

Asn Lys Ser Leu Lle Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys

35 40 45

Ile Ser Glu Cys Ala Glu Ser Gln Ala Lys Leu Gln Arg Asn Leu Met

50 55 60

Tyr Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Ser Met His

65 70 75 80

Ser Gln Leu Ala Ser Gly Gly Met Met Gln Gly Gly Ala His Tyr Met

85 90 95

Gln Gln Gln Gln Ala Gln Gln Leu Thr Thr Gln Ser Leu Met Ala Ala

100 105 110

Ala Arg Ser Ser Ser Ser Met Leu Tyr Gly Gln Gln Gln Gln Gln Gln

115 120 125

Gln Gln Gln Leu Ser Ser Leu Gln Gln Gln Gln Ala Ala Phe His Ser

130 135 140

Gln Gln Leu Gly Met Ser Ser Ser Gly Gly Gly Ser Ser Ser Gly Leu

145 150 155 160

His Met Leu Gln Ser Glu Asn Thr His Ser Ala Ser Thr Gly Gly Gly

165 170 175

Trp Phe Pro

<210>83

<211>684

<212>DNA

＜213〉Zea mays (Zea mays)

<400>83

atgcagcagc aacacctgat gcagatgaac cagaacatga tggggggcta cacctctcct 60

gccgccgtga ccaccgatct catccagcag cacctggacg agaacaagca gctgatcctg 120

gccatcctcg acaaccagaa caatggcaag gcggaggagt gcgaacggca ccaagctaag 180

ctccagcaca acctcatgta cctggccgcc atcgctgaca gccagccgcc acagaccgcg 240

ccactatcac agtacccgtc caacctgatg atgcagccgg gccctcggta catgccaccg 300

cagtccgggc agatgatgaa cccgcagtcg ctgatggcgg cgcggtcctc catgatgtac 360

gcgcacccgt ccctgtcgcc actccagcag cagcaggcgg cgcacggaca gctgggtatg 420

gctccagggg gcggcggtgg cggcacgacc agcgggttca gcatcctcca cggcgaggcc 480

agcatgggcg gtggtggtgc tggcgcaggc gccggcaaca acatgatgaa cgccggcatg 540

ttctcgggct ttggccgcag cggcagtggc gccaaggaag ggtcgacctc tctgtcggtt 600

gacgtccgag gtggaaccag ctccggcgcg cagagcgggg acggcgagta cctcaaagtc 660

ggcaccgagg aagaaggcag ttag 684

<210>84

<211>227

<212>PRT

＜213〉Zea mays

<400>84

Met Gln Gln Gln His Leu Met Gln Met Asn Gln Asn Met Met Gly Gly

1 5 10 15

Tyr Thr Ser Pro Ala Ala Val Thr Thr Asp Leu Ile Gln Gln His Leu

20 25 30

Asp Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Asn

35 40 45

Gly Lys Ala Glu Glu Cys Glu Arg His Gln Ala Lys Leu Gln His Asn

50 55 60

Leu Met Tyr Lau Ala Ala Ile Ala Asp Ser Gln Pro Pro Gln Thr Ala

65 70 75 80

Pro Leu Ser Gln Tyr Pro Ser Asn Leu Met Met Gln Pro Gly Pro Arg

85 90 95

Tyr Met Pro Pro Gln Ser Gly Gln Met Met Asn Pro Gln Ser Leu Met

100 105 110

Ala Ala Arg Ser Ser Met Met Tyr Ala His Pro Ser Leu Ser Pro Leu

115 120 125

Gln Gln Gln Gln Ala Ala His Gly Gln Leu Gly Met Ala Pro Gly Gly

130 135 140

Gly Gly Gly Gly Thr Thr Ser Gly Phe Ser Ile Leu His Gly Glu Ala

145 150 155 160

Ser Met Gly Gly Gly Gly Ala Gly Ala Gly Ala Gly Asn Asn Met Met

165 170 175

Asn Ala Gly Met Phe Ser Gly Phe Gly Arg Ser Gly Ser Gly Ala Lys

180 185 190

Glu Gly Ser Thr Ser Leu Ser Val Asp Val Arg Gly Gly Thr Ser Ser

195 200 205

Gly Ala Gln Ser Gly Asp Gly Glu Tyr Leu Lys Val Gly Thr Glu Glu

210 215 220

Glu Gly Ser

225

<210>85

<211>549

<212>DNA

＜213〉Zea mays

<400>85

atgcagcagc cgatgcacat gcagccacag gcgccggcga taaccccagc tgccggaatc 60

agcacggagc agatccaaaa gtatctggat gagaataagc agcttatttt ggctattttg 120

gaaaatcaga acctaggaaa attggcagaa tgtgctcagt atcaatcaca acttcagaag 180

aacctcttgt atctcgctgc aatcgcagat gctcaaccgc agactgctgt aagccgccct 240

cagatggcgc cgcctggtgg atcgcctgga gtagggcagt acatgtcaca ggtgcctatg 300

ttcccaccga ggacacctct tacaccccag cagatgcagg agcagcagct tcagcagcag 360

caggctcagt tgctaaactt cagtggccaa atggttgcta gaccaggcat ggtcaacggc 420

atggctcagt ccatgcaagc tcagctacca ccgggtgtga acaagcagga tgctggtggg 480

gtcgcctctg agccctcggg caccgagagc cacaggagca ctggtggtga cgatggtgga 540

agcgactag 549

<210>86

<211>182

<212>PRT

＜213〉Zea mays

<400>86

Met Gln Gln Pro Met His Met Gln Pro Gln Ala Pro Ala Ile Thr Pro

1 5 10 15

Ala Ala Gly Ile Ser Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn

20 25 30

Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys Leu

35 40 45

Ala Glu Cys Ala Gln Tyr Gln Ser Gln Leu Gln Lys Asn Leu Leu Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Ala Val Ser Arg Pro

65 70 75 80

Gln Met Ala Pro Pro Gly Gly Ser Pro Gly Val Gly Gln Tyr Met Ser

85 90 95

Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln Gln Met

100 105 110

Gln Glu Gln Gln Leu Gln Gln Gln Gln Ala Gln Leu Leu Asn Phe Ser

115 120 125

Gly Gln Met Val Ala Arg Pro Gly Met Val Asn Gly Met Ala Gln Ser

130 135 140

Met Gln Ala Gln Leu Pro Pro Gly Val Asn Lys Gln Asp Ala Gly Gly

145 150 155 160

Val Ala Ser Glu Pro Ser Gly Thr Glu Ser His Arg Ser Thr Gly Gly

165 170 175

Asp Asp Gly Gly Ser Asp

180

<210>87

<211>1173

<212>DNA

＜213〉people (Homo sapiens)

<400>87

atgggcggca acatgtctgt ggctttcgcg gccccgaggc agcgaggcaa gggggagatc 60

actcccgctg cgattcagaa gatgttggat gacaataacc atcttattca gtgtataatg 120

gactctcaga ataaaggaaa gacctcagag tgttctcagt atcagcagat gttgcacaca 180

aacttggtat accttgctac aatagcagat tctaatcaaa atatgcagtc tcttttacca 240

gcaccaccca cacagaatat gcctatgggt cctggaggga tgaatcagag cggccctccc 300

ccacctccac gctctcacaa catgccttca gatggaatgg taggtggggg tcctcctgca 360

ccgcacatgc agaaccagat gaacggccag atgcctgggc ctaaccatat gcctatgcag 420

ggacctggac ccaatcaact caatatgaca aacagttcca tgaatatgcc ttcaagtagc 480

catggatcca tgggaggtta caaccattct gtgccatcat cacagagcat gccagtacag 540

aatcagatga caatgagtca gggacaacca atgggaaact atggtcccag accaaatatg 600

agtatgcagc caaaccaagg tccaatgatg catcagcagc ctccttctca gcaatacaat 660

atgccacagg gaggcggaca gcattaccaa ggacagcagc cacctatggg aatgatgggt 720

caagttaacc aaggcaatca tatgatgggt cagagacaga ttcctcccta tagacctcct 780

caacagggcc caccacagca gtactcaggc caggaagact attacgggga ccaatacagt 840

catggtggac aaggtcctcc agaaggcatg aaccagcaat attaccctga tggaaattca 900

cagtatggcc aacagcaaga tgcataccag ggaccacctc cacaacaggg atatccaccc 960

cagcagcagc agtacccagg gcagcaaggt tacccaggac agcagcaggg ctacggtcct 1020

tcacagggtg gtccaggtcc tcagtatcct aactacccac agggacaagg tcagcagtat 1080

ggaggatata gaccaacaca gcctggacca ccacagccac cccagcagag gccttatgga 1140

tatgaccagg gacagtatgg aaattaccagcag 1173

<210>88

<211>391

<212>PRT

＜213〉people

<400>88

Met Gly Gly Asn Met Ser Val Ala Phe Ala Ala Pro Arg Gln Arg Gly

1 5 10 15

Lys Gly Glu Ile Thr Pro Ala Ala Ile Gln Lys Met Leu Asp Asp Asn

20 25 30

Asn His Leu Ile Gln Cys Ile Met Asp Ser Gln Asn Lys Gly Lys Thr

35 40 45

Ser Glu Cys Ser Gln Tyr Gln Gln Met Leu His Thr Asn Leu Val Tyr

50 55 60

Leu Ala Thr Ile Ala Asp Ser Asn Gln Asn Met Gln Ser Leu Leu Pro

65 70 75 80

Ala Pro Pro Thr Gln Asn Met Pro Met Gly Pro Gly Gly Met Asn Gln

85 90 95

Ser Gly Pro Pro Pro Pro Pro Arg Ser His Asn Met Pro Ser Asp Gly

100 105 110

Met Val Gly Gly Gly Pro Pro Ala Pro His Met Gln Asn Gln Met Asn

115 120 125

Gly Gln Met Pro Gly Pro Asn His Met Pro Met Gln Gly Pro Gly Pro

130 135 140

Asn Gln Leu Asn Met Thr Asn Ser Ser Met Asn Met Pro Ser Ser Ser

145 150 155 160

His Gly Ser Met Gly Gly Tyr Asn His Ser Val Pro Ser Ser Gln Ser

165 170 175

Met Pro Val Gln Asn Gln Met Thr Met Ser Gln Gly Gln Pro Met Gly

180 185 190

Asn Tyr Gly Pro Arg Pro Asn Met Ser Met Gln Pro Asn Gln Gly Pro

195 200 205

Met Met His Gln Gln Pro Pro Ser Gln Gln Tyr Asn Met Pro Gln Gly

210 215 220

Gly Gly Gln His Tyr Gln Gly Gln Gln Pro Pro Met Gly Met Met Gly

225 230 235 240

Gln Val Asn Gln Gly Asn His Met Met Gly Gln Arg Gln Ile Pro Pro

245 250 255

Tyr Arg Pro Pro Gln Gln Gly Pro Pro Gln Gln Tyr Ser Gly Gln Glu

260 265 270

Asp Tyr Tyr Gly Asp Gln Tyr Ser His Gly Gly Gln Gly Pro Pro Glu

275 280 285

Gly Met Asn Gln Gln Tyr Tyr Pro Asp Gly Asn Ser Gln Tyr Gly Gln

290 295 300

Gln Gln Asp Ala Tyr Gln Gly Pro Pro Pro Gln Gln Gly Tyr Pro Pro

305 310 315 320

Gln Gln Gln Gln Tyr Pro Gly Gln Gln Gly Tyr Pro Gly Gln Gln Gln

325 330 335

Gly Tyr Gly Pro Ser Gln Gly Gly Pro Gly Pro Gln Tyr Pro Asn Tyr

340 345 350

Pro Gln Gly Gln Gly Gln Gln Tyr Gly Gly Tyr Arg Pro Thr Gln Pro

355 360 365

Gly Pro Pro Gln Pro Pro Gln Gln Arg Pro Tyr Gly Tyr Asp Gln Gly

370 375 380

Gln Tyr Gly Asn Tyr Gln Gln

385 390

<210>89

<211>627

<212>DNA

＜213〉onion (Allium cepa)

<400>89

atgcagcagc cgcagccagc gatgggaacc atgggctcggtgccacctac tagcatcacc 60

accgaacaga ttcaaaggta cttggatgag aacaaacagt taatattggc aattttggat 120

aatcaaaatt taggaagact gaatgagtgt gctcaatatc aagctcagct tcaaaagaat 180

ctgctttacc tggcagcaat agctgatgct cagcctcagt ctcctgcggt gcgtctgcag 240

atgatgcctc aaggtgcagc tgccacgcct caagctggaa accaatttat gcagcagcag 300

agccctaatt tccctcccaa aacaggaatg caatttactc ctcaacaagt acaagaattg 360

cagcagcaac agctacaaca tcagccacat atgatgcctc catttcaagg tcaaatgggt 420

atgagaccta tgaatggaat gcaggcagca atgcatgcag attcatctct tgcttataac 480

actaacaata agcaagatgc aggaaacgca gcttatgaaa atactgctgc caacacagat 540

ggttccattc aaaagaaaac agcaaatgat gatttagacc cttctgcagc aaaccctaga 600

aggtctgaag atgccaaatc atcatga 627

<210>90

<211>208

<212>PRT

＜213〉onion

<400>90

Met Gln Gln Pro Gln Pro Ala Met Gly Thr Met Gly Ser Val Pro Pro

1 5 10 15

Thr Ser Ile Thr Thr Glu Gln Ile Gln Arg Tyr Leu Asp Glu Asn Lys

20 25 30

Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Arg Leu Asn

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Leu Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Ser Pro Ala Val Arg Leu Gln

65 70 75 80

Met Met Pro Gln Gly Ala Ala Ala Thr Pro Gln Ala Gly Asn Gln Phe

85 90 95

Met Gln Gln Gln Ser Pro Asn Phe Pro Pro Lys Thr Gly Met Gln Phe

100 105 110

Thr Pro Gln Gln Val Gln Glu Leu Gln Gln Gln Gln Leu Gln His Gln

115 120 125

Pro His Met Met Pro Pro Phe Gln Gly Gln Met Gly Met Arg Pro Met

130 135 140

Asn Gly Met Gln Ala Ala Met His Ala Asp Ser Ser Leu Ala Tyr Asn

145 150 155 160

Thr Asn Asn Lys Gln Asp Ala Gly Asn Ala Ala Tyr Glu Asn Thr Ala

165 170 175

Ala Asn Thr Asp Gly Ser Ile Gln Lys Lys Thr Ala Asn Asp Asp Leu

180 185 190

Asp Pro Ser Ala Ala Asn Pro Arg Arg Ser Glu Asp Ala Lys Ser Ser

195 200 205

<210>91

<211>633

<212>DNA

<213>Aquilegia formosa x Aquilegia pubescens

<400>91

atgcaacaca tgcagatgca gcccatgatg ccaccttata gtgccaacag cgtcactact 60

gatcatatcc aacagtactt ggatgaaaat aaggcgttga ttctgaagat acttgagaac 120

caaaattcgg gaaaagttag tgaatgtgca gagaaccaag caagacttca acgaaatctt 180

atgtatctgg ctgcaattgc tgattctcaa ccacagcctc ccaatatgca tgctcagtac 240

tctaatgcgg gtataccacc tggtgcacat tacctacaac accaacaggc ccaacagatg 300

acacaacagt cgctcatggc tgctcgatca aatatgctgt atgctcagcc aatcacagga 360

atgcagcaac agcaagcaat gcatagccag cttggcatga gctctggtgg taacagtgga 420

ctccacatga tgcacaatga gggcagcatg ggaggtagtg gggcacttgg aagctattct 480

gattatggcc gtggcagtgg tggtggagta actatcgcta gcaaacaaga tggtggaagt 540

ggttctggtg aaggacgagg tggaaactct ggaggccaaa gtgcagatgg aggtgaatct 600

ctttacctga aaaacagtga cgaagggaac taa 633

<210>92

<211>210

<212>PRT

<213>Aquilegia formosa x Aquilegia pubescens

<400>92

Met Gln His Met Gln Met Gln Pro Met Met Pro Pro Tyr Ser Ala Asn

1 5 10 15

Ser Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn Lys Ala

20 25 30

Leu Ile Leu Lys Ile Leu Glu Asn Gln Asn Ser Gly Lys Val Ser Glu

35 40 45

Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr Leu Ala

50 55 60

Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Asn Met His Ala Gln Tyr

65 70 75 80

Ser Asn Ala Gly Ile Pro Pro Gly Ala His Tyr Leu Gln His Gln Gln

85 90 95

Ala Gln Gln Met Thr Gln Gln Ser Leu Met Ala Ala Arg Ser Asn Met

100 105 110

Leu Tyr Ala Gln Pro Ile Thr Gly Met Gln Gln Gln Gln Ala Met His

115 120 125

Ser Gln Leu Gly Met Ser Ser Gly Gly Asn Ser Gly Leu His Met Met

130 135 140

His Asn Glu Gly Ser Met Gly Gly Ser Gly Ala Leu Gly Ser Tyr Ser

145 150 155 160

Asp Tyr Gly Arg Gly Ser Gly Gly Gly Val Thr Ile Ala Ser Lys Gln

165 170 175

Asp Gly Gly Ser Gly Ser Gly Glu Gly Arg Gly Gly Asn Ser Gly Gly

180 185 190

Gln Ser Ala Asp Gly Gly Glu Ser Leu Tyr Leu Lys Asn Ser Asp Glu

195 200 205

Gly Asn

210

<210>93

<211>615

<212>DNA

＜213〉two fringe false bromegrasses (Brachypodium distachyon)

<400>93

atgcagcagg cgatgtccat gtccccgggg tcggccggcg cggtgccgcc tccggccggc 60

atcaccacag agcagatcca aaagtatttg gatgaaaata agcaacttat tttggccatc 120

ctggaaaatc agaacctagg aaagttgact gaatgtgctc agtatcaagc tcaacttcag 180

aagaatctct tgtatctggc tgccattgcg gatgcccaac caccacagaa ccctggaagt 240

cgcccccaga tggtgcagcc tggtggtatg ccaggtgcag ggcattacat gtcgcaagta 300

ccaatgttcc ctccaagaac ccctttaacc ccacaacaga tgcaagagca acagcaccag 360

cagcttcagc agcagcaagc acaggctctt gctttcccca gccagatggt catgagacca 420

ggtactgtga acggcatgca gcctatgcaa gctgatctcc aagcagcagc agcagcacct 480

ggcctggcag acagccgagg aagtaagcag gacgcagcgg tagctggggc catctcggaa 540

ccttctggca ccgagagtca caagagtaca ggagcggatc atgaggcagg tggcgatgta 600

gctgagcaat cctaa 615

<210>94

<211>204

<212>PRT

＜213〉two fringe false bromegrasses

<400>94

Met Gln Gln Ala Met Ser Met Ser Pro Gly Ser Ala Gly Ala Val Pro

1 5 10 15

Pro Pro Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu

20 25 30

Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys

35 40 45

Leu Thr Glu Cys Ala GlnTyr Gln Ala Gln Leu Gln Lys Asn Leu Leu

50 55 60

Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Pro Gln Asn Pro Gly Ser

65 70 75 80

Arg Pro Gln Met Val Gln Pro Gly Gly Met Pro Gly Ala Gly His Tyr

85 90 95

Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln

100 105 110

Gln Met Gln Glu Gln Gln His Gln Gln Leu Gln Gln Gln Gln Ala Gln

115 120 125

Ala Leu Ala Phe Pro Ser Gln Met Val Met Arg Pro Gly Thr Val Asn

130 135 140

Gly Met Gln Pro Met Gln Ala Asp Leu Gln Ala Ala Ala Ala Ala Pro

145 150 155 160

Gly Leu Ala Asp Ser Arg Gly Ser Lys Gln Asp Ala Ala Val Ala Gly

165 170 175

Ala Ile Ser Glu Pro Ser Gly Thr Glu Ser His Lys Ser Thr Gly Ala

180 185 190

Asp His Glu Ala Gly Gly Asp Val Ala Glu Gln Ser

195 200

<210>95

<211>636

<212>DNA

＜213〉colea

<400>95

atgcagcagc agcagcagca gcagcagcag cctccgcaaa tgtttccgat ggctccttcg 60

atgccgccaa ctaacatcac caccgaacag atccaaaagt accttgagga gaacaagaag 120

ctgataatgg caatcatgga aaatcagaat cttggcaagc ttgcagagtg tgcacagtac 180

caagctcttc tccagaagaa cttaatgtac ctcgctgcta ttgctgatgc tcaacctcct 240

ccatctaccg ctggagctac accaccacca gctatggctt cccagatggg ggcaccgcat 300

cctgggatgc aaccgccgag ctactttatg caacacccac aagcttcagg gatggctcaa 360

caagcaccac ccgctggtat cttccctccg agaggtcctt tgcagtttgg tagcccacac 420

cagcttcagg atccgcaaca gcagcatatg catcaacagg ctatgcaagg acacatgggg 480

atgcgaccaa tgggtatcaa caacaacaat gggatgcagc atcagatgca gcaacaacaa 540

ccagaaacct ctcttggagg aagcgctgca aacgtggggc ttagaggtgg aaagcaagat 600

ggagcagatg gacaaggaaa agatgatggc aaatga 636

<210>96

<211>203

<212>PRT

＜213〉colea

<400>96

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Gly Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Ser Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Ser Val His Ser

65 70 75 80

Gln Tyr Gly Ser Ala Gly Gly Gly Leu Ile Gln Gly Glu Gly Ala Ser

85 90 95

His Tyr Leu Gln Gln Gln Gln Ala Thr Gln Gln Gln Gln Met Thr Gln

100 105 110

Gln Ser Leu Met Ala Ala Arg Ser Ser Met Met Tyr Gln Gln Gln Gln

115 120 125

Gln Pro Tyr Ala Thr Leu Gln His Gln Gln Leu His His Ser Gln Leu

130 135 140

Gly Met Ser Ser Ser Ser Gly Gly Gly Ser Ser Gly Leu His Ile Leu

145 150 155 160

Gln Gly Glu Ala Gly Gly Phe His Glu Phe Gly Arg Gly Lys Pro Glu

165 170 175

Met Gly Ser Gly Glu Gly Arg Gly Gly Ser Ser Gly Asp Gly Gly Glu

180 185 190

Thr Leu Tyr Leu Lys Ser Ser Asp Asp Gly Asn

195 200

<210>97

<211>636

<212>DNA

＜213〉sweet orange

<400>97

atgcagcagc caccgcaaat gatccctgtt atgccttcat ttccacccac caacatcacc 60

acagagcaga ttcaaaagta ccttgatgag aacaaaaagt tgattttggc aattttggac 120

aatcaaaatc ttggaaagct tacagaatgt gcccactatc aagctcagct tcaaaagaat 180

ttaatgtatt tagctgcaat tgctgatgca caaccacaag caccaacaat gcctcctcag 240

atggctccac atcctgcaat gcaagctagt gggtattaca tgcaacatcc tcaggcggca 300

gcaatggctc agcaacaagg aatctttccc caaaagatgc cattacaatt caataaccct 360

catcaactac aggatcctca acagcagcta caccaacatc aagccatgca agcacaaatg 420

ggaatgagac cgggtgccac taacaatggt atgcatccca tgcatgctga aagctctctt 480

ggaggtggca gcagtggagg acccccttca gcatcaggcc caggtgacat acgtggtgga 540

aataagcaag atgcctcgga ggctgggact actggtgctg atggccaggg cagttcggct 600

ggtgggcatg gtggggatgg agaggaggca aagtga 636

<210>98

<211>211

<212>PRT

＜213〉sweet orange

<400>98

Met Gln Gln Pro Pro Gln Met Ile Pro Val Met Pro Ser Phe Pro Pro

1 5 10 15

Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Thr

35 40 45

Glu Cys Ala His Tyr Gln Ala Gln Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Ala Pro Thr Met Pro Pro Gln

65 70 75 80

Met Ala Pro His Pro Ala Met Gln Ala Ser Gly Tyr Tyr Met Gln His

85 90 95

Pro Gln Ala Ala Ala Met Ala Gln Gln Gln Gly Ile Phe Pro Gln Lys

100 105 110

Met Pro Leu Gln Phe Asn Asn Pro His Gln Leu Gln Asp Pro Gln Gln

115 120 125

Gln Leu His Gln His Gln Ala Met Gln Ala Gln Met Gly Met Arg Pro

130 135 140

Gly Ala Thr Asn Asn Gly Met His Pro Met His Ala Glu Ser Ser Leu

145 150 155 160

Gly Gly Gly Ser Ser Gly Gly Pro Pro Ser Ala Ser Gly Pro Gly Asp

165 170 175

Ile Arg Gly Gly Asn Lys Gln Asp Ala Ser Glu Ala Gly Thr Thr Gly

180 185 190

Ala Asp Gly Gln Gly Ser Ser Ala Gly Gly His Gly Gly Asp Gly Glu

195 200 205

Glu Ala Lys

210

<210>99

<211>597

<212>DNA

＜213〉Euphorbia esula L (Euphorbia esula)

<400>99

atgcagcagc aaccgcagat gatgcctatg atgccttcat atccaccagc aaacattacc 60

acggagcaaa tccaaaagta tcttgatgaa aataaaaaat tgattttggc gatcttggat 120

aatcaaaatc ttggaaaact cgctgagtgt gcacagtatc aagccctgct gcaaaaaaat 180

ctgatgtatt tagccgcaat tgctgatgca caaccccaga ccccacccat gccacctcag 240

atgtccccac atccggctat gcaacaagga gcatattaca tgcaacatcc tcaggctgca 300

gcagcagcaa tggctcatca gtcgggtatt ttcccaccaa agatgtctcc gttacaattc 360

aataatcctc atcaaataca ggacccccag cagttacatc aagcagccct ccaagggcaa 420

atgggaatga ggcccatggg gcccaataac gggatgcatc cgatgcaccc cgaggcaaat 480

cttggaggat ctaatgatgg tcgtggagga aacaaacagg atgctccgga gacgggagca 540

tcgggaggtg atgggcaagg caattctggt ggtgatgggg ctgaagatgg gaaatga 597

<210>100

<211>198

<212>PRT

＜213〉Euphorbia esula L

<400>100

Met Gln Gln Gln Pro Gln Met Met Pro Met Met Pro Ser Tyr Pro Pro

1 5 10 15

Ala Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Ala

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Leu Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Pro Pro Met Pro Pro Gln

65 70 75 80

Met Ser Pro His Pro Ala Met Gln Gln Gly Ala Tyr Tyr Met Gln His

85 90 95

Pro Gln Ala Ala Ala Ala Ala Met Ala His Gln Ser Gly Ile Phe Pro

100 105 110

Pro Lys Met Ser Pro Leu Gln Phe Asn Asn Pro His Gln Ile Gln Asp

115 120 125

Pro Gln Gln Leu His Gln Ala Ala Leu Gln Gly Gln Met Gly Met Arg

130 135 140

Pro Met Gly Pro Asn Asn Gly Met His Pro Met His Pro Glu Ala Asn

145 150 155 160

Leu Gly Gly Ser Asn Asp Gly Arg Gly Gly Asn Lys Gln Asp Ala Pro

165 170 175

Glu Thr Gly Ala Ser Gly Gly Asp Gly Gln Gly Asn Ser Gly Gly Asp

180 185 190

Gly Ala Glu Asp Gly Lys

195

<210>101

<211>642

<212>DNA

＜213〉soybean

<400>101

atgcagcaga caccgccaat gattcctatg atgccttctt tcccacctac gaacataacc 60

accgagcaga ttcaaaaata ccttgatgag aacaagaagc tgattctggc aatattggac 120

aatcaaaatc ttggaaaact tgcagaatgt gcccagtacc aagctcagct tcaaaagaat 180

ttgatgtatt tagctgcaat tgctgatgcc cagcctcaaa ccccggccat gcctccgcag 240

atggcaccgc accctgccat gcaaccagga ttctatatgc aacatcctca ggctgctgca 300

gcagcaatgg ctcagcagca gcaaggaatg ttcccccaga aaatgccatt gcaatttggc 360

aatccacatc aaatgcagga acaacaacag cagctacacc agcaggccat ccaaggtcaa 420

atgggactta gacctggaga tataaataat ggcatgcatc caatgcacag tgaggctgct 480

cttggaggtg gaaacagcgg tggtccacct tcggctactg gtccaaacga tgcacgtggt 540

ggaagcaagc aagatgcctc tgaggctgga acagctggtg gagacggcca aggcagctcc 600

gcggctgc tc ataacagtgg agatggtgaa gaggcaaagt ga 642

<210>102

<211>213

<212>PRT

＜213〉soybean

<400>102

Met Gln Gln Thr Pro Pro Met Ile Pro Met Met Pro Ser Phe Pro Pro

1 5 10 15

Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Ala

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Pro Ala Met Pro Pro Gln

65 70 75 80

Met Ala Pro His Pro Ala Met Gln Pro Gly Phe Tyr Met Gln His Pro

85 90 95

Gln Ala Ala Ala Ala Ala Met Ala Gln Gln Gln Gln Gly Met Phe Pro

100 105 110

Gln Lys Met Pro Leu Gln Phe Gly Asn Pro His Gln Met Gln Glu Gln

115 120 125

Gln Gln Gln Leu His Gln Gln Ala Ile Gln Gly Gln Met Gly Leu Arg

130 135 140

Pro Gly Asp Ile Asn Asn Gly Met His Pro Met His Ser Glu Ala Ala

145 150 155 160

Leu Gly Gly Gly Asn Ser Gly Gly Pro Pro Ser Ala Thr Gly Pro Asn

165 170 175

Asp Ala Arg Gly Gly Ser Lys Gln Asp Ala Ser Glu Ala Gly Thr Ala

180 185 190

Gly Gly Asp Gly Gln Gly Ser Ser Ala Ala Ala His Asn Ser Gly Asp

195 200 205

Gly Glu Glu Ala Lys

210

<210>103

<211>633

<212>DNA

＜213〉wild soybean (Glycine soya)

<400>103

atgcagcaga caccgcctat gattcctatg atgccttcgt tcccacctac gaacataacc 60

accgagcaga ttcaaaaata ccttgatgag aacaagaagc tgattctggc aatattggac 120

aatcaaaatc ttggaaaact tgcagaatgt gcccagtacc aagctcagct tcaaaagaat 180

ttgatgtatt tagctgcaat tgctgatgcc cagcctcaaa caccagccat gcctccacag 240

atggcaccac accctgccat gcaaccagga ttctatatgc aacatcctca ggctgcagca 300

gcagcaatgg ctcagcagca gcagcaagga atgttccccc agaaaatgcc attgcaattt 360

ggcaatccac atcaaatgca ggaacaacag cagcagctac accagcaagc catccaaggt 420

caaatgggac tgagacctgg aggaataaat aatggcatgc atccaatgca caatgagggc 480

ggcaacagcg gtggtccacc ctcggctacc ggtccgaacg acgcacgtgg tggaagcaag 540

caagatgctt ctgaggctgg aacagctggt ggagatggcc aaggcagctc tgcagctgct 600

cataacagtg gagatggtga agaggcaaag tga 633

<210>104

<211>210

<212>PRT

＜213〉wild soybean

<400>104

Met Gln Gln Thr Pro Pro Met Ile Pro Met Met Pro Ser Phe Pro Pro

1 5 10 15

Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Ala

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Pro Ala Met Pro Pro Gln

65 70 75 80

Met Ala Pro His Pro Ala Met Gln Pro Gly Phe Tyr Met Gln His Pro

85 90 95

Gln Ala Ala Ala Ala Ala Met Ala Gln Gln Gln Gln Gln Gly Met Phe

100 105 110

Pro Gln Lys Met Pro Leu Gln Phe Gly Asn Pro His Gln Met Gln Glu

115 120 125

Gln Gln Gln Gln Leu His Gln Gln Ala Ile Gln Gly Gln Met Gly Leu

130 135 140

Arg Pro Gly Gly Ile Asn Asn Gly Met His Pro Met His Asn Glu Gly

145 150 155 160

Gly Asn Ser Gly Gly Pro Pro Ser Ala Thr Gly Pro Asn Asp Ala Arg

165 170 175

Gly Gly Ser Lys Gln Asp Ala Ser Glu Ala Gly Thr Ala Gly Gly Asp

180 185 190

Gly Gln Gly Ser Ser Ala Ala Ala His Asn Ser Gly Asp Gly Glu Glu

195 200 205

Ala Lys

210

<210>105

<211>690

<212>DNA

＜213〉upland cotton (Gossypium hirsutum)

<400>105

atgcagcagc acctgatgca gatgcagccc atgatggcag cttattatcc caacaacgtc 60

actactgatc atattcaaca gtatctcgat gagaacaagt cattgatctt aaagattgtt 120

gagagccaga attctgggaa attgagtgaa tgtgctgaga accaagcaag gctgcagcga 180

aacctcatgt acctggctgc cattgcggat tctcaacccc aaccacccac cgtgcatgca 240

cagtttccat ctggtggtat catgcagcca ggagctgggc actacatgca gcaccaacaa 300

gctcaacaaa tgacacaaca gtcgcttatg gctgctcggt cctcaatgtt gtattctcag 360

caaccatttt ctgcactgca acaacaacag cagcaagctt tgcacagtca gcttggcatg 420

agctctggcg gaagcacagg ccttcatatg ctgcaaactg aatctagtac tgcaggtggc 480

agtggagcac ttggggccgg agggtttcct gattttggac gtggttcttc tggagaaggc 540

atccatggtg gcaggccaat ggcaggtgga agcaagcaag atatcgggag tgccggctca 600

gctgaaggtc gtggaggaag ctctggtggt cagggtggtg gtgatggggg tgaaaccctt 660

tacttaaaag cagccgatga tgggaactga 690

<210>106

<211>229

<212>PRT

＜213〉upland cotton

<400>106

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Thr Val His Ala

65 70 75 80

Gln Phe Pro Ser Gly Gly Ile Met Gln Pro Gly Ala Gly His Tyr Met

85 90 95

Gln His Gln Gln Ala Gln Gln Met Thr Gln Gln Ser Leu Met Ala Ala

100 105 110

Arg Ser Ser Met Leu Tyr Ser Gln Gln Pro Phe Ser Ala Leu Gln Gln

115 120 125

Gln Gln Gln Gln Ala Leu His Ser Gln Leu Gly Met Ser Ser Gly Gly

130 135 140

Ser Thr Gly Leu His Met Leu Gln Thr Glu Ser Ser Thr Ala Gly Gly

145 150 155 160

Ser Gly Ala Leu Gly Ala Gly Gly Phe Pro Asp Phe Gly Arg Gly Ser

165 170 175

Ser Gly Glu Gly Ile His Gly Gly Arg Pro Met Ala Gly Gly Ser Lys

180 185 190

Gln Asp Ile Gly Ser Ala Gly Ser Ala Glu Gly Arg Gly Gly Ser Ser

195 200 205

Gly Gly Gln Gly Gly Gly Asp Gly Gly Glu Thr Leu Tyr Leu Lys Ala

210 215 220

Ala Asp Asp Gly Asn

225

<210>107

<211>642

<212>DNA

＜213〉upland cotton

<400>107

atgccgcagc caccgcaaat gattcctgtg atgccttcat atccacctac taatatcact 60

actgaacaga ttcagaagta ccttgatgag aataagaagt tgattttggc aattttggac 120

aatcagaatc ttggaaaact cgctgaatgc gcccagtatc aagctcagct gcaaaagaat 180

ttgatgtatt tagctgcaat tgcggatgct caacctcaat caacgccagc aatgtcgcct 240

cagatggcac cgcatccagc aatgcaaccc ggaggatatt ttatgcaaca tcctcaagct 300

gctgcaatgt cacagcaacc tggcatgtac cctcaaaagg tgccattgca attcaatagt 360

ccgcatcaaa tgcaggaccc tcagcacctc ctatatcagc agcatcaaca agcaatgcaa 420

ggtcaaatgg gaatcaggcc tgggggaccc aataatagca tgcatcccat gcattcagag 480

gctagccttg gaggcggcag cagtggtggt ccccctcaac cttcaggccc aagtgatgga 540

cgtgctggaa acaagcaaga gggctccgaa gctggtggta atgggcaggg cagcacaact 600

ggtgggcatg gtggcggtga tggagcggat gaggcaaagt ga 642

<210>108

<211>213

<212>PRT

＜213〉upland cotton

<400>108

Met Pro Gln Pro Pro Gln Met Ile Pro Val Met Pro Ser Tyr Pro Pro

1 5 10 15

Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Ala

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Ser Thr Pro Ala Met Ser Pro

65 70 75 80

Gln Met Ala Pro His Pro Ala Met Gln Pro Gly Gly Tyr Phe Met Gln

85 90 95

His Pro Gln Ala Ala Ala Met Ser Gln Gln Pro Gly Met Tyr Pro Gln

100 105 110

Lys Val Pro Leu Gln Phe Asn Ser Pro His Gln Met Gln Asp Pro Gln

115 120 125

His Leu Leu Tyr Gln Gln His Gln Gln Ala Met Gln Gly Gln Met Gly

130 135 140

Ile Arg Pro Gly Gly Pro Asn Asn Ser Met His Pro Met His Ser Glu

145 150 155 160

Ala Ser Leu Gly Gly Gly Ser Ser Gly Gly Pro Pro Gln Pro Ser Gly

165 170 175

Pro Ser Asp Gly Arg Ala Gly Asn Lys Gln Glu Gly Ser Glu Ala Gly

180 185 190

Gly Asn Gly Gln Gly Ser Thr Thr Gly Gly His Gly Gly Gly Asp Gly

195 200 205

Ala Asp Glu Ala Lys

210

<210>109

<211>561

<212>DNA

＜213〉barley

<400>109

atgcagcaag cgatgcccat gccgccggcg gcggcggcgc ctgggatgcc tccttctgcc 60

ggcctcagca ccgagcagat ccaaaagtac ctggatgaaa ataaacaact aattttggct 120

atcttggaaa atcagaacct gggaaagttg gcggaatgtg ctcagtatca agctcagctt 180

cagaagaatc ttttgtattt ggctgcgatt gctgatactc agccacagac ctctgtaagc 240

cgtcctcaga tggcaccacc tgctgcatcc ccaggggcag ggcattacat gtcacaggtg 300

ccaatgttcc ctccgaggac ccctctaacg cctcagcaga tgcaggagca gcaactacag 360

caacaacagg ctcagatgct tccgtttgct ggtcaaatgg ttgcgagacc cggggctgtc 420

aatggcattc cccaggcccc tcaagttgaa caaccagcct atgcagcagg tggggccagt 480

tccgagcctt ctggcaccga gagccacagg agcactggcg ccgataacga tggtgggagc 540

ggcttggctg accagtccta a 561

<210>110

<211>186

<212>PRT

＜213〉barley

<400>110

Met Gln Gln Ala Met Pro Met Pro Pro Ala Ala Ala Ala Pro Gly Met

1 5 10 15

Pro Pro Ser Ala Gly Leu Ser Thr Glu Gln Ile Gln Lys Tyr Leu Asp

20 25 30

Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly

35 40 45

Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu

50 55 60

Leu Tyr Leu Ala Ala Ile Ala Asp Thr Gln Pro Gln Thr Ser Val Ser

65 70 75 80

Arg Pro Gln Met Ala Pro Pro Ala Ala Ser Pro Gly Ala Gly His Tyr

85 90 95

Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln

100 105 110

Gln Met Gln Glu Gln Gln Leu Gln Gln Gln Gln Ala Gln Met Leu Pro

115 120 125

Phe Ala Gly Gln Met Val Ala Arg Pro Gly Ala Val Asn Gly Ile Pro

130 135 140

Gln Ala Pro Gln Val Glu Gln Pro Ala Tyr Ala Ala Gly Gly Ala Ser

145 150 155 160

Ser Glu Pro Ser Gly Thr Glu Ser His Arg Ser Thr Gly Ala Asp Asn

165 170 175

Asp Gly Gly Ser Gly Leu Ala Asp Gln Ser

180 185

<210>111

<211>555

<212>DNA

＜213〉fireweed (Lactuca serriola)

<220>

<221>misc_feature

<222>(253)..(253)

＜223〉n is a, c, g or t

<400>111

atgaagcagc cgatgatgcc gaatccaatg atgtcttctt cgtttcctcc tacaaacatc 60

accaccgatc agatccaaaa gttccttgat gaaaacaagc aactaattat agcaataatg 120

agcaacctaa atcttggaaa gcttgctgaa tgtgcccagt accaagctct actccaaaaa 180

aatttgatgt atctagcagc cattgcagat gctcaaccac ctacacctac accaacacta 240

aatatctctt atnagatggg cccggttcca catccaggga tgccacagca aggtggattt 300

tacatggcgc agcagcaccc tcaggcggct gtaatgacgg ctcagccacc ttctggtttt 360

ccacaaccga tgcctggtat gcaatttaac agcccacagg ctattcaagg gcagatgggc 420

gggaggtccg gtgggccgcc aagctcagcc gctagtgatg tctggagagg aagcatgcaa 480

gatggtggtg gtggtgctgc tgctgatggt ggtaaggatg gtcatgctgg cggtggacct 540

gaggaagcaa agtaa 555

<210>112

<211>184

<212>PRT

＜213〉fireweed

<220>

＜221〉uncertain

<222>(85)..(85)

＜223〉Xaa can make the amino acid of any natural generation

<400>112

Met Lys Gln Pro Met Met Pro Asn Pro Met Met Ser Ser Ser Phe Pro

1 5 10 15

Pro Thr Asn Ile Thr Thr Asp Gln Ile Gln Lys Phe Leu Asp Glu Asn

20 25 30

Lys Gln Leu Ile Ile Ala Ile Met Ser Asn Leu Asn Leu Gly Lys Leu

35 40 45

Ala Glu Cys Ala Gln Tyr Gln Ala Leu Leu Gln Lys Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ala Gln Pro Pro Thr Pro Thr Pro Thr Leu

65 70 75 80

Asn Ile Ser Tyr Xaa Met Gly Pro Val Pro His Pro Gly Met Pro Gln

85 90 95

Gln Gly Gly Phe Tyr Met Ala Gln Gln His Pro Gln Ala Ala Val Met

100 105 110

Thr Ala Gln Pro Pro Ser Gly Phe Pro Gln Pro Met Pro Gly Met Gln

115 120 125

Phe Asn Ser Pro Gln Ala Ile Gln Gly Gln Met Gly Gly Arg Ser Gly

130 135 140

Gly Pro Pro Ser Ser Ala Ala Ser Asp Val Trp Arg Gly Ser Met Gln

145 150 155 160

Asp Gly Gly Gly Gly Ala Ala Ala Asp Gly Gly Lys Asp Gly His Ala

165 170 175

Gly Gly Gly Pro Glu Glu Ala Lys

180

<210>113

<211>627

<212>DNA

＜213〉tomato (Lycopersicon esculentum)

<400>113

atgcagcagc acctgatgca gatgcagccc atgatggcag cttactatcc aacgaacgtc 60

actactgacc atattcaaca gtatttggat gaaaacaaat cactcattct gaagattgtt 120

gagagccaga actctgggaa actcagtgaa tgtgcggaga accaagctag gcttcagagg 180

aatctgatgt accttgctgc gattgctgat tcacaacctc aaccttctag catgcattct 240

cagttctctt ctggtgggat gatgcagcca gggacacaca gttacttgca gcagcagcag 300

cagcaacaac aagcgcaaca aatggcaaca caacaactca tggctgcaag atcctcgtcg 360

atgctctatg gacaacagca gcagcaatct cagttatcgc aatatcaaca aggcttgcat 420

agtagccaac tcggcatgag ttctggcagt ggcggaagca ctggacttca tcacatgctt 480

caaagtgaat catcacctca tggtggtggt ttctctcatg acttcggccg cgcaaataag 540

caagacattg ggagtagtat gtctgctgaa gggcgcggcg gaagttcagg tggtgagaat 600

ctttatc tga aagcttctga ggattga 627

<210>114

<211>208

<212>PRT

＜213〉tomato

<400>114

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Thr Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Ser Ser Met His Ser

65 70 75 80

Gln Phe Ser Ser Gly Gly Met Met Gln Pro Gly Thr His Ser Tyr Leu

85 90 95

Gln Gln Gln Gln Gln Gln Gln Gln Ala Gln Gln Met Ala Thr Gln Gln

100 105 110

Leu Met Ala Ala Arg Ser Ser Ser Met Leu Tyr Gly Gln Gln Gln Gln

115 120 125

Gln Ser Gln Leu Ser Gln Tyr Gln Gln Gly Leu His Ser Ser Gln Leu

130 135 140

Gly Met Ser Ser Gly Ser Gly Gly Ser Thr Gly Leu His His Met Leu

145 150 155 160

Gln Ser Glu Ser Ser Pro His Gly Gly Gly Phe Ser His Asp Phe Gly

165 170 175

Arg Ala Asn Lys Gln Asp Ile Gly Ser Ser Met Ser Ala Glu Gly Arg

180 185 190

Gly Gly Ser Ser Gly Gly Glu Asn Leu Tyr Leu Lys Ala Ser Glu Asp

195 200 205

<210>115

<211>624

<212>DNA

＜213〉domestication apple (Malus domestica)

<400>115

atgcagcagc caccacaaat gatccccgtc atgccttcat ttcctcccac caacatcacc 60

accgaacaaa ttcagaagta ccttgatgac aacaaaaagt tgattctggc aatattggat 120

aatcaaaatc ttggaaaact tgctgagtgt gctcagtacc aggctctgct tcaaaagaat 180

ctgatgtatt tagcagcaat tgccgatgcg caaccacagg caccagctgc ccctccccag 240

atggccccac atcctgctat gcaacaggca ggatattaca tgcaacatcc tcaggcagca 300

gcaatggctc agcaacaggg tattttctcc ccaaagatgc cgatgcaatt caataacatg 360

catcaaatgc acgatccaca gcagcaccaa caagccatgc aagggcaaat gggaatgaga 420

cctggagggc ctaacggcat gccttccatg cttcatactg aggccacaca tggtggtggt 480

agtggcggcc caaattcagc tggagaccca aatgatgggc gtggaggaag caagcaagac 540

gcc tctgagt ctggggcagg tggtgatggc caggggacct cagccggcgg gcgtggaact 600

ggtgatggag aggacggcaa gtga 624

<210>116

<211>207

<212>PRT

＜213〉domestication apple

<400>116

Met Gln Gln Pro Pro Gln Met Ile Pro Val Met Pro Ser Phe Pro Pro

1 5 10 15

Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Asp Asn Lys

20 25 30

Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu Ala

35 40 45

Glu Cys Ala Gln Tyr Gln Ala Leu Leu Gln Lys Asn Leu Met Tyr Leu

50 55 60

Ala Ala Ile Ala Asp Ala Gln Pro Gln Ala Pro Ala Ala Pro Pro Gln

65 70 75 80

Met Ala Pro His Pro Ala Met Gln Gln Ala Gly Tyr Tyr Met Gln His

85 90 95

Pro Gln Ala Ala Ala Met Ala Gln Gln Gln Gly Ile Phe Ser Pro Lys

100 105 110

Met Pro Met Gln Phe Asn Asn Met His Gln Met His Asp Pro Gln Gln

115 120 125

His Gln Gln Ala Met Gln Gly Gln Met Gly Met Arg Pro Gly Gly Pro

130 135 140

Asn Gly Met Pro Ser Met Leu His Thr Glu Ala Thr His Gly Gly Gly

145 150 155 160

Ser Gly Gly Pro Asn Ser Ala Gly Asp Pro Asn Asp Gly Arg Gly Gly

165 170 175

Ser Lys Gln Asp Ala Ser Glu Ser Gly Ala Gly Gly Asp Gly Gln Gly

180 185 190

Thr Ser Ala Gly Gly Arg Gly Thr Gly Asp Gly Glu Asp Gly Lys

195 200 205

<210>117

<211>639

<212>DNA

＜213〉puncture vine clover

<400>117

atgcagcaga cacctcaaat gattcctatg atgccttcat tcccacaaca aacaaacata 60

accactgagc agattcaaaa atatcttgat gagaacaaga agctgatcct ggcaatattg 120

gacaatcaaa atcttggaaa acttgcagaa tgtgcccagt accaagctca gcttcagaag 180

aatttgatgt atttagctgc aattgctgac gcgcagccac aaacaccggc cttgcctcca 240

cagatggccc cgcaccctgc gatgcaacaa ggattctata tgcaacatcc tcaggctgca 300

gcaatggctc agcaacaagg aatgttcccc caaaaaatgc caatgcagtt cggtaatccg 360

catcaaatgc aggatcagca gcatcagcag caacaacagc agctacatca gcaagctatg 420

caaggtcaaa tgggacttag acctggaggg ataaataacg gcatgcatcc aatgcacaac 480

gaggctgctc tcggaggtag cggcagtggt ggtcaaatga cgggcgtggt ggtggagcaa 540

gcaagatgct tcggagctgg gacagccggc ggtgatggtc aaggaacctc tgccgcagct 600

gcgcacaaca gtggagatgc ttcagaagaa ggaaagtaa 639

<210>118

<211>213

<212>PRT

＜213〉puncture vine clover

<400>118

Met Gln Gln Thr Pro Gln Met Ile Pro Met Met Pro Ser Phe Pro Gln

1 5 10 15

Gln Thr Asn Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn

20 25 30

lys Lys Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu

35 40 45

Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Pro Ala Leu Pro Pro

65 70 75 80

Gln Met Ala Pro His Pro Ala Met Gln Gln Gly Phe Tyr Met Gln His

85 90 95

Pro Gln Ala Ala Ala Met Ala Gln Gln Gln Gly Met Phe Pro Gln Lys

100 105 110

Met Pro Met Gln Phe Gly Asn Pro His Gln Met Gln Asp Gln Gln His

115 120 125

Gln Gln Gln Gln Gln Gln Leu His Gln Gln Ala Met Gln Gly Gln Met

130 135 140

Gly Leu Arg Pro Gly Gly Ile Asn Asn Gly Met His Pro Met His Asn

145 150 155 160

Glu Ala Ala Leu Gly Gly Ser Gly Ser Gly Gly Pro Asn Asp Gly Arg

165 170 175

Gly Gly Gly Ser Lys Gln Asp Ala Ser Glu Ala Gly Thr Ala Gly Gly

180 185 190

Asp Gly Gln Gly Thr Ser Ala Ala Ala Ala His Asn Ser Gly Asp Ala

195 200 205

Ser Glu Glu Gly Lys

210

<210>119

<211>624

<212>DNA

＜213〉switchgrass (Panicum virgatum)

<400>119

atgcagcagc agatgcccat gcagtcggcg cccccggcga ccggcatcac caccgagcag 60

atccaaaagt atttggatga aaataagcag cttattttgg ccatcctgga aaatcagaac 120

ttaggaaagt tggctgaatg tgctcagtat caagctcagc ttcaaaagaa tctcttgtac 180

ctggctgcga ttgcagatgc ccaaccccaa ccaccacaga accctgcaag tcgcccacag 240

atgatgcaac ctggcatggt accaggtgca gggcattaca tgtcccaagt accaatgttc 300

ccgccaagaa caccattaac cccgcaacag atgcaagaac agcagcagca gcagcagcag 360

cttcaacagc agcaagcaca ggctcttgct ttcccgggac agatggtcat gagacctacc 420

attaatggca tgcagcctat gcaagccgac cctgctgccg ccgccgccag cctacagcag 480

tcagcacctg gccctactga tgggcgagga ggcaagcaag atgcaactgc tggggtgagc 540

acagagcctt ctggcaccga gagccacaag agcacaaccg cagcagatca cgatgtgggc 600

actgatgtcg cggagaaatc ctaa 624

<210>120

<211>207

<212>PRT

＜213〉switchgrass

<400>120

Met Gln Gln Gln Met Pro Met Gln Ser Ala Pro Pro Ala Thr Gly Ile

1 5 10 15

Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn Lys Gln Leu Ile

20 25 30

Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys Leu Ala Glu Cys Ala

35 40 45

Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Leu Tyr Leu Ala Ala Ile

50 55 60

Ala Asp Ala Gln Pro Gln Pro Pro Gln Asn Pro Ala Ser Arg Pro Gln

65 70 75 80

Met Met Gln Pro Gly Met Val Pro Gly Ala Gly His Tyr Met Ser Gln

85 90 95

Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln Gln Met Gln

100 105 110

Glu Gln Gln Gln Gln Gln Gln Gln Leu Gln Gln Gln Gln Ala Gln Ala

115 120 125

Leu Ala Phe Pro Gly Gln Met Val Met Arg Pro Thr Ile Asn Gly Met

130 135 140

Gln Pro Met Gln Ala Asp Pro Ala Ala Ala Ala Ala Ser Leu Gln Gln

145 150 155 160

Ser Ala Pro Gly Pro Thr Asp Gly Arg Gly Gly Lys Gln Asp Ala Thr

165 170 175

Ala Gly Val Ser Thr Glu Pro Ser Gly Thr Glu Ser His Lys Ser Thr

180 185 190

Thr Ala Ala Asp His Asp Val Gly Thr Asp Val Ala Glu Lys Ser

195 200 205

<210>121

<211>747

<212>DNA

＜213〉picea sitchensis (Picea sitchensis)

<400>121

atgcagcagc atctcatgca aatgcagccc atgatggcgg catacgcctc caacaacatc 60

accactgatc acatccagaa gtacctggat gagaacaagc agttgattct ggcaattctg 120

gacaaccaaa atcttggaaa gctcaatgag tgtgctcagt accaagcaaa acttcagcag 180

aatttgatgt atctggctgc gattgctgat tctcaaccac aagcacaaac tgcacatgct 240

cagattcctc ctaatgcagt gatgcagtct ggtgggcatt acatgcagca ccagcaggca 300

cagcaacaag tgactcctca gtctctgatg gcagctagat cttccatgct gtattctcag 360

cagccgatgg ctgctttgca tcaagctcag caacaacagc agcagcagca tcagcagcaa 420

caacaatctc ttcacagcca gcttggcata aattctggag gaagcagtgg attgcatatg 480

ttgcatggtg agacaaacat gggatgtaat gggcctctct catctggggg cttccctgaa 540

tttgggcgtg ggtctgctac ctctgctgaa ggtatgcagg ccaacagggg cttcactata 600

gatcgtggtt caaataagca ggatggagta ggatcagaga atgcccatcc aggtgctggt 660

gatggaagag ggagttcaac tggagggcag aatgcagatg agtcagaacc atcatacctg 720

aaagcctccg aagaagaagg aaac tag 747

<210>122

<211>248

<212>PRT

＜213〉picea sitchensis

<40>122

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Ala

1 5 10 15

Ser Asn Asn Ile Thr Thr Asp His Ile Gln Lys Tyr Leu Asp Glu Asn

20 25 30

Lys Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu

35 40 45

Asn Glu Cys Ala Gln Tyr Gln Ala Lys Leu Gln Gln Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Ala Gln Thr Ala His Ala

65 70 75 80

Gln Ile Pro Pro Asn Ala Val Met Gln Ser Gly Gly His Tyr Met Gln

85 90 95

His Gln Gln Ala Gln Gln Gln Val Thr Pro Gln Ser Leu Met Ala Ala

100 105 110

Arg Ser Ser Met Leu Tyr Ser Gln Gln Pro Met Ala Ala Leu His Gln

115 120 125

Ala Gln Gln Gln Gln Gln Gln Gln His Gln Gln Gln Gln Gln Ser Leu

130 135 140

His Ser Gln Leu Gly Ile Asn Ser Gly Gly Ser Ser Gly Leu His Met

145 150 155 160

Leu His Gly Glu Thr Asn Met Gly Cys Asn Gly Pro Leu Ser Ser Gly

165 170 175

Gly Phe Pro Glu Phe Gly Arg Gly Ser Ala Thr Ser Ala Glu Gly Met

180 185 190

Gln Ala Asn Arg Gly Phe Thr Ile Asp Arg Gly Ser Asn Lys Gln Asp

195 200 205

Gly Val Gly Ser Glu Asn Ala His Pro Gly Ala Gly Asp Gly Arg Gly

210 215 220

Ser Ser Thr Gly Gly Gln Asn Ala Asp Glu Ser Glu Pro Ser Tyr Leu

225 230 235 240

Lys Ala Ser Glu Glu Glu Gly Asn

245

<210>123

<211>735

<212>DNA

＜213〉torch pine (Pinus taeda)

<400>123

atgcagcagc acctcatgca aatgcagccc atgatggcgg cctacgcctc caacaatatc 60

accactgatc acatccagaa gtacctggat gagaacaagc agttgattct ggcaattttg 120

gacaaccaaa atctcggaaa gctcaatgag tgtgctcaat accaagcaaa acttcagcag 180

aatttgatgt atctggctgc tattgctgat tctcaacctc aagcacaaac tgcacatgct 240

cagattcctc caaatgcggt gatgcagtct ggtgggcatt acatgcagca tcaacaggca 300

cagcaacaag ttactcctca gtctctgatg gcagctagat cttccatact gtatgctcag 360

caacaacagc agcagcagca tcagcagcat cagcagcaac agcagcaaca acagtctctt 420

cacagccagc ttggcataaa ttctggagga agcagcggtt tgcatatgtt gcatggtgag 480

acaaacatgg gatgtaatgg gcctctgtca tctgggggat tccctgaatt tgggcgtggg 540

tctgctacct ctgctgatgg tatgcaggtg aacaggggct ttgctataga tcgtggttca 600

aacaagcagg atggagttgg atcagagaat gcccatgctg gtgctggtga tggaagaggg 660

agttcaactg gagggcagaa tgcagatgag tcagaaccat catacctgaa ggcctccgag 720

gaagaaggaa ac tag 735

<210>124

<211>244

<212>PRT

＜213〉torch pine

<400>124

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Ala

1 5 10 15

Ser Asn Asn Ile Thr Thr Asp His Ile Gln Lys Tyr Leu Asp Glu Asn

20 25 30

Lys Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Leu Gly Lys Leu

35 40 45

Asn Glu Cys Ala Gln Tyr Gln Ala Lys Leu Gln Gln Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Ala Gln Thr Ala His Ala

65 70 75 80

Gln Ile Pro Pro Asn Ala Val Met Gln Ser Gly Gly His Tyr Met Gln

85 90 95

His Gln Gln Ala Gln Gln Gln Val Thr Pro Gln Ser Leu Met Ala Ala

100 105 110

Arg Ser Ser Ile Leu Tyr Ala Gln Gln Gln Gln Gln Gln Gln His Gln

115 120 125

Gln His Gln Gln Gln Gln Gln Gln Gln Gln Ser Leu His Ser Gln Leu

130 135 140

Gly Ile Asn Ser Gly Gly Ser Ser Gly Leu His Met Leu His Gly Glu

145 150 155 160

Thr Asn Met Gly Cys Asn Gly Pro Leu Ser Ser Gly Gly Phe Pro Glu

165 170 175

Phe Gly Arg Gly Ser Ala Thr Ser Ala Asp Gly Met Gln Val Asn Arg

180 185 190

Gly Phe Ala Ile Asp Arg Gly Ser Asn Lys Gln Asp Gly Val Gly Ser

195 200 205

Glu Asn Ala His Ala Gly Ala Gly Asp Gly Arg Gly Ser Ser Thr Gly

210 215 220

Gly Gln Asn Ala Asp Glu Ser Glu Pro Ser Tyr Leu Lys Ala Ser Glu

225 230 235 240

Glu Glu Gly Asn

<210>125

<211>663

<212>DNA

＜213〉trembling poplar (Populus tremula)

<400>125

atgcaacagc acctgatgca gatgcagccc atgatggcag cctattaccc cagcaacgtc 60

actactgatc atattcaaca gtatctggac gaaaacaagt cattgatttt gaagattgtt 120

gagagccaga attcagggaa actcagtgag tgtgcagaga accaagcaag actgcaacaa 180

aatctcatgt acttggctgc aattgctgat tgtcagcccc aaccacctac catgcatgcc 240

cagttccctt ccagcggcat tatgcagcca ggagcacatt acatgcagca tcaacaagct 300

caacagatga caccacaagc ccttatggct gcacgctctt ctatgctgca gtatgctcaa 360

cagccattct cagcgcttca acaacagcaa gccttacaca gccagctcgg catgagctct 420

ggtggaagcg caggacttca tatgatgcaa agcgaggcta acactgcagg aggcagtgga 480

gctcttggtg ctggacgatt tcctgatttt ggcatggatg cctccagtag aggaatcgca 540

agtgggagca agcaagatat tcggagtgca gggtctagtg aagggcgagg aggaagctct 600

ggaggccagg gtggtgatgg aggtgaaacc ctttacttga aatctgctga tgatgggaac 660

tga 663

<210>126

<211>220

<212>PRT

＜213〉trembling poplar

<400>126

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Gln Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Cys Gln Pro Gln Pro Pro Thr Met His Ala

65 70 75 80

Gln Phe Pro Ser Ser Gly Ile Met Gln Pro Gly Ala His Tyr Met Gln

85 90 95

His Gln Gln Ala Gln Gln Met Thr Pro Gln Ala Leu Met Ala Ala Arg

100 105 110

Ser Ser Met Leu Gln Tyr Ala Gln Gln Pro Phe Ser Ala Leu Gln Gln

115 120 125

Gln Gln Ala Leu His Ser Gln Leu Gly Met Ser Ser Gly Gly Ser Ala

130 135 140

Gly Leu His Met Met Gln Ser Glu Ala Asn Thr Ala Gly Gly Ser Gly

145 150 155 160

Ala Leu Gly Ala Gly Arg Phe Pro Asp Phe Gly Met Asp Ala Ser Ser

165 170 175

Arg Gly Ile Ala Ser Gly Ser Lys Gln Asp Ile Arg Ser Ala Gly Ser

180 185 190

Ser Glu Gly Arg Gly Gly Ser Ser Gly Gly Gln Gly Gly Asp Gly Gly

195 200 205

Glu Thr Leu Tyr Leu Lys Ser Ala Asp Asp Gly Asn

210 215 220

<210>127

<211>678

<212>DNA

＜213〉sugarcane (Saccharum officinarum)

<400>127

atgcagcagc aacacctgat gcagatgaac cagaacatga ttgggggcta cacctctcct 60

gccgctgtga caaccgatct catccagcag tacctggatg agaacaagca gctgatcctg 120

gccatcctcg acaaccagaa caatggcaag gtggaggagt gcgaacggca ccaagctaag 180

ctccagcaca acctcatgta cctggccgcc atcgccgaca gccagccacc acagactgca 240

ccactatcac aatacccgtc caacctgatg atgcagccgg gccctcggta catgccaccg 300

cagtccgggc agatgatgag cccgcagtcg ctaatggcgg cgcggtcctc catgatgtac 360

gcgcacccgt ccatgtcacc actccagcag cagcaggcag cgcacgggca gctgggcatg 420

gcttcagggg gcggcggtgg cacgaccagt gggttcaaca tcctccatgg cgaggccagt 480

atgggcggtg ctggtggcgc ttgtgccggc aacaacatga tgaacgccgg catgttctca 540

ggctttggcc gcagcggcag tggcgccaag gagggatcga cctcgctgtc ggttgacgtc 600

cgtggtggca ccagctccgg cgcgcaaagc ggggacggcg agtacctgaa agcaggcacc 660

gaggaagaag gcagt taa 678

<210>128

<211>225

<212>PRT

＜213〉sugarcane

<400>128

Met Gln Gln Gln His Leu Met Gln Met Asn Gln Asn Met Ile Gly Gly

1 5 10 15

Tyr Thr Ser Pro Ala Ala Val Thr Thr Asp Leu Ile Gln Gln Tyr Leu

20 25 30

Asp Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Asp Asn Gln Asn Asn

35 40 45

Gly Lys Val Glu Glu Cys Glu Arg His Gln Ala Lys Leu Gln His Asn

50 55 60

Leu Met Tyr Leu Ala Ala Ile Ala Asp Ser Gln Pro Pro Gln Thr Ala

65 70 75 80

Pro Leu Ser Gln Tyr Pro Ser Asn Leu Met Met Gln Pro Gly Pro Arg

85 90 95

Tyr Met Pro Pro Gln Ser Gly Gln Met Met Ser Pro Gln Ser Leu Met

100 105 110

Ala Ala Arg Ser Ser Met Met Tyr Ala His Pro Ser Met Ser Pro Leu

115 120 125

Gln Gln Gln Gln Ala Ala His Gly Gln Leu Gly Met Ala Ser Gly Gly

130 135 140

Gly Gly Gly Thr Thr Ser Gly Phe Asn Ile Leu His Gly Glu Ala Ser

145 150 155 160

Met Gly Gly Ala Gly Gly Ala Cys Ala Gly Asn Asn Met Met Asn Ala

165 170 175

Gly Met Phe Ser Gly Phe Gly Arg Ser Gly Ser Gly Ala Lys Glu Gly

180 185 190

Ser Thr Ser Leu Ser Val Asp Val Arg Gly Gly Thr Ser Ser Gly Ala

195 200 205

Gln Ser Gly Asp Gly Glu Tyr Leu Lys Ala Gly Thr Glu Glu Glu Gly

210 215 220

Ser

225

<210>129

<211>561

<212>DNA

＜213〉sugarcane

<400>129

atgcagcagc cgatgcccat gcagccgcag gcgccggaga tgaccccggc cgccggaatc 60

accacggagc agatccaaaa gtatctggat gagaataagc agcttatttt ggctattttg 120

gaaaatcaga acctaggaaa attggcagaa tgtgctcagt atcaatcaca acttcagaag 180

aacctcttgt atctcgctgc aatcgcagat gcccaaccac agactgctgt aagccgccct 240

cagatggcgc cgcctggtgc attgcctgga gtagggcagt acatgtcaca ggtgcctatg 300

ttcccaccga ggacacctct aacaccccag cagatgcagg agcagcaact tcagcagcag 360

caggctcagc tgctaaattt cagtggccta atggttgcta gacctggcat ggtcaacggc 420

atgcctcagt ccattcaagt tcagcaagct cagccaccac cagcagggaa caaacaggat 480

gctggtgggg tcgcctcgga gccctcgggc attgagaacc acaggagcac tggtggtgat 540

aatgatggtg gaagcgacta g 561

<210>130

<211>186

<212>PRT

＜213〉sugarcane

<400>130

Met Gln Gln Pro Met Pro Met Gln Pro Gln Ala Pro Glu Met Thr Pro

1 5 10 15

Ala Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu Asn

20 25 30

Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys Leu

35 40 45

Ala Glu Cys Ala Gln Tyr Gln Ser Gln Leu Gln Lys Asn Leu Leu Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Thr Ala Val Ser Arg Pro

65 70 75 80

Gln Met Ala Pro Pro Gly Ala Leu Pro Gly Val Gly Gln Tyr Met Ser

85 90 95

Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln Gln Met

100 105 110

Gln Glu Gln Gln Leu Gln Gln Gln Gln Ala Gln Leu Leu Asn Phe Ser

115 120 125

Gly Leu Met Val Ala Arg Pro Gly Met Val Asn Gly Met Pro Gln Ser

130 135 140

lle Gln Val Gln Gln Ala Gln Pro Pro Pro Ala Gly Asn Lys Gln Asp

145 150 155 160

Ala Gly Gly Val Ala Ser Glu Pro Ser Gly Ile Glu Asn His Arg Ser

165 170 175

Thr Gly Gly Asp Asn Asp Gly Gly Ser Asp

180 185

<210>131

<211>642

<212>DNA

＜213〉sugarcane

<400>131

atgcagcagc agatgcccat gccgccggcg cccgctgcgg cggcggcgcc cccggcggcc 60

ggcatcacca ccgagcagat ccaaaagtat ttggacgaaa ataagcaact tattttggcc 120

atcctggaaa atcagaactt aggaaagttg gctgaatgtg ctcagtatca agctcaactt 180

caaaagaacc tcttgtacct ggctgcgatt gctgatgccc aaccccagcc accacaaaac 240

cctgcaggtc gccctcagat gatgcaacct ggtatagtgc caggtgcggg gcattacatg 300

tcacaagtac caatgttccc tccaagaact ccattaaccc cacagcagat gcaagagcag 360

cagcagcaac agcttcagca gcagcaagcg caggctctta cattccctgg acagatggtc 420

atgagaccag ctaccatcaa cggcatacag cagcctatgc aagctgaccc tgcccgggca 480

gcggagctgc aacaaccacc acctatccca gctgacgggc gagtaagcaa gcagcaggac 540

acaacggctg gcgtgagctc agagccttct gccaatgaga gccacaagac cacaactgga 600

gcagatagtg aggcaggtgg tgacgtggcg gagaaatcct aa 642

<210>132

<211>213

<212>PRT

＜213〉sugarcane

<400>132

Met Gln Gln Gln Met Pro Met Pro Pro Ala Pro Ala Ala Ala Ala Ala

1 5 10 15

Pro Pro Ala Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp

20 25 30

Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly

35 40 45

Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu

50 55 60

Leu Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Pro Pro Gln Asn

65 70 75 80

Pro Ala Gly Arg Pro Gln Met Met Gln Pro Gly Ile Val Pro Gly Ala

85 90 95

Gly His Tyr Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu

100 105 110

Thr Pro Gln Gln Met Gln Glu Gln Gln Gln Gln Gln Leu Gln Gln Gln

115 120 125

Gln Ala Gln Ala Leu Thr Phe Pro Gly Gln Met Val Met Arg Pro Ala

130 135 140

Thr Ile Asn Gly Ile Gln Gln Pro Met Gln Ala Asp Pro Ala Arg Ala

145 150 155 160

Ala Glu Leu Gln Gln Pro Pro Pro Ile Pro Ala Asp Gly Arg Val Ser

165 170 175

Lys Gln Gln Asp Thr Thr Ala Gly Val Ser Ser Glu Pro Ser Ala Asn

180 185 190

Glu Ser His Lys Thr Thr Thr Gly Ala Asp Ser Glu Ala Gly Gly Asp

195 200 205

Val Ala Glu Lys Ser

210

<210>133

<211>645

<212>DNA

＜213〉potato

<400>133

atgcagcagc acctgatgca gatgcagccc atgatggcag cttactatcc aacgaacgtc 60

actactgacc atattcaaca gtatttggat gagaacaaat cactcattct gaaaattgtt 120

gagagccaaa actcgggaaa actcagtgaa tgtgcagaga accaagctag gcttcagagg 180

aatctgatgt accttgctgc tattgctgat tcacaacctc agccttctag catgcattct 240

cagttctctt ctggtgggat gatgcagcca gggacacaca gttacctgca gcagcagcag 300

cagcaacaac aagcgcaaca aatggcaaca caacaactca tggctgcaag atcctcatca 360

atgctctatg gacaacaaca gcagcagcag cagcagtctc agttatcaca atttcaacaa 420

ggcttgcata gtagccaact tggcatgagt tctggcagtg gtggaagcac tggacttcat 480

cacatgcttc aaagtgaatc atcacctcat ggtggtggtt tctctcatga cttcggccgt 540

gcaaataagc aagacattgg gagtagtatg tctgctgaag ggcgcggcgg aagctcaggt 600

ggtgatggtg gtgagaatct ttatctgaaa gcttctgagg attga 645

<210>134

<211>214

<212>PRT

＜213〉potato

<400>134

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Thr Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Ser Ser Met His Ser

65 70 75 80

Gln Phe Ser Ser Gly Gly Met Met Gln Pro Gly Thr His Ser Tyr Leu

85 90 95

Gln Gln Gln Gln Gln Gln Gln Gln Ala Gln Gln Met Ala Thr Gln Gln

100 105 110

Leu Met Ala Ala Arg Ser Ser Ser Met Leu Tyr Gly Gln Gln Gln Gln

115 120 125

Gln Gln Gln Gln Ser Gln Leu Ser Gln Phe Gln Gln Gly Leu His Ser

130 135 140

Ser Gln Leu Gly Met Ser Ser Gly Ser Gly Gly Ser Thr Gly Leu His

145 150 155 160

His Met Leu Gln Ser Glu Ser Ser Pro His Gly Gly Gly Phe Ser His

165 170 175

Asp Phe Gly Arg Ala Asn Lys Gln Asp Ile Gly Ser Ser Met Ser Ala

180 185 190

Glu Gly Arg Gly Gly Ser Ser Gly Gly Asp Gly Gly Glu Asn Leu Tyr

195 200 205

Leu Lys Ala Ser Glu Asp

210

<210>135

<211>645

<212>DNA

＜213〉dichromatism chinese sorghum (Sorghum bicolor)

<400>135

atgcagcagc agatgcccat gccgccggcg cccgctgcgg cggcggcgac ggcgcccccg 60

gcggccggca tcaccaccga gcagatccag aagtatttgg acgaaaataa gcaacttatt 120

ttggccatcc tagaaaatca gaacttagga aagttggctg aatgtgctca gtatcaagct 180

caacttcaaa agaacctctt gtacctggct gcgattgctg atgcccaacc ccgaccaccg 240

caaaaccctg caggtcgccc tcagatgatg caacctggta tagtgccagg tgcagggcat 300

tacatgtcac aagtaccaat gttccctcca agaactccat taaccccaca gcaaatgcaa 360

gagcagcagc agcaacagct tcagcagcag caagcgcagg ctcttgcatt ccctgggcag 420

atggtcatga gaccagctac catcaacggc atgcagcagc ctatgcaggc tgaccctgcc 480

cgggcagcgg agctgcaaca gccagcatct gtcccagccg acgggcgagt aagcaagcag 540

gacacagcgg ctggggtgag ctcagagcct tctgccaatg agagccacaa gaccacaacc 600

ggagcagata gtgaggcagg tggagacgtg gcggagaaat cctaa 645

<210>136

<211>214

<212>PRT

＜213〉dichromatism chinese sorghum

<400>136

Met Gln Gln Gln Met Pro Met Pro Pro Ala Pro Ala Ala Ala Ala Ala

1 5 10 15

Thr Ala Pro Pro Ala Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr

20 25 30

Leu Asp Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn

35 40 45

Leu Gly Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys

50 55 60

Asn Leu Leu Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Arg Pro Pro

65 70 75 80

Gln Asn Pro Ala Gly Arg Pro Gln Met Met Gln Pro Gly Ile Val Pro

85 90 95

Gly Ala Gly His Tyr Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr

100 105 110

Pro Leu Thr Pro Gln Gln Met Gln Glu Gln Gln Gln Gln Gln Leu Gln

115 120 125

Gln Gln Gln Ala Gln Ala Leu Ala Phe Pro Gly Gln Met Val Met Arg

130 135 140

Pro Ala Thr Ile Asn Gly Met Gln Gln Pro Met Gln Ala Asp Pro Ala

145 150 155 160

Arg Ala Ala Glu Leu Gln Gln Pro Ala Ser Val Pro Ala Asp Gly Arg

165 170 175

Val Ser Lys Gln Asp Thr Ala Ala Gly Val Ser Ser Glu Pro Ser Ala

180 185 190

Asn Glu Ser His Lys Thr Thr Thr Gly Ala Asp Ser Glu Ala Gly Gly

195 200 205

Asp Val Ala Glu Lys Ser

210

<210>137

<211>558

<212>DNA

＜213〉common wheat

<400>137

atgcagcaag cgatgcccat gccgccggcg gcggcggcgc cggggatgcc tccgtctgct 60

ggcctcagca ccgagcagat ccaaaagtac ctggatgaaa ataagcaact aattttggct 120

atcttggaaa atcagaacct gggaaagttg gcggaatgtg ctcagtatca agctcagctt 180

cagaagaatc ttttgtattt ggctgcaatc gctgatactc agccacagac cactgtaagc 240

cgtcctcaga tggcaccacc tagtgcatcc ccaggggcag ggcattacat gtcacaggtg 300

ccaatgttcc ctccgaggac ccctctaacg cctcagcaga tgcaggagca gcaactacag 360

cagcaacagg ctcagatgct tccgtttgct ggtcaaatgg ttgcgagacc tggggctgtc 420

aatggcatgc ctcaggcccc tcaagttgaa ccagcctatg cagcaggtgg ggccagttct 480

gagccttctg gcactgagag ccacaggagc actggtgccg ataatgacgg ggggagcggc 540

tgggctgatc agtcctaa 558

<210>138

<211>185

<212>PRT

＜213〉common wheat

<400>138

Met Gln Gln Ala Met Pro Met Pro Pro Ala Ala Ala Ala Pro Gly Met

1 5 10 15

Pro Pro Ser Ala Gly Leu Ser Thr Glu Gln Ile Gln Lys Tyr Leu Asp

20 25 30

Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly

35 40 45

Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu

50 55 60

Leu Tyr Leu Ala Ala Ile Ala Asp Thr Gln Pro Gln Thr Thr Val Ser

65 70 75 80

Arg Pro Gln Met Ala Pro Pro Ser Ala Ser Pro Gly Ala Gly His Tyr

85 90 95

Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln

100 105 110

Gln Met Gln Glu Gln Gln Leu Gln Gln Gln Gln Ala Gln Met Leu Pro

115 120 125

Phc Ala Gly Gln Met Val Ala Arg Pro Gly Ala Val Asn Gly Met Pro

130 135 140

Gln Ala Pro Gln Val Glu Pro Ala Tyr Ala Ala Gly Gly Ala Ser Ser

145 150 155 160

Glu Pro Ser Gly Thr Glu Ser His Arg Ser Thr Gly Ala Asp Asn Asp

165 170 175

Gly Gly Ser Gly Trp Ala Asp Gln Ser

180 185

<210>139

<211>603

<212>DNA

＜213〉common wheat

<400>139

atgcagcagg cgatgtcctt gcccccggga gcggtcggcg cggtgtcctc gccggccggc 60

atcaccaccg agcagatcca aaagtatttg gatgaaaata agcaacttat tttggccatc 120

cttgaaaatc agaacctagg aaagttggct gaatgtgctc agtatcaagc tcaactccaa 180

aagaatctct tgtatctagc tgctatcgcg gatgcccaac caccacagaa ccctacaagt 240

caccctcaga tggtgcagcc tggtagtatg caaggtgcag ggcattacat gtcacaagta 300

ccaatgttcc ctccaagaac gcctttaacc ccacagcaga tgcaagagca gcagcaccag 360

cagcttcagc agcagcaagc ccaggccctt tctttccccg cccaggtggt catgagacca 420

ggcaccgtca acggcatgca gcagcctatg caagcagccg gcgacctcca gccagcagca 480

gcacctggag ggagcaagca ggacgccgca gtggctgggg ccagctcgga accatctggc 540

accaagagcc acaagaacgc gggagcagag gaggtgggcg ctgatgtagc agaacaatcc 600

taa 603

<210>140

<211>200

<212>PRT

＜213〉common wheat

<400>140

Met Gln Gln Ala Met Ser Leu Pro Pro Gly Ala Val Gly Ala Val Ser

1 5 10 15

Ser Pro Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr Leu Asp Glu

20 25 30

Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn Leu Gly Lys

35 40 45

Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys Asn Leu Leu

50 55 60

Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Pro Gln Asn Pro Thr Ser

65 70 75 80

His Pro Gln Met Val Gln Pro Gly Ser Met Gln Gly Ala Gly His Tyr

85 90 95

Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr Pro Leu Thr Pro Gln

100 105 110

Gln Met Gln Glu Gln Gln His Gln Gln Leu Gln Gln Gln Gln Ala Gln

115 120 125

Ala Leu Ser Phe Pro Ala Gln Val Val Met Arg Pro Gly Thr Val Asn

130 135 140

Gly Met Gln Gln Pro Met Gln Ala Ala Gly Asp Leu Gln Pro Ala Ala

145 150 155 160

Ala Pro Gly Gly Ser Lys Gln Asp Ala Ala Val Ala Gly Ala Ser Ser

165 170 175

Glu Pro Ser Gly Thr Lys Ser His Lys Asn Ala Gly Ala Glu Glu Val

180 185 190

Gly Ala Asp Val Ala Glu Gln Ser

195 200

<210>141

<211>672

<212>DNA

＜213〉grape (Vitis vinifera)

<400>141

atgcagcagc acctgatgca gatgcagccc atgatggcag cctattaccc cagcaacgtc 60

accactgatc acattcagca gtatcttgat gaaaacaagt cattgattct gaagattgtt 120

gagagccaga attcaggaaa attgactgaa tgtgcagaga accaggcaag actacagaga 180

aacctcatgt acctggctgc aattgctgat tctcaacccc aaccacccac catgcatgct 240

cagttccctc ctagtggcat tgttcagcca ggagctcact acatgcaaca ccaacaagct 300

caacaaatga caccacagtc gctcctggct gcacgctcct ccatgctgta cacccaacaa 360

ccattttcgg ccctgcaaca acaacaagcc atccatagcc agcttggcat gggctctggt 420

ggaagtgcag gacttcacat gctgcaaagc gaggggagta atccaggagg caatggaaca 480

ctggggactg gtgggtttcc tgatttcagc cgtggaactt ctggagaagg cctgcaggct 540

gcaggcaggg gaatggctgg tgggagcaag caagatatgg gaaatgcaga agggcgagga 600

gggaactcag gaggtcaggg tggggatgga ggtgagactc tttacttgaa agctgctgaa 660

gatgggaatt ga 672

<210>142

<211>223

<212>PRT

＜213〉grape

<400>142

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Thr Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Thr Met His Ala

65 70 75 80

Gln Phe Pro Pro Ser Gly Ile Val Gln Pro Gly Ala His Tyr Met Gln

85 90 95

His Gln Gln Ala Gln Gln Met Thr Pro Gln Ser Leu Leu Ala Ala Arg

100 105 110

Ser Ser Met Leu Tyr Thr Gln Gln Pro Phe Ser Ala Leu Gln Gln Gln

115 120 125

Gln Ala Ile His Ser Gln Leu Gly Met Gly Ser Gly Gly Ser Ala Gly

130 135 140

Leu His Met Leu Gln Ser Glu Gly Ser Asn Pro Gly Gly Asn Gly Thr

145 150 155 160

Leu Gly Thr Gly Gly Phe Pro Asp Phe Ser Arg Gly Thr Ser Gly Glu

165 170 175

Gly Leu Gln Ala Ala Gly Arg Gly Met Ala Gly Gly Ser Lys Gln Asp

180 185 190

Met Gly Asn Ala Glu Gly Arg Gly Gly Asn Ser Gly Gly Gln Gly Gly

195 200 205

Asp Gly Gly Glu Thr Leu Tyr Leu Lys Ala Ala Glu Asp Gly Asn

210 215 220

<210>143

<211>663

<212>DNA

＜213〉Zea mays

<400>143

atgcagcagc agatgcccat gccgccggcg cccgctgccg ccgcggcggc ggcgcccccg 60

gcggcaggca tcactaccga gcagatccag aagtatttgg acgaaaataa gcaacttatt 120

ttggccatcc tggaaaatca gaacttaggg aagttggctg aatgtgctca gtatcaagct 180

caacttcaaa agaacctctt gtacctggct gcgattgctg atgcccaacc ccagcctccg 240

caaaaccctg caggtcgccc tcagatgatg cagcctggta tagtgccagg tgcggggcat 300

tacatgtcac aagtaccaat gttccctcca agaaccccat taaccccaca gcagatgcag 360

gagcagcagc aacaacaaca gtttcagcag cagcagcagc aagtgcaggc tcttacattt 420

cctggacaga tggtcatgag accaggcacc atcaacggca tgcagcagca gcagcctatg 480

caggctgacc ctgcccgggc agcagcggag ctgcagcagg cagcacctat cccagctgac 540

gggcgaggaa gcaagcagga caccgcgggt ggggcgagct cagagccttc tgccaatgag 600

agccacaaga gcgccaccgg agcagatacc gaggcaggtg gcgacgtggc cgagaaatcc 660

taa 663

<210>144

<211>220

<212>PRT

＜213〉Zea mays

<400>144

Met Gln Gln Gln Met Pro Met Pro Pro Ala Pro Ala Ala Ala Ala Ala

1 5 10 15

Ala Ala Pro Pro Ala Ala Gly Ile Thr Thr Glu Gln Ile Gln Lys Tyr

20 25 30

Leu Asp Glu Asn Lys Gln Leu Ile Leu Ala Ile Leu Glu Asn Gln Asn

35 40 45

Leu Gly Lys Leu Ala Glu Cys Ala Gln Tyr Gln Ala Gln Leu Gln Lys

50 55 60

Asn Leu Leu Tyr Leu Ala Ala Ile Ala Asp Ala Gln Pro Gln Pro Pro

65 70 75 80

Gln Asn Pro Ala Gly Arg Pro Gln Met Met Gln Pro Gly Ile Val Pro

85 90 95

Gly Ala Gly His Tyr Met Ser Gln Val Pro Met Phe Pro Pro Arg Thr

100 105 110

Pro Leu Thr Pro Gln Gln Met Gln Glu Gln Gln Gln Gln Gln Gln Phe

115 120 125

Gln Gln Gln Gln Gln Gln Val Gln Ala Leu Thr Phe Pro Gly Gln Met

130 135 140

Val Met Arg Pro Gly Thr Ile Asn Gly Met Gln Gln Gln Gln Pro Met

145 150 155 160

Gln Ala Asp Pro Ala Arg Ala Ala Ala Glu Leu Gln Gln Ala Ala Pro

165 170 175

Ile Pro Ala Asp Gly Arg Gly Ser Lys Gln Asp Thr Ala Gly Gly Ala

180 185 190

Ser Ser Glu Pro Ser Ala Asn Glu Ser His Lys Ser Ala Thr Gly Ala

195 200 205

Asp Thr Glu Ala Gly Gly Asp Val Ala Glu Lys Ser

210 215 220

<210>145

<211>2193

<212>DNA

＜213〉rice

<400>145

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140

cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200

tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260

gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320

ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380

gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440

aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500

ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560

atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620

acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680

cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740

ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800

gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860

tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920

attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980

tgtcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040

cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100

agetgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160

tggtgtagct tgccactttc accagcaaag ttc 2193

<210>146

<211>12

<212>PRT

＜213〉artificial sequence

<220>

＜223〉frame I

<220>

＜221〉variant

<222>(3)..(3)

＜223 〉/replace=" Lys "

<220>

＜221〉variant

<222>(4)..(4)

＜223 〉/replace=" Met "/replace=" Phe "/replace=" His "

<220>

＜221〉variant

<222>(6)..(6)

＜223 〉/replace=" Glu "

<220>

＜221〉variant

<222>(7)..(7)

＜223 〉/replace=" Asp "

<220>

＜221〉variant

<222>(9)..(9)

＜223 〉/replace=" Asn "

<220>

＜221〉variant

<222>(10)..(10)

＜223〉Xaa can make the amino acid of any natural generation

<400>146

Ile Gln Gln Tyr Leu Asp Glu Asn Lys Xaa Leu Ile

1 5 10

<210>147

<211>10

<212>PRT

＜213〉artificial sequence

<220>

＜223〉frame II

<220>

＜221〉variant

<222>(3)..(3)

＜223 〉/replace=" Leu "/replace=" Val "

<220>

＜221〉variant

<222>(7)..(7)

＜223 〉/replace=" Thr "

<400>147

Asn Leu Met Tyr Leu Ala Ala Ile Ala Asp

1 5 10

<210>148

<211>53

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm06681

<400>148

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgca acagcacctg atg 53

<210>149

<211>50

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm06682

<400>149

ggggaccac tttgtacaaga aagctgggtc atcattaaga ttccttgtgc 50

<210>150

<211>615

<212>DNA

＜213〉colea

<400>150

atgcaacagc acctgatgca gatgcagccc atgatggctg gttactaccc cagcaatgtc 60

acctctgatc atattcagca gtacttggac gagaacaaat cgttgattct gaagatagtt 120

gaatctcaaa actcgggaaa gctcagcgag tgtgccgaga accaggcaag gcttcaacgc 180

aacttaatgt acttagctgc aattgcagat tctcagcctc aacctccaag catgcatagc 240

cagtatggaa ctgctggtgg tggtgggttg atgcagggag aaggagggtc acactatttg 300

caacagcaac aggcaattca acagcagcag agtcagcagt ctctaatggc ggctcgatct 360

tcaatgttgt atgctcagca gcagcaacag ccttatgcaa cgcttcagca gcagcaattg 420

caccatagcc agcttgggat gagctcaagc agcggaggag gaagcagcgg tctccatatg 480

ctacagggag aggctggtgg gtttcatgat tttggccgtg agaagttgga aatgggaagt 540

ggtgaaggca gaggaggaag ctcaggggat ggtggagaaa ccctttactt gaagtcatca 600

gatgatggga ac tga 615

<210>151

<211>204

<212>PRT

＜213〉colea

<400>151

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Gly Tyr Tyr

1 5 10 15

Pro Ser Asn Val Thr Ser Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ala Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Pro Ser Met His Ser

65 70 75 80

Gln Tyr Gly Thr Ala Gly Gly Gly Gly Leu Met Gln Gly Glu Gly Gly

85 90 95

Ser His Tyr Leu Gln Gln Gln Gln Ala Ile Gln Gln Gln Gln Ser Gln

100 105 110

Gln Ser Leu Met Ala Ala Arg Ser Ser Met Leu Tyr Ala Gln Gln Gln

115 120 125

Gln Gln Pro Tyr Ala Thr Leu Gln Gln Gln Gln Leu His His Ser Gln

130 135 140

Leu Gly Met Ser Ser Ser Ser Gly Gly Gly Ser Ser Gly Leu His Met

145 150 155 160

Leu Gln Gly Glu Ala Gly Gly Phe His Asp Phe Gly Arg Glu Lys Leu

165 170 175

Glu Met Gly Ser Gly Glu Gly Arg Gly Gly Ser Ser Gly Asp Gly Gly

180 185 190

Glu Thr Leu Tyr Leu Lys Ser Ser Asp Asp Gly Asn

195 200

<210>152

<211>639

<212>DNA

＜213〉soybean

<400>152

atgcagcagc acctgatgca gatgcagccc atgatggctg cctactaccc caacaacgtc 60

accactgatc acattcaaca gtacctggat gagaacaagt ccttgattct gaagattgtt 120

gaaagccaga attctggcaa gctgagcgag tgtgccgaga accaatcaag gctgcagaga 180

aatctcatgt acctagctgc aatagctgat tctcaaccac aaccatctcc attggctggt 240

cagtatcctt ctagtggact tgtgcagcag ggagcacact acatgcaggc tcaacaggct 300

cagcagatgt cacaacaaca gctaatggct tcgcgctcct cgctcctgta ctcccaacag 360

cctttctcag tgcttcaaca gcagcaaggc atgcacagcc aacttggcat gagctccagt 420

ggaagtcaag gcctccacat gctgcaaagt gaagccacta atgttggagg caatgcaacc 480

ataggaaccg gaggagggtt tccggacttt gtacgcattg gtagtggcaa gcaagatatt 540

ggaatctctg gtgaaggcag aggaggaaac tctagtggcc actctggtga tggtggtgag 600

acacttaatt acctgaaagc tgctggtgat ggaaactga 639

<210>153

<211>212

<212>PRT

＜21〉soybean

<400>153

Met Gln Gln His Leu Met Gln Met Gln Pro Met Met Ala Ala Tyr Tyr

1 5 10 15

Pro Asn Asn Val Thr Thr Asp His Ile Gln Gln Tyr Leu Asp Glu Asn

20 25 30

Lys Ser Leu Ile Leu Lys Ile Val Glu Ser Gln Asn Ser Gly Lys Leu

35 40 45

Ser Glu Cys Ala Glu Asn Gln Ser Arg Leu Gln Arg Asn Leu Met Tyr

50 55 60

Leu Ala Ala Ile Ala Asp Ser Gln Pro Gln Pro Ser Pro Leu Ala Gly

65 70 75 80

Gln Tyr Pro Ser Ser Gly Leu Val Gln Gln Gly Ala His Tyr Met Gln

85 90 95

Ala Gln Gln Ala Gln Gln Met Ser Gln Gln Gln Leu Met Ala Ser Arg

100 105 110

Ser Ser Leu Leu Tyr Ser Gln Gln Pro Phe Ser Val Leu Gln Gln Gln

115 120 125

Gln Gly Met His Ser Gln Leu Gly Met Ser Ser Ser Gly Ser Gln Gly

130 135 140

Leu His Met Leu Gln Ser Glu Ala Thr Asn Val Gly Gly Asn Ala Thr

145 150 155 160

Ile Gly Thr Gly Gly Gly Phe Pro Asp Phe Val Arg Ile Gly Ser Gly

165 170 175

Lys Gln Asp Ile Gly Ile Ser Gly Glu Gly Arg Gly Gly Asn Ser Ser

180 185 190

Gly His Ser Gly Asp Gly Gly Glu Thr Leu Asn Tyr Leu Lys Ala Ala

195 200 205

Gly Asp Gly Asn

210

<210>154

<211>1248

<212>DNA

＜213〉dish mattress chlamydomonas (Chlamydomonas reinhardtii)

<400>154

atggccgcca ccatgctccg ctccagcacc cagtcgggca ttgccgctaa ggccggccgc 60

aaggaggctg tcagcgtccg cgcggtcgcc cagccccagc gccaggctgg tgctgccagc 120

gtcttctcct cgtcgtcgtc gggcgctgct gctcgccgcg gggtcgtcgc tcaggccacc 180

gctgttgcca cccccgcggc caagcctgcg gccaagacca gccagtatga gctgttcacg 240

ctcaccacct ggctgctgaa ggaggagatg aagggcacaa tcgatggcga gcttgtgacc 300

gtcatctcgt cggtgtcgct ggcctgcaag caaatcgcgt cgctggtgaa ccgcgctggt 360

atctccaacc tgaccggtgt ggctggcaac cagaacgtgc agggtgagga ccagaagaag 420

ctggacgtgg tgtccaacga ggtcttcaag aactgcctgg cctcctgcgg ccgcacgggt 480

gtgatcgcct ccgaggagga ggaccagccc gtggccgtgg aggagaccta ctcgggcaac 540

tacatcgtgg tgttcgaccc cctggacggc tcgtccaaca tcgacgccgg catctccgtc 600

ggctccatct tcggcatcta cgagcccagc gaggagtgcc ccattgacgc catggacgac 660

ccccagaaga tgatggagca gtgcgtcatg aacgtgtgcc agcccggctc gcgcctcaag 720

tgcgccggct actgcctgta cagcagcagc accatcatgg tgctgaccat cggcaacggt 780

gtgttcggct tcacgctgga ccccctggtc ggcgagttcg tgctgaccca ccccaacgtg 840

cagatccccg aggtgggcaa gatctacccg ttcaacgagg gcaactacgg cctgtgggac 900

gacagcgtta aggcttacat ggacagcctg aaggacccca agaagtggga cggcaagccc 960

tactcggccc gctacatcgg ctccctggtc ggtgacttcc accgcaccct gctgtacgga 1020

ggcatctacg gctaccccgg cgacgccaag aacaagaacg gcaagctccg cctgctgtac 1080

gagtgcgcgc ccatgtcgtt cattgccgag caggccggcg gcctcggctc caccggccag 1140

gagcgcgtgc tggacgtgaa ccccgagaag gtgcaccagc gcgtgccgct gttcatcggc 1200

tccaagaagg aggtcgagta cctggagtcc ttcaccaaga agcactaa 1248

<210>155

<211>415

<212>PRT

＜213〉Chlamydomonas reinhardtii

<400>155

Met Ala Ala Thr Met Leu Arg Ser Ser Thr Gln Ser Gly Ile Ala Ala

1 5 10 15

Lys Ala Gly Arg Lys Glu Ala Val Ser Val Arg Ala Val Ala Gln Pro

20 25 30

Gln Arg Gln Ala Gly Ala Ala Ser Val Phe Ser Ser Ser Ser Ser Gly

35 40 45

Ala Ala Ala Arg Arg Gly Val Val Ala Gln Ala Thr Ala Val Ala Thr

50 55 60

Pro Ala Ala Lys Pro Ala Ala Lys Thr Ser Gln Tyr Glu Leu Phe Thr

65 70 75 80

Leu Thr Thr Trp Leu Leu Lys Glu Glu Met Lys Gly Thr Ile Asp Gly

85 90 95

Glu Leu Val Thr Val Ile Ser Ser Val Ser Leu Ala Cys Lys Gln Ile

100 105 110

Ala Ser Leu Val Asn Arg Ala Gly Ile Ser Asn Leu Thr Gly Val Ala

115 120 125

Gly Asn Gln Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val

130 135 140

Ser Asn Glu Val Phe Lys Asn Cys Leu Ala Ser Cys Gly Arg Thr Gly

145 150 155 160

Val Ile Ala Ser Glu Glu Glu Asp Gln Pro Val Ala Val Glu Glu Thr

165 170 175

Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser

180 185 190

Asn Ile Asp Ala Gly Ile Ser Val Gly Ser Ile Phe Gly Ile Tyr Glu

195 200 205

Pro Ser Glu Glu Cys Pro Ile Asp Ala Met Asp Asp Pro Gln Lys Met

210 215 220

Mct Glu Gln Cys Val Met Asn Val Cys Gln Pro Gly Ser Arg Leu Lys

225 230 235 240

Cys Ala Gly Tyr Cys Leu Tyr Ser Ser Ser Thr Ile Met Val Leu Thr

245 250 255

Ile Gly Asn Gly Val Phe Gly Phe Thr Leu Asp Pro Leu Val Gly Glu

260 265 270

Phe Val Leu Thr His Pro Asn Val Gln Ile Pro Glu Val Gly Lys Ile

275 280 285

Tyr Pro Phe Asn Glu Gly Asn Tyr Gly Leu Trp Asp Asp Ser Val Lys

290 295 300

Ala Tyr Met Asp Ser Leu Lys Asp Pro Lys Lys Trp Asp Gly Lys Pro

305 310 315 320

Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr

325 330 335

Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Gly Asp Ala Lys Asn Lys

340 345 350

Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile

355 360 365

Ala Glu Gln Ala Gly Gly Leu Gly Ser Thr Gly Gln Glu Arg Val Leu

370 375 380

Asp Val Asn Pro Glu Lys Val His Gln Arg Val Pro Leu Phe Ile Gly

385 390 395 400

Ser Lys Lys Glu Val Glu Tyr Leu Glu Ser Phe Thr Lys Lys His

405 410 415

<210>156

<211>1239

<212>DNA

<213>Bigelowiella natans

<400>156

atggctgccg tgctctttcg cggaggtgtg ctgtcttcca ccatgactgc ggctcgcaca 60

ttcgcagcgc catctgtgca cacccgtggc atgcacatgt cagtcaagag cagtaaccct 120

ttctctcagg ctggtcgtcg tgctgctgtt agatccagtg tcgcaccctc ccctgtccaa 180

gactctgctg gtactatcgt tgatgacgga actgtcacat taacaagatt catgattgag 240

gaggcaatga agagcaagac tcctgggcag gaggacatgg ttcgtttgat ttcttccatc 300

tcagttgcat gcaaacgcat tgcatccatg gtgcaaactg caggaatctc cggatctaca 360

ggtctggctg aaggtggtgg atcagtcaac gttcaaggcg aggaacagaa gaaacttgat 420

gtcatttcta acgatgtact aaagtctgcc cttcgtcctt ctggaaaact tggagtaatt 480

gcctctgagg aggaagataa tccagttgtc gttgatgaac tttactccgg cgaatatgtt 540

gctactttcg atccgctcga tggatcttcc aatattgatg ctgcaatttc cactggaact 600

atcttcggag tgttcaaagc tccagaagag tgcttgatcg gggattctga taatctcagt 660

attgcagagc agcaatgttt ggaggcaaca cttcaacctg gaactaacct tgttgctgct 720

ggatactgca tgtattcgtc ctccaccatc cttgttctca ccaccggaga cgggctcaat 780

ggatttactc ttgacccttc cattggagag ttcatcctca cccatcctaa tatccagatt 840

cctagccgtg gaaagatcta ttctatgaac gaagcaaact acttcgattg ggaccctaag 900

cttcagacct acgttgataa ccttaagaag gcagaaggtc aaactggaga aaagtacagc 960

tctcgctaca tcggatccat ggtcggagat gtgcaccgta cccttctcta tggaggcatc 1020

ttcgcttacc ctggagataa gaagaacgtc aacggaaaac ttcgccttct gtacgaagct 1080

gctccaatgt cccttatctt cgaacaagcg ggtggaaaat ctattactgg acctggtgga 1140

cgtgtgttgg acttagttcc tgataaggtt caccagcgtt gtcctgtgtt cattggatcc 1200

cctgatgatg tggatgaagt tgaaaaggct ctcgcataa 1239

<210>157

<211>412

<212>PRT

<213>Bigelowiella natans

<400>157

Met Ala Ala Val Leu Phe Arg Gly Gly Val Leu Ser Ser Thr Met Thr

1 5 10 15

Ala Ala Arg Thr Phe Ala Ala Pro Ser Val His Thr Arg Gly Met His

20 25 30

Met Ser Val Lys Ser Ser Asn Pro Phe Ser Gln Ala Gly Arg Arg Ala

35 40 45

Ala Val Arg Ser Ser Val Ala Pro Ser Pro Val Gln Asp Ser Ala Gly

50 55 60

Thr Ile Val Asp Asp Gly Thr Val Thr Leu Thr Arg Phe Met Ile Glu

65 70 75 80

Glu Ala Met Lys Ser Lys Thr Pro Gly Gln Glu Asp Met Val Arg Leu

85 90 95

Ile Ser Ser Ile Ser Val Ala Cys Lys Arg Ile Ala Ser Met Val Gln

100 105 110

Thr Ala Gly Ile Ser Gly Ser Thr Gly Leu Ala Glu Gly Gly Gly Ser

115 120 125

Val Asn Val Gln Gly Glu Glu Gln Lys Lys Leu Asp Val Ile Ser Asn

130 135 140

Asp Val Leu Lys Ser Ala Leu Arg Pro Ser Gly Lys Leu Gly Val Ile

145 150 155 160

Ala Ser Glu Glu Glu Asp Asn Pro Val Val Val Asp Glu Leu Tyr Ser

165 170 175

Gly Glu Tyr Val Ala Thr Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile

180 185 190

Asp Ala Ala Ile Ser Thr Gly Thr Ile Phe Gly Val Phe Lys Ala Pro

195 200 205

Glu Glu Cys Leu Ile Gly Asp Ser Asp Asn Leu Ser Ile Ala Glu Gln

210 215 220

Gln Cys Leu Glu Ala Thr Leu Gln Pro Gly Thr Asn Leu Val Ala Ala

225 230 235 240

Gly Tyr Cys Met Tyr Ser Ser Ser Thr Ile Leu Val Leu Thr Thr Gly

245 250 255

Asp Gly Leu Asn Gly Phe Thr Leu Asp Pro Ser Ile Gly Glu Phe Ile

260 265 270

Leu Thr His Pro Asn Ile Gln Ile Pro Ser Arg Gly Lys Ile Tyr Ser

275 280 285

Met Asn Glu Ala Asn Tyr Phe Asp Trp Asp Pro Lys Leu Gln Thr Tyr

290 295 300

Val Asp Asn Leu Lys Lys Ala Glu Gly Gln Thr Gly Glu Lys Tyr Ser

305 310 315 320

Ser Arg Tyr Ile Gly Ser Met Val Gly Asp Val His Arg Thr Leu Leu

325 330 335

Tyr Gly Gly Ile Phe Ala Tyr Pro Gly Asp Lys Lys Asn Val Asn Gly

340 345 350

Lys Leu Arg Leu Leu Tyr Glu Ala Ala Pro Met Ser Leu Ile Phe Glu

355 360 365

Gln Ala Gly Gly Lys Ser Ile Thr Gly Pro Gly Gly Arg Val Leu Asp

370 375 380

Leu Val Pro Asp Lys Val His Gln Arg Cys Pro Val Phe Ile Gly Ser

385 390 395 400

Pro Asp Asp Val Asp Glu Val Glu Lys Ala Leu Ala

405 410

<210>158

<211>1230

<212>DNA

<213>Aquilegia formosa x Aquilegia pubescens

<400>158

atggttgcag cagctatccc tacttcttgt caactgctct tctccacctc ttcttcgacc 60

acttctcgtc tatatcctcc ttatcttgat gccaaaactc tcttctcatt tcctacaaac 120

aagagacatg taagcattgt tagaactccg gcaggtgtac ggtgtcaagc attaggagca 180

gaggtagtag tgaccaagag gagtgcattt gagatacaaa ctctaacagg atggttattg 240

aaacaagaac aaacaggggt tatagatgca gagttaacta tagttttgtc tagcatttca 300

atggcctgta agcagattgc ttcattggtg cagagggcaa gtatttccaa cttaactgga 360

gttcaaggag ctgttaatat tcaaggtgaa gatcagaaga agcttgatgt catctctaac 420

gaggtgttct ctaattgcct tagatcaagt ggaagaacag ggattatagc atcagaagaa 480

gaggatgtac cagttgcagt ggaggaaagc tactctggca actatattgt cgtttttgat 540

cctcttgacg ggtcatcaaa cattgatgcc gctgtgtcaa ctggatccat atttggaatt 600

tatagtccga acgatgagtg tcttactgaa gtcgatgata atgccacagt gcttcagcaa 660

gtggaacaga agtgcatcat caatgtgtgt caacctggca acaacttgtt ggcagctggc 720

tactgcatgt actcaagctc tgtaatcttt gtgctttcca ttggacaagg ggttttctca 780

tttaccttag accctatgta cggagaattc gtcttgacac aggaaaacat tcaaatacct 840

aaatcaggta aaatctactc attcaacgaa ggcaactacc aattatggga tgataagttg 900

aagaagtata tcgatgacct taaggaccct ggtcctagtg gcaaaccata ctcttccaga 960

tacattggaa gcttggttgg cgatttccac cggacccttc tttatggagg gatatatgga 1020

taccctaggg ataagaatag aaaaaatggg aagctaaggc tactgtatga gtgtgcacct 1080

atgagttatt tagttgaaca agcaggagga aaaggatcag atggacacca aagagtactc 1140

gatattgaac cggtggagat tcatcagcgt gttccacttt tcattggcag cgttgaagaa 1200

gttgagaaac tagagaagtt cttggcttga 1230

<210>159

<211>409

<212>PRT

<213>Aquilegia formosa x Aquilegia pubescens

<400>159

Met Val Ala Ala Ala Ile Pro Thr Ser Cys Gln Leu Leu Phe Ser Thr

1 5 10 15

Ser Ser Ser Thr Thr Ser Arg Leu Tyr Pro Pro Tyr Leu Asp Ala Lys

20 25 30

Thr Leu Phe Ser Phe Pro Thr Asn Lys Arg His Val Ser Ile Val Arg

35 40 45

Thr Pro Ala Gly Val Arg Cys Gln Ala Leu Gly Ala Glu Val Val Val

50 55 60

Thr Lys Arg Ser Ala Phe Glu Ile Gln Thr Leu Thr Gly Trp Leu Leu

65 70 75 80

Lys Gln Glu Gln Thr Gly Val Ile Asp Ala Glu Leu Thr Ile Val Leu

85 90 95

Ser Ser Ile Ser Met Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg

100 105 110

Ala Ser Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val Asn Ile Gln

115 120 125

Gly Glu Asp Gln Lys Lys Leu Asp Val Ile Ser Asn Glu Val Phe Ser

130 135 140

Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu

145 150 155 160

Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile

165 170 175

Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val

180 185 190

Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu

195 200 205

Thr Glu Val Asp Asp Asn Ala Thr Val Leu Gln Gln Val Glu Gln Lys

210 215 220

Cys Ile Ile Asn Val Cys Gln Pro Gly Asn Asn Leu Leu Ala Ala Gly

225 230 235 240

Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val Leu Ser Ile Gly Gln

245 250 255

Gly Val Phe Ser Phe Thr Leu Asp Pro Met Tyr Gly Glu Phe Val Leu

260 265 270

Thr Gln Glu Asn Ile Gln Ile Pro Lys Ser Gly Lys Ile Tyr Ser Phe

275 280 285

Asn Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys Leu Lys Lys Tyr Ile

290 295 300

Asp Asp Leu Lys Asp Pro Gly Pro Ser Gly Lys Pro Tyr Ser Ser Arg

305 310 315 320

Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu Tyr Gly

325 330 335

Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Asn Arg Lys Asn Gly Lys Leu

340 345 350

Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Tyr Leu Val Glu Gln Ala

355 360 365

Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile Glu Pro

370 375 380

Val Glu Ile His Gln Arg Val Pro Leu Phe Ile Gly Ser Val Glu Glu

385 390 395 400

Val Glu Lys Leu Glu Lys Phe Leu Ala

405

<210>160

<211>1254

<212>DNA

＜213〉Arabidopis thaliana

<400>160

atggcagcat cggccgcaac aacgacgtcg tctcaccttc ttctctcaag ctcacgtcac 60

gtggcttcat catcccagcc ttcaatcctt tcaccgagat ctctcttctc caacaacggg 120

aaacgagcac caactggagt gagaaaccat cagtatgcga gtggagtgag gtgtatggcc 180

gtagcggcgg atgcttctga gacgaaaacg gcggcgagga agaagagtgg atacgaactt 240

caaacgttga cgggctggtt gctgagacaa gagatgaaag gagagataga tgcagagctg 300

acgatagtga tgtcgagtat atcattggct tgtaagcaga tcgcttcact tgttcaacgt 360

gccggaatat ctaacctgac cggagttcaa ggcgccatta atattcaggg agaggatcag 420

aagaagcttg acgtcatctc taatgaggtg ttttccaact gtttgagatc aagtggaaga 480

acgggaatca tagcctcgga ggaagaggac gtgccagttg cggtggagga gagttactcc 540

ggcaactacg tcgtcgtgtt tgaccctctt gatggttcct ccaacattga cgctgccgtc 600

tctactggtt ctatcttcgg tatctatagc cccaatgacg aatgcattgt cgacgactcc 660

gacgatatct cagctcttgg gtcagaagaa caaaggtgta tagtaaacgt gtgccagcca 720

gggaacaact tgttagcagc cggctactgt atgtactcga gctcggtcat cttcgttctt 780

actctaggca aaggcgtttt ctccttcacg cttgatccaa tgtacggtga gtttgtcctc 840

acgcaagaaa acattgagat ccccaaagcc gggagaatct actctttcaa cgaagggaat 900

taccagatgt gggacgataa actaaagaag tacattgatg accttaagga ccctggtcca 960

actgggaagc cttactcggc aaggtacatt ggaagtttgg ttggagattt tcacaggact 1020

ttgttgtacg gtgggattta cgggtaccct cgtgacgcaa agagcaaaaa tggaaagctt 1080

aggcttttgt atgagtgtgc accaatgagt ttcattgttg aacaagctgg agggaaaggt 1140

tctgatggac attcgagagt actagatatc caaccgactg agatacatca gagggttcct 1200

ctatacattg gaagcacaga ggaagtagag aagttggaga agtacttggc ttga 1254

<210>161

<211>417

<212>PRT

＜213〉Arabidopis thaliana

<400>161

Met Ala Ala Ser Ala Ala Thr Thr Thr Ser Ser His Leu Leu Leu Ser

1 5 10 15

Ser Ser Arg His Val Ala Ser Ser Ser Gln Pro Ser Ile Leu Ser Pro

20 25 30

Arg Ser Leu Phe Ser Asn Asn Gly Lys Arg Ala Pro Thr Gly Val Arg

35 40 45

Asn His Gln Tyr Ala Ser Gly Val Arg Cys Met Ala Val Ala Ala Asp

50 55 60

Ala Ser Glu Thr Lys Thr Ala Ala Arg Lys Lys Ser Gly Tyr Glu Leu

65 70 75 80

Gln Thr Leu Thr Gly Trp Leu Leu Arg Gln Glu Met Lys Gly Glu Ile

85 90 95

Asp Ala Glu Leu Thr Ile Val Met Ser Ser Ile Ser Leu Ala Cys Lys

100 105 110

Gln Ile Ala Ser Leu Val Gln Arg Ala Gly Ile Ser Asn Leu Thr Gly

115 120 125

Val Gln Gly Ala Ile Asn Ile Gln Gly Glu Asp Gln Lys Lys Leu Asp

130 135 140

Val Ilc Ser Asn Glu Val Phe Ser Asn Cys Leu Arg Ser Ser Gly Arg

145 150 155 160

Thr Gly Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu

165 170 175

Glu Ser Tyr Ser Gly Asn Tyr Val Val Val Phe Asp Pro Leu Asp Gly

180 185 190

Ser Ser Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile

195 200 205

Tyr Ser Pro Asn Asp Glu Cys Ile Val Asp Asp Ser Asp Asp Ile Ser

210 215 220

Ala Leu Gly Ser Glu Glu Gln Arg Cys Ile Val Asn ValCys Gln Pro

225 230 235 240

Gly Asn Asn Leu Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val

245 250 255

Ile Phe Val Leu Thr Leu Gly Lys Gly Val Phe Ser Phe Thr Leu Asp

260 265 270

Pro Met Tyr Gly Glu Phe Val Leu Thr Gln Glu Asn Ile Glu Ile Pro

275 280 285

Lys Ala Gly Arg Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Gln Met Trp

290 295 300

Asp Asp Lys Leu Lys Lys Tyr Ile Asp Asp Leu Lys Asp Pro Gly Pro

305 310 315 320

Thr Gly Lys Pro Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp

325 330 335

Phe His Arg Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp

340 345 350

Ala Lys Ser Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro

355 360 365

Met Ser Phe Ile Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His

370 375 380

Ser Arg Val Leu Asp Ile Gln Pro Thr Glu Ile His Gln Arg Val Pro

385 390 395 400

Leu Tyr Ile Gly Ser Thr Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu

405 410 415

Ala

210>162

<211>1254

<212>DNA

＜213〉colea

<400>162

atggcagcaa ccgccgcgac aacgtcatcg tctcatcttc ttctctcaag ctctcgccac 60

gtggcagcct catctcagcc acaaatcctt ttccagagat ctctgttttc cggcgggaaa 120

cgatcggcag ctggaaaaaa ccatcatgct agtggtggag tgaggtgtat ggcggttgca 180

gcggatgctt cggcggaggc taagccggcg gcagcgagga agaagagtgg gtacgagctt 240

cagacgctga cgggttggtt gctgagacaa gagcagaaag gagagataga tacagagctg 300

acgatagtga tgtcgagcat agcgatggct tgtaagcaga tcgcttcgct tgtacagcgc 360

gccggaatct ctaatctgac cggcgttcaa ggagccgtta acattcaggg agaggatcag 420

aaaaagctcg acgtcgtctc taatgaggta ttttcaaact gtttgagatc aagtggaagg 480

acagggatca ttgcgtcaga ggaagaggac gtccccgtag cagttgaaga gagttactcc 540

ggaaactaca ttgtcgtctt tgatcctctc gacggttcct ccaacatcga cgctgcagtc 600

tccactggtt caatcttcgg tatctacagc cccaatgacg agtgtatcgt tgacgactcc 660

gacgatatct cctctcttgg ttcagaagaa caaaggtgta tagtaaacgt gtgtcaacca 720

gggaacaact tgctcgcagc tggctactgt atgtactcga gctcagtcat cttcgttctc 780

accttaggca agggcgtttt ctccttcaca ctcgacccaa tgtacggcga gttcgtcctc 840

actcaagaga acattgagat ccccaaagca gggaaaatct actctttcaa cgaagggaac 900

taccagatgt gggacgagaa actgaagaag tacattgatg atcttaagga ccctggtcca 960

agtgggaagc cttactctgc aaggtacatt ggtagtttgg tcggagactt tcacagaact 1020

ttgttgtacg gtgggattta cgggtaccct cgtgacgcca agagcaaaaa cggtaagctt 1080

aggcttttgt atgagtgtgc accgatgagt ttcatcgttg aacaagctgg aggaaaagga 1140

tcagatggcc accaaagagt actagatatc caacccaccg agatacatca gagggttcca 1200

ctttacattg gaagcaaaga agaagtagag aagctggaga agtacttggc ttga 1254

<210>163

<211>417

<212>PRT

＜213〉colea

<400>163

Met Ala Ala Thr Ala Ala Thr Thr Ser Ser Ser His Leu Leu Leu Ser

1 5 10 15

Ser Ser Arg His Val Ala Ala Ser Ser Gln Pro Gln Ile Leu Phe Gln

20 25 30

Arg Ser Leu Phe Ser Gly Gly Lys Arg Ser Ala Ala Gly Lys Asn His

35 40 45

His Ala Ser Gly Gly Val Arg Cys Met Ala Val Ala Ala Asp Ala Ser

50 55 60

Ala Glu Ala Lys Pro Ala Ala Ala Arg Lys Lys Ser Gly Tyr Glu Leu

65 70 75 80

Gln Thr Leu Thr Gly Trp Leu Leu Arg Gln Glu Gln Lys Gly Glu Ile

85 90 95

Asp Thr Glu Leu Thr Ile Val Met Ser Ser Ile Ala Met Ala Cys Lys

100 105 110

Gln Ile Ala Ser Leu Val Gln Arg Ala Gly Ile Ser Asn Leu Thr Gly

115 120 125

Val Gln Gly Ala Val Asn Ile Gln Gly Glu Asp Gln Lys Lys Leu Asp

130 135 140

Val Val Ser Asn Glu Val Phe Ser Asn Cys Leu Arg Ser Ser Gly Arg

145 150 155 160

Thr Gly Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu

165 170 175

Glu Ser Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly

180 185 190

Ser Ser Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile

195 200 205

Tyr Ser Pro Asn Asp Glu Cys Ile Val Asp Asp Ser Asp Asp Ile Ser

210 215 220

Ser Leu Gly Ser Glu Glu Gln Arg Cys Ile Val Asn Val Cys Gln Pro

225 230 235 240

Gly Asn Asn Leu Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val

245 250 255

Ile Phe Val Leu Thr Leu Gly Lys Gly Val Phe Ser Phe Thr Leu Asp

260 265 270

Pro Met Tyr Gly Glu Phe Val Leu Thr Gln Glu Asn Ile Glu Ile Pro

275 280 285

Lys Ala Gly Lys Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Gln Met Trp

290 295 300

Asp Glu Lys Leu Lys Lys Tyr Ile Asp Asp Leu Lys Asp Pro Gly Pro

305 310 315 320

Ser Gly Lys Pro Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp

325 330 335

Phe His Arg Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp

340 345 350

Ala Lys Ser Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro

355 360 365

Met Ser Phe Ile Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His

370 375 380

Gln Arg Val Leu Asp Ile Gln Pro Thr Glu Ile His Gln Arg Val Pro

385 390 395 400

Leu Tyr Ile Gly Ser Lys Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu

405 410 415

Ala

<210>164

<211>1130

<212>DNA

<213>Cyanidioschyzon merolae

<400>164

atggcgaagc aagcaccgca cgctttcgtt agccttgggg ctcctaagct gcgctcaaca 60

aacatatggg gaagggtatc ggtgtcggtc ttgccgctgg aaacaaggca tcaacgtcgc 120

tttggtgcag tgagactgcg cgcagcttta gacctaccca tgacactcgg acagtgggtt 180

ctccaacagg agaggcagca tccagagacc gctgatttgg ctcctttgat aacagccaca 240

gcgacagctg gcaagcagat tgcctcactg gtcgcccgag cgggcgtgag caacttgact 300

ggccttcagg gctcggtcaa cgtgcagggc gaggagcaga aaaagctgga cgtgttgacg 360

aacgaggtgc tcaagaacgc gctacgctca acagggaaac taggtgtctt ggcttcggaa 420

gaggaaaacg agccagtgtc gctcatggaa gaggcgtata caggtgactt tattgctgcg 480

tttgacccgt tggacggatc ctcgaatttg gatgcagcga tcgccacggg tacgattttt 540

ggtgtaatgc gcagcggcga gcactgtctg acgggacccg aggacgcgga tataagtcag 600

cgacaaatgc agtgcctcgt gcagacgctg caaccgggtg cgaatctggt ggcagcgggc 660

tacattctct actcctcatc ggtgattttc atgctgtcta ttggtcacgg agtgcagggt 720

ttttctctgg acccagcgat tggcgagttt gttttgacgc acccagatgt tcgtgttccg 780

gagcgtggca agatctactc cttcaacgag gcaaactcgg ataactggga ccctgttctt 840

caagaatata ttcgatcgat gaagaaaaag ggctattctt cgcgctacat aggatctctc 900

gttggcgatg tccatcgcac gctgatttac ggcggcgttt ttggataccc tggtgataag 960

aagaacccga acggcaagtt gcgtcttctg tacgagtgcg cacctatggc gtatctcatg 1020

gagcaggccg gcggtatagc gaccacggga aagcaaagaa ttctggatat tgttccgcga 1080

gatgtccatc agcgagagcc gttcatttgc ggaagcccca gggatgttga 1130

<210>165

<211>391

<212>PRT

<213>Cyanidioschyzon merolae

<400>165

Met Ala Lys Gln Ala Pro His Ala Phe Val Ser Leu Gly Ala Pro Lys

1 5 10 15

Leu Arg Ser Thr Asn Ile Trp Gly Arg Val Ser Val Ser Val Leu Pro

20 25 30

Leu Glu Thr Arg His Gln Arg Arg Phe Gly Ala Val Arg Leu Arg Ala

35 40 45

Ala Leu Asp Leu Pro Met Thr Leu Gly Gln Trp Val Leu Gln Gln Glu

50 55 60

Arg Gln His Pro Glu Thr Ala Asp Leu Ala Pro Leu Ile Thr Ala Thr

65 70 75 80

Ala Thr Ala Gly Lys Gln Ile Ala Ser Leu Val Ala Arg Ala Gly Val

85 90 95

Ser Asn Leu Thr Gly Leu Gln Gly Ser Val Asn Val Gln Gly Glu Glu

100 105 110

Gln Lys Lys Leu Asp Val Leu Thr Asn Glu Val Leu Lys Asn Ala Leu

115 120 125

Arg Ser Thr Gly Lys Leu Gly Val Leu Ala Ser Glu Glu Glu Asn Glu

130 135 140

Pro Val Ser Leu Met Glu Glu Ala Tyr Thr Gly Asp Phe Ile Ala Ala

145 150 155 160

Phe Asp Pro Leu Asp Gly Ser Ser Asn Leu Asp Ala Ala Ile Ala Thr

165 170 175

Gly Thr Ile Phe Gly Val Met Arg Ser Gly Glu His Cys Leu Thr Gly

180 185 190

Pro Glu Asp Ala Asp Ile Ser Gln Arg Gln Met Gln Cys Leu Val Gln

195 200 205

Thr Leu Gln Pro Gly Ala Asn Leu Val Ala Ala Gly Tyr Ile Leu Tyr

210 215 220

Ser Ser Ser Val Ile Phe Met Leu Ser Ile Gly His Gly Val Gln Gly

225 230 235 240

Phe Ser Leu Asp Pro Ala Ile Gly Glu Phe Val Leu Thr His Pro Asp

245 250 255

Val Arg Val Pro Glu Arg Gly Lys Ile Tyr Ser Phe Asn Glu Ala Asn

260 265 270

Ser Asp Asn Trp Asp Pro Val Leu Gln Glu Tyr Ile Arg Ser Met Lys

275 280 285

Lys Lys Gly Tyr Ser Ser Arg Tyr Ile Gly Ser Leu Val Gly Asp Val

290 295 300

His Arg Thr Leu Ile Tyr Gly Gly Val Phe Gly Tyr Pro Gly Asp Lys

305 310 315 320

Lys Asn Pro Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met

325 330 335

Ala Tyr Leu Met Glu Gln Ala Gly Gly Ile Ala Thr Thr Gly Lys Gln

340 345 350

Arg Ile Leu Asp Ile Val Pro Arg Asp Val His Gln Arg Glu Pro Phe

355 360 365

Ile Cys Gly Ser Pro Arg Asp Val Glu Asp Leu Leu Lys Ile Tyr Gln

370 375 380

Gly Ser Met Ala Met Arg Gly

385 390

<210>166

<211>1218

<212>DNA

＜213〉soybean

<400>166

atggttgcaa tggcagcagc aacagcatcc acccagttga ttttctcaaa gccttgttcc 60

ccttcacgtc tatgcccctt ccaactatgt gtctttgaca ctaaacaagt gctatcaagt 120

ggcaggagaa ggcatgtggg gggttctgga gttaggtgca tggctgtggg ggaagcagca 180

accactggga caaagaagag aagtggatat gagcttcaaa cactcactag ctggttgctg 240

aagcaggagc aagctggggt gattgatgca gaactcacta ttgtgctgtc tagcatttcc 300

atggcatgca aacagattgc ttctttggtg caaagagcta acatttccaa cctcactggg 360

gttcaaggtg ctgtcaatgt tcaaggggaa gaccagaaaa agcttgatgt tgtttcaaat 420

gaggttttct caaactgctt gaggtcaagt gggaggacag ggataatagc atcagaggag 480

gaagatgtgc cagtggcagt agaagagagt tattctggaa actacattgt ggtgtttgac 540

ccacttgatg ggtcatccaa tattgatgct gcagcgtcaa ctgggtccaa tttttggata 600

tacagcccca atgatgagtg tcttgctgac attgatgatg accccaccct tgacacaaca 660

gaacaaagat gtattgtgaa cgtgtgccaa cctggaagca accttcttgc agctggttac 720

tgcatgtatt ctagctcaat aatctttgtt ctcacacttg gaaatggagt gtttgtgttt 780

acattggacc cgatgtatgg cgaattcgtt ttgactcagg aaaacctcca gatacctaga 840

gcaggcaaaa tctatgcatt caatgaaggt aattatcagt tgtgggatga gaagctaaag 900

aaatatattg atgatctcaa ggacccaggt caaagcggca agccttattc tgcaaggtac 960

attggtagct tggtaggaga tttccacagg acactgctat atggtggcat ttacgggtac 1020

cccagggaca agaaaagcaa gaatgggaaa ctaaggctcc tgtatgaatg tgctcctatt 1080

aacttcattg tagaacaagc tggtggaaaa ggtacagatg gccttcaagt actccggctt 1140

caagggacag agattcatca acgtgtgcca ctgtacattg gggaagaggt agagaaggtg 1200

gaaaagtact tggcttaa 1218

<210>167

<211>405

<212>PRT

＜213〉soybean

<400>167

Met Val Ala Met Ala Ala Ala Thr Ala Ser Thr Gln Leu Ile Phe Ser

1 5 10 15

Lys Pro Cys Ser Pro Ser Arg Leu Cys Pro Phe Gln Leu Cys Val Phe

20 25 30

Asp Thr Lys Gln Val Leu Ser Ser Gly Arg Arg Arg His Val Gly Gly

35 40 45

Ser Gly Val Arg Cys Met Ala Val Gly Glu Ala Ala Thr Thr Gly Thr

50 55 60

Lys Lys Arg Ser Gly Tyr Glu Leu Gln Thr Leu Thr Ser Trp Leu Leu

65 70 75 80

Lys Gln Glu Gln Ala Gly Val Ile Asp Ala Glu Leu Thr Ile Val Leu

85 90 95

Ser Ser Ile Ser Met Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg

100 105 110

Ala Asn Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val Asn Val Gln

115 120 125

Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser

130 135 140

Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu

145 150 155 160

Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile

165 170 175

Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Ala

180 185 190

Ser Thr Gly Ser Asn Phe Trp Ile Tyr Ser Pro Asn Asp Glu Cys Leu

195 200 205

Ala Asp Ile Asp Asp Asp Pro Thr Leu Asp Thr Thr Glu Gln Arg Cys

210 215 220

Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ala Ala Gly Tyr

225 230 235 240

Cys Met Tyr Ser Ser Ser Ile Ile Phe Val Leu Thr Leu Gly Asn Gly

245 250 255

Val Phe Val Phe Thr Leu Asp Pro Met Tyr Gly Glu Phe Val Leu Thr

260 265 270

Gln Glu Asn Leu Gln Ile Pro Arg Ala Gly Lys Ile Tyr Ala Phe Asn

275 280 285

Glu Gly Asn Tyr Gln Leu Trp Asp Glu Lys Leu Lys Lys Tyr Ile Asp

290 295 300

Asp Leu Lys Asp Pro Gly Gln Ser Gly Lys Pro Tyr Ser Ala Arg Tyr

305 310 315 320

Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu Tyr Gly Gly

325 330 335

Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys Asn Gly Lys Leu Arg

340 345 350

Leu Leu Tyr Glu Cys Ala Pro Ile Asn Phe Ile Val Glu Gln Ala Gly

355 360 365

Gly Lys Gly Thr Asp Gly Leu Gln Val Leu Arg Leu Gln Gly Thr Glu

370 375 380

Ile His Gln Arg Val Pro Leu Tyr Ile Gly Glu Glu Val Glu Lys Val

385 390 395 400

Glu Lys Tyr Leu Ala

405

<210>168

<211>1212

<212>DNA

＜213〉tomato

<400>168

atggcagcaa cagcaacaac ttcatattta agtgctctag acaaaaagac tccattttta 60

tttgccttgg acaaaaagac tccattttta tgcccaaaaa acagcacaaa gagaaggtca 120

tttaatggag gagttaagtg catggcaata gagacagctt caggtgttac acaaaccaag 180

aaaaagagtg gctatgagtt acaaacttta acaagttggc tattgagaca agaacaagct 240

ggagttattg atgctgaact taccatagta atttcaagta tttcaatggc ttgtaaacag 300

attgcttctt tggtccaaag agctggaatt tctaacctta ctggagttca aggtgctgtc 360

aatattcaag gagaagatca gaagaaactt gatgttgtct ctaatgaggt tttctcgaat 420

tgtctaagat caagtggaag gactgggatt atagcatcag aagaagagga tgtacctgtg 480

gcagtggaag agagttactc aggcaactac attgtggtgt ttgatcctct tgatggatca 540

tcaaacattg atgctgctgt atctaccggt tctatctttg gaatatacag cccgaatgat 600

gagtgcctag ctgatcttgg agatgattcc acgcttgaca atatagagca aaagtgcatc 660

gtgaatgtat gtcaaccagg gacaaacctt cttgcagcag gatactgcat gtactcaagc 720

tctgtgattt tcgtactcac cttgggaaat ggcgtttttt cctttaactt ggatccaatg 780

tatggagaat ttgttctgac tcaagaaaat gtccaaatac caaagtctgg aaagatctat 840

tcattcaatg aaggaaacta ccagctctgg gatgacaagt tgaaaaaata tatcgatgac 900

ttgaaagacc ctggtcctag tggcaagcct tactctgcaa ggtacattgg tagtttggta 960

ggtgacttcc atagaactct tctatatggt ggcatttatg gttatcctag agacagaaag 1020

agcaagaatg gaaagttgag gcttttgtac gaatgtgctc ccatgagctt cattgtggaa 1080

caagctggtg gcaaaggatc cgatggtcac caaagagttc tcgatatcca accaactgag 1140

atacatcaac gagttccatt gtacattgga agcacagaag aagttgaaaa attggagaag 1200

tacttgtctt aa 1212

<210>169

<211>403

<212>PRT

＜213〉tomato

<400>169

Met Ala Ala Thr Ala Thr Thr Ser Tyr Leu Ser Ala Leu Asp Lys Lys

1 5 10 15

Thr Pro Phe Leu Phe Ala Leu Asp Lys Lys Thr Pro Phe Leu Cys Pro

20 25 30

Lys Asn Ser Thr Lys Arg Arg Ser Phe Asn Gly Gly Val Lys Cys Met

35 40 45

Ala Ile Glu Thr Ala Ser Gly Val Thr Gln Thr Lys Lys Lys Ser Gly

50 55 60

Tyr Glu Leu Gln Thr Leu Thr Ser Trp Leu Leu Arg Gln Glu Gln Ala

65 70 75 80

Gly Val Ile Asp Ala Glu Leu Thr Ile Val Ile Ser Ser Ile Ser Met

85 90 95

Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg Ala Gly Ile Ser Asn

100 105 110

Leu Thr Gly Val Gln Gly Ala Val Asn Ile Gln Gly Glu Asp Gln Lys

115 120 125

Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser Asn Cys Leu Arg Ser

130 135 140

Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val

145 150 155 160

Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro

165 17 0175

Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile

180 185 190

Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu Ala Asp Leu Gly Asp

195 200 205

Asp Ser Thr Leu Asp Asn Ile Glu Gln Lys Cys Ile Val Asn Val Cys

210 215 220

Gln Pro Gly Thr Asn Leu Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser

225 230 235 240

Ser Val Ile Phe Val Leu Thr Leu Gly Asn Gly Val Phe Ser Phe Asn

245 250 255

Leu Asp Pro Met Tyr Gly Glu Phe Val Leu Thr Gln Glu Asn Val Gln

260 265 270

Ile Pro Lys Ser Gly Lys Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Gln

275 28 0285

Leu Trp Asp Asp Lys Leu Lys Lys Tyr Ile Asp Asp Leu Lys Asp Pro

290 295 300

Gly Pro Ser Gly Lys Pro Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val

305 310 315 320

Gly Asp Phe His Arg Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro

325 330 335

Arg Asp Arg Lys Ser Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys

340 345 350

Ala Pro Met Ser Phe Ile Val Glu Gln Ala Gly Gly Lys Gly Ser Asp

355 360 365

Gly His Gln Arg Val Leu Asp Ile Gln Pro Thr Glu Ile His Gln Arg

370 375 380

Val Pro Leu Tyr Ile Gly Ser Thr Glu Glu Val Glu Lys Leu Glu Lys

385 390 395 400

Tyr Leu Ser

<210>170

<211>1236

<212>DNA

＜213〉puncture vine clover

<400>170

atggttgcaa tggcagcagc aacagcatca tcacaattaa tattctcaaa accttgttct 60

ccctcacgtc tatgtccctt ccaactttgt gttttcgaca cgaaatcagt gttatcaagt 120

tcaagaagaa agcatgtgag tggctctgga ggagtgagat gcatggctgt tggtgaagta 180

gctgctgaga caaagaaaag aagtagttat gagcttataa cattgactag ttggttgttg 240

aagcaagaac aaacaggggt tattgatgct gaacttacta ttgttttgaa tagtatttcg 300

ttggcatgta aacagattgc ttctttggtt caaagagcta atatttctaa ccttactggt 360

gttcaaggtg ctgtaaatat tcaaggggag gatcagaaaa aacttgatgt tgtctcaaat 420

gaggtattct caaattgttt gaggtcaagt gggaggacag ggattatagc atcagaggaa 480

gaggatgtgc cagtggcagt agaagagagt tattcaggaa actacattgt ggtctttgat 540

ccacttgatg gttcatcgaa tattgatgct gcagtttcaa ctggttctat ttttgggatt 600

tacagcccca atgatgagtg tcttgctgac gtaggcaatg agtccgatga ccccacactt 660

ggcacagaag aacaacgatg cattgtgaat gtgtgccaac caggaagcaa ccttctagca 720

gccggttact gcatgtattc tagctcagta atcttcgttc taacaatcgg caaaggagtt 780

ttcgtattca cattagatcc aatgtatggg gaatttgttt tgactcaaga aaatctccaa 840

ataccaaaat cagggaaaat ttattctttc aatgaaggaa attacaagtt atgggatgac 900

aacttgaaga aatacatcga tgatctcaag gaaccgggtg ctaatggcaa accttattca 960

gcaaggtata ttggtagttt ggtaggtgat ttccatagga cactgctata tggtggcatt 1020

tatggttacc ctagggacaa gaaaagtaag aatggaaggc ttaggctttt atatgagtgt 1080

gctccaatga gtttcattgt agaacaggct ggtggaaaag gttcagatgg tcatcaaaga 1140

gtacttgaca ttcaacccac cgaaattcat caacgtgttc cactgtacat tgggagcaca 1200

gaggaagtgg agaaggttga aaagtacttg gcttaa 1236

<210>171

<211>411

<212>PRT

＜213〉puncture vine clover

<400>171

Met Val Ala Met Ala Ala Ala Thr Ala Ser Ser Gln Leu Ile Phe Ser

1 5 10 15

Lys Pro Cys Ser Pro Ser Arg Leu Cys Pro Phe Gln Leu Cys Val Phe

20 25 30

Asp Thr Lys Ser Val Leu Ser Ser Ser Arg Arg Lys His Val Ser Gly

35 40 45

Ser Gly Gly Val Arg Cys Met Ala Val Gly Glu Val Ala Ala Glu Thr

50 55 60

Lys Lys Arg Ser Ser Tyr Glu Leu Ile Thr Leu Thr Ser Trp Leu Leu

65 70 75 80

Lys Gln Glu Gln Thr Gly Val Ile Asp Ala Glu Leu Thr Ile Va1 Leu

85 90 95

Asn Ser Ile Ser Leu Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg

100 105 110

Ala Asn Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val Asn Ile Gln

115 120 125

Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser

130 135 140

Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu

145 150 155 160

Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile

165 170 175

Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val

180 185 190

Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu

195 200 205

Ala Asp Val Gly Asn Glu Ser Asp Asp Pro Thr Leu Gly Thr Glu Glu

210 215 220

Gln Arg Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ala

225 230 235 240

Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val Leu Thr Ile

245 250 255

Gly Lys Gly Val Phe Val Phe Thr Leu Asp Pro Met Tyr Gly Glu Phe

260 265 270

Val Leu Thr Gln Glu Asn Leu Gln Ile Pro Lys Ser Gly Lys Ile Tyr

275 280 285

Ser Phe Asn Glu Gly Asn Tyr Lys Leu Trp Asp Asp Asn Leu Lys Lys

290 295 300

Tyr Ile Asp Asp Leu Lys Glu Pro Gly Ala Asn Gly Lys Pro Tyr Ser

305 310 315 320

Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu

325 330 335

Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys Asn Gly

340 345 350

Arg Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile Val Glu

355 36 0365

Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile

370 375 380

Gln Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile Gly Ser Thr

385 390 395 400

Glu Glu Val Glu Lys Val Glu Lys Tyr Leu Ala

405 410

<210>172

<211>1236

<212>DNA

＜213〉tobacco (Nicotiana tabacum)

<400>172

atggcagcat catcagcaac agcaacagca acaacttcat ttttatgtgc tttagacaaa 60

aagactccat ttttatgtac tctagacaaa aaaggtactc catttttatg cccaaaaggc 120

agcagcacaa caaagagaag gtcatttaat ggaggagtga agtgcatggc aatagagaca 180

acagcaggag ctacagagac caagaaaaga agtggctatg agttacaaac tttaacaagc 240

tggctattaa ggcaagaaca agctgggagt attgatgctg aacttaccat agtgatttca 300

agtatttcta tggcttgtaa gcagattgct tctttggttc agagagctgg aatttctaac 360

cttactggag ttcaaggtgc tgtcaatatt caaggagaag accagaagaa gcttgatgtt 420

gtctctaacg aggttttctc gaattgtcta aggtcgagtg gaaggactgg gattatagca 480

tcagaggaag aggatgtacc agtggcagtg gaagagagtt actcaggaaa ctacattgtg 540

gtgtttgacc ctcttgatgg atcatcaaac attgatgctg ctgtttctac tggttctatt 600

ttcggaatat acagcccaaa tgatgagtgc ctcgcagatc atggagatga ttccacgctt 660

gacaatgttg aacagaggtg tattgtgaat gtatgccaac cagggagcaa ccttcttgca 720

gcaggctact gcatgtactc aagctctgtg atttttgtgg tcaccttggg aaacggagtc 780

tttgccttca acttggatcc gatgtatgga gaattcgttc tgacccaaga aaacatccaa 840

ataccaaaat ctggaaagat ctattcgttc aacgaaggaa actaccagct ttgggatgac 900

aaactgaaga aatacatcga tgacttgaag gaccctggtc ctagcggcaa gccttactct 960

gctaggtaca ttggtagttt ggttggtgac ttccatagaa ctcttctata tggtggcatt 1020

tatggctatc ctagagataa aaagagtaag aatggaaagt tgaggctttt gtatgagtgt 1080

gcccctatga gcttcctcgt ggaacaagct ggtggcaaag gatccgatgg ccaccaaaga 1140

gttcttgata tccaaccaac tgagatacac cagcgagtcc cattgtacat tggaagcaca 1200

gaagaagttg aaaagttgga gaagtattta tcttaa 1236

<210>173

<211>411

<212>PRT

＜213〉tobacco

<400>173

Met Ala Ala Ser Ser Ala Thr Ala Thr Ala Thr Thr Ser Phe Leu Cys

1 5 10 15

Ala Leu Asp Lys Lys Thr Pro Phe Leu Cys Thr Leu Asp Lys Lys Gly

20 25 30

Thr Pro Phe Leu Cys ProLys Gly Ser Ser Thr Thr Lys Arg Arg Ser

35 40 45

Phe Asn Gly Gly Val Lys Cys Met Ala Ile Glu Thr Thr Ala Gly Ala

50 55 60

Thr Glu Thr Lys Lys Arg Ser Gly Tyr Glu Leu Gln Thr Leu Thr Ser

65 70 75 80

Trp Leu Leu Arg Gln Glu Gln Ala Gly Ser Ile Asp Ala Glu Leu Thr

85 90 95

Ile Val Ile Ser Ser Ile Ser Met Ala Cys Lys Gln Ile Ala Ser Leu

100 105 110

yal Gln Arg Ala Gly Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val

115 120 125

Asn Ile Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser Asn Glu

130 135 140

Val Phe Ser Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala

145 150 155 160

Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly

165 170 175

Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp

180 185 190

Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro Asn Asp

195 200 205

Glu Cys Leu Ala Asp His Gly Asp Asp Ser Thr Leu Asp Asn Val Glu

210 215 220

Gln Arg Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ala

225 230 235 240

Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val Val Thr Leu

245 250 255

Gly Asn Gly Val Phe Ala Phe Asn Leu Asp Pro Met Tyr Gly Glu Phe

260 265 270

Val Leu Thr Gln Glu Asn Ile Gln Ile Pro Lys Ser Gly Lys Ile Tyr

275 280 285

Ser Phe Asn Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys Leu Lys Lys

290 295 300

Tyr Ile Asp Asp Leu Lys Asp Pro Gly Pro Ser Gly Lys Pro Tyr Ser

305 310 315 320

Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu

325 330 335

Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys Asn Gly

340 345 350

lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Leu Val Glu

355 360 365

Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile

370 375 380

Gln Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile Gly Ser Thr

385 390 395 400

Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu Ser

405 410

<210>174

<211>1221

<212>DNA

＜213〉rice

<400>174

atggccgccg cagccacgac atcctcccac ctgctcctgc tctcccgcca gcaggcggcg 60

gcctcgctcc aatgcggcct ctccttccgg aggcagcccg gcaggctcgc cggtgggtcg 120

tcggccccga gcgtgcggtg catggcggcc gtcgacacgg cctcggcgcc cgccgcgacg 180

gaggctagca agaagagcag ctacgagatc accacgctga cgacgtggct gctgaagcag 240

gagcaggccg ggaccatcga cggcgagatg accatcgtgc tggccagcat ctccacggcg 300

tgcaagcaga tcgcctcgct ggtgcagcgc gcgcccatct ccaacctcac cggcgtccag 360

ggcgccgtca acgtgcaggg cgaggaccag aaaaaactcg acgtcgtctc caacgaggtg 420

ttctccaact gcctcaaatc gagcgggcgc accggcgtga tcgcgtcgga ggaggaggac 480

gtgccggtgg ccgtggagga gagctactcg ggcaactaca tcgtggtgtt cgacccgctc 540

gacggctcct ccaacatcga cgccgccgtc tccaccggct ccatcttcgg catctacagc 600

cccaacgacg agtgcctagc cgacatcgcc gacgaccaaa atcttgacca ggtggagcag 660

aggtgcatcg tgagcgtgtg ccagccgggg agcaacctgc tcgccgccgg ctactgcatg 720

tactcgagct cggtcatctt cgtgctcacc atcgggacag gggtgtacgt gttcacgctg 780

gacccgatgt acggcgagtt cgtgctgacg caggagaagg tgcagatccc caaggcaggc 840

aagatctatg ccttcaacga gggcaactac gcgctctggg acgacaagct caagagctac 900

atggacagcc tcaaggagcc cgggccgtcc gggaagccat actccgcgcg ctacatcggc 960

agcctcgtcg gcgacttcca ccgcacgctg ctctacggcg gcatctacgg ctaccccagg 1020

gaccagaaga gcaagaacgg caagctgcgg ctgctgtacg agtgcgcgcc gatgagcttc 1080

atcgtcgagc aggccggcgg caagggctcc gatggccacc agaggatact tgacatcatg 1140

cctacggaga tccatcagag agtgccgctg tacatcggga gcgtggagga agtggagaag 1200

gtcgagaagt tcttggcttg a 1221

<210>175

<211>406

<212>PRT

＜213〉rice

<400>175

Met Ala Ala Ala Ala Thr Thr Ser Ser His Leu Leu Leu Leu Ser Arg

1 5 10 15

Gln Gln Ala Ala Ala Ser Leu Gln Cys Gly Leu Ser Phe Arg Arg Gln

20 25 30

Pro Gly Arg Leu Ala Gly Gly Ser Ser Ala Pro Ser Val Arg Cys Met

35 40 45

Ala Ala Val Asp Thr Ala Ser Ala Pro Ala Ala Thr Glu Ala Ser Lys

50 55 60

Lys Ser Ser Tyr Glu Ile Thr Thr Leu Thr Thr Trp Leu Leu Lys Gln

65 70 75 80

Glu Gln Ala Gly Thr Ile Asp Gly Glu Met Thr Ile Val Leu Ala Ser

85 90 95

Ile Ser Thr Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg Ala Pro

100 105 110

Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val Asn Val Gln Gly Glu

115 120 125

Asp Gln Lys Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser Asn Cys

130 135 140

Leu Lys Ser Ser Gly Arg Thr Gly Val Ile Ala Ser Glu Glu Glu Asp

145 150 155 160

Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile Val Val

165 170 175

Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val Ser Thr

180 185 190

Gly Ser Ile Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu Ala Asp

195 200 205

Ile Ala Asp Asp Gln Asn Leu Asp Gln Val Glu Gln Arg Cys Ile Val

210 215 220

Ser Val Cys Gln Pro Gly Ser Asn Leu Leu Ala Ala Gly Tyr Cys Met

225 230 235 240

Tyr Ser Ser Ser Val Ile Phe Val Leu Thr Ile Gly Thr Gly Val Tyr

245 250 255

Val Phe Thr Leu Asp Pro Met Tyr Gly Glu Phe Val Leu Thr Gln Glu

260 265 270

Lys Val Gln Ile Pro Lys Ala Gly Lys Ile Tyr Ala Phe Asn Glu Gly

275 280 285

Asn Tyr Ala Leu Trp Asp Asp Lys Leu Lys Ser Tyr Met Asp Ser Leu

290 295 300

Lys Glu Pro Gly Pro Ser Gly Lys Pro Tyr Ser Ala Arg Tyr Ile Gly

305 310 315 320

Ser Leu Val Gly Asp Phe Hi s Arg Thr Leu Leu Tyr Gly Gly Ile Tyr

325 330 335

Gly Tyr Pro Arg Asp Gln Lys Ser Lys Asn Gly Lys Leu Arg Leu Leu

340 345 350

Tyr Glu Cys Ala Pro Met Ser Phe Ile Val Glu Gln Ala Gly Gly Lys

355 360 365

Gly Ser Asp Gly His Gln Arg Ile Leu Asp Ile Met Pro Thr Glu Ile

370 375 380

His Gln Arg Val Pro Leu Tyr Ile Gly Ser Val Glu Glu Val Glu Lys

385 390 395 400

Val Glu Lys Phe Leu Ala

405

<210>176

<211>1143

<212>DNA

<213>Ostreococcus lucimarinus

<400>176

atgtctgccg ccacctccgc ctgcgctcgc gcgatcgtcg cgacgaagaa gaccaccgcg 60

gcgcgccgcg cgggcaagtc ggcgacgcgg gcgacgccgc gatccgtcgc ggcgcgcgcg 120

gggggcgcgg gcacgccgtt cacgacgtgg atcttgcaac aagagatgga agaaaagatc 180

gacggcgaac tcgcggtcgt gttgtcgagc atcggcttgg cgtgcaagca aatcgcgagc 240

ttggtgcaac gcgccgggct tcaaggcatg acgggtttgg cgggcgagac gaacgtgcaa 300

ggtgaagacc aaaagaagct cgacgtcatc tccaacgatg tgttctgcga tgtcttgcgt 360

caaaccggcc gcacgggcgt gattgcgtcc gaggaagaag acgtgccggt cgccgtcgaa 420

gaaaccttcg gcggtaacta cgtcgtcgtc ttcgatccgc tcgatggctc ttccaacatc 480

gacgccgccg tttccacggg ttccatctgg ggcatctacg agtccgactc cacgtgcatc 540

ccggattttg gcaccgaaga tgcggccaag attgaagaga agtgcgtcat gaacgtgtgc 600

caaccgggga acaacttgtt gtgcgcgggc tactgcatgt actcctcttc caccatcctc 660

gtcttgaccc tcggtgaggg tgtgtacggt ttcaccctcg acccgaccgt cggcgagttc 720

atcatgtccc acgacaacat caaggttccg gaatctggta agatttactc cttcaacgaa 780

ggcaactacg acatgtggac tccgggcttg aagaagtaca tggactccct caagactggc 840

ggcgcggaac aaggcaccaa gccgtacagc gcccgttaca tcggttccct cgtcggtgac 900

ttccacagaa ccatcttgta cggaggcatc tacggctacc cgggcgactc caagaacccg 960

aacggtaagc ttcgcctctt gtacgagtgc gcgccgatgt ccttcatcgc cgaacaagcc 1020

ggcggtatgg gttccaccgg taaggagcgc gtccttgacg ttgtcccgga aaagtttcac 1080

caacgtgtgc cgttcttcac cggctccaag aaggaagtgc agtacttgga atccttcatg 1140

tag 1143

<210>177

<211>380

<212>PRT

<213>Ostreococcus lucimarinus

<40>177

Met Ser Ala Ala Thr Ser Ala Cys Ala Arg Ala Ile Val Ala Thr Lys

1 5 10 15

Lys Thr Thr Ala Ala Arg Arg Ala Gly Lys Ser Ala Thr Arg Ala Thr

20 25 30

Pro Arg Ser Val Ala Ala Arg Ala Gly Gly Ala Gly Thr Pro Phe Thr

35 40 45

Thr Trp Ile Leu Gln Gln Glu Met Glu Glu Lys Ile Asp Gly Glu Leu

50 55 60

Ala Val Val Leu Ser Ser Ile Gly Leu Ala Cys Lys Gln Ile Ala Ser

65 70 75 80

Leu Val Gln Arg Ala Gly Leu Gln Gly Met Thr Gly Leu Ala Gly Glu

85 90 95

Thr Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Ile Ser Asn

100 105 110

Asp Val Phe Cys Asp Val Leu Arg Gln Thr Gly Arg Thr Gly Val Ile

115 120 125

Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Thr Phe Gly

130 135 140

Gly Asn Tyr Val Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile

145 150 155 160

Asp Ala Ala Val Ser Thr Gly Ser Ile Trp Gly Ile Tyr Glu Ser Asp

165 170 175

Ser Thr Cys Ile Pro Asp Phe Gly Thr Glu Asp Ala Ala Lys Ile Glu

180 185 190

Glu Lys Cys Val Met Asn Val Cys Gln Pro Gly Asn Asn Leu Leu Cys

195 200 205

Ala Gly Tyr Cys Met Tyr Ser Ser Ser Thr Ile Leu Val Leu Thr Leu

210 215 220

Gly Glu Gly Val Tyr Gly Phe Thr Leu Asp Pro Thr Val Gly Glu Phe

225 230 235 240

Ile Met Ser His Asp Asn Ile Lys Val Pro Glu Ser Gly Lys Ile Tyr

245 250 255

Ser Phe Asn Glu Gly Asn Tyr Asp Met Trp Thr Pro Gly Leu Lys Lys

260 265 270

Tyr Met Asp Ser Leu Lys Thr Gly Gly Ala Glu Gln Gly Thr Lys Pro

275 280 285

Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr

290 295 300

Ile Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Gly Asp Ser Lys Asn Pro

305 310 315 320

Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile

325 330 335

Ala Glu Gln Ala Gly Gly Met Gly Ser Thr Gly Lys Glu Arg Val Leu

340 345 350

Asp Val Val Pro Glu Lys Phe His Gln Arg Val Pro Phe Phe Thr Gly

355 360 365

Ser Lys Lys Glu Val Gln Tyr Leu Glu Ser Phe Met

370 375 380

<210>178

<211>1134

<212>DNA

<213>Ostreococcus tauri

<400>178

atgtctgcgt gtatttctgc ctctgtgacg cgcgcgcgaa cggcgcgcgc gcccgcggct 60

cgccgcgcgg cgagtaaggc gcgggcgtcg ccgcgcgccg tcgcggcgcg cgcggggggc 120

gttggtacgc cgtacacgac gtggattctc cagcaagaga tgcaagagaa cattgatggt 180

gaattggcgg tggttttgtc ctccatcggt ctcgcgtgca agcaaatcgc gagcctcgtc 240

cagcgcgcgg gtctcgcggg catgactggt ttggcgggcg agcaaaacgt gcaaggcgag 300

gaccagaaga agctcgacgt catctccaac gatgttttct gcaacgtatt gcgccaatcc 360

ggccgcacgg gcgtcatcgc ctccgaagaa gaagacgttc cggtcgccgt cgaggaaacg 420

tacggcggta actacgtcgt cgtcttcgat ccgcttgatg gttcctccaa tatcgacgcc 480

gccgtctcca cgggatccat ctggggtatc tacgagtccg actcctcgtg tattcccgac 540

ttcggctccg atgactccgc caaggttgaa gagaagtgcg tcatgaacgt ttgccaaccg 600

ggcagcaact tgctctgcgc gggttactgc atgtactcct cgtccaccat cctcgtcatc 660

accatcggcc aaggcgtgtt cggttttacc ctcgacccga ccgtcggtga gttcatcatg 720

tcccacgaga acatcaaggt tccggactct ggcaagattt actctttcaa cgaaggcaac 780

tacgccatgt ggtccgacgg tttgaagaag tacatggact ccctcaagac gggcggcaag 840

gacggtggca agccgtacag cgcccgctac atcggttcgc tcgtcggtga cttccaccgc 900

acgatccttt acggaggcat ctacggttac ccgggtgacg cgaagaaccc gaacggtaag 960

ctccgcctcc tgtacgagtg cgcgccgatg tccatgatcg ctgaacaagc cggtggtaag 1020

ggctccaccg gtgtcgcgcg tgtcctggac atcgttccgg aaaaggttca ccagcgcgtg 1080

ccgttcttcg ttggctccaa gaacgaggtt gcctacttgg agtccttcat gtga 1134

<210>179

<211>377

<212>PRT

<213>Ostreococcus tauri

<400>179

Met Ser Ala Cys Ile Ser Ala Ser Val Thr Arg Ala Arg Thr Ala Arg

1 5 10 15

Ala Pro Ala Ala Arg Arg Ala Ala Ser Lys Ala Arg Ala Ser Pro Arg

20 25 30

Ala Val Ala Ala Arg Ala Gly Gly Val Gly Thr Pro Tyr Thr Thr Trp

35 40 45

Ile Leu Gln Gln Glu Met Gln Glu Asn Ile Asp Gly Glu Leu Ala Val

50 55 60

Val Leu Ser Ser Ile Gly Leu Ala Cys Lys Gln Ile Ala Ser Leu Val

65 70 75 80

Gln Arg Ala Gly Leu Ala Gly Met Thr Gly Leu Ala Gly Glu Gln Asn

85 90 95

Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Ile Ser Asn Asp Val

100 105 110

Phe Cys Asn Val Leu Arg Gln Ser Gly Arg Thr Gly Val Ile Ala Ser

115 120 125

Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Thr Tyr Gly Gly Asn

130 135 140

Tyr Val Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala

145 150 155 160

Ala Val Ser Thr Gly Ser Ile Trp Gly Ile Tyr Glu Ser Asp Ser Ser

165 170 175

Cys Ile Pro Asp Phe Gly Ser Asp Asp Ser Ala Lys Val Glu Glu Lys

180 185 190

Cys Val Met Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Cys Ala Gly

195 200 205

Tyr Cys Met Tyr Ser Ser Ser Thr Ile Leu Val Ile Thr Ile Gly Gln

210 215 220

Gly Val Phe Gly Phe Thr Leu Asp Pro Thr Val Gly Glu Phe Ile Met

225 230 235 240

Ser His Glu Asn Ile Lys Val Pro Asp Ser Gly Lys Ile Tyr Ser Phe

245 250 255

Asn Glu Gly Asn Tyr Ala Met Trp Ser Asp Gly Leu Lys Lys Tyr Met

260 265 270

Asp Ser Leu Lys Thr Gly Gly Lys Asp Gly Gly Lys Pro Tyr Ser Ala

275 280 285

Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Ile Leu Tyr

290 295 300

Gly Gly Ile Tyr Gly Tyr Pro Gly Asp Ala Lys Asn Pro Asn Gly Lys

305 310 315 320

Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Met Ile Ala Glu Gln

325 330 335

Ala Gly Gly Lys Gly Ser Thr Gly Val Ala Arg Val Leu Asp Ile Val

340 345 350

Pro Glu Lys Val His Gln Arg Val Pro Phe Phe Val Gly Ser Lys Asn

355 360 365

Glu Val Ala Tyr Leu Glu Ser Phe Met

370 375

<210>180

<211>1254

<212>DNA

＜213〉Phaeodactylum tricornutum (Phaeodactylum tricornutum)

<400>180

atgtttattt tgaagtcccc ggcgctttgg ttgcttcttt acccagttgt tgcttttacg 60

gcggcgaggg cgaactcgat tcgtccagcg gccgcgttat ctgtcttcga tctgtcgagc 120

gtggaggcag taccgtctag aaagaccaag gccccaattt tcgatgaagt ttgcgacaca 180

actggagtca ctctcaaacg cttcatgacc gaggtttctc tactgaatcc tgaaatcgaa 240

gagctgacga cgctgtttgg tgcgattgaa accgcttgca aagccattgc aaatcttgtc 300

aagcgatcgc cgcttccctc cagtgacacg ttgggtcttc agggagaaat caacgttcaa 360

ggcgaagacc aaaagaaatt agatgtgatc gccaacgata ttttgaagcg agcactccgc 420

ttcacgggcc gcctcggagt cttagcctcg gaagaagagg atactcccgt cgatttgatg 480

ccaagggatc ctagtaccaa aaaagttcta atcgatgagg gagaaaagta tgtcgctgtc 540

ttcgatccgc tcgatggtag ctcaaacgtt gatgcaggca taccgacagg cacaataatt 600

gggatatacg agcacgacga aacttgcaag attgatcctg atgctttgga agaggatcgg 660

accaaacaag aaaacctatg cctcgcaaat actctgcagc ccggcaccaa cttggtagca 720

gcggcgtact gtttatattc ttcgtcaaca tttttggtgt tgacgctggg agctggaaca 780

tatggattca cgctagatga gactatcggt gaatttgtct tgagccatcc aaacattaag 840

attcctgaat gctcatccat tatgtcgttc aacgaggcaa atactcccag ctgggatcgt 900

ccgcttcaag acactttcgc aaagtggagg acagggacag gaaagagcgg caagaaattt 960

tcaagtcgct acattggttc tatggtaggg gatgtccatc ggacgctcct gtacggagga 1020

gtttttgggt atcctggcga caaaaagaat cccaacggga agctacgcct gctttatgaa 1080

ggagctccaa tgtcattcat catggaacag gcgggtggat tgtcgaccac tggcacgcag 1140

cgtgtcatgg aaatctcccc agatacggtc catcaacgtg tgccgattat catgggatcc 1200

agacaagatg tcgaagaggt catggacgcc tataaaaact ttggcatcga ataa 1254

<210>181

<211>417

<212>PRT

＜213〉Phaeodactylum tricornutum

<400>181

Met Phe Ile Leu Lys Ser Pro Ala Leu Trp Leu Leu Leu Tyr Pro Val

1 5 10 15

Val Ala Phe Thr Ala Ala Arg Ala Asn Ser Ile Arg Pro Ala Ala Ala

20 25 30

Leu Ser Val Phe Asp Leu Ser Ser Val Glu Ala Val Pro Ser Arg Lys

35 40 45

Thr Lys Ala Pro Ile Phe Asp Glu Val Cys Asp Thr Thr Gly Val Thr

50 55 60

Leu Lys Arg Phe Met Thr Glu Val Ser Leu Leu Asn Pro Glu Ile Glu

65 70 75 80

Glu Leu Thr Thr Leu Phe Gly Ala Ile Glu Thr Ala Cys Lys Ala Ile

85 90 95

Ala Asn Leu Val Lys Arg Ser Pro Leu Pro Ser Ser Asp Thr Leu Gly

100 105 110

Leu Gln Gly Glu Ile Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp

115 120 125

Val Ile Ala Asn Asp Ile Leu Lys Arg Ala Leu Arg Phe Thr Gly Arg

130 135 140

Leu Gly Val Leu Ala Ser Glu Glu Glu Asp Thr Pro ValAsp Leu Met

145 150 155 160

Pro Arg Asp Pro Ser Thr Lys Lys Val Leu Ile Asp Glu Gly Glu Lys

165 170 175

Tyr Val Ala Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Val Asp Ala

180 185 190

Gly Ile Pro Thr Gly Thr Ile Ile Gly Ile Tyr Glu His Asp Glu Thr

195 200 205

Cys Lys Ile Asp Pro Asp Ala Leu Glu Glu Asp Arg Thr Lys Gln Glu

210 215 220

Asn Leu Cys Leu Ala Asn Thr Leu Gln Pro Gly Thr Asn Leu Val Ala

225 230 235 240

Ala Ala Tyr Cys Leu Tyr Ser Ser Ser Thr Phe Leu Val Leu Thr Leu

245 250 255

Gly Ala Gly Thr Tyr Gly Phe Thr Leu Asp Glu Thr Ile Gly Glu Phe

260 265 270

Val Leu Ser His Pro Asn Ile Lys Ile Pro Glu Cys Ser Ser Ile Met

275 280 285

Ser Phe Asn Glu Ala Asn Thr Pro Ser Trp Asp Arg Pro Leu Gln Asp

290 295 300

Thr Phe Ala Lys Trp Arg Thr Gly Thr Gly Lys Ser Gly Lys Lys Phe

305 310 315 320

Ser Ser Arg Tyr Ile Gly Ser Met Val Gly Asp Val His Arg Thr Leu

325 330 335

Leu Tyr Gly Gly Val Phe Gly Tyr Pro Gly Asp Lys Lys Asn Pro Asn

340 345 350

Gly Lys Leu Arg Leu Leu Tyr Glu Gly Ala Pro Met Ser Phe Ile Met

355 360 365

Glu Gln Ala Gly Gly Leu Ser Thr Thr Gly Thr Gln Arg Val Met Glu

370 375 380

Ile Ser Pro Asp Thr Val His Gln Arg Val Pro Ile Ile Met Gly Ser

385 390 395 400

Arg Gln Asp Val Glu Glu Val Met Asp Ala Tyr Lys Asn Phe Gly Ile

405 410 415

Glu

<210>182

<211>1233

<212>DNA

＜213〉ripe pea (Pisum sativa)

<400>182

atggttgcaa tggcagcagc aacagcctca tctcagttaa tattctcaaa accttactct 60

ccttcacgtc tttgcccctt ccaactctgt gtctttgatg caaaatcagt gttatcaagt 120

tcaaggagaa agcatgtgaa tggctctggt gttagatgta tggctgtgaa ggaagcaact 180

agtgagacaa agaaaagaag tggatatgag attataacac tgactagttg gttgttgcag 240

caagaacaaa aagggattat tgatgcagaa cttactattg tactttctag tatttctatg 300

gcatgtaaac aaattgcttc tttggttcaa agagccaata tttctaacct cactggtact 360

caaggtgctg taaatattca aggggaagac cagaaaaaac ttgatgttat ctcaaatgag 420

gtattctcaa attgcttgag gtcaagtggg aggacaggga taattgcgtc ggaggaagag 480

gatgtcgcgg tggcagtaga agagagttat tcaggaaact acattgttgt atttgatcca 540

cttgatggtt catccaatct tgatgctgca gtctcaaccg gttccatttt cgggatttac 600

agccccaatg acgagtgtct tcctgatttt ggtgatgact ctgatgacaa cacacttggc 660

acagaagaac aaaggtgcat tgtgaatgtg tgtcaaccag gaagcaacct tctagcagct 720

ggctactgca tgtattctag ttcagtagct tttgttctta ccataggcaa aggagtgttt 780

gtattcacat tagatccatt gtacggagaa ttcgttttga ctcaagagaa tctccaaata 840

ccgaaatcag gggaaatcta ttctttcaat gaagggaatt acaagttgtg ggatgaaaac 900

ttgaagaaat atattgatga tcttaaggaa ccaggtccta gtggcaagcc ttattcagca 960

aggtatattg gtagtttggt tggtgatttt cacaggacac tgttatatgg tggcatttat 1020

ggatacccta gggacaagaa aagtaagaat gggaagctta ggcttttata tgaatgtgct 1080

ccaatgagct tcattgttga acaggctggt ggaaaaggtt cagatggtca tcaaagagta 1140

cttgacattc aacccacaga gattcatcaa cgtgttccac tttacattgg gagcacagaa 1200

gaggtggaga aggttgaaaa gtacttagct taa 1233

<210>183

<211>410

<212>PRT

＜213〉ripe pea

<400>183

Met Val Ala Met Ala Ala Ala Thr Ala Ser Ser Gln Leu Ile Phe Ser

1 5 10 15

Lys Pro Tyr Ser Pro Ser Arg Leu Cys Pro Phe Gln Leu Cys Val Phe

20 25 30

Asp Ala Lys Ser Val Leu Ser Ser Ser Arg Arg Lys His Val Asn Gly

35 40 45

Ser Gly Val Arg Cys Met Ala Val Lys Glu Ala Thr Ser Glu Thr Lys

50 55 60

Lys Arg Ser Gly Tyr Glu Ile Ile Thr Leu Thr Ser Trp Leu Leu Gln

65 70 75 80

Gln Glu Gln Lys Gly Ile Ile Asp Ala Glu Leu Thr Ile Val Leu Ser

85 90 95

Sey Ile Ser Met Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg Ala

100 105 110

Asn Ile Ser Asn Leu Thr Gly Thr Gln Gly Ala Val Asn Ile Gln Gly

115 120 125

Glu Asp Gln Lys Lys Leu Asp Val Ile Ser Asn Glu Val Phe Ser Asn

130 135 140

Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu Glu

145 150 155 160

Asp Val Ala Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile Val

165 170 175

Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Leu Asp Ala Ala Val Ser

180 185 190

Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu Pro

195 200 205

Asp Phe Gly Asp Asp Ser Asp Asp Asn Thr Leu Gly Thr Glu Glu Gln

210 215 220

Arg Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ala Ala

225 230 235 240

Gly Tyr Cys Met Tyr Ser Ser Ser Val Ala Phe Val Leu Thr Ile Gly

245 250 255

Lys Gly Val Phe Val Phe Thr Leu Asp Pro Leu Tyr Gly Glu Phe Val

260 265 270

Leu Thr Gln Glu Asn Leu Gln Ile Pro Lys Ser Gly Glu Ile Tyr Ser

275 280 285

Phe Asn Glu Gly Asn Tyr Lys Leu Trp Asp Glu Asn Leu Lys Lys Tyr

290 295 300

Ile Asp Asp Leu Lys Glu Pro Gly Pro Ser Gly Lys Pro Tyr Ser Ala

305 310 315 320

Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu Tyr

325 330 335

Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys Asn Gly Lys

340 345 350

Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile Val Glu Gln

355 360 365

Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile Gln

370 375 380

Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile Gly Ser Thr Glu

385 390 395 400

Glu Val Glu Lys Val Glu Lys Tyr Leu Ala

405 410

<210>184

<211>1242

<212>DNA

＜213〉trifoliate orange (Poncirus trifoliata)

<400>184

atggttgcag cagcagcgac aacatcatca cagcttctct tttcaagctc tcactctttc 60

tctcggctct ctccttacca aatatgtgtc ttcgattcca aagcactcgt gtcgtcatgt 120

cccagcaacg ttatgaagag aagacatgtt ggtgttgctg ctggggttcg gtgcatggct 180

gttgggacaa catcggaggt tgcaaccaag aagagaagtt cgtatgagat tgaaacgctg 240

acaaactggc tgttgaagca agaacagtct ggcgttattg atgctgagct cactattgtg 300

ctttccagca tttcaacggc gtgcaagcag attgcttctt tggtgcaaag agctggcatt 360

tccaacttga ctggaattca gggtgctgtc aatgttcaag gcgaggacca gaagaagctc 420

gacgtcgttt caaatgaggt gttctcaaac tgtttgagat caagtgggcg aactgggatt 480

atagcatcag aggaagagga tgtaccagtg gcggtagaag agagctactc tggaaactat 540

attgtagttt ttgatccact tgatggatca tccaacattg atgctgcagt gtctactgga 600

tccatctttg gcatatacag cccaaatgat gagtgtcttg ccgatattgg tgatgattct 660

actctgggca atactgaaca aagatgtgta gtgaacgtgt gccagccagg aagcaacctt 720

cttgctgcag gctattgcat gtattcaagc tctgtaatct ttgtgataac tttaggcaac 780

ggagtctttg cattcaccct ggatcccatg tatggagaat ttgttttgac acaagagaac 840

attcagatac ccaaaaccgg aaagatctat gcttttaatg aaggcaacta ccagctttgg 900

gatgacaagt tgaagaagta cattgacgat cttaaggatc caggacccag cggcaagccc 960

tattctgcta ggtacattgg cagcttggtt ggtgatttcc atcgaactct gctctacggc 1020

ggcatttatg gctatccaag agacaagaag agcaagaatg gaaagctgag gctcttgtat 1080

gaatgtgcac caatgagctt catagtggaa caagcaggag gcaaaggatc tgacggccat 1140

caaagagtac ttgacattca acctactgag attcaccagc gtattccttt aaagattgga 1200

agccaggagg aagtggagaa actggagaag tatttggcct aa 1242

<210>185

<211>413

<212>PRT

＜213〉trifoliate orange

<400>185

Met Val Ala Ala Ala Ala Thr Thr Ser Ser Gln Leu Leu Phe Ser Ser

1 5 10 15

Ser His Ser Phe Ser Arg Leu Ser Pro Tyr Gln Ile Cys Val Phe Asp

20 25 30

Ser Lys Ala Leu Val Ser Ser Cys Pro Ser Asn Val Met Lys Arg Arg

35 40 45

His Val Gly Val Ala Ala Gly Val Arg Cys Met Ala Val Gly Thr Thr

50 55 60

Ser Glu Val Ala Thr Lys Lys Arg Ser Ser Tyr Glu Ile Glu Thr Leu

65 70 75 80

Thr Asn Trp Leu Leu Lys Gln Glu Gln Ser Gly Val Ile Asp Ala Glu

85 90 95

Leu Thr Ile Val Leu Ser Ser Ile Ser Thr Ala Cys Lys Gln Ile Ala

100 105 110

Ser Leu Val Gln Arg Ala Gly Ile Ser Asn Leu Thr Gly Ile Gln Gly

115 120 125

Ala Val Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser

130 135 140

Asn Glu Val Phe Ser Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile

145 150 155 160

Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr

165 170 175

Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn

180 185 190

Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro

195 200 205

Asn Asp Glu Cys Leu Ala Asp Ile Gly Asp Asp Ser Thr Leu Gly Asn

210 215 220

Thr Glu Gln Arg Cys Val Val Asn Val Cys Gln Pro Gly Ser Asn Leu

225 230 235 240

Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val Ile

245 250 255

Thr Leu Gly Asn Gly Val Phe Ala Phe Thr Leu Asp Pro Met Tyr Gly

260 265 270

Glu Phe Val Leu Thr Gln Glu Asn Ile Gln Ile Pro Lys Thr Gly Lys

275 280 285

Ile Tyr Ala Phe Asn Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys Leu

290 295 300

Lys Lys Tyr Ile Asp Asp Leu Lys Asp Pro Gly Pro Ser Gly Lys Pro

305 310 315 320

Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr

325 330 335

Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys

340 345 350

Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile

355 360 365

Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu

370 375 380

Asp Ile Gln Pro Thr Glu Ile His Gln Arg Ile Pro Leu Lys Ile Gly

385 390 395 400

Ser Gln Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu Ala

405 410

<210>186

<211>1245

<212>DNA

＜213〉Populus tremuloides (Populus tremuloides)

<400>186

atggttgcac aagcagcagc aataacaact tcatcatcac atcttctctt ctcaacctca 60

cgctcactgt ctcgcccatc tccttcccag ttatgtgtct ttgactcaaa aacacttgtg 120

tcatacccca acagcactag tacttacaag aaaaaacgtg gtggtgatgg gctcaagtgc 180

atggctgtga gcacagcctc ggatgctaaa acaaagaaga gtacgtttga gattcaaaca 240

ctgactggtt ggctgttgaa gcaagaacaa gctggtgtta ttgatgctga gctcactatt 300

gttatatcaa gcat ttcaat ggcatgtaagcagattgctt ctttggtgca aagagctagc 360

atttctaact taactggagt tcaaggttct gttaacgttc aaggagaaga tcagaagaag 420

cttgacgtgg tctctaatga ggtgttctct agctgcttga gatcaagtgg gaggacagga 480

atcatagcat cagaggaaga ggacgtgcca gtggcagtgg aggagagtta ctctggaaac 540

tatatagtgg tttttgaccc actcgatgga tcatccaaca ttgatgctgc agtgtctact 600

ggttccatct ttggaatata cagccccaat gacgaatgcc tggccgatat tggagatgac 660

tccactcttg atcaaacgga acagaggtgt attgtgaatg tgtgccagcc aggaaataac 720

ctccttgttg ctggctactg catgtattca agctcagtga tttttgtgct aactattgga 780

aaaggcgtgt tctctttcag cttggatcca atgtatggag agtttgtttt aactcaagaa 840

aacatccaga taccaaaggc tggaaagatt tattcattta atgaaggaaa ctaccagttg 900

tgggatgaca agctgaagaa gtacattgat gaccttaaag accctggtcc gagtggcaag 960

ccctactccg ctagatacat tggaagcttg gtcggtgact tccaccggac gctgctgtac 1020

ggtggcattt atgggtaccc cagggacaag aagagcaaga atgggaagct gaggcttctg 1080

tatgagtgtg caccgatgag ctttatagtg gaacaagctg gtgggaaagg atcagacggg 1140

catcagagag tactggatat cactcctact gagatacacc agcgtgttcc gctttacata 1200

gggagcgtgg aggaagtgga gaaattggag aagtatttgg cttga 1245

<210>187

<211>414

<212>PRT

＜213〉Populus tremuloides

<400>187

Met Val Ala Gln Ala Ala Ala Ile Thr Thr Ser Ser Ser His Leu Leu

1 5 10 15

Phe Ser Thr Ser Arg Ser Leu Ser Arg Pro Ser Pro Ser Gln Leu Cys

20 25 30

Val Phe Asp Ser Lys Thr Leu Val Ser Tyr Pro Asn Ser Thr Ser Thr

35 40 45

Tyr Lys Lys Lys Arg Gly Gly Asp Gly Leu Lys Cys Met Ala Val Ser

50 55 60

Thr Ala Ser Asp Ala Lys Thr Lys Lys Ser Thr Phe Glu Ile Gln Thr

65 70 75 80

Leu Thr Gly Trp Leu Leu Lys Gln Glu Gln Ala Gly Val Ile Asp Ala

85 90 95

Glu Leu Thr Ile Val Ile Ser Ser Ile Ser Met Ala Cys Lys Gln Ile

100 105 110

Ala Ser Leu Val Gln Arg Ala Ser Ile Ser Asn Leu Thr Gly Val Gln

115 120 125

Gly Ser Val Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val

130 135 140

Ser Asn Glu Val Phe Ser Ser Cys Leu Arg Ser Ser Gly Arg Thr Gly

145 150 155 160

Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser

165 170 175

Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser

180 185 190

Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser

195 200 205

Pro Asn Asp Glu Cys Leu Ala Asp Ile Gly Asp Asp Ser Thr Leu Asp

210 215 220

Gln Thr Glu Gln Arg Cys Ile Val Asn Val Cys Gln Pro Gly Asn Asn

225 230 235 240

Leu Leu Val Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val

245 250 255

Leu Thr Ile Gly Lys Gly Val Phe Ser Phe Ser Leu Asp Pro Met Tyr

260 265 270

Gly Glu Phe Val Leu Thr Gln Glu Asn Ile Gln Ile Pro Lys Ala Gly

275 280 285

Lys Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys

290 295 300

Leu Lys Lys Tyr Ile Asp Asp Leu Lys Asp Pro Gly Pro Ser Gly Lys

305 310 315 320

Pro Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg

325 330 335

Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser

340 345 350

Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe

355 360 365

Ile Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val

370 375 380

Leu Asp Ile Thr Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile

385 390 395 400

Gly Ser Val Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu Ala

405 410

<210>188

<211>1227

<212>DNA

＜213〉potato

<400>188

atggcagcat cagcagccac agcaacagca acaacttcat atttaagtgc tctagacaaa 60

aagactccat ttttatttgc cttagacaaa aagactccat ttttatgccc aaaaagcagc 120

acgaagagaa ggtcatttaa tggaggagta aagtgcatgg caatagagac aacttcaggt 180

tttacagcaa ccaagaaaag gagtggctat gagctgcaaa ctttaacaag ttggctatta 240

agacaagaac aagctggagt gattgatgct gaacttacca tagttatttc aagtatttca 300

atggcttgta aacagattgc ttccttagtc caaagagctg gaatttctaa ccttactgga 360

gttcaaggtg ctgtcaatat tcaaggagaa gatcagaaga aacttgatgt tgtctctaat 420

gaggttttct caaattgtct aagatcaagt ggaaggactg ggattatagc atcagaagaa 480

gaggatgtac ctgtggcagt ggaagagagt tactcaggca actacattgt ggtgtttgat 540

cctcttgatg gatcatcaaa cattgatgct gctgtgtcta ccggttctat ctttggaata 600

tacaacccga atgatgagtg cctcgctgat catggagatg attccacgct tgacaatata 660

gagcagaagt gcattgtgaa tgtatgtcaa ccagggacaa accttcttgc agcaggatac 720

tgcatgtact caagctctgt gatattcgta ctcaccttgg gaaatggcgt tttttccttt 780

aacttggatc cgatgtacgg agaatttgtt ctgactcaag aaaatgtcca aataccaaag 840

tctggaaaga tctattcatt caatgaagga aactaccagc tctgggatga caagttgaag 900

aaatatatcg atgacttgaa ggaccctggc cctagtggca agccttactc tgcaaggtac 960

attggtagtt tggttggtga cttccataga actcttctat atggtggcat ttatggttat 1020

cctagagacc aaaagagcaa gaatggaaag ttgaggcttt tgtacgagtg tgctcccatg 1080

agcttcattg tggaacaagc tggtggtaaa ggatccgatg gtcaccaaag agttctcgat 1140

atccaaccaa ctgaggtaca tcaacgagtt ccattgtaca ttggaagcac agaagaagtt 1200

gaaaaattgg agaagtactt gtcttaa 1227

<210>189

<211>408

<212>PRT

＜213〉potato

<400>189

Met Ala Ala Ser Ala Ala Thr Ala Thr Ala Thr Thr Ser Tyr Leu Ser

1 5 10 15

Ala Leu Asp Lys Lys Thr Pro Phe Leu Phe Ala Leu Asp Lys Lys Thr

20 25 30

Pro Phe Leu Cys Pro Lys Ser Ser Thr Lys Arg Arg Ser Phe Asn Gly

35 40 45

Gly Val Lys Cys Met Ala Ile Glu Thr Thr Ser Gly Phe Thr Ala Thr

50 55 60

Lys Lys Arg Ser Gly Tyr Glu Leu Gln Thr Leu Thr Ser Trp Leu Leu

65 70 75 80

Arg Gln Glu Gln Ala Gly Val Ile Asp Ala Glu Leu Thr Ile Val Ile

85 90 95

Ser Ser Ile Ser Met Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg

100 105 110

Ala Gly Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Val Asn Ile Gln

115 120 125

Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser

130 135 140

Asn Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu

145 150 155 160

Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile

165 170 175

Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val

180 185 190

Ser Thr Gly Ser Ile Phe Gly Ile Tyr Asn Pro Asn Asp Glu Cys Leu

195 200 205

Ala Asp His Gly Asp Asp Ser Thr Leu Asp Asn Ile Glu Gln Lys Cys

210 215 220

Ile Val Asn Val Cys Gln Pro Gly Thr Asn Leu Leu Ala Ala Gly Tyr

225 230 235 240

Cys Met Tyr Ser Ser Ser Val Ile Phe Val Leu Thr Leu Gly Asn Gly

245 250 255

Val Phe Ser Phe Asn Leu Asp Pro Met Tyr Gly Glu Phe Val Leu Thr

260 265 270

Gln Glu Asn Val Gln Ile Pro Lys Ser Gly Lys Ile Tyr Ser Phe Asn

275 280 285

Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys Leu Lys Lys Tyr Ile Asp

290 295 300

Asp Leu Lys Asp Pro Gly Pro Ser Gly Lys Pro Tyr Ser Ala Arg Tyr

305 310 315 320

Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Leu Leu Tyr Gly Gly

325 330 335

Ile Tyr Gly Tyr Pro Arg Asp Gln Lys Ser Lys Asn Gly Lys Leu Arg

340 345 350

Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile Val Glu Gln Ala Gly

355 360 365

Gly Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile Gln Pro Thr

370 375 380

Glu Val His Gln Arg Val Pro Leu Tyr Ile Gly Ser Thr Glu Glu Val

385 390 395 400

Glu Lys Leu Glu Lys Tyr Leu Ser

405

<210>190

<211>1248

<212>DNA

＜213〉spinach (Spinacia oleracea)

<400>190

atggcatcaa taggaccagc aacaacaact gcagtaaaac tgcgtagctc aatcttcaat 60

cctcagtcat ctactctttc cccgtctcaa caatgcatta catttacaaa gtcccttcac 120

tcgtttccta ctgccacccg acataatgtg gcttccgggg ttcgttgtat ggcagccgta 180

ggagaggcgg ctacagaaac aaaggcaagg actagaagta agtacgaaat tgaaacacta 240

acaggctggc tgcttaaaca agaaatggca ggtgttattg atgctgaact taccatcgtt 300

ctttctagca tttcattggc ttgtaaacaa attgcttcct tggttcaacg agctggtatt 360

tctaacttga ctggaattca aggtgctgtc aatatccaag gagaggatca gaagaaactt 420

gatgttgt ctccaatgaggt gttttcgagc tgcttgagat cgagtggaag aacaggaata 480

atagcatcag aagaagagga tgtaccagtg gcagtggaag agagttactc tggaaactat 540

attgttgtgt ttgatccact tgatggttca tccaacattg atgcagctgt ctccactggt 600

tccatctttg gcatttatag ccctaacgat gagtgcattg ttgactctga tcacgacgat 660

gagtcacagc taagtgcaga agaacagagg tgtgtagtga atgtatgtca accaggggat 720

aacctattag cagcagggta ttgtatgtac tcaagctctg ttatcttcgt acttacaatt 780

ggtaaaggtg tgtatgcatt cacattagat ccaatgtatg gtgaattcgt actcacttca 840

gagaaaatcc aaatcccaaa agctgggaag atctattcat tcaatgaagg taactacaaa 900

atgtgggatg ataaattgaa gaagtacatg gatgatctta aagagccagg agagtcacag 960

aaaccgtact cgtctcgtta catagggagt ttagttgggg actttcatag aacactttta 1020

tatggtggga tttatggtta cccaagagat gcaaagagta agaatgggaa attgaggctt 1080

ttgtatgaat gtgcacctat gagttttatt gttgaacaag ctggtggtaa aggttctgat 1140

ggtcatcaaa gaattcttga cattcaaccc accgagatac atcaacgtgt gccactgtac 1200

atcgggagtg tggaggaagt agagaaatta gagaagtact tagcataa 1248

<210>191

<211>415

<212>PRT

＜213〉spinach

<400>191

Met Ala Ser Ile Gly Pro Ala Thr Thr Thr Ala Val Lys Leu Arg Ser

1 5 10 15

Ser Ile Phe Asn Pro Gln Ser Ser Thr Leu Ser Pro Ser Gln Gln Cys

20 25 30

Ile Thr Phe Thr Lys Ser Leu His Ser Phe Pro Thr Ala Thr Arg His

35 40 45

Asn Val Ala Ser Gly Val Arg Cys Met Ala Ala Val Gly Glu Ala Ala

50 55 60

Thr Glu Thr Lys Ala Arg Thr Arg Ser Lys Tyr Glu Ile Glu Thr Leu

65 70 75 80

Thr Gly Trp Leu Leu Lys Gln Glu Met Ala Gly Val Ile Asp Ala Glu

85 90 95

Leu Thr Ile Val Leu Ser Ser Ile Ser Leu Ala Cys Lys Gln Ile Ala

100 105 110

Ser Leu Val Gln Arg Ala Gly Ile Ser Asn Leu Thr Gly Ile Gln Gly

115 120 125

Ala Val Asn Ile Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser

130 135 140

Asn Glu Val Phe Ser Ser Cys Leu Arg Ser Ser Gly Arg Thr Gly Ile

145 150 155 160

Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr

165 170 175

Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn

180 185 190

Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro

195 200 205

Asn Asp Glu Cys Ile Val Asp Ser Asp His Asp Asp Glu Ser Gln Leu

210 215 220

Ser Ala Glu Glu Gln Arg Cys Val Val Asn Val Cys Gln Pro Gly Asp

225 230 235 240

Asn Leu Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe

245 250 255

Val Leu Thr Ile Gly Lys Gly Val Tyr Ala Phe Thr Leu Asp Pro Met

260 265 270

Tyr Gly Glu Phe Val Leu Thr Ser Glu Lys Ile Gln Ile Pro Lys Ala

275 280 285

Gly Lys Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Lys Met Trp Asp Asp

290 295 300

Lys Leu Lys Lys Tyr Met Asp Asp Leu Lys Glu Pro Gly Glu Ser Gln

305 310 315 320

Lys Pro Tyr Ser Ser Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His

325 330 335

Arg Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Ala Lys

340 345 350

Ser Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser

355 360 365

Phe Ile Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg

370 375 380

Ile Leu Asp Ile Gln Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr

385 390 395 400

Ile Gly Ser Val Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu Ala

405 410 415

<210>192

<211>1230

<212>DNA

＜213〉common wheat

<400>192

atggccgccg cgaccaccac cacctcccgc ccgcttctgc tgtcccgcca gcaggcggcg 60

gctagctccc tccaatgccg cctccccagg aggcccggaa gcagcctctt tgccggccag 120

ggccaggcgt cgactccgaa tgtgcggtgc atggcagtcg tggacacggc ctcggcgccg 180

gcgccggcgg cggctaggaa gaggagcagc tacgacatga tcacgctgac gacgtggctg 240

ctgaagcagg agcaggaggg ggtcatcgac aacgagatga ccatcgtgct gtccagcata 300

tccacggcgt gcaagcagat cgcctcgttg gtgcagcgcg cgcccatctc caacctcacc 360

ggcgtccagg gcgccaccaa cgtgcagggc gaggaccaga agaagctcga cgtcatctcc 420

aacgaggtgt tctcgaactg cctgaggtgg agtggccgca ccggcgtgat cgcatcggag 480

gaggaggacg tgccggtggc ggtggaggag agctactcgg gcaactacat cgtggtgttc 540

gacccgctcg acggctcctc caacatcgac gccgccgtct ccaccggctc catcttcggc 600

atctacagcc catccgacga gtgccacatt ggcgacgacg caacccttga cgaagtgacg 660

cagatgtgca tagtgaacgt gtgccagcca gggagcaacc tgctcgccgc cggctactgc 720

atgtactcga gctcggtcat cttcgtgctc accatcggca ccggggtgta cgtgttcacg 780

ctggacccga tgtacggcga gttcgtgctg acgcaggaga aggtgcagat cccaaagtcg 840

ggcaagatct actccttcaa cgagggcaac tacgcgctct gggacgacaa gctcaagaag 900

tacatggaca gcctcaagga gcccggcacc tccggcaagc cctactccgc gcgctacatc 960

ggcagcctcg tcggcgactt ccaccgcacc atgctctacg gcggcatcta cgggtacccc 1020

agcgaccaga agagcaagaa cggcaagctg cggctgctct acgagtgcgc gcccatgagc 1080

ttcatcgccg agcaggccgg cggcaaaggc tccgacggcc accagagggt actcgacatc 1140

atgcccacag cggtccatca gagagtgcct ctgtacgtcg ggagcgtgga ggaagtggag 1200

aaggtggaga aattcttgtc ttcagagtag 1230

<210>193

<211>409

<212>PRT

＜213〉common wheat

<400>193

Met Ala Ala Ala Thr Thr Thr Thr Ser Arg Pro Leu Leu Leu Ser Arg

1 5 10 15

Gln Gln Ala Ala Ala Ser Ser Leu Gln Cys Arg Leu Pro Arg Arg Pro

20 25 30

Gly Ser Ser Leu Phe Ala Gly Gln Gly Gln Ala Ser Thr Pro Asn Val

35 40 45

Arg Cys Met Ala Val Val Asp Thr Ala Ser Ala Pro Ala Pro Ala Ala

50 55 60

Ala Arg Lys Arg Ser Ser Tyr Asp Met Ile Thr Leu Thr Thr Trp Leu

65 70 75 80

Leu Lys Gln Glu Gln Glu Gly Val Ile Asp Asn Glu Met Thr Ile Val

85 90 95

Leu Ser Ser Ile Ser Thr Ala Cys Lys Gln Ile Ala Ser Leu Val Gln

100 105 110

Arg Ala Pro Ile Ser Asn Leu Thr Gly Val Gln Gly Ala Thr Asn Val

115 120 125

Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Ile Ser Asn Glu Val Phe

130 135 140

Ser Asn Cys Leu Arg Trp Ser Gly Arg Thr Gly Val Ile Ala Ser Glu

145 150 155 160

Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr

165 170 175

Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala

180 185 190

Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser Pro Ser Asp Glu Cys

195 200 205

His Ile Gly Asp Asp Ala Thr Leu Asp Glu Val Thr Gln Met Cys Ile

210 215 220

Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ala Ala Gly Tyr Cys

225 230 235 240

Met Tyr Ser Ser Ser Val Ile Phe Val Leu Thr Ile Gly Thr Gly Val

245 250 255

Tyr Val Phe Thr Leu Asp Pro Met Tyr Gly Glu Phe Val Leu Thr Gln

260 265 270

Glu Lys Val Gln Ile Pro Lys Ser Gly Lys Ile Tyr Ser Phe Asn Glu

275 280 285

Gly Asn Tyr Ala Leu Trp Asp Asp Lys Leu Lys Lys Tyr Met Asp Ser

290 295 300

Leu Lys Glu Pro Gly Thr Ser Gly Lys Pro Tyr Ser Ala Arg Tyr Ile

305 310 315 320

Gly Ser Leu Val Gly Asp Phe His Arg Thr Met Leu Tyr Gly Gly Ile

325 330 335

Tyr Gly Tyr Pro Ser Asp Gln Lys Ser Lys Asn Gly Lys Leu Arg Leu

340 345 350

Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile Ala Glu Gln Ala Gly Gly

355 360 365

Lys Gly Ser Asp Gly His Gln Arg Val Leu Asp Ile Met Pro Thr Ala

370 375 380

Val His Gln Arg Val Pro Leu Tyr Val Gly Ser Val Glu Glu Val Glu

385 390 395 400

Lys Val Glu Lys Phe Leu Ser Ser Glu

405

<210>194

<211>1242

<212>DNA

＜213〉Zea mays

<400>194

atggccgccg ccgccaccac ctcctcatcc tcccacttgc tcctcctctc ccgccagcag 60

gcggcctccc tacgatgccg cctctccttc ctcggccagc ccgccagaag gtccggcagg 120

gtcacggccc aggcgccggc cgctaaggac gtgcggtgca tggcggccgt ggacactgcg 180

gcgtccgcgg cggcggcgga gacgagcccc aagtcgagca gctacgagat cgtgacgctc 240

acgacgtggc tgctgcagca ggagcggacc ggcgcgatcg acaacgagat gaccatcgtg 300

ctggccagca tatccacggc gtgcaagcag atcgccgcgc tggtgcagcg cgcgcccatc 360

tccaacctca cgggcgttca gggcgccgtc aacgtgcagg gcgaggacca gaagaagctc 420

gatgtcgtct ccaacgaggt gttctccaac tgcctcaagt cgagcgggcg caccggcgtg 480

atcgcctcgg aggaggagga cgtgcccgta gcggtggagc agagctactc cggcaactac 540

atcgtcgtgt tcgaccctct cgacggctcc tccaacatcg acgccgccgt ctccactggc 600

tccatcttcg gcatctacaa ccccaacgac gagtgcctcg ccgacgtcga cgacaacgac 660

acccttgatt cggtggagca gaggtgcatc gtgaacgtgt gccagccggg gagcaacctg 720

ctggccgccg gctactgcat gtactcgagc tcggtgatct tcgtgctcac cgtcggcacc 780

ggggtgtacg tgttcacgct ggaccccatg tacggcgagt tcgtgctgac gcaggagaag 840

gtgcagatcc ccaaggcggg caagatctac gccttcaacg agggcaacta cgcgctctgg 900

gacgacaagc tgaagctgta catggacagc ctcaaggagc ccggcgactc ggggaagccc 960

tactccgcgc ggtacatcgg cagcctcgtc ggcgacttcc accgcactct gctctacgga 1020

gggatctacg ggtaccccag ggacaagaag agcaagaacg gcaagctgcg gcttctctac 1080

gagtgcgccc ccatgagctt catcgtcgag caggccggtg gcaagggctc tgacggccac 1140

cagagaattc ttgacatcac acctacagag atccaccaaa gagtgcctct gtacattggg 1200

agcgtggagg aagtggacaa ggtggagaaa ttcctggctt ga 1242

<210>195

<211>413

<212>PRT

＜213〉Zea mays

<400>195

Met Ala Ala Ala Ala Thr Thr Ser Ser Ser Ser His Leu Leu Leu Leu

1 5 10 15

Ser Arg Gln Gln Ala Ala Ser Leu Arg Cys Arg Leu Ser Phe Leu Gly

20 25 30

Gln Pro Ala Arg Arg Ser Gly Arg Val Thr Ala Gln Ala Pro Ala Ala

35 40 45

Lys Asp Val Arg Cys Met Ala Ala Val Asp Thr Ala Ala Ser Ala Ala

50 55 60

Ala Ala Glu Thr Ser Pro Lys Ser Ser Ser Tyr Glu Ile Val Thr Leu

65 70 75 80

Thr Thr Trp Leu Leu Gln Gln Glu Arg Thr Gly Ala Ile Asp Asn Glu

85 90 95

Met Thr Ile Val Leu Ala Ser Ile Ser Thr Ala Cys Lys Gln Ile Ala

100 105 110

Ala Leu Val Gln Arg Ala Pro Ile Ser Asn Leu Thr Gly Val Gln Gly

115 120 125

Ala Val Asn Val Gln Gly Glu Asp Gln Lys Lys Leu Asp Val Val Ser

130 135 140

Asn Glu Val Phe Ser Asn Cys Leu Lys Ser Ser Gly Arg Thr Gly Val

145 150 155 160

Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Gln Ser Tyr

165 170 175

Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn

180 185 190

Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Asn Pro

195 200 205

Asn Asp Glu Cys Leu Ala Asp Val Asp Asp Asn Asp Thr Leu Asp Ser

210 215 220

Val Glu Gln Arg Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu

225 230 235 240

Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val Leu

245 250 255

Thr Val Gly Thr Gly Val Tyr Val Phe Thr Leu Asp Pro Met Tyr Gly

260 265 270

Glu Phe Val Leu Thr Gln Glu Lys Val Gln Ile Pro Lys Ala Gly Lys

275 280 285

Ile Tyr Ala Phe Asn Glu Gly Asn Tyr Ala Leu Trp Asp Asp Lys Leu

290 295 300

Lys Leu Tyr Met Asp Ser Leu Lys Glu Pro Gly Asp Ser Gly Lys Pro

305 310 315 320

Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr

325 330 335

Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser Lys

340 345 350

Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Phe Ile

355 360 365

Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Ile Leu

370 375 380

Asp Ile Thr Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile Gly

385 390 395 400

Ser Val Glu Glu Val Asp Lys Val Glu Lys Phe Leu Ala

405 410

<210>196

<211>1266

<212>DNA

＜213〉small liwan moss (Physicomitrella patens)

<400>196

atggcgacca cacaagcgat tctctctgcc acccttgcca tagccccggc ttccagctgc 60

gagacttcgt cacggagccc ggcgtccacc aaaacttgtc tctcagtggc aggatcgtcg 120

ctgcatggct cagtggccgg actcggagct gggaaacaga ttgtgagcgt gcagaggaag 180

agcgttgccg tgagggccgc cgttgcagct gagactgccg ctcccaagca gcaggcgaag 240

agccagtatg acatcactac cctgacgacg tggttgctga agaaagagca ggcgggcgtc 300

atcgatggcg agctcaccat tgtgctctcc agcatcgccc tggcttgcaa gcaaattgcg 360

tctctggtgc agagggctgg catctccaac atgactgggt tgcaaggagc tgctaacatt 420

caaggggagg accagaagaa gctagacgtt atttcgaacg aggtgttctc aagctgtctg 480

cgctcaagcg gacggacagg catcatcgct tctgaggaag aagacacccc ggttgcagtg 540

gaggagagct actctggcaa ctacattgtg gtgttcgacc ctcttgacgg ctcttccaac 600

atcgatgctg ctgtttccac ggggtcaatc tggggaatct acaagcccaa cgaggaatgc 660

ctgaccaatc tcggagagga gccaactatt gatgagatcg ccgaaaactg cgtcgtcaat 720

gtctgtcaac cagggagcaa tttgttgtcc gccgggtact gcatgtactc gagctccgtc 780

atcctcgttc tctcagtcgg tgatggagtc tacggcttca ctctggaccc tctctacgga 840

gaattcgtcc tctcgcacga caacatccaa attccaaaat ctggtaagat ctattccatg 900

aatgaaggca actacgccct ctgggatgac aacctcaaga agtacgtcga cagcttgaag 960

gaccccggtc ccagcggcaa gccctactcc gctcgttaca tcggcagtct ggtgggcgac 1020

ttccacagga caatgctgta tggtggcatc tatggctacc ccagggattc caagagcaag 1080

aacgggaagt tgaggttgct ttacgagtgt gctcccatga gctacctcgc tgaacaagca 1140

ggtgggaagg gctccgacgg tcaccagagg attctggaca tccaacctga gcaggttcac 1200

caacgtgtgc cattgtacgt tggaagcacg gaggaggtgg agaagttgga gaagttctta 1260

gcttaa 1266

<210>197

<211>421

<212>PRT

＜213〉small liwan moss

<400>197

Met Ala Thr Thr Gln Ala Ile Leu Ser Ala Thr Leu Ala Ile Ala Pro

1 5 10 15

Ala Ser Ser Cys Glu Thr Ser Ser Arg Ser Pro Ala Ser Thr Lys Thr

20 25 30

Cys Leu Ser Val Ala Gly Ser Ser Leu His Gly Ser Val Ala Gly Leu

35 40 45

Gly Ala Gly Lys Gln Ile Val Ser Val Gln Arg Lys Ser Val Ala Val

50 55 60

Arg Ala Ala Val Ala Ala Glu Thr Ala Ala Pro Lys Gln Gln Ala Lys

65 70 75 80

Ser Gln Tyr Asp Ile Thr Thr Leu Thr Thr Trp Leu Leu Lys Lys Glu

85 90 95

Gln Ala Gly Val Ile Asp Gly Glu Leu Thr Ile Val Leu Ser Ser Ile

100 105 110

Ala Leu Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg Ala Gly Ile

115 120 125

Ser Asn Met Thr Gly Leu Gln Gly Ala Ala Asn Ile Gln Gly Glu Asp

130 135 140

Gln Lys Lys Leu Asp Val Ile Ser Asn Glu Val Phe Ser Ser Cys Leu

145 150 155 160

Arg Ser Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu Glu Asp Thr

165 170 175

Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile Val Val Phe

180 185 190

Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val Ser Thr Gly

195 200 205

Ser Ile Trp Gly Ile Tyr Lys Pro Asn Glu Glu Cys Leu Thr Asn Leu

210 215 220

Gly Glu Glu Pro Thr Ile Asp Glu Ile Ala Glu Asn Cys Val Val Asn

225 230 235 240

Val Cys Gln Pro Gly Ser Asn Leu Leu Ser Ala Gly Tyr Cys Met Tyr

245 250 255

Ser Ser Ser Val Ile Leu Val Leu Ser Val Gly Asp Gly Val Tyr Gly

260 265 270

Phe Thr Leu Asp Pro Leu Tyr Gly Glu Phe Val Leu Ser His Asp Asn

275 280 285

Ile Gln Ile Pro Lys Ser Gly Lys Ile Tyr Ser Met Asn Glu Gly Asn

290 295 300

Tyr Ala Leu Trp Asp Asp Asn Leu Lys Lys Tyr Val Asp Ser Leu Lys

305 310 315 320

Asp Pro Gly Pro Ser Gly Lys Pro Tyr Ser Ala Arg Tyr Ile Gly Ser

325 330 335

Leu Val Gly Asp Phe His Arg Thr Met Leu Tyr Gly Gly Ile Tyr Gly

340 345 350

Tyr Pro Arg Asp Ser Lys Ser Lys Asn Gly Lys Leu Arg Leu Leu Tyr

355 360 365

Glu Cys Ala Pro Met Ser Tyr Leu Ala Glu Gln Ala Gly Gly Lys Gly

370 375 380

Ser Asp Gly His Gln Arg Ile Leu Asp Ile Gln Pro Glu Gln Val His

385 390 395 400

Gln Arg Val Pro Leu Tyr Val Gly Ser Thr Glu Glu Val Glu Lys Leu

405 410 415

Glu Lys Phe Leu Ala

420

<210>198

<211>1263

<212>DNA

<213>Galderia sulphuraria

<400>198

ggcacgagga aagatgtgca agtatacttt attcaagtcc tactatatct ttacaaagaa 60

gagtgtcaag aaataaccac tgcaaaagat atcttcctta taagatggat atccatcaaa 120

atgatagctg ttcaacccca acccaaaaag actcccgtag cagaatattt acaaacctca 180

caagacacac caacttcact cactcgttac ttgttggaag tagccaaaca aaataaggat 240

atgggagata tggtggcatt gataaacggt attcaatttg cttgcaaaaa gatagcttca 300

ttggttggta aagcaggggt cactgacttg atgggaatct atcaacaagg catagtcaac 360

gttcacggag aagaacagaa aaagttggat gttctttcta atgaagtatt gaagaacgct 420

ctcaaatatt cgggaaaaat ggctgtcata gcttcggaag aagaagacgt tcctatcatg 480

gtagaagaaa gttattccgg taactatgta gtcgtgtttg atccattaga tggttcttcc 540

aatttggatg ctggattgcc tactggaact atatttggag tgtttcaaca acaattctcg 600

tgtctcattc atgactatga ggagtccatc gataacatgg aattggcttg tttacaaaac 660

actttacaac caggacgtag attgattgcc gctggatatt gtatctattc ttcttctacc 720

atgttggtac tctcattggg taatggtctt cattgtttta ctttggatac ggaagttggt 780

gaatttgtat tgactcgtgc taacatccaa attceacaaa gaggtaatat ttattctttc 840

aacgagtcca acttttatca atgggataaa ggagttcaag attatataga gaggttaaaa 900

aaaggaaaca atcaaacaaa ttgtcgttat tccgcaagat atgttggctc catggttgct 960

gatgtacatc gtacaatatt gtatggcgga atatttggtt atccagcaga taaaaagaat 1020

gtctctggta aattgagatt ggtatatgaa tgtgcaccga tggcatattt ggttgaacaa 1080

gcaggaggta aagcaaccac gggaatagaa aatattttag atttgacacc aaaagatatt 1140

cacgaaagga agcctcttat attaggttct ccagcagata tcgaagaatt tctgcaagta 1200

tatggaatgt cacgagttga tagagaagat atgcatatgt gggataactt gagtagttta 1260

taa 1263

<210>199

<211>420

<212>PRT

<213>Galderia sulphuraria

<400>199

Gly Thr Arg Lys Asp Val Gln Val Tyr Phe Ile Gln Val Leu Leu Tyr

1 5 10 15

Leu Tyr Lys Glu Glu Cys Gln Glu Ile Thr Thr Ala Lys Asp Ile Phe

20 25 30

Leu Ile Arg Trp Ile Ser Ile Lys Met Ile Ala Val Gln Pro Gln Pro

35 40 45

Lys Lys Thr Pro Val Ala Glu Tyr Leu Gln Thr Ser Gln Asp Thr Pro

50 55 60

Thr Ser Leu Thr Arg Tyr Leu Leu Glu Val Ala Lys Gln Asn Lys Asp

65 70 75 80

Met Gly Asp Met Val Ala Leu Ile Asn Gly Ile Gln Phe Ala Cys Lys

85 90 95

Lys Ile Ala Ser Leu Val Gly Lys Ala Gly Val Thr Asp Leu Met Gly

100 105 110

Ile Tyr Gln Gln Gly Ile Val Asn Val His Gly Glu Glu Gln Lys Lys

115 120 125

Leu Asp Val Leu Ser Asn Glu Val Leu Lys Asn Ala Leu Lys Tyr Ser

130 135 140

Gly Lys Met Ala Val Ile Ala Ser Glu Glu Glu Asp Val Pro Ile Met

145 150 155 160

Val Glu Glu Ser Tyr Ser Gly Asn Tyr Val Val Val Phe Asp Pro Leu

165 170 175

Asp Gly Ser Ser Asn Leu Asp Ala Gly Leu Pro Thr Gly Thr Ile Phe

180 185 190

Gly Val Phe Gln Gln Gln Phe Ser Cys Leu Ile His Asp Tyr Glu Glu

195 200 205

Ser Ile Asp Asn Met Glu Leu Ala Cys Leu Gln Asn Thr Leu Gln Pro

210 215 220

Gly Arg Arg Leu Ile Ala Ala Gly Tyr Cys Ile Tyr Ser Ser Ser Thr

225 230 235 240

Met Leu Val Leu Ser Leu Gly Asn Gly Leu His Cys Phe Thr Leu Asp

245 250 255

Thr Glu Val Gly Glu Phe Val Leu Thr Arg Ala Asn Ile Gln Ile Pro

260 265 270

Gln Arg Gly Asn Ile Tyr Ser Phe Asn Glu Ser Asn Phe Tyr Gln Trp

275 280 285

Asp Lys Gly Val Gln Asp Tyr Ile Glu Arg Leu Lys Lys Gly Asn Asn

290 295 300

Gln Thr Asn Cys Arg Tyr Ser Ala Arg Tyr Val Gly Ser Met Val Ala

305 310 315 320

Asp Val His Arg Thr Ile Leu Tyr Gly Gly Ile Phe Gly Tyr Pro Ala

325 330 335

Asp Lys Lys Asn Val Ser Gly Lys Leu Arg Leu Val Tyr Glu Cys Ala

340 345 350

Pro Met Ala Tyr Leu Val Glu Gln Ala Gly Gly Lys Ala Thr Thr Gly

355 360 365

Ile Glu Asn Ile Leu Asp Leu Thr Pro Lys Asp Ile His Glu Arg Lys

370 375 380

Pro Leu Ile Leu Gly Ser Pro Ala Asp Ile Glu Glu Phe Leu Gln Val

385 390 395 400

Tyr Gly Met Ser Arg Val Asp Arg Glu Asp Met His Met Trp Asp Asn

405 410 415

Leu Ser Ser Leu

420

<210>200

<211>710

<212>DNA

＜213〉marchantia (Marchantia polymorpha)

<400>200

gaggaggata ccccagtcgc cgtcgaagag agctactcag gaaactacgt cgttgtcttc 60

gatcctctcg atggatcctc caacatcgac gctgcggtat ctaccggatc catctttgga 120

atctacaggc ccactgagga gtgtcttgca gacatggacg acgactcaca gctcggtatg 180

gtggaacaaa actgcatcgt gaacgtatgc cagcccggta gcaacctcct atccgctggg 240

tactgcatgt actccagctc cgtcattttg gtgttgtcag tcggagacgg tgtctatgga 300

ttcacactgg atcccctgta cggtgaattc gtcatgaccc acgacaacat caagatccca 360

aagaagggat cgatctactc gttcaatgaa ggtaactacg cactctggga tgacaagctc 420

aagaagtaca tcgactcact gaaggacccc gaacccaccg gcaagcctta ctctgctcgt 480

tacatcggta gtctggtcgg tgacttccac agaaccatgc tctatggtgg catctacgga 540

taccctgccg acaagaaaag caagaacgga aagttgagac ttctgtacga gtgtgctcct 600

atgagctact tggcagagca ggccggagga aagggttctg atggttaccg cagagttctg 660

gaaatcgagc ctgagcaggt acatcagcga gtgccacttt tcgtcggaag 710

<210>201

<211>236

<212>PRT

＜213〉marchantia

<400>201

Glu Glu Asp Thr Pro Val Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr

1 5 10 15

Val Val Val Phe Asp Pro Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala

20 25 30

Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Arg Pro Thr Glu Glu Cys

35 40 45

Leu Ala Asp Met Asp Asp Asp Ser Gln Leu Gly Met Val Glu Gln Asn

50 55 60

Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn Leu Leu Ser Ala Gly

65 70 75 80

Tyr Cys Met Tyr Ser Ser Ser Val Ile Leu Val Leu Ser Val Gly Asp

85 90 95

Gly Val Tyr Gly Phe Thr Leu Asp Pro Leu Tyr Gly Glu Phe Val Met

100 105 110

Thr His Asp Asn Ile Lys Ile Pro Lys Lys Gly Ser Ile Tyr Ser Phe

115 120 125

Asn Glu Gly Asn Tyr Ala Leu Trp Asp Asp Lys Leu Lys Lys Tyr Ile

130 135 140

Asp Ser Leu Lys Asp Pro Glu Pro Thr Gly Lys Pro Tyr Ser Ala Arg

145 150 155 160

Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg Thr Met Leu Tyr Gly

165 170 175

Gly Ile Tyr Gly Tyr Pro Ala Asp Lys Lys Ser Lys Asn Gly Lys Leu

180 185 190

Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Tyr Leu Ala Glu Gln Ala

195 200 205

Gly Gly Lys Gly Ser Asp Gly Tyr Arg Arg Val Leu Glu Ile Glu Pro

210 215 220

Glu Gln Val His Gln Arg Val Pro Leu Phe Val Gly

225 230 235

<210>202

<211>765

<212>DNA

＜213〉Herba Tagetis Patulae (Tagetes patula)

<400>202

atcattgcat cggaggaaga agacgtgccg gtggctgtgg aagagagtta ctccggaaac 60

tacattgtcg tgtttgatcc ccttgatggg tcatctaata ttgatgctgc tgtttcaacc 120

gattctattt tcggaatcta tagccccaat gatgagtgtc tcgctgatat tagtgacgac 180

tccacgctag acagtgtgga acaaaaatgt attgtcaacg tatgccagcc cggaagcaat 240

ctactagcag ccggttactg tatgtactca agctccgtaa tcttcgtgct ctcgatcggt 300

actggtgtct acgcgttcac attagaccca atgtacggtg agtttgtact cactcaagaa 360

aagattcaaa tcccgaaatc ggggaagatt tactcgttta acgaaggaaa ctatcaacta 420

tgggacgata aattgaagaa gtacatggat gatctaaagg acccgggccc cacgggcaag 480

ccgtattcgg ctcgctacat tggtagcttg gttggtgatt ttcataggac attattgtac 540

ggagggattt acgggtaccc acgtgacaaa aagagcaaga acgggaagct tcggttgttg 600

tatgagtgtg caccgatgag ttacttggtt gaacaagcgg gtggaaaggg gtcggatgga 660

catcaacgag tgctcgacat tcaaccaacc gagattcatc agcgcgttcc gctatacatt 720

gggagcgtag aggaagtgga aaaattggag aagtatttgg cttga 765

<210>203

<211>254

<212>PRT

＜213〉Herba Tagetis Patulae

<400>203

Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val Ala Val Glu Glu Ser

1 5 10 15

Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro Leu Asp Gly Ser Ser

20 25 30

Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile Phe Gly Ile Tyr Ser

35 40 45

Pro Asn Asp Glu Cys Leu Ala Asp Ile Ser Asp Asp Ser Thr Leu Asp

50 55 60

Ser Val Glu Gln Lys Cys Ile Val Asn Val Cys Gln Pro Gly Ser Asn

65 70 75 80

Leu Leu Ala Ala Gly Tyr Cys Met Tyr Ser Ser Ser Val Ile Phe Val

85 90 95

Leu Ser Ile Gly Thr Gly Val Tyr Ala Phe Thr Leu Asp Pro Met Tyr

100 105 110

Gly Glu Phe Val Leu Thr Gln Glu Lys Ile Gln Ile Pro Lys Ser Gly

115 120 125

Lys Ile Tyr Ser Phe Asn Glu Gly Asn Tyr Gln Leu Trp Asp Asp Lys

130 135 140

Leu Lys Lys Tyr Met Asp Asp Leu Lys Asp Pro Gly Pro Thr Gly Lys

145 150 155 160

Pro Tyr Ser Ala Arg Tyr Ile Gly Ser Leu Val Gly Asp Phe His Arg

165 170 175

Thr Leu Leu Tyr Gly Gly Ile Tyr Gly Tyr Pro Arg Asp Lys Lys Ser

180 185 190

Lys Asn Gly Lys Leu Arg Leu Leu Tyr Glu Cys Ala Pro Met Ser Tyr

195 200 205

Leu Val Glu Gln Ala Gly Gly Lys Gly Ser Asp Gly His Gln Arg Val

210 215 220

Leu Asp Ile Gln Pro Thr Glu Ile His Gln Arg Val Pro Leu Tyr Ile

225 230 235 240

Gly Ser Val Glu Glu Val Glu Lys Leu Glu Lys Tyr Leu Ala

245 250

<210>204

<211>795

<212>DNA

＜213〉flax (Linum usitatissimum)

<220>

<221>misc_feature

<222>(738)..(738)

＜223〉n is a, c, g or t

<400>204

gaggaggaac aacgtcgccg ggggttctgg tttcaaatgc tctgccgtcg ggacgcgccg 60

tcggaggcgg caacgacgac gaagaagagg agtagctacg agattctgac gctgacgacc 120

tggcttctgc agcaggaaca ggccggagtg atcgacgccg agctcacgat tgtgctctcc 180

agcatctcca cggcgtgtaa gcagatcgct tctctagttc agcgatccgg gatttctaac 240

ctcaccggcg tccagggcgc cgtcaatgtc cagggtgagg atcagaagaa gctcgacgtc 300

gtttcgaacg aggtgttttc aaattgcttg aggtcgagcg gcaggacggg gatcatagcg 360

tcggaggaag aagacgtgcc ggtcgccgtg gaggaaagct actccggtaa ctatatcgta 420

gtgttcgatc cacttgatgg atcgtcaaac atcgatgccg cagtctccac cggctcaatc 480

ttcggaatct acagtcccaa cgatgagtgc ctggccgaca tcggagacgg agacgagtca 540

aatctagata ctcaggagca gaagtgtgtg gtgagcgtgt gccagccggg gagcaaccta 600

ctcgccgccg gctactgcat gtactcaagc tcggtgatct tcgtcctcac gatcggaaac 660

ggcgttttcg ccttcaatct ggacccaatg tacggcgagt tggtgttgac tcaagagaac 720

attcagatcc cgaaagcngg aaagatctac tcattcaacg aaggggacta ccaaatgtgg 780

gatgagaaat tgaag 795

<210>205

<211>265

<212>PRT

＜213〉flax

<400>205

Glu Glu Glu Gln Arg Arg Arg Gly Phe Trp Phe Gln Met Leu Cys Arg

1 5 10 15

Arg Asp Ala Pro Ser Glu Ala Ala Thr Thr Thr Lys Lys Arg Ser Ser

20 25 30

Tyr Glu Ile Leu Thr Leu Thr Thr Trp Leu Leu Gln Gln Glu Gln Ala

35 40 45

Gly Val Ile Asp Ala Glu Leu Thr Ile Val Leu Ser Ser Ile Ser Thr

50 55 60

Ala Cys Lys Gln Ile Ala Ser Leu Val Gln Arg Ser Gly Ile Ser Asn

65 70 75 80

Leu Thr Gly Val Gln Gly Ala Val Asn Val Gln Gly Glu Asp Gln Lys

85 90 95

Lys Leu Asp Val Val Ser Asn Glu Val Phe Ser Asn Cys Leu Arg Ser

100 105 110

Ser Gly Arg Thr Gly Ile Ile Ala Ser Glu Glu Glu Asp Val Pro Val

115 120 125

Ala Val Glu Glu Ser Tyr Ser Gly Asn Tyr Ile Val Val Phe Asp Pro

130 135 140

Leu Asp Gly Ser Ser Asn Ile Asp Ala Ala Val Ser Thr Gly Ser Ile

145 150 155 160

Phe Gly Ile Tyr Ser Pro Asn Asp Glu Cys Leu Ala Asp Ile Gly Asp

165 170 175

Gly Asp Glu Ser Asn Leu Asp Thr Gln Glu Gln Lys Cys Val Val Ser

180 185 190

Val Cys Gln Pro Gly Ser Asn Leu Leu Ala Ala Gly Tyr Cys Met Tyr

195 200 205

Ser Ser Ser Val Ile Phe Val Leu Thr Ile Gly Asn Gly Val Phe Ala

210 215 220

Phe Asn Leu Asp Pro Met Tyr Gly Glu Leu Val Leu Thr Gln Glu Asn

225 230 235 240

Ile Gln Ile Pro Lys Ala Gly Lys Ile Tyr Ser Phe Asn Glu Gly Asp

245 250 255

Tyr Gln Met Trp Asp Glu Lys Leu Lys

260 265

<210>206

<211>50

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm08448

<400>206

ggggacaagt ttgtacaaaa aagcaggctt aaacaatggc cgccaccatg 50

<210>207

<211>50

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm08449

<400>207

ggggaccact ttgtacaaga aagctgggta gctgcttagt gcttcttggt 50

<210>208

<211>1167

<212>DNA

＜213〉rice

<400>208

atccaagcca agaagaggga gagcaccaag gacacgcgac tagcagaagc cgagcgaccg 60

ccttcttcga tccatatctt ccggtcgagt tcttggtcga tctcttccct cctccacctc 120

ctcctcacag ggtatgtgcc cttcggttgt tcttggattt attgttctag gttgtgtagt 180

acgggcgttg atgttaggaa aggggatctg tatctgtgat gattcctgtt cttggatttg 240

ggatagaggg gttcttgatg ttgcatgtta tcggttcggt ttgattagta gtatggtttt 300

caatcgtctg gagagctcta tggaaatgaa atggtttagg gtacggaatc ttgcgatttt 360

gtgagtacct tttgtttgag gtaaaatcag agcaccggtg attttgcttg gtgtaataaa 420

agtacggttg tttggtcctc gattctggta gtgatgcttc tcgatttgac gaagctatcc 480

tttgtttatt ccctattgaa caaaaataat ccaactttga agacggtccc gttgatgaga 540

ttgaatgatt gattcttaag cctgtccaaa atttcgcagc tggcttgttt agatacagta 600

gtccccatca cgaaattcat ggaaacagtt ataatcctca ggaacagggg attccctgtt 660

cttccgattt gctttagtcc cagaattttt tttcccaaat atcttaaaaa gtcactttct 720

ggttcagttc aatgaattga ttgctacaaa taatgctttt atagcgttat cctagctgta 780

gttcagttaa taggtaatac ccctatagtt tagtcaggag aagaacttat ccgatttctg 840

atctccattt ttaattatat gaaatgaact gtagcataag cagtattcat ttggattatt 900

ttttttatta gctctcaccc cttcattatt ctgagctgaa agtctggcat gaactgtcct 960

caattttgtt ttcaaattca catcgattat ctatgcatta tcctcttgta tctacctgta 1020

gaagtttctt tttggttatt ccttgactgc ttgattacag aaagaaattt atgaagctgt 1080

aatcgggata gttatactgc ttgttcttat gattcatttc ctttgtgcag ttcttggtgt 1140

agcttgccac tttcaccagc aaagttc 1167

<210>209

<211>1629

<212>DNA

＜213〉rice

<400>209

atgatgggtt gcttcactgt cctgagatcc aagaagaaga agcctcttgc tctcaccaag 60

aaatcggttg atgcaaggga aagcacgtcc tcaagactcc cagagccaga agcgcatgtg 120

ccatcgttac aatctgctcc tcctagtttt aggaacaagg ctaaaatcca ccaatcggaa 180

aagaaagctt cttacagcag agcgcgcgtg ctgtctgctc cttccagcct aattgtggtt 240

gatcaggatg gtcttccata tgccgaattc gatgatcaag atgactccag gggcaaggga 300

ggttctataa agggccaccg tttctctaat ccactgcccc ttcctctccc atcaccagaa 360

ggaaaatcat tgaggaactt tggcagcttc aaagccatca atgcaagtgg accactcgat 420

gcttcaggcc ctctgccact tcctccaaag aagtgtgatg ggcttaagaa tttctcctat 480

gaggaacttt catcagcttg ccaatggttt tctggtgacc agtgtgtttc cgaaagtttg 540

acatcaacat catacaaggc gtcctttagg gatgatttta ccgacccaaa gaccattgaa 600

gcaatagtat ctcggttgct ctcctccact cagagtttga aagagtttaa aacacaagtg 660

aataccttgg catcacttca gcatcccaac ttatgtaaac taatcggctt tcacgcaaga 720

gaagaatcta atgaaaggat gttggtctat gagcgactcc atcatggcag cttagataaa 780

ctactctttg gaagatcgga tggtcgtttc atggactggt cagcacgttt gaaggttgct 840

cttggtgctg ctagaggcct ggctttccta catgatgaag ggccttttca ggccatgtac 900

aatgacttct caacctcaaa catccaaatt gacaaagatt tcactgcaaa gctatcagga 960

tatggatgtg ttggattcaa taccgaggag gaaatatcaa atgcatctgt ggctgctgca 1020

aacctctcag tggaaacctt ggagaaaggt gtactgactc ccaagagcaa cgtatggtgc 1080

tttggagttg tcctgctgga gctaataaca ggaaggaaga accttgatgt ccgttcctca 1140

aaagaagaac gcaatattgt caagtggagt aggcctttcc tcaccgatga tagtcgccta 1200

tcgttaatca tggactcccg tataaaagga cgcttcccta ccaaggctgc tcggattgta 1260

gcagatatca tattgagatg ccttaataaa gatccatcag agaggcctac catgagggcc 1320

gttgtggagt ccctagcaag cgtccaggac ataaaggttc catgtcgata tcctttgcaa 1380

gagccatctg ctgccccaag aaaggtgatg ttaaaatcta caagtctcaa tggcatcatt 1440

catcaccatc ctgtcgtaac cttctcaccg tcacctcctt cgcgaaacca acatttgctc 1500

tcgccaaggt catccacgtc tgcactgctt cctccaagga ccagctgtgc tctggatgac 1560

cctagagtaa gctctatcaa gaaatcgcct tcccctattt tacggagatc tggtgttgag 1620

ggtttttga 1629

<210>210

<211>542

<212>PRT

＜213〉rice

<400>210

Met Met Gly Cys Phe Thr Val Leu Arg Ser Lys Lys Lys Lys Pro Leu

1 5 10 15

Ala Leu Thr Lys Lys Ser Val Asp Ala Arg Glu Ser Thr Ser Ser Arg

20 25 30

Leu Pro Glu Pro Glu Ala His Val Pro Ser Leu Gln Ser Ala Pro Pro

35 40 45

Ser Phe Arg Asn Lys Ala Lys Ile His Gln Ser Glu Lys Lys Ala Ser

50 55 60

Tyr Ser Arg Ala Arg Val Leu Ser Ala Pro Ser Ser Leu Ile Val Val

65 70 75 80

Asp Gln Asp Gly Leu Pro Tyr Ala Glu Phe Asp Asp Gln Asp Asp Ser

85 90 95

Arg Gly Lys Gly Gly Ser Ile Lys Gly His Arg Phe Ser Asn Pro Leu

100 105 110

Pro Leu Pro Leu Pro Ser Pro Glu Gly Lys Ser Leu Arg Asn Phe Gly

115 120 125

Ser Phe Lys Ala Ile Asn Ala Ser Gly Pro Leu Asp Ala Ser Gly Pro

130 135 140

Leu Pro Leu Pro Pro Lys Lys Cys Asp Gly Leu Lys Asn Phe Ser Tyr

145 150 155 160

Glu Glu Leu Ser Ser Ala Cys Gln Trp Phe Ser Gly Asp Gln Cys Val

165 170 175

Ser Glu Ser Leu Thr Ser Thr Ser Tyr Lys Ala Ser Phe Arg Asp Asp

180 185 190

Phe Thr Asp Pro Lys Thr Ile Glu Ala Ile Val Ser Arg Leu Leu Ser

195 200 205

Ser Thr Gln Ser Leu Lys Glu Phe Lys Thr Gln Val Asn Thr Leu Ala

210 215 220

Ser Leu Gln His Pro Asn Leu Cys Lys Leu Ile Gly Phe His Ala Arg

225 230 235 240

Glu Glu Ser Asn Glu Arg Met Leu Val Tyr Glu Arg Leu His His Gly

245 250 255

Ser Leu Asp Lys Leu Leu Phe Gly Arg Ser Asp Gly Arg Phe Met Asp

260 265 270

Trp Ser Ala Arg Leu Lys Val Ala Leu Gly Ala Ala Arg Gly Leu Ala

275 280 285

Phe Leu His Asp Glu Gly Pro Phe Gln Ala Met Tyr Asn Asp Phe Ser

290 295 300

Thr Ser Asn Ile Gln Ile Asp Lys Asp Phe Thr Ala Lys Leu Ser Gly

305 310 315 320

Tyr Gly Cys Val Gly Phe Asn Thr Glu Glu Glu Ile Ser Asn Ala Ser

325 330 335

Val Ala Ala Ala Asn Leu Ser Val Glu Thr Leu Glu Lys Gly Val Leu

340 345 350

Thr Pro Lys Ser Asn Val Trp Cys Phe Gly Val Val Leu Leu Glu Leu

355 360 365

Ile Thr Gly Arg Lys Asn Leu Asp Val Arg Ser Ser Lys Glu Glu Arg

370 375 380

Asn Ile Val Lys Trp Ser Arg Pro Phe Leu Thr Asp Asp Ser Arg Leu

385 390 395 400

Ser Leu Ile Met Asp Ser Arg Ile Lys Gly Arg Phe Pro Thr Lys Ala

405 410 415

Ala Arg Ile Val Ala Asp Ile Ile Leu Arg Cys Leu Asn Lys Asp Pro

420 425 430

Ser Glu Arg Pro Thr Met Arg Ala Val Val Glu Ser Leu Ala Ser Val

435 440 445

Gln Asp Ile Lys Val Pro Cys Arg Tyr Pro Leu Gln Glu Pro Ser Ala

450 455 460

Ala Pro Arg Lys Val Met Leu Lys Ser Thr Ser Leu Asn Gly Ile Ile

465 470 475 480

His His His Pro Val Val Thr Phe Ser Pro Ser Pro Pro Ser Arg Asn

485 490 495

Gln His Leu Leu Ser Pro Arg Ser Ser Thr Ser Ala Leu Leu Pro Pro

500 505 510

Arg Thr Ser Cys Ala Leu Asp Asp Pro Arg Val Ser Ser Ile Lys Lys

515 520 525

Ser Pro Ser Pro Ile Leu Arg Arg Ser Gly Val Glu Gly Phe

530 535 540

<210>211

<211>1674

<212>DNA

＜213〉Arabidopis thaliana

<400>211

atggttttgg ggtgtttccc tttgaaaagc aagaagaaac gtggctctgt ttctatgaag 60

cggttggatc ttgaagaaag caagccaact gctttacctg agccaccaaa gattccaagt 120

cgtaatttac aatcaggtcc tccgagtttc agaaatcgtg tgaagccaat tcaatcaaac 180

aacggtggaa acggagagat gagtagccga gcaagagtca tgtttgctcc gtcaagcatc 240

cacggtgcag cggaacggga tttgcttgct ggtgtttacc acgacgagca agatgaacaa 300

ccaagagatc cacgtacttc tactaaagaa tctagccctc aaccacttcc gttaccgtca 360

ccaagaactg gttcttcatt gaagaattgg ggaagcttta agtcgtttaa cggaagcagc 420

ggtcggttat catcatccgc agctgtatct ggacctttac ctttgccacc tagcgggtca 480

gttaggagct tttcatatga tgaagtaatg gctgcgtgta acgctttttc ttcagaccga 540

tgtgtcatgg aaggtctttc atctgttatg tacatggctt cctttggtga tgaggcttcg 600

acctcaggtt taaagaaggt tgacgcaact gttgtacgac ttcacgtaac tactcagagt 660

attagggagt tcattaatga agtcaacaca ttggcgtcgc tgcaacacca gaacctttgt 720

aagctggtag gctatcatgc tcgtgacggt tctgacacaa gaatgttggt gtacgagagg 780

cttgctctgg gcagcttgga ccgtttactg catgggagat cagatgggcc tcctcttgat 840

tggaacacta gaatgaagat tgcactatgt gcagctcagg gtctaacctt cttgcacgaa 900

gaaggccctt ttcaggcaat gtacaatgaa ttttcgacgg caaatatcca agtcgataaa 960

gatttcagcg ccaagctatc aggatacggt tgtgcaggcc atgcgcctga gacagagaca 1020

tctaatagtt cggcacttgc taatctctct gtcgagactc tagagagagg gcttttgacc 1080

ccgaagagca atgtgtggag ctatggaata gttcttcttg agatgttaac gggtcggaaa 1140

aatatggacg ggtcttaccc gaaagaagag aggaacttag tgaaatggag cagagctttt 1200

ctagcagatg attgcaggct ctcgcttata atggatcctc agcttaaagg tcggtttccg 1260

gcaaaagcgg cgaggagcat agcagatata gcacagaaat gtctgcaggt ggagccatca 1320

gagcgtccaa ccatgagaaa catcgtggat caactcaaga tcatacagga catgaagtac 1380

tcgtgtaggt tcccgttaag agaacccgca ccagtcgcgg caaggaaaca tatgggaaga 1440

tcaagcagtc tcaacacgat tatttggacc ccggcatcag tgccaccaag gtcaagtttt 1500

tcaccgtcac ctccaccacg acgaccgtct gtttcaccca caaggggacg gacgctcgtg 1560

tttcccccag tgtttccgcc gcgagcgtgt tcatctttgg aggaaatggc tcgggaagag 1620

gttcgaagat cgtcttcagc cagtggtagg agaactagcc tcgaagggtt ttga 1674

<210>212

<211>557

<212>PRT

＜213〉Arabidopis thaliana

<400>212

Met Val Leu Gly Cys Phe Pro Leu Lys Ser Lys Lys Lys Arg Gly Ser

1 5 10 15

Val Ser Met Lys Arg Leu Asp Leu Glu Glu Ser Lys Pro Thr Ala Leu

20 25 30

Pro Glu Pro Pro Lys Ile Pro Ser Arg Asn Leu Gln Ser Gly Pro Pro

35 40 45

Ser Phe Arg Asn Arg Val Lys Pro Ile Gln Ser Asn Asn Gly Gly Asn

50 55 60

Gly Glu Met Ser Ser Arg Ala Arg Val Met Phe Ala Pro Ser Ser Ile

65 70 75 80

His Gly Ala Ala Glu Arg Asp Leu Leu Ala Gly Val Tyr His Asp Glu

85 90 95

Gln Asp Glu Gln Pro Arg Asp Pro Arg Thr Ser Thr Lys Glu Ser Ser

100 105 110

Pro Gln Pro Leu Pro Leu Pro Ser Pro Arg Thr Gly Ser Ser Leu Lys

115 120 125

Asn Trp Gly Ser Phe Lys Ser Phe Asn Gly Ser Ser Gly Arg Leu Ser

130 135 140

Ser Ser Ala Ala Val Ser Gly Pro Leu Pro Leu Pro Pro Ser Gly Ser

145 150 155 160

Val Arg Ser Phe Ser Tyr Asp Glu Val Met Ala Ala Cys Asn Ala Phe

165 170 175

Ser Ser Asp Arg Cys Val Met Glu Gly Leu Ser Ser Val Met Tyr Met

180 185 190

Ala Ser Phe Gly Asp Glu Ala Ser Thr Ser Gly Leu Lys Lys Val Asp

195 200 205

Ala Thr Val Val Arg Leu His Val Thr Thr Gln Ser Ile Arg Glu Phe

210 215 220

Ile Asn Glu Val Asn Thr Leu Ala Ser Leu Gln His Gln Asn Leu Cys

225 230 235 240

Lys Leu Val Gly Tyr His Ala Arg Asp Gly Ser Asp Thr Arg Met Leu

245 250 255

Val Tyr Glu Arg Leu Ala Leu Gly Ser Leu Asp Arg Leu Leu His Gly

260 265 270

Arg Ser Asp Gly Pro Pro Leu Asp Trp Asn Thr Arg Met Lys Ile Ala

275 280 285

Leu Cys Ala Ala Gln Gly Leu Thr Phe Leu His Glu Glu Gly Pro Phe

290 295 300

Gln Ala Met Tyr Asn Glu Phe Ser Thr Ala Asn Ile Gln Val Asp Lys

305 310 315 320

Asp Phe Ser Ala Lys Leu Ser Gly Tyr Gly Cys Ala Gly His Ala Pro

325 330 335

Glu Thr Glu Thr Ser Asn Ser Ser Ala Leu Ala Asn Leu Ser Val Glu

340 345 350

Thr Leu Glu Arg Gly Leu Leu Thr Pro Lys Ser Asn Val Trp Ser Tyr

355 360 365

Gly Ile Val Leu Leu Glu Met Leu Thr Gly Arg Lys Asn Met Asp Gly

370 375 380

Ser Tyr Pro Lys Glu Glu Arg Asn Leu Val Lys Trp Ser Arg Ala Phe

385 390 395 400

Leu Ala Asp Asp Cys Arg Leu Ser Leu Ile Met Asp Pro Gln Leu Lys

405 410 415

Gly Arg Phe Pro Ala Lys Ala Ala Arg Ser Ile Ala Asp Ile Ala Gln

420 425 430

Lys Cys Leu Gln Val Glu Pro Ser Glu Arg Pro Thr Met Arg Asn Ile

435 440 445

Val Asp Gln Leu Lys Ile Ile Gln Asp Met Lys Tyr Ser Cys Arg Phe

450 455 460

Pro Leu Arg Glu Pro Ala Pro Val Ala Ala Arg Lys His Met Gly Arg

465 470 475 480

Ser Ser Ser Leu Asn Thr Ile Ile Trp Thr Pro Ala Ser Val Pro Pro

485 490 495

Arg Ser Ser Phe Ser Pro Ser Pro Pro Pro Arg Arg Pro Ser Val Ser

500 505 510

Pro Thr Arg Gly Arg Thr Leu Val Phe Pro Pro Val Phe Pro Pro Arg

515 520 525

Ala Cys Ser Ser Leu Glu Glu Met Ala Arg Glu Glu Val Arg Arg Ser

530 535 540

Ser Ser Ala Ser Gly Arg Arg Thr Ser Leu Glu Gly Phe

545 550 555

<210>213

<211>2069

<212>DNA

＜213〉dichromatism chinese sorghum

<400>213

ctactggatt aaccccccgc gtgcgttccc tccctctgtc agtctgtctc tctcttggcg 60

tcgactgggc gaccgtcgca gccgccctaa gccaggtacc cagaccatct tcgggccctt 120

cgccggcgac gagcaccacc aggaattttc ccagctgtag caatgatggg ttgcttcact 180

gttctcagat ccaagaagaa gaaaatccct tttgataatc ctcttcttcc aagcaagaaa 240

tcggttgatg caagggaaag tacgtcgtct agacttccgg aaccagaagt tcatgtgcca 300

tctttgcaat cagctctctc ctagttttag gaacaggact aagatatccc agtcgtcaaa 360

taaagtttca aacagcagag ctcgcgtgtt gtctgctccg tcaaccctta ttgtggttga 420

tcagtttggc tttccatatg ctgaataccg agatcaagac gactccagag ataaggaagg 480

ttcaacaaag gggcatcgct tttcaaatcc tttgcccctt cctcttcctt caccggaagg 540

acattctttt aggaactctg gtagcttcaa agctagcaac gtaagtgggc cattggagat 600

gtccagccct ctcccattgc ctccaaagaa atgtgatgga cttagaatct tttcttacga 660

ggaggttttg tctgcttgcc aatggttttc aagtgatcag tgtgtttcag aagcacttgg 720

ttcaacatca tacaaggcaa catttaggga tgaatttatt gatacaaaga ccactgaagc 780

aacggtagct cgattactcc cctccactca gagtttgaag gagtttaaaa cacaagcaac 840

tacattggca ttgcttcagc atcccaattt atgtaaactg attggctatt atgcaaaaga 900

agattctaat gaaaggatgt tggtctatga acggcttcat catgggagct tagataagct 960

actctttgga agaccagatg gtcgtttcat ggactggtct aaacgtttga aggttgccct 1020

tggtgcggcc agaggtcttg ctttcttgca tgatgaagga ccctttcagg ccatgtacag 1080

cgagttctca actttgaaca tccaaataga taaagatttc actgctaagc tttcaggata 1140

tggttgtgtt ggcttcaata ctgaggagat atctaatgca cctgcgtctg cagcaaacct 1200

ttcggtggag accttggaga agggtttact cactcccaag agcaatgtct ggagctttgg 1260

agttgtgtta ctggagctaa taactggaag gaagaacctt gatgccaatt cttctaaaga 1320

agaacgcaat attgtcaggt ggagtaggcc tttccttact gatgatagtc gcctatctct 1380

gatcatggac tcccgtataa aagggcgctt tcctactaag gctgctcgga tagtagcaga 1440

catcattctg aaatgcttgc ataatgatcc atccgagagg ccgaccatga gggatgttgt 1500

ggaggctcta gcaagagtcc aggagataaa ggttccgtgc agatatcctt tgcaagaacc 1560

ttctgctgca ccaagaaaag taatgctaaa atctacatcc ctcaatggaa ttgtgcctca 1620

tcatcctgtc ataaccttct ctccatcgcc gccttcgcat aaccagcact tgatttcacc 1680

aaggtcatca acatctgcct tgtttcatcc aaggacatgc tcttctactc tggatgatcc 1740

cggtgtaagc tctataaaga aaacacctcc tattatgcgt aggtctagcg ttgaaggctt 1800

ttgattgtcc atattgttct tttatttctt tttctcggat ttatttgttt ctttgttagc 1860

ctagtgattt tagctgtgtt ctgtattgta agacaagtgg ccttatgtag tgcccttgag 1920

tctcgagtaa taactaagca gagcaggatg aattgtaaat agtatcaggt tgtagatacc 1980

tttttgtgct ctaattgggt ggaagcagcg tgctgagtct gagtaggcct tgtaacctca 2040

taaataaact cgtttcccag tttctgtcc 2069

<210>214

<211>1360

<212>DNA

＜213〉sugarcane

<400>214

agaaacactt ggttcaacat catacaaggc aacatttagg gatgaattta ttgatacaaa 60

gaccactgaa gcaacggtag ctcgattact cccctccact cagagtttga aggagtttaa 120

aacacaagcg accacattgg catcacttca gcatcccaat ttgtgtaaac tgattggcta 180

ttatgcaaaa gaagattcta atgaaaggat gttggtctat gaacggcttc atcatgggag 240

cttagataag ctactctttg gaagaccaga tggtcgtttc atggactggt ctaaacgttt 300

gaaggtttcc cttggtgctg cccgaggtct agctttcttg catgatgaag gaccctttca 360

ggccatgtac agtgagttct caactttgaa catccaaata gataaagatt tcactgctaa 420

gctttcagga tatggttgtg ttggcttcaa tactgaggag atatctaatg cacctgcgtc 480

tgcagcaaac ctttcggtgg agaccttgga gaagggttta ctcactccca agagcaacgt 540

ctggagcttt ggagttgtgt tactggagct aataactgga aggaagaacc ttgatgccaa 600

ttcttccaaa gaagaacgca atattgtcag gtggagtagg cctttcctta ctgatgatag 660

tcgcctatct ctgatcatgg actcccgtat aaaagggcgc tttccaacta aggctgctcg 720

gatagtagca gacatcattc tgaaatgctt gcataaagat ccatcagaga ggccgaccat 780

gagggatgtt gtggaggccc tagcaagagt ccaggaaata aaggttccat gcagatatcc 840

tctgcaagaa ccttttgctg caccaagaaa aataatgtta aaatctacat ccctcaatgg 900

aattgtgcct catcatcctg tcataacctt ctccccttcg ccgccttcac ataaccaaca 960

cttgatttca ccaaggtcat caacatctgc cttgtttcat ccaaggacat gctcttctac 1020

tctggatgat cccggtgtaa gctctataaa gaaaacacct cctattatgc gtaggtctag 1080

cgtcgaaggc ttttgattgt ccatattgtt cttttatttc cttttcttgg atttattagt 1140

ttctttggta gcctagtgat tctagctatg ttctgtattg caagacaagc ggccttaatg 1200

tagtgccctt gggtctagag taataactaa gcagagcagg atgaattgta aataggatca 1260

ggttgtagat acctttttgt gctctaattg ggtggaagca gcgtgctaag tgagccttgt 1320

aacctcccaa tgcaataaac tcgtttttca gtttttgttc 1360

<210>215

<211>2062

<212>DNA

＜213〉puncture vine clover

<400>215

acgaggctat tctctctctt tctctctcta catttctcaa cattcttcaa atttctctct 60

ccaaaacctt cacttcgcag ctttttcaat ctgctttctt tgactctgta acagtttttt 120

tcgtggaaga atcagaacca caaaaaagtt tcgattttta cccatttctt caacccgtga 180

tcgcttcact gtaattggca tgagcttcca tttctcgttc tgacaaaaac aggtatctat 240

aacatcctag ttctgtacat agaatggggt gttttactat attgaagaga aagaagaaaa 300

agcctgatca gattgtgtat gtaaaacgcg taagccctgg cgaagattca cctacagtac 360

tgcccgaacc ccaaactcat acccgctcac tccagtctgc gcctcctagt tttaagatca 420

gagtgaaacc cattcaacct agcaataaag ccactaacaa tagaatacga gcattgtctg 480

ctccatcgag tcttgatgat gcagaacaag atgcattggc caccattgag tatgaagaac 540

aagaagggtc aaaataccga actggatcat ggaaggagca gcgttcgcct agtccacaac 600

cattaccgct tccatctcct aagggtggtg gtacattgaa gactgttgga agctttaagt 660

tggggatagc tagtagtcct ttatatgctt ctggaccttt gccgcttcca ccaactgggt 720

cactgagaaa cttttcttat gatgaacttg ctgctgcttg cctcaatttc tcttcagatc 780

gatacatgtc agaatctctt tcatccacca tgtataaagc ttcctttggt gatgatacct 840

caacttcaag ttcaaagaag tttgaagcta ctgtcacacg ccttcgccca tcatctcagg 900

gcctgaagga attcataaat gaggttaata ctcttgcatc attgcagcat ccgaacctct 960

gtagattgct aggatttcac gcaggtgatg gttcagagca tagaatgttg gtttatgaga 1020

ggctatacca tggaagcttg gaccgcttat tgtatgggag atctgatggg ccatcaattg 1080

attggaatac aagaatgaaa attgcaatat gtgctgcact aggtctttct ttcttgcatg 1140

aagaagggcc ttttcaggca atgtataatg aattttcaac agctaacatt cagattgaca 1200

aagatttcag tgcaaagctt tcaggatatg gttgtgttgg acatgttccg aaggaagaga 1260

tttcaagcag ttcatctgcc gttggaaacc tatcgatgga gacactggag aaaggaatgc 1320

ttactccgaa aagcaatgta tggagtttcg gaattttcct tctagagcta cttacaggaa 1380

gaaagaatct cgatagcccg tcacccaaag gaagagagaa atttggtgaa gtggagcagg 1440

cctttcctgt ctgataatca tcgtttgtca atgatcatgg atcctcaact taaaggtcgc 1500

tttccttcta aagcggcgag tacaatagct aacattgcac aaagatgcct tcaaatggag 1560

ccatcagaac gaccaaccat gggaacagtt gttgagcagc tgaaaaagat acaagatttg 1620

aagcattcta gcagattccc gctgcaagaa cctgcacaaa tgtcgagatc accaagtctt 1680

aacggtatca accaccc tgc accaaggccg agtttctctc catcaccatc atccagagcc 1740

ctggtatctg tctcacctcc aagatggtcg ggagtgtcaa ttcaacttcc acctcgcgcg 1800

ttttcttcaa ccctctattt ggaggagctt gataggcaag aaagccgcaa gtcagcttca 1860

gcctctagga aggctagtgt tgaaggattt tgattgtcga tagtgttcat tttttatttg 1920

ttattctttt tttggtgtag ttcaacagag atttatagga gataaagatt tgtaatcatc 1980

cctcagtgtg atgcagagag gatgctcttt tgtacatagt gcaaaggttg gtccccttgg 2040

ttcaattagt tactccattt tg 2062

<210>216

<211>2039

<212>DNA

＜213〉Zea mays

<400>216

ctcccccggc gtgcgctccc tcctgtctct cttggcgacg actgggcaac ggtcgcagct 60

gtatgttcga gcccttcgcc gacgacgttc accaccagga attttctcag ctgtgccaat 120

gatgggttgc ttcactgttc tcagatccaa gaagaagaaa agccattttg ataatcctct 180

tgtcccaagc aagaaatcag ttgatgcaag ggaaagtacg tcctctagac ttccggagcc 240

agaagttcat gtgccatctt tgcaatcagc tcctcctagt tttaggaaca gggtcaagat 300

atccgagtca tcaaatgaag tttcaaacag aaacagcagg gctcgcgtgc tgtctgctcc 360

atcagccctt attgtggttg atcagtttgg ctttccatat tcggaatacc gagatcaaga 420

tgactccagg gataaggaag gtttgacaaa ggggcatcgc ttttcaaatc ctttgcccct 480

tcctcttcct tcacgcgaag gacattcttt taggaactct ggtagcttca aagccagcaa 540

cgtaagcggg ccattggaga tgtctggccc tctcccattg cctctaaaga aatgtgatgg 600

acttagaatc ttctcttatg aggagatttc atctgcctgc caacgatttt ctagtgatca 660

gtgtgtttca gaaacacttg gttcaacatc atacaaggca acatttagag atgaatttat 720

tgatacgagg accactgaag caacagtagc tcgattactc ccctccactc agagtttgaa 780

ggagtttaaa acgcaagcga ccacattggc atcgcttcag catcccaatt tatgtaaact 840

gattggctat tatgcaaaag aagattctaa tgaaaggatg ttggtctatg aacggcttta 900

tcatggaagc ttagataagc tactctttgg aagatcagat ggtcgtttca tggactggtc 960

taaacgtttg aaggttgccc tgggtgctgc cagaggtcta gctttcttgc atgatgaagg 1020

accctttcag gccatgtaca gcgagttctc aactttgaac atccaaatag ataaaggttt 1080

cactgctaag ctttcaggat atggttgtgt tggcttcaat accgaggaga tatctaatgc 1140

acctgtgtct gcagcaaacc tttcggtgga gaccttggag aagggtttac tcactcccaa 1200

gagcaacgtc tggagctttg gagtcgtgtt actggagcta ataactggaa ggaagaacct 1260

tgatcccaat tattccaaag aagaacgcaa tattgtcaac tggagtaggc ctttccttac 1320

tgatgacagt cgcttgtctc ttatcatgga ctcccgtata aaagggcgct ttcccactaa 1380

ggctgctcgg atagtagcag acatcatttt gaaatgcctg cgtaaggatc catcggagag 1440

gcctaccatg agggatgttg tggaggccct agcaagagtc caggaaataa aggttccgtg 1500

cagatatcct ctgcaagaac cttctgctgc accaagaaaa gtgatgttga aatctacatc 1560

cctcaatggg attgtgcctc accatcctgt cataaccttc tctccatcgc cgccttcaca 1620

taaccagcac ttgatttcac caaggtcgtc gacatctgcc ttgtttcatc caagggcatg 1680

ctcttctacc ctggacgatc cgagtgtaag ttctataaag aaaacaccac ctattatgcg 1740

aaggtctagc gtcgaaggct tttgattgtc catattgttc ctttatttcc ttttcttgga 1800

tttatttgtt tctttgttag tagtgtagcg gtttttagct gtgttctgta ttgtaagaca 1860

agtggcctca tatgtagtgc ccttgggtct ggagtaaagc agaacaggat gaattgtaaa 1920

taggaccagg ttgtagatac cttttttttg tgctcgaatt gggtggaagc agcgtgccga 1980

gtgggccttg taacctcata atgcaataaa ctccgtttcc cagtttctgt aaaaaaaaa 2039

<210>217

<211>1479

<212>DNA

＜213〉barley

<400>217

tgccgaattc ggcacgagaa agaggtccga ggggcttaag aacttctcgt acgatgagat 60

ttccactgct tgccagtggt tttctggcga tcatcgtgtc tcagaaactc tgacttcgac 120

atcatacaag gcattgttca gggatgattt cgttgaacca aagaagatgg aagcaatagt 180

agccaggtta ctcccttcca atcagagttt caaagaattt aaggcacaag taaatacgtt 240

ggcatcactt caacatccca atttatgtaa actcatcggc tatcacgcaa gagaagaatc 300

taatgaaagg atgttggtct atgagcggct ccatcatggc agtttagaca ggttactctt 360

tggaagaccg gaaggtcgtt tcatggactg gtctacacgt ttgaaggttg cccttggtgc 420

tgctaaaggt ctagctttcc tacacgatga aggacccttt caggcaatgt atgatgactt 480

ctcaacgtca aacatccaaa tagagaaaga tttcactgca aagctgtcgg gatatggttg 540

tgttggcttc aattctgacg aggaaagatc caaggcatct gtggctgcaa acctctcaga 600

ggaaaccttg gagaaaggcg tactgactcc gaagagcaac gtatggagct ttggagttgt 660

cttgctcgag ctaataacag gacggaagaa cctcgatgta cgttccacca aagaagaacg 720

taatattgtc aagtggggta ggcctttcct caccgacgat agtcgtctat ccctcatcat 780

ggatccccgt ataaaaggac gctttcctac caaggctgct cgaactgtgg cagacataat 840

tctgaagtgc cttcaaagag atccatcaga aaggcctacg atgagggccg tcgtggaggc 900

cctaacaagc gttcaggaca taaaggtccc ctgccggtat cctcttcaag taccgtcccg 960

ccgcacccga ggaaagtgat gctaaagtct acgagcctca atggcattgt tcctcagcat 1020

cccgtcatga ccttctcgcc gtcgcctcct tcgcgcaacc agcacctggc gtcgccaaga 1080

tcatcgactt ccgcgcttct tcccccaagg acctgctctt ccatcatcac cctggattac 1140

cccagggtaa gctcggtcaa gaagtcgcct tccggtatcc tgcggagacc tggcgtcgag 1200

ggtttttgat tgtccataca tggtcggatt tcttctcttt gccttggatt tatttgttgt 1260

tttggttagc ctagcgagtg gtctcagtgc tatgtgatat gtatatgttg gaaagacatc 1320

tgaaaaaagg gcagagtgaa gtgtagatag tagaaccagc tttgtagata cttgtcgttc 1380

tctgattgga tgggggtcat ggaagcagtg tgtgttagag tggggcttgt aaccccatta 1440

ttatgcaata aacgtgggtt ggtcggtcgt atttgctcg 1479

<210>218

<211>769

<212>DNA

＜213〉soybean

<400>218

gtaaattgct aggatttcat gcacgagagg gttcagaaca tagaatgttg gtttatgaaa 60

ggctatacca tggaagcttg gatcgcttat tgtatgggag atctgatggg ccatcaattg 120

attggaatac aagaatgaaa attgctatat gtgctgcaca aggtctaacc ttcttgcacg 180

aagaggggcc ttttcaggct atgtataatg agttctcaac agccaacata cagattgaca 240

aagatttcag cgcaaagctt tcaggatatg gttgtgttgg acatattccc gaagaagaga 300

tttcaagcag ctcatctgct gttggaaacc tatcaatgga aacactggag aaaggaatgc 360

tcactccaaa gagcaacgta tggagttttg gaatttttct tctggagcta cttactggaa 420

gaaagaatct tgatagccgt caccccaagg aagagaggaa tttagtcaag tggagccggc 480

ctttcttagc cgataactac cgtctgtcgt tgatcatgga tcctcaactc aaaggccgct 540

ttccttctaa agcagcaaga acaatagctg atattgcaca aagatgcctt caaaaggagc 600

catcagacag acctaccatg aggactgttg ttgagcatct caagataata caggatttga 660

aatattcgtg ccggttccct ctgcaagaac cagcatcaaa ctctggaaaa catatgtcaa 720

gatcaccaag tctcaatggc atcatctgcc ctgcaccaag ggtgagttt 769

<210>219

<211>1150

<212>DNA

＜213〉potato

<400>219

ttcttagctc atcgaaacgt taaaccttac aatacttcaa acatcgatca atacaatttg 60

ctgaagcacg caatttttca aggactgtat aaacaacaaa gatgggttgt ttcacagttt 120

taaaaagtaa gaagaagaag tctgaacaga gtattcacat caaacgtgtg aatcctcagg 180

aacattctcc tactgcattg cccgagcctc aagtgcagac acggtcgttg cagtcggcac 240

ctccaagttt tagaactaga gtaaaacctg tacagtcgag taatagagtt acaagcagta 300

gggcgcgggc actctctgcc ccatctagcc tggactcagc agaacaagat gtagcatcaa 360

atgaatgtga ggaacatgat gagttcaaga gtcgtattgg ttcaatcaag gagtaccaat 420

caccaagtcc tcagcctctt cctcttccat ctccacagag tgccgctgcc actctcaaga 480

ctatgggaag ctttaaagtt ggcaacgcca gcggtccgtt aaatgcctct ggacccctgc 540

cactgcctcc tacactgcct tcaacattgc cttctactgg agcactcagg aacttctcat 600

ttgaagaact tgctgctgcc tgccaccgct tctctcctga acggtgtatg tcagaaggtc 660

tctcttctgt tatttacaga gcttcttttg gagatgatgc aactggtaca aagaagcttg 720

aagccactgt aacccggctt catccttctt cgcaggggtt gaaggaattt gtgactgagg 780

tgaacacact agcttctttg caacatccat cactttgtaa actgattggt ttccatgcca 840

cggggaaggt tccgagcaca gaatgttggt ttatgaaagg ctttttcatg gaagcttaga 900

ccggcttttg tttgggaggt ccgatggtcc cccgatagac tggaatgctc gaacgaaaat 960

tgctttatgt gctgctcaag gtctcacatt cctgcatgag gaaggacctt ttcaggcaat 1020

gttccatgaa ttttccactg caaatataca aattgataag gattgcagtg caaagctctc 1080

gggatatgga tgcattactc ctataccaga gacagacata tcatgcagtt cagctgccct 1140

ggcaatctct 1150

<210>220

<211>910

<212>DNA

＜213〉tomato

<400>220

catggaagct tacaccggct tttatttggg aggtcggatg gtcccccaat agactggaat 60

gctcgaataa aaattgcttt atgtgctgct caaggtctca cattcctgca tgaggaagga 120

ccttttcagg caatgttcca tgaattttcc actgcaaata tacaaattga taaggattgc 180

agtgcaaagc tctcgggata tggatgcatt actcatatac aagagacaga catatcatgc 240

agttcagctg ccctggcaaa tctctctgag gagactctgg agcgaggatt ggtaactcca 300

aagagtaatg tttggagttt tgggattgtt cttcttgagc tgctgactgg ccggaagaat 360

ttaagcagtc ggcatccaaa ggaagagagg aatttagtga agtggagcaa gctttttcta 420

gctgatgaca gtagattatc gctaatcatg gaccctcagt taaaaggtcg gttccctgcc 480

aaagcagcaa gaactgtggc tgatattgct caaagatgtc tgcaaaagga tccatctgaa 540

aggcccacca tgagaactgt agtggagcaa ctcaagactg tacaagttat gaagtaccct 600

tctcggtttc ctcttcaaga gccaagggca gttggtgtaa aacacatgtc aaagtgtccg 660

agcctaaatg gaattattac cccaacatca agattgagct tctccccttc accaccaacc 720

caccctatat ctatttctcc gacaaggaca gctgctccac tgctgtctct accttcatgt 780

tcctccatcc tctctatgga ggactttgat cgattggaaa atcgaaggcc atcatcttca 840

tctgttcgga ggtctagtgt cgaaggattt tgatcgattg tagagcactt ttttcttcaa 900

attgcttttc 910

<210>221

<211>681

<212>DNA

＜213〉tomato

<400>221

gagaggcttt ttcatggaag cttagacaga cttttgttcg gaagatcaga tggcccctct 60

atagattgga atgcgagaac caaaattgcc ctatgtgctg cacaaggtct tacatttcta 120

catgaggagg gaccttttca ggcaatgttc caagaatttt caactggaaa tatacaaatc 180

gacaaggatt ttagtgcaaa gctctcgggg tatggatgca ttacgaatat acaagagacg 240

gagatatctt gcaattcaat cgccctggcg aatctctcac aggagacact ggagagaggg 300

ttgataactc ctaagagcaa tgtttggagt ttcgggattg ttcttttaga actgctcact 360

ggccggaaga atcttgatgg tcggtattca aaggaagaga ggaatctagt caagtggagt 420

aggcctttcc ttgctgatga tggtagatta tcgcttatca tggatcctca gcttaaagga 480

cggtttcccg caaaagcagc ccgtacagtg gctgatattg cccaaagatg cctgcaaaag 540

gatccatctg aaaggcccac catgagaact atcttggacc aactcaaatc cgtacaagtg 600

atgaagtgcc cttcacggtt tcctctgcaa gaaccagcgg ttgttggtgg taaacacatg 660

tcaaagtctc caagcatgaa t 681

<210>222

<211>741

<212>DNA

＜213〉common wheat

<400>222

tttcctccca aggctgctag gactgtggca gacatcattc tgaagtgcct tcatagggat 60

ccatccgaaa ggcctacgat gagggccgtc gtggagtctc tagcaagcgt ccaggacata 120

aaggtcccct gccggtatcc tcttcaagta ccgtccgccg caccgaggaa agtgatgcta 180

aaatccacga gtctcaacgg cattgttcct cagcatcctg tcatgacctt ctcgccgtcg 240

cctccttcgc gcaaccagca cctggtgtca ccgagatcat cgacgtctgc gctgcttccc 300

ccaaggaact gctcttccac catcaccctg gactacccca gggtaagctc ggtcaagaaa 360

tcgcccccca acatcatgcg gagacctggc gtcgagggtt tttgattgtc catatagtgt 420

cggattttct tctctttgcc ttgatttatt tgttgttttg gttagcctag cgagtgatct 480

cagtgctatg ggatatacta tgttgcaagg acatgtgaaa aagggcagag tgaagcgtag 540

atagtagaac cagcttgtag atacttgtcg ttctctgatt ggatgggggt catggaagaa 600

gtgtgtgttg agtggggctt gtaaccccac tatgcaataa acgtgggttg gttggtcggt 660

cggtatttgc tcgccgtcaa acatttcagc tgtttctttc ttctcttgaa tgctaagttt 720

tgttgctgct agtcgtggaa t 741

<210>223

<211>2039

212>DNA

＜213〉Zea mays

<400>223

ctcccccggc gtgcgctccc tcctgtctct cttggcgacg actgggcaac ggtcgcagct 60

gtatgttcga gcccttcgcc gacgacgttc accaccagga attttctcag ctgtgccaat 120

gatgggttgc ttcactgttc tcagatccaa gaagaagaaa agccattttg ataatcctct 180

tgtcccaagc aagaaatcag ttgatgcaag ggaaagtacg tcctctagac ttccggagcc 240

agaagttcat gtgccatctt tgcaatcagc tcctcctagt tttaggaaca gggtcaagat 300

atccgagtca tcaaatgaag tttcaaacag aaacagcagg gctcgcgtgc tgtctgctcc 360

atcagccctt attgtggttg atcagtttgg ctttccatat tcggaatacc gagatcaaga 420

tgactccagg gataaggaag gtttgacaaa ggggcatcgc ttttcaaatc ctttgcccct 480

tcctcttcct tcacgcgaag gacattcttt taggaactct ggtagcttca aagccagcaa 540

cgtaagcggg ccattggaga tgtctggccc tctcccattg cctctaaaga aatgtgatgg 600

acttagaatc ttctcttatg aggagatttc atctgcctgc caacgatttt ctagtgatca 660

gtgtgtttca gaaacacttg gttcaacatc atacaaggca acatttagag atgaatttat 720

tgatacgagg accactgaag caacagtagc tcgattactc ccctccactc agagtttgaa 780

ggagtttaaa acgcaagcga ccacattggc atcgcttcag catcccaatt tatgtaaact 840

gattggctat tatgcaaaag aagattctaa tgaaaggatg ttggtctatg aacggcttta 900

tcatggaagc ttagataagc tactctttgg aagatcagat ggtcgtttca tggactggtc 960

taaacgtttg aaggttgccc tgggtgctgc cagaggtcta gctttcttgc atgatgaagg 1020

accctttcag gccatgtaca gcgagttctc aactttgaac atccaaatag ataaaggttt 1080

cactgctaag ctttcaggat atggttgtgt tggcttcaat accgaggaga tatctaatgc 1140

acctgtgtct gcagcaaacc tttcggtgga gaccttggag aagggtttac tcactcccaa 1200

gagcaacgtc tggagctttg gagtcgtgtt actggagcta ataactggaa ggaagaacct 1260

tgatcccaat tattccaaag aagaacgcaa tattgtcaac tggagtaggc ctttccttac 1320

tgatgacagt cgcttgtctc ttatcatgga ctcccgtata aaagggcgct ttcccactaa 1380

ggctgctcgg atagtagcag acatcatttt gaaatgcctg cgtaaggatc catcggagag 1440

gcctaccatg agggatgttg tggaggccct agcaagagtc caggaaataa aggttccgtg 1500

cagatatcct ctgcaagaac cttctgctgc accaagaaaa gtgatgttga aatctacatc 1560

cctcaatggg attgtgcctc accatcctgt cataaccttc tctccatcgc cgccttcaca 1620

taaccagcac ttgatttcac caaggtcgtc gacatctgcc ttgtttcatc caagggcatg 1680

ctcttctacc ctggacgatc cgagtgtaag ttctataaag aaaacaccac ctattatgcg 1740

aaggtctagc gtcgaaggct tttgattgtc catattgttc ctttatttcc ttttcttgga 1800

tttatttgtt tctttgttag tagtgtagcg gtttttagct gtgttctgta ttgtaagaca 1860

agtggcctca tatgtagtgc ccttgggtct ggagtaaagc agaacaggat gaattgtaaa 1920

taggaccagg ttgtagatac cttttttttg tgctcgaatt gggtggaagc agcgtgccga 1980

gtgggccttg taacctcata atgcaataaa ctccgtttcc cagtttctgt aaaaaaaaa 2039

<210>224

<211>768

<212>DNA

＜213〉lettuce (Lactuca sativa)

<400>224

accgactgat ggccgctagt cggggaagaa cacgaagaat ccaaaacccg tggcagcggc 60

ggaggcggag gcgggttgcc gcctgtcccg cagccgctgc cgcttcctgc accgcattca 120

cgccaacttc gaaaaccatc gcggaatgct tttaaagatt gagtggcggt gagttgggca 180

tgctgccagc ctcttaatac ttccggaccc tctgccactt ccgccatcgg gaaccaccca 240

tcttccgcca atgattccgg catctttacc gacatctgga acacttaaga acttcacata 300

ttgaagaaat cgcagctgct tgccacaaca ttttcaccct gacagatgcg tgtctgaagg 360

tctttcttct gttatgtata gagcttcttt tggagaagat acttctaatt taaagaatct 420

tcaagccact gttaccagtc tccatccttc aactcagggt ttgaaggaat ttgtgagtga 480

agtgaacacg ctagcatcat tgcaacaccc ttatctctgt aaatagattg gtttccatgc 540

gcgtgaggga tctgatagaa gaatgttggt ttacgagagg ctttttcatg gaagcttaga 600

tcggctttta tatggtacaa cagatggccc acctattgat tgcaatgcaa gaatgaaagt 660

cgcactctgt gctgctcaag ggcttacttt tttgcatgag gaaggaccat ttcaggcgat 720

gtttcatgaa ttttccactg ctaatataca aattgataaa gattttag 768

<210>225

<211>1637

<212>DNA

＜213〉Gossypium certain (Gossypium spp.)

<400>225

ctttttcctc tcaaacctct ctcacactga taaacttttc tctccattac ctttctttga 60

cttgcaaagt tgttctcctc aaactaacca aacaagggat tctgggtttt cagctttctc 120

tgttttccta ctctctttcg cctttgcatg acctgcttac atagaaacat aaaagtccga 180

aatacaattt aggtggtttg tatataagat ggggtgtttt acagttttga gaagcaacaa 240

gaaaaagtcc aaacagtcag tttttgttaa acacattgct cataaagagc atattcctac 300

catgctgcct gagccccaaa ttcagacacg atcactgcaa tctgcacccc caagttttat 360

aaccagagta aaaccaattc aatctaataa caaggcaagc tgcaatagga cacgtgcatt 420

atctgctccg tcaagtcttg atgctgctga gcaagatgac cttgcgtcag ttgaatttga 480

agaacaagaa gagttgaaga gtcgtgttgg tttagtcaag gaacagaagt catcaagtcc 540

acagcctctt ccacttccat ctccccacag tactgcgctg aagacaatgg gaagttttaa 600

agcagggaat gttagtggcc ctctttttgc ttcgggacca ttacccctgc ctccctctgg 660

aacactacgt aacttcgcct acgaagaaat tgcagctgct tgccatcatt tctcttctga 720

tcgatgcaca tctgagggtc tatcttctgt tatgtacaag gcatccttcg gagatgacac 780

atcaagttca aagaagtttg aagctactgt tactcgcctt cacccatcca ctcagggttt 840

aagggaattt ataaacgaag taaacactct tgcatcattg caacatccaa atctctgtaa 900

attgcttggg taccatgcac gtgataattc agaacaacga atgttggtct atgagaggtt 960

atttcatgga agcttagacc ggcttttata cgggagatcg gatggaccgc cacttgattg 1020

gaatactcgc atgaaaattg ctttatgttc tgcacagggt cttactttct tgcatgagga 1080

aggaccattt caggcaatgt acaatgaatt ttcgactgcc aacatacaga tcgacaagga 1140

tttcagtgca aagctctcag gatatgggtg cgttggtcat atcccagaga cagaagagat 1200

ctccagtaat tcagttgctg tggcaaatat atccgtagag actctggaga gagggcgact 1260

aactccaaag agcaatgttt ggagttttgg gatcattcta cttgaattac tcactggccg 1320

gaagaacctt gacaaccgtt atcccaagga agagaggaac ctagtgaagt ggagccggcc 1380

attcctagcc gacaattgta gattgtcact catcatggat cctcagctca aaggtcgctt 1440

tcctatgaaa gctgcccgta cggtggctga cattgcacaa aggtgtctcc agatggaccc 1500

atcagagagg ccaaccatga gaaccatcgt tgagcatctc aaaatcatcc aagacctgaa 1560

atactcttgt cggtttcctc tgcaagatcc agcagcaatt gctggaaaac agatgtcaag 1620

gtcaccgagt ctcaacg 1637

<210>226

<211>2193

<212>DNA

＜213〉rice

<400>226

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140

cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200

tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260

gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320

ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380

gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440

aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500

ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560

atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620

acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680

cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740

ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800

gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860

tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920

attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980

tgtcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040

cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100

agctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160

tggtgtagct tgccactttc accagcaaag ttc 2193

<210>227

<211>56

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm06729

<400>227

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgat gggttgcttc actgtc 56

<210>228

<211>50

<212>DNA

＜213〉artificial sequence

<220>

＜223〉thing: prm06730

<400>228

ggggaccact ttgtacaaga aagctgggta tggacaatca aaaaccctca 50

<210>229

<211>1176

<212>DNA

＜213〉Arabidopis thaliana

<400>229

cttcaactgc tgcgtctagc tgtccttcct tcttcatttg cttcttcttc tctagctcag 60

cgcggatcgc tgcagtagta ccctgacgta gcctttcttc ttcctttact ttctcatctt 120

ctatctctca aaagaaaagc agacaacttt atttgcaaaa acagagtttt tttttcttat 180

cttgagaaag ttcaacagaa gatgatgttc gagaaagacg atctgggtct aagcttaggc 240

ttgaattttc caaagaaaca gatcaatctc aaatcaaatc catctgtttc tgttactcct 300

tcttcttctt cttttggatt attcagaaga tcttcatgga acgagagttt tacttcttca 360

gttccaaact cagattcgtc acaaaaagaa acaagaactt tcatccgagg aatcgacgtg 420

aacagaccac cgtctacagc ggaatacggc gacgaagacg ctggagtatc ttcacctaac 480

agtacagtct caagctctac agggaaaaga agcgagagag aagaagacac agatccacaa 540

ggctcaagag gaatcagtga cgatgaagat ggtgataact ccaggaaaaa gcttagactt 600

tccaaagatc aatctgctat tcttgaagag accttcaaag atcacagtac tctcaatccg 660

aagcagaagc aagcattggc taaacaatta gggttacgag caagacaagt ggaagtttgg 720

tttcagaaca gacgagcaag aacaaagctg aagcaaacgg aggtagactg cgagttctta 780

cggagatgct gcgagaatct aacggaagag aaccgtcggc tacaaaaaga agtaacggaa 840

ttgagagtac ttaagctctc tcctcagttc tacatgcaca tgagcccacc cactactttg 900

accatgtgcc cttcatgtga acacgtgtcg gtcccgccac cacaacctca ggctgctacg 960

tcagcgcacc accggtcgtt gccggtcaat gcgtgggctc ctgctacgag gatatctcac 1020

ggcttgactt ttgacgctct tcgtcctagg tcctaagtct ttttacttgc aaccaaaggg 1080

cattttggtc gttttttaag tttcatggac cagatatgca tgtagttgtt aacatgtatg 1140

tattttctta gaaagaaaga aaaacagattaatatt 1176

<210>230

<211>284

<212>PRT

＜213〉Arabidopis thaliana

<400>230

Met Met Phe Glu Lys Asp Asp Leu Gly Leu Ser Leu Gly Leu Asn Phe

1 5 10 15

Pro Lys Lys Gln Ile Asn Leu Lys Ser Asn Pro Ser Val Ser Val Thr

20 25 30

Pro Ser Ser Ser Ser Phe Gly Leu Phe Arg Arg Ser Ser Trp Asn Glu

35 40 45

Ser Phe Thr Ser Ser Val Pro Asn Ser Asp Ser Ser Gln Lys Glu Thr

50 55 60

Arg Thr Phe Ile Arg Gly Ile Asp Val Asn Arg Pro Pro Ser Thr Ala

65 70 75 80

Glu Tyr Gly Asp Glu Asp Ala Gly Val Ser Ser Pro Asn Ser Thr Val

85 90 95

Ser Ser Ser Thr Gly Lys Arg Ser Glu Arg Glu Glu Asp Thr Asp Pro

100 105 110

Gln Gly Ser Arg Gly Ile Ser Asp Asp Glu Asp Gly Asp Asn Ser Arg

115 120 125

Lys Lys Leu Arg Leu Ser Lys Asp Gln Ser Ala Ile Leu Glu Glu Thr

130 135 140

Phe Lys Asp His Ser Thr Leu Asn Pro Lys Gln Lys Gln Ala Leu Ala

145 150 155 160

Lys Gln Leu Gly Leu Arg Ala Arg Gln Val Glu Val Trp Phe Gln Asn

165 170 175

Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Phe

180 185 190

Leu Arg Arg Cys Cys Glu Asn Leu Thr Glu Glu Asn Arg Arg Leu Gln

195 200 205

Lys Glu Val Thr Glu Leu Arg Ala Leu Lys Leu Ser Pro Gln Phe Tyr

210 215 220

Met His Met Ser Pro Pro Thr Thr Leu Thr Met Cys Pro Ser Cys Glu

225 230 235 240

His Val Ser Val Pro Pro Pro Gln Pro Gln Ala Ala Thr Ser Ala His

245 250 255

His Arg Ser Leu Pro Val Asn Ala Trp Ala Pro Ala Thr Arg Ile Ser

260 265 270

His Gly Leu Thr Phe Asp Ala Leu Arg Pro Arg Ser

275 280

<210>231

<211>3171

<212>DNA

＜213〉Arabidopis thaliana

<400>231

gacaatccga aattaggtaa ccatgttcat tgaatttatc gtttcaacac attcgaatca 60

ctgagtccta cgtttcagtt taattccctg atgcacattt tgaattcggt ttgcgtattg 120

gtcttcgagt tttccgtcgg cggccatgga cgcgttcaag gaggagatag aaatcgggag 180

ctcggtggag tctttaatgg agctattgga ttcgcagaag gtgctttttc atagccagat 240

cgatcagctc caagatgtcg tcgttgcgca atgcaaactt accggcgtta atccccttgc 300

gcaagaaatg gctgctggtg ctttgtccat taaaattgga aagcggccaa gagacttgtt 360

gaatcctaag gctgttaagt atctacaagc agttttcgca attaaagatg ctattagtaa 420

gagggaatct cgggagataa gtgctttatt tggcatcaca gtcgcccagg ttcgagaatt 480

ttttgttact caaaagacaa gagtgaggaa acaggtgagg ctttcaaggg agaaagtagt 540

tatgtccaat acgcatgctt tacaagatga tggtgttccg gaaaataaca atgccacaaa 600

tcatgttgaa cccgttccct tgaactctat acatccggag gcatgttcta taagctgggg 660

tgaaggtgaa acagtggcac ttattccacc tgaagatatt ccacctgaca tcagcgattc 720

agacaaatac tttgttgaga atatattttc tctgctgcgt aaagaggaaa cattttcagg 780

ccaggtgaaa ctaatggagt ggatcatgca gatacaagat gcttctgtgc tgatctggtt 840

tttatcaaaa ggaggggttt tgatacttac aacatggtta agtcaagctg ctagtgaaga 900

gcaaacaagt gtcttacttc ttatcctgaa ggttctttgt catttacctc tccacaaagc 960

atctcctgaa aatatgtctg ccatattaca aagcgttaat ggacttcgtt tctataggat 1020

atcagacata tcaaacaggg caaaaggttt gttatcaagg tggaccaagt tatttgcgaa 1080

aatccaagct atgaagaaac aaaatcgtaa cagttcgcaa attgattcgc agagtcaatt 1140

gcttctgaaa cagagtattg ctgaaatcat gggtgatagt agcaatcctg aagatattct 1200

tagtctctca aatggaaagt cagagaatgt caggaggatt gaatcgtcac agggtccaaa 1260

actgttgctt acttctgcag atgattccac caagaaacac atgcttggtt caaatccatc 1320

gtataacaaa gaacgcagga aagtacagat ggtggaacaa ccaggccaaa aagctgctgg 1380

aaagagtccg cagacagtaa gaataggaac ttcaggtcga agccgcccaa tgtctgctga 1440

tgatattcag aaagcaaaga tgcgtgccct ttatatgcag agcaagaaca gtaaaaagga 1500

tcctttacca agtgccattg gtgattcgaa aatcgttgct cctgagaagc ccttggctct 1560

tcattcagcc aaggattctc cacctattca gaacaatgaa gctaagactg aagacacacc 1620

tgtactctcg actgttcagc ccgtcaatgg attttcaact attcagcccg tcaatggacc 1680

ttcagctgtt cagcccgtca atggaccttt ggctgttcag cctgtcaatg gaccttcggc 1740

tcttcagccc gtcaatggac cttcggccgt aattgtcccg gtacaagctg atgaaattaa 1800

aaaaccttca acacctccta aaagcatttc tagtaaggtg ggagttatga tgaaaatgag 1860

ttcacaaact attctcaaga attgcaagag aaaacagatt gattggcatg taccaccagg 1920

aatggaactt gacgaactct ggagagtagc cgctggtggt aatagcaagg aggctgatgt 1980

tcagagaaac agaaaccggc gagaaagaga aacaacatat cagtctcttc aaactatacc 2040

gttgaaccct aaagaaccat gggataggga aatggactat gatgacagtt tgacccctga 2100

aattccatct caacagccac cagaagaaag tttaacggaa ccacaggatt cacttgatga 2160

acgaagaatt gctgctggtg ctgccacaac ctcttcatct ctaagcagtc ctgaacctga 2220

tctcgagtta ttagctgcgt tacttaagaa cccagatctt gtttatgcac taacttcggg 2280

aaaacccagt aatttagccg gccaagatat ggtaaaactg cttgatgtga ttaagactgg 2340

tgcaccaaac tcaagcagta gctcaaataa acaggttgaa gaaagggtcg aagtttccct 2400

tccatctccc actccatcaa ctaatcctgg aatgagtgga tggggacaag aagggattcg 2460

gaatccattt tcaaggcaaa accaagttgg tactgcagtt gctagatcgg gtacacagct 2520

tcgtgttggt tcaatgcaat ggcatcaaac aaatgaacaa tcaatcccac gacatgctcc 2580

atcagcatac agtaactcga tcacattggc tcacacagaa agagaacagc aacaatatat 2640

gcaaccaaaa cttcatcaca atttacattt tcaacaacaa caacaacaac caatctcaac 2700

aacctcgtat gcagttaggg aaccagtagg acaaatggga acaggtacat cgagttcatg 2760

gaggagtcag cagagtcaga acagttacta ctcacatcaa gaaaacgaga ttgcatcggc 2820

ttcacaagtt acttcatacc aagggaatag ccagtacatg agtagcaatc caggatatga 2880

atcatggagt cctgataata gcccaagtag gaaccagctt aacatgaggg gacaacaaca 2940

acaagcatca aggaaacatg attcttctac tcatccatat tggaaccaaa acaaaagatg 3000

gcgttaaacc atatataaga ctatggttct ttggtcctca atatttgctt gtcatggttg 3060

aattagcatt atactctgtt cagacacaga ttttcatttt taactcacaa cagattcatc 3120

agagataata aactaaaaga aagaatgaat ggataaagtt attactgtct c 3171

<210>232

<211>953

<212>PRT

＜213〉Arabidopis thaliana

<400>232

Met Asp Ala Phe Lys Glu Glu Ile Glu Ile Gly Ser Ser Val Glu Ser

1 5 10 15

Leu Met Glu Leu Leu Asp Ser Gln Lys Val Leu Phe His Ser Gln Ile

20 25 30

Asp Gln Leu Gln Asp Val Val Val Ala Gln Cys Lys Leu Thr Gly Val

35 40 45

Asn Pro Leu Ala Gln Glu Met Ala Ala Gly Ala Leu Ser Ile Lys Ile

50 55 60

Gly Lys Arg Pro Arg Asp Leu Leu Asn Pro Lys Ala Val Lys Tyr Leu

65 70 75 80

Gln Ala Val Phe Ala Ile Lys Asp Ala Ile Ser Lys Arg Glu Ser Arg

85 90 95

Glu Ile Ser Ala Leu Phe Gly Ile Thr Val Ala Gln Val Arg Glu Phe

100 105 110

Phe Val Thr Gln Lys Thr Arg Val Arg Lys Gln Val Arg Leu Ser Arg

115 120 125

Glu Lys Val Val Met Ser Asn Thr His Ala Leu Gln Asp Asp Gly Val

130 135 140

Pro Glu Asn Asn Asn Ala Thr Asn His Val Glu Pro Val Pro Leu Asn

145 150 155 160

Ser Ile His Pro Glu Ala Cys Ser Ile Ser Trp Gly Glu Gly Glu Thr

165 170 175

Val Ala Leu Ile Pro Pro Glu Asp Ile Pro Pro Asp Ile Ser Asp Ser

180 185 190

Asp Lys Tyr Phe Val Glu Asn Ile Phe Ser Leu Leu Arg Lys Glu Glu

195 200 205

Thr Phe Ser Gly Gln Val Lys Leu Met Glu Trp Ile Met Gln Ile Gln

210 215 220

Asp Ala Ser Val Leu Ile Trp Phe Leu Ser Lys Gly Gly Val Leu Ile

225 230 235 240

Leu Thr Thr Trp Leu Ser Gln Ala Ala Ser Glu Glu Gln Thr Ser Val

245 250 255

Leu Leu Leu Ile Leu Lys Val Leu Cys His Leu Pro Leu His Lys Ala

260 265 270

Ser Pro Glu Asn Met Ser Ala Ile Leu Gln Ser Val Asn Gly Leu Arg

275 280 285

Phe Tyr Arg Ile Ser Asp Ile Ser Asn Arg Ala Lys Gly Leu Leu Ser

290 295 300

Arg Trp Thr Lys Leu Phe Ala Lys Ile Gln Ala Met Lys Lys Gln Asn

305 310 315 320

Arg Asn Ser Ser Gln Ile Asp Ser Gln Ser Gln Leu Leu Leu Lys Gln

325 330 335

Ser Ile Ala Glu Ile Met Gly Asp Ser Ser Asn Pro Glu Asp Ile Leu

340 345 350

Ser Leu Ser Asn Gly Lys Ser Glu Asn Val Arg Arg Ile Glu Ser Ser

355 360 365

Gln Gly Pro Lys Leu Leu Leu Thr Ser Ala Asp Asp Ser Thr Lys Lys

370 375 380

His Met Leu Gly Ser Asn Pro Ser Tyr Asn Lys Glu Arg Arg Lys Val

385 390 395 400

Gln Met Val Glu Gln Pro Gly Gln Lys Ala Ala Gly Lys Ser Pro Gln

405 410 415

Thr Val Arg Ile Gly Thr Ser Gly Arg Ser Arg Pro Met Ser Ala Asp

420 425 430

Asp Ile Gln Lys Ala Lys Met Arg Ala Leu Tyr Met Gln Ser Lys Asn

435 440 445

Ser Lys Lys Asp Pro Leu Pro Ser Ala Ile Gly Asp Ser Lys Ile Val

450 455 460

Ala Pro Glu Lys Pro Leu Ala Leu His Ser Ala Lys Asp Ser Pro Pro

465 470 475 480

Ile Gln Asn Asn Glu Ala Lys Thr Glu Asp Thr Pro Val Leu Ser Thr

485 490 495

Val Gln Pro Val Asn Gly Phe Ser Thr Ile Gln Pro Val Asn Gly Pro

500 505 510

Ser Ala Val Gln Pro Val Asn Gly Pro Leu Ala Val Gln Pro Val Asn

515 520 525

Gly Pro Ser Ala Leu Gln Pro Val Asn Gly Pro Ser Ala Val Ile Val

530 535 540

Pro Val Gln Ala Asp Glu Ile Lys Lys Pro Ser Thr Pro Pro Lys Ser

545 550 555 560

Ile Ser Ser Lys Val Gly Val Met Met Lys Met Ser Ser Gln Thr Ile

565 570 575

Leu Lys Asn Cys Lys Arg Lys Gln Ile Asp Trp His Val Pro Pro Gly

580 585 590

Met Glu Leu Asp Glu Leu Trp Arg Val Ala Ala Gly Gly Asn Ser Lys

595 600 605

Glu Ala Asp Val Gln Arg Asn Arg Asn Arg Arg Glu Arg Glu Thr Thr

610 615 620

Tyr Gln Ser Leu Gln Thr Ile Pro Leu Asn Pro Lys Glu Pro Trp Asp

625 630 635 640

Arg Glu Met Asp Tyr Asp Asp Ser Leu Thr Pro Glu Ile Pro Ser Gln

645 650 655

Gln Pro Pro Glu Glu Ser Leu Thr Glu Pro Gln Asp Ser Leu Asp Glu

660 665 670

Arg Arg Ile Ala Ala Gly Ala Ala Thr Thr Ser Ser Ser Leu Ser Ser

675 680 685

Pro Glu Pro Asp Leu Glu Leu Leu Ala Ala Leu Leu Lys Asn Pro Asp

690 695 700

Leu Val Tyr Ala Leu Thr Ser Gly Lys Pro Ser Asn Leu Ala Gly Gln

705 710 715 720

Asp Met Val Lys Leu Leu Asp Val Ile Lys Thr Gly Ala Pro Asn Ser

725 730 735

Ser Ser Ser Ser Asn Lys Gln Val Glu Glu Arg Val Glu Val Ser Leu

740 745 750

Pro Ser Pro Thr Pro Ser Thr Asn Pro Gly Met Ser Gly Trp Gly Gln

755 760 765

Glu Gly Ile Arg Asn Pro Phe Ser Arg Gln Asn Gln Val Gly Thr Ala

770 775 780

Val Ala Arg Ser Gly Thr Gln Leu Arg Val Gly Ser Met Gln Trp His

785 790 795 800

Gln Thr Asn Glu Gln Ser Ile Pro Arg His Ala Pro Ser Ala Tyr Ser

805 810 815

Asn Ser Ile Thr Leu Ala His Thr Glu Arg Glu Gln Gln Gln Tyr Met

820 825 830

Gln Pro Lys Leu His His Asn Leu His Phe Gln Gln Gln Gln Gln Gln

835 840 845

Pro Ile Ser Thr Thr Ser Tyr Ala Val Arg Glu Pro Val Gly Gln Met

850 855 860

Gly Thr Gly Thr Ser Ser Ser Trp Arg Ser Gln Gln Ser Gln Asn Ser

865 870 875 880

Tyr Tyr Ser His Gln Glu Asn Glu Ile Ala Ser Ala Ser Gln Val Thr

885 890 895

Ser Tyr Gln Gly Asn Ser Gln Tyr Met Ser Ser Asn Pro Gly Tyr Glu

900 905 910

Ser Trp Ser Pro Asp Asn Ser Pro Ser Arg Asn Gln Leu Asn Met Arg

915 920 925

Gly Gln Gln Gln Gln Ala Ser Arg Lys His Asp Ser Ser Thr His Pro

930 935 940

Tyr Trp Asn Gln Asn Lys Arg Trp Arg

945 950

<210>233

<211>501

<212>DNA

＜213〉rice

<400>233

atggagcagc agcttcctct tcttgcacct gactctaagg ctgccacgtc gtcccccttg 60

tgcctcacct tggacaaccc aacctccaca tccacctcgc cggcggtgcc gtcgtcggcg 120

ccgccgccgg cagccgcctt ggaaccttct agacaatctt tccatgagag ggaaacggat 180

gcaatcaaag ccaagatcat gtcgcacccc ctctacccgg ctctcctcag agccttcata 240

gattgccaga aggtcggagc tccgccggag gtcgtcggcc ggctttccgc cctcgccggc 300

gagctcgact cgcgtgcaga agacaggtac cttcaagggc agtcgtcaga cccggagctc 360

gacgagttta tggaaaccta cattgatatg ctggtgagct acaggcagga gctgacaaga 420

ccaattcaag aggccgacca gttcttcaga aacatggagg cacagatcga ctcgtttaca 480

ctagagatgt gcagtttctg a 501

<210>234

<211>166

<212>PRT

＜213〉rice

<400>234

Met Glu Gln Gln Leu Pro Leu Leu Ala Pro Asp Ser Lys Ala Ala Thr

1 5 10 15

Ser Ser Pro Leu Cys Leu Thr Leu Asp Asn Pro Thr Ser Thr Ser Thr

20 25 30

Ser Pro Ala Val Pro Ser Ser Ala Pro Pro Pro Ala Ala Ala Leu Glu

35 40 45

Pro Ser Arg Gln Ser Phe His Glu Arg Glu Thr Asp Ala Ile Lys Ala

50 55 60

Lys Ile Met Ser His Pro Leu Tyr Pro Ala Leu Leu Arg Ala Phe Ile

65 70 75 80

Asp Cys Gln Lys Val Gly Ala Pro Pro Glu Val Val Gly Arg Leu Ser

85 90 95

Ala Leu Ala Gly Glu Leu Asp Ser Arg Ala Glu Asp Arg Tyr Leu Gln

100 105 110

Gly Gln Ser Ser Asp Pro Glu Leu Asp Glu Phe Met Glu Thr Tyr Ile

115 120 125

Asp Met Leu Val Ser Tyr Arg Gln Glu Leu Thr Arg Pro Ile Gln Glu

130 135 140

Ala Asp Gln Phe Phe Arg Asn Met Glu Ala Gln Ile Asp Ser Phe Thr

145 150 155 160

Leu Glu Met Cys Ser Phe

165

<210>235

<211>879

<212>DNA

＜213〉rice

<400>235

atggctcagg aggacgtcgg tcacctgagc gacgccggcc tggcgctggg cctgtccctc 60

ggcgggggag gaggagggac gaccgacgcg gcggcggcgc accgtggcgg ctgccggcgg 120

ccgtcgccgt cgtcgcagtg cccgccgctg gagccgtcgc tgaccctgag cttgcccgac 180

gacgcggcgg ccggcgcggc cgcgaccgcg accgcgaccg cgtccggcgg gggcggccct 240

gcgcacagcg tgtcgtcgct gtccgtcggc gcggcggcgg cggcggccgt gaagagggag 300

cgcgcggagg aggccgacgg cgagagggtg tcgtcgacgg cggccgggcg tgacgacgac 360

gacgacggga gcacccgcaa gaagctccgg ctgaccaagg agcagtccgc gctcctggag 420

gaccgcttcc gggagcacag cacgctcaac ccgaagcaga aagtcgcttt agcgaagcaa 480

ctgaacctca ggccaaggca ggtggaggtc tggttccaaa acagaagagc aaggacaaag 540

ctgaagcaga cggaggtgga ctgcgagttc ctgaagcgct gctgcgagac gctcaccgag 600

gagaaccgtc ggctgcagcg cgagctgcag gagctccgcg cgctcaagtt cgccccgccg 660

ccgccgtcct cggcggccca ccagccgtcg ccggcgccac cggcgccgtt ctacatgcaa 720

ctcccggccg ccacgctcac catctgcccg tcctgcgagc gcgtcggcgg gcccgcgtcc 780

gccgccaagg tcgtcgccgc cgacgggacc aaggccggcc ccggccggac caccacccac 840

cacttcttca accccttcac ccactccgcc gcctgctga 879

<210>236

<211>292

<212>PRT

＜213〉rice

<400>236

Met Ala Gln Glu Asp Val Gly His Leu Ser Asp Ala Gly Leu Ala Leu

1 5 10 15

Gly Leu Ser Leu Gly Gly Gly Gly Gly Gly Thr Thr Asp Ala Ala Ala

20 25 30

Ala His Arg Gly Gly Cys Arg Arg Pro Ser Pro Ser Ser Gln Cys Pro

35 40 45

Pro Leu Glu Pro Ser Leu Thr Leu Ser Leu Pro Asp Asp Ala Ala Ala

50 55 60

Gly Ala Ala Ala Thr Ala Thr Ala Thr Ala Ser Gly Gly Gly Gly Pro

65 70 75 80

Ala His Ser Val Ser Ser Leu Ser Val Gly Ala Ala Ala Ala Ala Ala

85 90 95

Val Lys Arg Glu Arg Ala Glu Glu Ala Asp Gly Glu Arg Val Ser Ser

100 105 110

Thr Ala Ala Gly Arg Asp Asp Asp Asp Asp Gly Ser Thr Arg Lys Lys

115 120 125

Leu Arg Leu Thr Lys Glu Gln Ser Ala Leu Leu Glu Asp Arg Phe Arg

130 135 140

Glu His Ser Thr Leu Asn Pro Lys Gln Lys Val Ala Leu Ala Lys Gln

145 150 155 160

Leu Asn Leu Arg Pro Arg Gln Val Glu ValTrp Phe Gln Asn Arg Arg

165 170 175

Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Phe Leu Lys

180 185 190

Arg Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu Gln Arg Glu

195 200 205

Leu Gln Glu Leu Arg Ala Leu Lys Phe Ala Pro Pro Pro Pro Ser Ser

210 215 220

Ala Ala His Gln Pro Ser Pro Ala Pro Pro Ala Pro Phe Tyr Met Gln

225 230 235 240

Leu Pro Ala Ala Thr Leu Thr Ile Cys Pro Ser Cys Glu Arg Val Gly

245 250 255

Gly Pro Ala Ser Ala Ala Lys Val Val Ala Ala Asp Gly Thr Lys Ala

260 265 270

Gly Pro Gly Arg Thr Thr Thr His His Phe Phe Asn Pro Phe Thr His

275 280 285

Ser Ala Ala Cys

290

<210>237

<211>744

<212>DNA

＜213〉rice

<400>237

atggcgcaag atgatgagga cgtgggtcta gctctgggcc tctccctggg ctccggcggc 60

cacaggcggc agagagaaag tagagacgaa gcgccgtcgt cggcggcggc gtccctgctg 120

acgctgaggc tcccggcaga gagcgggggg cagccgcagg tggtggtgaa gagggaggtg 180

gtgcgggcgg aagaggagga gtacgaatac gagtacgaga gggcgctcta ctcgtcgtcg 240

gctgcggcgg cggacgacga cgagggctgc aacagccgga agaagctgag gctgagcaag 300

gagcagtcgg ctctgctgga ggatcgcttc aaggagcata gcactctcaa ccctaagcaa 360

aaggttgctt tggcgaagca gctaaacctg aggccaaggc aagttgaggt gtggttccaa 420

aacagaagag ccaggacgaa gctgaagcag acggaggtgg attgcgagct tctgaagcgc 480

tgctgcgaga cgctgacgga ggagaaccga cgcctccatc gtgagctcca gcagctcagg 540

gctctgaccc actccaccgc cgccggcttc ttcatggcca ccaccctccc cgtccccgcc 600

gccacgctct ccatctgccc ctcctgcgag cgcctcgcca ccgccgccgc cgccggagct 660

tcccccaccg ccgccgccga ccgcaccaac aagcccaccg ccccgcactt gttcagccct 720

ttcgccaagt ccgccgcctg ctag 744

<210>238

<211>247

<212>PRT

＜213〉rice

<400>238

Met Ala Gln Asp Asp Glu Asp Val Gly Leu Ala Leu Gly Leu Ser Leu

1 5 10 15

Gly Ser Gly Gly His Arg Arg Gln Arg Glu Ser Arg Asp Glu Ala Pro

20 25 30

Ser Ser Ala Ala Ala Ser Leu Leu Thr Leu Arg Leu Pro Ala Glu Ser

35 40 45

Gly Gly Gln Pro Gln Val Val Val Lys Arg Glu Val Val Arg Ala Glu

50 55 60

Glu Glu Glu Tyr Glu Tyr Glu Tyr Glu Arg Ala Leu Tyr Ser Ser Ser

65 70 75 80

Ala Ala Ala Ala Asp Asp Asp Glu Gly Cys Asn Ser Arg Lys Lys Leu

85 90 95

Arg Leu Ser Lys Glu Gln Ser Ala Leu Leu Glu Asp Arg Phe Lys Glu

100 105 110

His Ser Thr Leu Asn Pro Lys Gln Lys Val Ala Leu Ala Lys Gln Leu

115 120 125

Asn Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg Arg Ala

130 135 140

Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Leu Leu Lys Arg

145 150 155 160

Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu His Arg Glu Leu

165 170 175

Gln Gln Leu Arg Ala Leu Thr His Ser Thr Ala Ala Gly Phe Phe Met

180 185 190

Ala Thr Thr Leu Pro Val Pro Ala Ala Thr Leu Ser Ile Cys Pro Ser

195 200 205

Cys Glu Arg Leu Ala Thr Ala Ala Ala Ala Gly Ala Ser Pro Thr Ala

210 215 220

Ala Ala Asp Arg Thr Asn Lys Pro Thr Ala Pro His Leu Phe Ser Pro

225 230 235 240

Phe Ala Lys Ser Ala Ala Cys

245

<210>239

<211>1065

<212>DNA

＜213〉rice

<400>239

atggagctgg ggctgagctt gggggatgcg gtgaccgtgg cggatggcgg gaggctggag 60

ctggttcttg ggctcggggt tggggttggg gctggggtga ggagaggaga ggaggaggag 120

agggggagga gggaggatgt ggtgggagct gggaggtggg cggcgatggc ggcggcctcg 180

ccggagccgt cggtgcggct cagcctcgtg tcgagcctcg ggctccactg gccttccgag 240

accgggcgtt cggaggcggc ggcgcgtggg ttcgacgtga accgggcgcc gtcggtggcg 300

gcgggcgcgc cggggatgga ggacgacgag gagggcccgg gcgccgcgcc ggcgttgtcg 360

tcgtcgccca acgacagcgg gggatccttc ccgctggacc tctccgggca aggcctccgt 420

ggccacgccg aggcggcggc gcagggtggc ggcggcggcg gcggcggcga gcggtcgtcc 480

tcgcgcgcga gcgacgacga cgagggcgcg tccgcgcgca agaagctgcg actctccaag 540

gagcagtcgg cgttcctcga ggagagcttc aaggagcaca gcacgctgaa tcccaagcag 600

aaggtggcgc tggcgaagca gctcaacctc cggccgcggc aagtggaggt ctggttccag 660

aaccgccgag ccaggacgaa gctgaagcag acggaggtgg actgcgagta cctgaagcgc 720

tgctgcgaga cgctcaccga ggagaaccgg cggctgcaca aggagctcgc cgagctgcgc 780

gcgctcaaga cggcgcgccc cttctacatg cacctcccgg ccacgaccct ctccatgtgc 840

ccctcctgcg agcgtgtcgc ctccaacccg gccaccgctt cgacctccgc ccccgccgcg 900

gccacgtccc cggcggcggc gccgaccgcg gccgcaagaa ccgccgtcgc gtcgcccgag 960

ccgcaccggc cgtcgtcctt cgccgcgctg ttcgcggcac ccctcggctt cccgctgacc 1020

gccgcccagc cgcggccgcc gccgccggcg agcaactgcc tgtaa 1065

<210>240

<211>354

<212>PRT

＜213〉rice

<400>240

Met Glu Leu Gly Leu Ser Leu Gly Asp Ala Val Thr Val Ala Asp Gly

1 5 10 15

Gly Arg Leu Glu Leu Val Leu Gly Leu Gly Val Gly Val Gly Ala Gly

20 25 30

Val Arg Arg Gly Glu Glu Glu Glu Arg Gly Arg Arg Glu Asp Val Val

35 40 45

Gly Ala Gly Arg Trp Ala Ala Met Ala Ala Ala Ser Pro Glu Pro Ser

50 55 60

Val Arg Leu Ser Leu Val Ser Ser Leu Gly Leu His Trp Pro Ser Glu

65 70 75 80

Thr Gly Arg Ser Glu Ala Ala Ala Arg Gly Phe Asp Val Asn Arg Ala

85 90 95

Pro Ser Val Ala Ala Gly Ala Pro Gly Met Glu Asp Asp Glu Glu Gly

100 105 110

Pro Gly Ala Ala Pro Ala Leu Ser Ser Ser Pro Asn Asp Ser Gly Gly

115 120 125

Ser Phe Pro Leu Asp Leu Ser Gly Gln Gly Leu Arg Gly His Ala Glu

130 135 140

Ala Ala Ala Gln Gly Gly Gly Gly Gly Gly Gly Gly Glu Arg Ser Ser

145 150 155 160

Ser Arg Ala Ser Asp Asp Asp Glu Gly Ala Ser Ala Arg Lys Lys Leu

165 170 175

Arg Leu Ser Lys Glu Gln Ser Ala Phe Leu Glu Glu Ser Phe Lys Glu

180 185 190

His Ser Thr Leu Asn Pro Lys Gln Lys Val Ala Leu Ala Lys Gln Leu

195 200 205

Asn Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg Arg Ala

210 215 220

Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Tyr Leu Lys Arg

225 230 235 240

Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu His Lys Glu Leu

245 250 255

Ala Glu Leu Arg Ala Leu Lys Thr Ala Arg Pro Phe Tyr Met His Leu

260 265 270

Pro Ala Thr Thr Leu Ser Met Cys Pro Ser Cys Glu Arg Val Ala Ser

275 280 285

Asn Pro Ala Thr Ala Ser Thr Ser Ala Pro Ala Ala Ala Thr Ser Pro

290 295 300

Ala Ala Ala Pro Thr Ala Ala Ala Arg Thr Ala Val Ala Ser Pro Glu

305 310 315 320

Pro His Arg Pro Ser Ser Phe Ala Ala Leu Phe Ala Ala Pro Leu Gly

325 330 335

Phe Pro Leu Thr Ala Ala Gln Pro Arg Pro Pro Pro Pro Ala Ser Asn

340 345 350

Cys Leu

<210>241

<211>1089

<212>DNA

＜213〉rice

<400>241

atggagctgg ggttgagctt gggggaggct atggcggatg ctgggaggga gctggttctt 60

gggcttggga tggggaggag ggaggaggcg gcggaggcgg ggaggaggga tcatgaggtg 120

aggagggagc tggagttcgg gtcgatgtcg agcaggtgcg gtggttcttc gccggagccg 180

acggtgcggc tcacgcttct gcccatggtg cccggccttg gcctcccatg gccgccgccg 240

ccgccgccgt cgtccgagag caggcatttg gaggcgtcga cgcggggatt cgacgtgaac 300

cgcccgccgt cgtccggcgg cggcggcggc ggcggcggcg cggaggagga gcaggacgac 360

gtggccgggg cggcactctc gtcgtccccc aacaacagcg ccggatcctt cccgatggat 420

gacttctccg ggcacggcct cggcggcaac gacgcggccc ctggcggtgg cggcggcgac 480

cgctcgtgct cccgcgccag cgacgaggac gacggcggct ccgcgcgcaa gaagctccgc 540

ctctccaagg agcagtccgc gttcctcgag gagagcttca aggagcacag caccctaaac 600

cccaagcaga agctggcgct ggcgaagcag ctcaacctcc ggccgcgcca ggtggaggtg 660

tggttccaga accgccgcgc caggacgaag ctgaagcaga cggaggtgga ctgcgagtac 720

ctgaagcggt gctgcgagac gctgacggag gagaaccggc ggctgcagaa ggagctcgcc 780

gagctccggg cgctcaagac ggtgcacccc ttctacatgc acctcccggc gacgacactc 840

tccatgtgcc cctcctgcga gcgcgtcgcc tccaactccg ccccggccac cgcctcctcc 900

gccgcaacat cgtcgacggc cgcgccgccc gcggcaccct catccggcgg cattgcggcc 960

acctcctcct ccgccgccgc cgccgcggcg ccggaccaca ggccgtcgtc gttcgccgcg 1020

ctgttctcgt cgccgcgtgg cttcccgcta tccgtagccc cgcaggcgca gccgccgacg 1080

agctcgtga 1089

<210>242

<211>362

<212>PRT

＜213〉rice

<400>242

Met Glu Leu Gly Leu Ser Leu Gly Glu Ala Met Ala Asp Ala Gly Arg

1 5 10 15

Glu Leu Val Leu Gly Leu Gly Met Gly Arg Arg Glu Glu Ala Ala Glu

20 25 30

Ala Gly Arg Arg Asp His Glu Val Arg Arg Glu Leu Glu Phe Gly Ser

35 40 45

Met Ser Ser Arg Cys Gly Gly Ser Ser Pro Glu Pro Thr Val Arg Leu

50 55 60

Thr Leu Leu Pro Met Val Pro Gly Leu Gly Leu Pro Trp Pro Pro Pro

65 70 75 80

Pro Pro Pro Ser Ser Glu Ser Arg His Leu Glu Ala Ser Thr Arg Gly

85 90 95

PheAsp Val Asn Arg Pro Pro Ser Ser Gly Gly Gly Gly Gly Gly Gly

100 105 110

Gly Ala Glu Glu Glu Gln Asp Asp Val Ala Gly Ala Ala Leu Ser Ser

115 120 125

Ser Pro Asn Asn Ser Ala Gly Ser Phe Pro Met Asp Asp Phe Ser Gly

130 135 140

His Gly Leu Gly Gly Asn Asp Ala Ala Pro Gly Gly Gly Gly Gly Asp

145 150 155 160

Arg Ser Cys Ser Arg Ala Ser Asp Glu Asp Asp Gly Gly Ser Ala Arg

165 170 175

Lys Lys Leu Arg Leu Ser Lys Glu Gln Ser Ala Phe Leu Glu Glu Ser

180 185 190

Phe Lys Glu His Ser Thr Leu Asn Pro Lys Gln Lys Leu Ala Leu Ala

195 200 205

Lys Gln Leu Asn Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn

210 215 220

Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Tyr

225 230 235 240

Leu Lys Arg Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu Gln

245 250 255

Lys Glu Leu Ala Glu Leu Arg Ala Leu Lys Thr Val His Pro Phe Tyr

260 265 270

Met His Leu Pro Ala Thr Thr Leu Ser Met Cys Pro Ser Cys Glu Arg

275 280 285

Val Ala Ser Asn Ser Ala Pro Ala Thr Ala Ser Ser Ala Ala Thr Ser

290 295 300

Ser Thr Ala Ala Pro Pro Ala Ala Pro Ser Ser Gly Gly Ile Ala Ala

305 310 315 320

Thr Ser Ser Ser Ala Ala Ala Ala Ala Ala Pro Asp His Arg Pro Ser

325 330 335

Ser Phe Ala Ala Leu Phe Ser Ser Pro Arg Gly Phe Pro Leu Ser Val

340 345 350

Ala Pro Gln Ala Gln Pro Pro Thr Ser Ser

355 360

<210>243

<211>771

<212>DNA

＜213〉rice

<400>243

atggagaggc aaggcttgga tcttggcctg agcctcgggc taggcttgac gacggcggcg 60

acatggccgg ctgctgggtt ctgtctgaac tccggcatgg cggagcagga agtgatcagg 120

cgtgatgatg tggttgcggc gacggcggcg gaggatgaga ggttcgcgtg ctcacccggc 180

agcccggtgt cgagcggcag cgggaagcga ggcagcggca gcggcagcgg cgacgaggtc 240

gacgacgccg gctgcgacgt cggcggcggc ggcgcgcgca agaagctgcg gttgtccaag 300

gaccaggccg ccgtcctcga ggagtgcttc aagacgcacc acaccctcac tccgaagcag 360

aaggtggcgc tggcgaagag cttgaacctg cggccgcggc aggtggaggt gtggttccag 420

aaccgccgcg cgaggacgaa gctgaagcag acggaggtgg actgcgagca cctcaagcgg 480

tggtgcgacc agctcgccga cgacaaccgc cgcctccaca aggagctcgc cgagctcagg 540

gcgctcaagg ccacgcccac accgcccgcc gccgcgccgc cattgaccac cctcacaatg 600

tgcctctcct gcaagcgcgt cgccaatgcc ggcgtgccct cgccggcggc ggcgatattc 660

cccggccacc cccagttctt gtgcggattc agagatcacg ccggagcagc gtcgtcgtcg 720

tacggcggcg catcatctgg actcgcgaag gcggtcaggg cggcgaggta g 771

<210>244

<211>256

<212>PRT

＜213〉rice

<400>244

Met Glu Arg Gln Gly Leu Asp Leu Gly Leu Ser Leu Gly Leu Gly Leu

1 5 10 15

Thr Thr Ala Ala Thr Trp Pro Ala Ala Gly Phe Cys Leu Asn Ser Gly

20 25 30

Met Ala Glu Gln Glu Val Ile Arg Arg Asp Asp Val Val Ala Ala Thr

35 40 45

Ala Ala Glu Asp Glu Arg Phe Ala Cys Ser Pro Gly Ser Pro Val Ser

50 55 60

Ser Gly Ser Gly Lys Arg Gly Ser Gly Ser Gly Ser Gly Asp Glu Val

65 70 75 80

Asp Asp Ala Gly Cys Asp Val Gly Gly Gly Gly Ala Arg Lys Lys Leu

85 90 95

Arg Leu Ser Lys Asp Gln Ala Ala Val Leu Glu Glu Cys Phe Lys Thr

100 105 110

His His Thr Leu Thr Pro Lys Gln Lys Val Ala Leu Ala Lys Ser Leu

115 120 125

Asn Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg Arg Ala

130 135 140

Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu His Leu Lys Arg

145 150 155 160

Trp Cys Asp Gln Leu Ala Asp Asp Asn Arg Arg Leu His Lys Glu Leu

165 170 175

Ala Glu Leu Arg Ala Leu Lys Ala Thr Pro Thr Pro Pro Ala Ala Ala

180 185 190

Pro Pro Leu Thr Thr Leu Thr Met Cys Leu Ser Cys Lys Arg Val Ala

195 200 205

Asn Ala Gly Val Pro Ser Pro Ala Ala Ala Ile Phe Pro Gly His Pro

210 215 220

Gln Phe Leu Cys Gly Phe Arg Asp His Ala Gly Ala Ala Ser Ser Ser

225 230 235 240

Tyr Gly Gly Ala Ser Ser Gly Leu Ala Lys Ala Val Arg Ala Ala Arg

245 250 255

<210>245

<211>927

<212>DNA

＜213〉rice

<400>245

atgatggatc tcggcctcag cctcggcctc ggcctcgcct cgcagggcag cctcacctcc 60

tccaccacca ccacctcctc ccccggcgcc ggatcatcct ccccgtgggc cgccgcgctc 120

aactccatcg tcggcgacgt gaggcgggat caggccgcgg cgcatgctgc cgcggcggtg 180

ggggttgggg ttgggggcga ggagatgtac caggggaggg cgtccacgtc gccggacagc 240

gcggcggcgc tgtcgagcgc gagcgggaag agggagaggg agctggagcg gtcgggatcc 300

ggggttgacg acgacgacgg cgcggacggc gccggcgggc ggaagaagct caggctgtcc 360

aaggaccagg ccgccgtgct cgaggagtgc ttcaagacgc actccactct caaccccaag 420

cagaaggtgg cgctggcgaa caggctgggg ctgcggccgc ggcaggtgga ggtgtggttc 480

cagaaccgga gggcgaggac gaagctgaag cagacggagg tggactgcga gtacctgaag 540

cggtggtgcg agcgcctcgc cgacgagaac aagcgcctcg agaaggagct cgccgacctc 600

agggcgctca aggccgcgcc ctcgccggcg tccgcgtccg cgatgcagcc ctcctcctcc 660

gccgccgcca cgctcaccat gtgcccctcc tgccgccgcg tcgccaccgc cggcgcgccg 720

caccagccta accaccaaca atgccatccc aaatctaaca ccaccatctc ctcctcctcc 780

accgccgccg ccgccgtcgc cgtcgccggc ggcaacgtgc tgcccagcca ctgccagttc 840

ttcccggccg ccgccgccgc cgccgaccgg acaagccaga gcacgtggaa cgccgccgcg 900

ccgctcgtca ccagagagct cttctag 927

<210>246

<211>308

<212>PRT

＜213〉rice

<400>246

Met Met Asp Leu Gly Leu Ser Leu Gly Leu Gly Leu Ala Ser Gln Gly

1 5 10 15

Ser Leu Thr Ser Ser Thr Thr Thr Thr Ser Ser Pro Gly Ala Gly Ser

20 25 30

Ser Ser Pro Trp Ala Ala Ala Leu Asn Ser Ile Val Gly Asp Val Arg

35 40 45

Arg Asp Gln Ala Ala Ala His Ala Ala Ala Ala Val Gly Val Gly Val

50 55 60

Gly Gly Glu Glu Met Tyr Gln Gly Arg Ala Ser Thr Ser Pro Asp Ser

65 70 75 80

Ala Ala Ala Leu Ser Ser Ala Ser Gly Lys Arg Glu Arg Glu Leu Glu

85 90 95

Arg Ser Gly Ser Gly Val Asp Asp Asp Asp Gly Ala Asp Gly Ala Gly

100 105 110

Gly Arg Lys Lys Leu Arg Leu Ser Lys Asp Gln Ala Ala Val Leu Glu

115 120 125

Glu Cys Phe Lys Thr His Ser Thr Leu Asn Pro Lys Gln Lys Val Ala

130 135 140

Leu Ala Asn Arg Leu Gly Leu Arg Pro Arg Gln Val Glu Val Trp Phe

145 150 155 160

Gln Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys

165 170 175

Glu Tyr Leu Lys Arg Trp Cys Glu Arg Leu Ala Asp Glu Asn Lys Arg

180 185 190

Leu Glu Lys Glu Leu Ala Asp Leu Arg Ala Leu Lys Ala Ala Pro Ser

195 200 205

Pro Ala Ser Ala Ser Ala Met Gln Pro Ser Ser Ser Ala Ala Ala Thr

210 215 220

Leu Thr Met Cys Pro Ser Cys Arg Arg Val Ala Thr Ala Gly Ala Pro

225 230 235 240

His Gln Pro Asn His Gln Gln Cys His Pro Lys Ser Asn Thr Thr Ile

245 250 255

Ser Ser Ser Ser Thr Ala Ala Ala Ala Val Ala Val Ala Gly Gly Asn

260 265 270

Val Leu Pro Ser His Cys Gln Phe Phe Pro Ala Ala Ala Ala Ala Ala

275 280 285

Asp Arg Thr Ser Gln Ser Thr Trp Asn Ala Ala Ala Pro Leu Val Thr

290 295 300

Arg Glu Leu Phe

305

<210>247

<211>936

<212>DNA

＜213〉rice

<400>247

atggagatga tggttcatgg gaggagagac gagcagtatg gcgggctcag gctcgggctt 60

gggcttgggc tcagcctcgg cgtcgccggt ggtgcagccg acgacgagca gccgccgccg 120

cgccgtggtg ccgccccgcc gccgcagcag cagctgtgcg gctggaacgg cggcggtctc 180

ttctcctcgt cttcctccga tcatcggggg aggtcggcga tgatggcgtg ccacgacgtc 240

atcgagatgc cgttcctgcg ggggatcgac gtgaaccgtg cgccggcggc agagacgacc 300

acgacgacgg cgagggggcc cagctgcagc gaggaagacg aggagcccgg cgcgtcctcc 360

cccaacagca cgctctccag cctcagcggc aagcgcggcg caccatctgc cgccaccgcc 420

gccgccgccg ccgccagcga cgacgaggac tccggcggcg gatcccgcaa gaagctccgc 480

ctctccaagg accaagccgc cgtcctcgag gacaccttca aagagcacaa caccctcaat 540

cccaagcaga aggcggcgct ggcgaggcag ctgaatctga agccgcggca ggtggaggtg 600

tggttccaga acaggagggc gaggacgaag ctgaagcaga cggaggtgga ctgcgagctg 660

ctcaagcgct gctgcgagac gctcaccgac gagaaccgcc gcctccaccg cgagctccag 720

gagctccgcg ccctcaagct cgccaccgcc gccgccgcgc cgcaccacct ctacggcgcc 780

cgcgtcccgc cgcccaccac cctcaccatg tgcccctcct gcgagcgcgt cgcctccgca 840

gccaccacca cccgcaacaa ctccggcgcc gcccccgcgc ggccggtgcc cacccgcccg 900

tggccgccgg cggcggcgca gaggtcgtcg gcgtag 936

<210>248

<211>311

<212>PRT

＜213〉rice

<400>248

Met Glu Met Met Val His Gly Arg Arg Asp Glu Gln Tyr Gly Gly Leu

1 5 10 15

Arg Leu Gly Leu Gly Leu Gly Leu Ser Leu Gly Val Ala Gly Gly Ala

20 25 30

Ala Asp Asp Glu Gln Pro Pro Pro Arg Arg Gly Ala Ala Pro Pro Pro

35 40 45

Gln Gln Gln Leu Cys Gly Trp Asn Gly Gly Gly Leu Phe Ser Ser Ser

50 55 60

Ser Ser Asp His Arg Gly Arg Ser Ala Met Met Ala Cys His Asp Val

65 70 75 80

Ile Glu Met Pro Phe Leu Arg Gly Ile Asp Val Asn Arg Ala Pro Ala

85 90 95

Ala Glu Thr Thr Thr Thr Thr Ala Arg Gly Pro Ser Cys Ser Glu Glu

100 105 110

Asp Glu Glu Pro Gly Ala Ser Ser Pro Asn Ser Thr Leu Ser Ser Leu

115 120 125

Ser Gly Lys Arg Gly Ala Pro Ser Ala Ala Thr Ala Ala Ala Ala Ala

130 135 140

Ala Ser Asp Asp Glu Asp Ser Gly Gly Gly Ser Arg Lys Lys Leu Arg

145 150 155 160

Leu Ser Lys Asp Gln Ala Ala Val Leu Glu Asp Thr Phe Lys Glu His

165 170 175

Asn Thr Leu Asn Pro Lys Gln Lys Ala Ala Leu Ala Arg Gln Leu Asn

180 185 190

Leu Lys Pro Arg Gln Val Glu Val Trp Phe Gln Asn Arg Arg Ala Arg

195 200 205

Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Leu Leu Lys Arg Cys

210 215 220

Cys Glu Thr Leu Thr Asp Glu Asn Arg Arg Leu His Arg Glu Leu Gln

225 230 235 240

Glu Leu Arg Ala Leu Lys Leu Ala Thr Ala Ala Ala Ala Pro His His

245 250 255

Leu Tyr Gly Ala Arg Val Pro Pro Pro Thr Thr Leu Thr Met Cys Pro

260 265 270

Ser Cys Glu Arg Val Ala Ser Ala Ala Thr Thr Thr Arg Asn Asn Ser

275 280 285

Gly Ala Ala Pro Ala Arg Pro Val Pro Thr Arg Pro Trp Pro Pro Ala

290 295 300

Ala Ala Gln Arg Ser Ser Ala

305 310

<210>249

<211>1674

<212>DNA

＜213〉Arabidopis thaliana

<400>249

aaaatggaaa actcagacac tgattcagaa gttttctttt ggtttcagaa ccaaaaccaa 60

aaccatagcc ataagtttcc ttcttcttgt tttcctccga gctctcactc tgctttttat 120

ggatctagct caatgatcaa cacagagact gcaactatgg acgaagaaga cgtatgtgag 180

agctacatga tgcgtgagat aaccaagaaa cggaagctaa caccgatcca attgcgttta 240

ctagaggaga gtttcgaaga agagaaaaga ctcgaaccag acaggaaact ctggctagcc 300

gagaagctag ggctgcagcc aagccaagtc gctgtctggt tccaaaacag aagagctaga 360

tacaagacca aacagctcga acacgactgt gactctctta aagctagcta tgctaagctc 420

aagactgact gggacattct ctttgtacaa aaccaaaccc tcaagagcaa ggtacagttt 480

cttaatagat taacaagcca ctattttcaa gagtctgttc aaaattttga tgacacattc 540

aaacaggtcg atcttctcaa agagaagctg aaaatgcaag agaatcttga aactcagtct 600

attgagagga aaagacttgg ggaagaaggt agttcggtga aaagcgataa cacgcagtac 660

agtgaagaag aaggtttgga gaatcaatac agtttcccgg agcttgcagt tctcggattt 720

tattatgatc caaccttaac tgcttcaaat ctaagacaag aacctttgaa agtcacgtgt 780

gcggatcaga tgactcagat ccaaatatct gacgtcacag aacctgcgag ctccgcacat 840

aaaaaaattg aagtgacaca gcgaagttcg agtatgagcc gcaagagaga taaaccctac 900

acaaaccgtc acacaccggc gcgaatttcg aaacgccggc gaccatgggc gccttcgtca 960

tcggagcacg atgagattat cgacaaaccg atcaccaaac ctccgccgcc accggcgttg 1020

gtagttatgg gacttcccgc caactgttca gttctggagc taaaatctcg attcgagatc 1080

tacggttcaa tctcccgaat ccgaatccat aaagacggaa ttggctccgt ttcgtatcga 1140

accgccgagt cagcagaagc cgccattgct ggtagtcacg agccttcttt cggtatctcc 1200

atcgattcca aaaagttgga ggtggtttgg gcgacggatc cattggtgaa gtggaaggaa 1260

ggtgtgacgg cgggagaagg aaaggagaga acgtcgtcgt tttcgtcgaa gcttttacga 1320

cctgtgatgc ctttgagaaa acatgggaga agcagtaggt tagcgtcagc tattgttaat 1380

ccgagaagtg ataatactaa aggaattagt ggagatggag ggatatcatc tccggcgacg 1440

acaagtgaag ttaaacagag aaatatagta acctacgacg atatcgttta acaccattct 1500

actacaatta tagtaacttt gattctttta catattttgg caattttatg agtttagcta 1560

atcctgttgg taatctcagt tgtcgaatgt tgtgaaatga gttgagtttt gtttatttgt 1620

atgattgtaa cttataagaa agagaaggat tccaaaatgg gtagaagatt caaa 1674

<210>250

<211>495

<212>PRT

＜213〉Arabidopis thaliana

<400>250

Met Glu Asn Ser Asp Thr Asp Ser Glu Val Phe Phe Trp Phe Gln Asn

1 5 10 15

Gln Asn Gln Asn His Ser His Lys Phe Pro Ser Ser Cys Phe Pro Pro

20 25 30

Ser Ser His Ser Ala Phe Tyr Gly Ser Ser Ser Met Ile Asn Thr Glu

35 40 45

Thr Ala Thr Met Asp Glu Glu Asp Val Cys Gl u SerTyr Met Met Arg

50 55 60

Glu Ile Thr Lys Lys Arg Lys Leu Thr Pro Ile Gln Leu Arg Leu Leu

65 70 75 80

Glu Glu Ser Phe Glu Glu Glu Lys Arg Leu Glu Pro Asp Arg Lys Leu

85 90 95

Trp Leu Ala Glu Lys Leu Gly Leu Gln Pro Ser Gln Val Ala Val Trp

100 105 110

Phe Gln Asn Arg Arg Ala Arg Tyr Lys Thr Lys Gln Leu Glu His Asp

115 120 125

Cys Asp Ser Leu Lys Ala Ser Tyr Ala Lys Leu Lys Thr Asp Trp Asp

130 135 140

Ile Leu Phe Val Gln Asn Gln Thr Leu Lys Ser Lys Val Gln Phe Leu

145 150 155 160

Asn Arg Leu Thr Ser His Tyr Phe Gln Glu Ser Val Gln Asn Phe Asp

165 170 175

AspThr Phe Lys Gln Val Asp Leu Leu Lys Glu Lys Leu Lys Met Gln

180 185 190

Glu Asn Leu Glu Thr Gln Ser Ile Glu Arg Lys Arg Leu Gly Glu Glu

195 200 205

Gly Ser Ser Val Lys Ser Asp Asn Thr Gln Tyr Ser Glu Glu Glu Gly

210 215 220

Leu Glu Asn Gln Tyr Ser Phe Pro Glu Leu Ala Val Leu Gly Phe Tyr

225 230 235 240

Tyr Asp Pro Thr Leu Thr Ala Ser Asn Leu Arg Gln Glu Pro Leu Lys

245 250 255

Val Thr Cys Ala Asp Gln Met Thr Gln Ile Gln Ile Ser Asp Val Thr

260 265 270

Glu Pro Ala Ser Ser Ala His Lys Lys Ile Glu ValThr Gln Arg Ser

275 280 285

Ser Ser Met Ser Arg Lys Arg Asp Lys Pro Tyr Thr Asn Arg His Thr

290 295 300

Pro Ala Arg Ile Ser Lys Arg Arg Arg Pro Trp Ala Pro Ser Ser Ser

305 310 315 320

Glu His Asp Glu Ile Ile Asp Lys Pro Ile Thr Lys Pro Pro Pro Pro

325 330 335

Pro Ala Leu Val Val Met Gly Leu Pro Ala Asn Cys Ser Val Leu Glu

340 345 350

Leu Lys Ser Arg Phe Glu Ile Tyr Gly Ser Ile Ser Arg Ile Arg Ile

355 360 365

His Lys Asp Gly Ile Gly Ser Val Ser Tyr Arg Thr Ala Glu Ser Ala

370 375 380

Glu Ala Ala Ile Ala Gly Ser His Glu Pro Ser Phe Gly Ile Ser Ile

385 390 395 400

Asp Ser Lys Lys Leu Glu Val Val Trp Ala Thr Asp Pro Leu Val Lys

405 410 415

Trp Lys Glu Gly Val Thr Ala Gly Glu Gly Lys Glu Arg Thr Ser Ser

420 425 430

Phe Ser Ser Lys Leu Leu Arg Pro Val Met Pro Leu Arg Lys His Gly

435 440 445

Arg Ser Ser Arg Leu Ala Ser Ala Ile Val Asn Pro Arg Ser Asp Asn

450 455 460

Thr Lys Gly Ile Ser Gly Asp Gly Gly Ile Ser Ser Pro Ala Thr Thr

465 470 475 480

Ser Glu Val Lys Gln Arg Asn Ile Val Thr Tyr Asp Asp Ile Val

485 490 495

<210>251

<211>1428

<212>DNA

＜213〉Arabidopis thaliana

<400>251

aaggccacaa cagatcttct tgttcctctc tctcaatctc tctctctaaa actcttattt 60

tcttcgtaag agatggaact agcgttgtct ctaggcgaca atactaagaa acagttctct 120

tttatggaga agaactcgaa gattaataat ccctctgtct cgtcgacatc tacttccgag 180

aaggatcttg ggttctgcat ggctttagat gttgcttttg gtggtcacag atcgttgtca 240

tcctcttcgt ctccgtcggt agaggatgag aagaagaaac cggcgcccag agcaaaaaaa 300

tctgacgaat ttagggtttc gtcttctgta gatccaccat tacagcttca gcttcacttc 360

cctaattggc tccctgagaa cagtaaaggt cgacaaggag gaagaatgcc cttaggagca 420

gctacggttg tggaggagga agaggaggag gaggaagcgg tgcctagtat gtcagtatcg 480

ccgccggata gtgtaacgtc gtcgtttcaa ttggactttg ggattaaaag ttatggttat 540

gagagaagaa gcaataagag agatattgat gatgaagtgg agagatcagc ttcaagagcc 600

agcaacgaag acaacgatga cgagaatgga tccactagga agaaacttag actctccaaa 660

gaccaatctg cttttcttga agacagcttc aaagaacaca gtacccttaa tcctgttcgt 720

gtcccattct ttacagtttt tatttattta aaatttgtct ttcttgaatt tattctattt 780

ttttagtacc aaattttagc cttaatgctt ttttttttct ttctaaaaca ttgcagaaac 840

agaagattgc attggcgaag cagttgaatc ttcgtcctcg tcaggttgaa gtctggtttc 900

aaaacagacg agccaggaca aagctgaagc aaacggaagt ggactgtgaa tacctaaaga 960

gatgctgtga gtcactaacc gaagaaaacc ggaggcttca aaaagaggtt aaagaattga 1020

gaaccttgaa gacttccaca cccttttaca tgcaacttcc ggccactact ctcactatgt 1080

gcccttcttg tgaacgtgtt gccacttcag cagcacagcc ctccacgtca gctgcccaca 1140

acctctgttt gtccacgtca tcattgattc cggttaagcc tcggccggcc aaacaagttt 1200

catgaaagca cctgcgaaat acagtttgag caaacggtcg agctagagtg gttttaaaag 1260

ttgtcttctt gtgtatatat ttattttact tttcatattt tattagagac cgctattttg 1320

aaagacgaat agattgatta tccggttagt gttttgtttt tcttagatag gaccggataa 1380

aaaacagatg gagcaaaagg gttgacatgt tttattgaat gatagagg 1428

<210>252

<211>237

<212>PRT

＜213〉Arabidopis thaliana

<400>252

Met Glu Leu Ala Leu Ser Leu Gly Asp Asn Thr Lys Lys Gln Phe Ser

1 5 10 15

Phe Met Glu Lys Asn Ser Lys Ile Asn Asn Pro Ser Val Ser Ser Thr

20 25 30

Ser Thr Ser Glu Lys Asp Leu Gly Phe Cys Met Ala Leu Asp Val Ala

35 40 45

Phe Gly Gly His Arg Ser Leu Ser Ser Ser Ser Ser Pro Ser Val Glu

50 55 60

Asp Glu Lys Lys Lys Pro Ala Pro Arg Ala Lys Lys Ser Asp Glu Phe

65 70 75 80

Arg Val Ser Ser Ser Val Asp Pro Pro Leu Gln Leu Gln Leu His Phe

85 90 95

Pro Asn Trp Leu Pro Glu Asn Ser Lys Gly Arg Gln Gly Gly Arg Met

100 105 110

Pro Leu Gly Ala Ala Thr Val Val Glu Glu Glu Glu Glu Glu Glu Glu

115 120 125

Ala Val Pro Ser Met Ser Val Ser Pro Pro Asp Ser Val Thr Ser Ser

130 135 140

Phe Gln Leu Asp Phe Gly Ile Lys Ser Tyr Gly Tyr Glu Arg Arg Ser

145 150 155 160

Asn Lys Arg Asp Ile Asp Asp Glu Val Glu Arg Ser Ala Ser Arg Ala

165 170 175

Ser Asn Glu Asp Asn Asp Asp Glu Asn Gly Ser Thr Arg Lys Lys Leu

180 185 190

Arg Leu Ser Lys Asp Gln Ser Ala Phe Leu Glu Asp Ser Phe Lys Glu

195 200 205

His Ser Thr Leu Asn Pro Val Arg Val Pro Phe Phe Thr Val Phe Ile

210 215 220

Tyr Leu Lys Phe Val Phe Leu Glu Phe Ile Leu Phe Phe

225 230 235

<210>253

<211>1167

<212>DNA

＜213〉Arabidopis thaliana

<400>253

tgcccccagc tagtcacata ctcatgattg caaaatctct ctctctctct gcctctctat 60

atattaacct ttcttcttcc tttactttct catcttctat ctctcaaaag aaaagcagac 120

aactttattt gcaaaaacag agtttttttt tcttatcttg agaaagttca acagaagatg 180

atgttcgaga aagacgatct gggtctaagc ttaggcttga attttccaaa gaaacagatc 240

aatctcaaat caaatccatc tgtttctgtt actccttctt cttcttcttt tggattattc 300

agaagatctt catggaacga gagttttact tcttcagttc caaactcaga ttcgtcacaa 360

aaagaaacaa gaactttcat ccgaggaatc gacgtgaaca gaccaccgtc tacagcggaa 420

tacggcgacg aagacgctgg agtatcttca cctaacagta cagtctcaag ctctacaggg 480

aaaagaagcg agagagaaga agacacagat ccacaaggct caagaggaat cagtgacgat 540

gaagatggtg ataactccag gaaaaagctt agactttcca aagatcaatc tgctattctt 600

gaagagacct tcaaagatca cagtactctc aatccgaagc agaagcaagc attggctaaa 660

caattagggt tacgagcaag acaagtggaa gtttggtttc agaacagacg agcaagaaca 720

aagctgaagc aaacggaggt agactgcgag ttcttacgga gatgctgcga gaatctaacg 780

gaagagaacc gtcggctaca aaaagaagta acggaattga gagcacttaa gctctctcct 840

cagttctaca tgcacatgag cccacccact actttgacca tgtgcccttc atgtgaacac 900

gtgtcggtcc cgccaccaca acctcaggct gctacgtcag cgcaccaccg gtcgttgccg 960

gtcaatgcgt gggctcctgc gacgaggata tctcacggct tgacttttga cgctcttcgt 1020

cctaggtcct aagtcttttt acttgcaacc aaagggcatt ttggtcgttt tttaagtttc 1080

atggaccaga tatgcatgta gttgttaaca tgtatgtatt ttcttagaaa gaaagaaaaa 1140

cagattaata tttttctagc ttaaacc 1167

<210>254

<211>284

<212>PRT

＜213〉Arabidopis thaliana

<400>254

Met Met Phe Glu Lys Asp Asp Leu Gly Leu Ser Leu Gly Leu Asn Phe

1 5 10 15

Pro Lys Lys Gln Ile Asn Leu Lys Ser Asn Pro Ser Val Ser Val Thr

20 25 30

Pro Ser Ser Ser Ser Phe Gly Leu Phe Arg Arg Ser Ser Trp Asn Glu

35 40 45

Ser Phe Thr Ser Ser Val Pro Asn Ser Asp Ser Ser Gln Lys Glu Thr

50 55 60

Arg Thr Phe Ile Arg Gly Ile Asp Val Asn Arg Pro Pro Ser Thr Ala

65 70 75 80

Glu Tyr Gly Asp Glu Asp Ala Gly Val Ser Ser Pro Asn Ser Thr Val

85 90 95

Ser Ser Ser Thr Gly Lys Arg Ser Glu Arg Glu Glu Asp Thr Asp Pro

100 105 110

Gln Gly Ser Arg Gly Ile Ser Asp Asp Glu Asp Gly Asp Asn Ser Arg

115 120 125

Lys Lys Leu Arg Leu Ser Lys Asp Gln Ser Ala Ile Leu Glu Glu Thr

130 135 140

Phe Lys Asp His Ser Thr Leu Asn Pro Lys Gln Lys Gln Ala Leu Ala

145 150 155 160

Lys Gln Leu Gly Leu Arg Ala Arg Gln Val Glu Val Trp Phe Gln Asn

165 170 175

Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu Phe

180 185 190

Leu Arg Arg Cys Cys Glu Asn Leu Thr Glu Glu Asn Arg Arg Leu Gln

195 200 205

Lys Glu Val Thr Glu Leu Arg Ala Leu Lys Leu Ser Pro Gln Phe Tyr

210 215 220

Met His Met Ser Pro Pro Thr Thr Leu Thr Met Cys Pro Ser Cys Glu

225 230 235 240

His Val Ser Val Pro Pro Pro Gln Pro Gln Ala Ala Thr Ser Ala His

245 250 255

His Arg Ser Leu Pro Val Asn Ala Trp Ala Pro Ala Thr Arg Ile Ser

260 265 270

His Gly Leu Thr Phe Asp Ala Leu Arg Pro Arg Ser

275 280

<210>255

<211>1194

<212>DNA

＜213〉Arabidopis thaliana

<400>255

atttcacatc tctctctctc tataagaacc ctagaagaag gtttctcttg tcctccatac 60

acttagcaca actgataaat cttttgaggt aaaatcagct ttagatcaag gtttttctag 120

tcatctctac tcataaagat caaagctttt gctattctca ttttctacca agagacaata 180

tcatgatgat gggtaaagag gatttgggtt taagtcttag cttgggattt gcacaaaacc 240

atcctctcca gctaaatctt aaacccactt cttcaccaat gtccaatctc cagatgtttc 300

catggaacca aacccttgtt tcttcctcag atcaacaaaa gcaacagttt cttaggaaaa 360

tcgacgtgaa cagcttgcca acaacggtgg atttggaaga ggagacagga gtttcgtctc 420

caaacagtac gatctcgagc acagtgagtg gaaagaggag gagtactgaa agagaaggta 480

cctccggtgg tggttgcgga gatgaccttg acatcactct agatagatct tcctcacgtg 540

gaacctccga tgaagaggaa gattacggag gtgagacttg taggaagaag cttagactat 600

ccaaagatca atccgcagtt ctcgaagaca ctttcaaaga gcacaatact ctcaatccca 660

aacagaagct ggctttggct aagaagctag gtttaacagc aagacaagtg gaagtgtggt 720

tccaaaacag aagagcaagg acaaagttaa agcagaccga agtggattgc gagtatttga 780

aaagatgtgt tgagaaatta acggaagaga atcggcggct cgagaaagag gcagcggaac 840

taagagcatt aaagctttca ccgcggttgt atggtcagat gagtccaccg accacacttt 900

tgatgtgtcc atcgtgtgaa cgtgtggccg gaccatcctc atctaaccac aaccagcgat 960

ctgtctcatt gagtccatgg ctccaaatgg cccatgggtc aacctttgat gtgatgcgtc 1020

ctaggtctta actttaatgc tgcttctatg ggttgtgtgt gggtcattgt actttttaga 1080

ttattgactc tcagctaatg tatccttaaa agcctttttc tacttttaaa tttactttaa 1140

tctaattaaa ttagttattc ttgtcttctt gataacaaac aaatttataa taat 1194

<210>256

<211>282

<212>PRT

＜213〉Arabidopis thaliana

<400>256

Met Met Met Gly Lys Glu Asp Leu Gly Leu Ser Leu Ser Leu Gly Phe

1 5 10 15

Ala Gln Asn His Pro Leu Gln Leu Asn Leu Lys Pro Thr Ser Ser Pro

20 25 30

Met Ser Asn Leu Gln Met Phe Pro Trp Asn Gln Thr Leu Val Ser Ser

35 40 45

Ser Asp Gln Gln Lys Gln Gln Phe Leu Arg Lys Ile Asp Val Asn Ser

50 55 60

Leu Pro Thr Thr Val Asp Leu Glu Glu Glu Thr Gly Val Ser Ser Pro

65 70 75 80

Asn Ser Thr Ile Ser Ser Thr Val Ser Gly Lys Arg Arg Ser Thr Glu

85 90 95

Arg Glu Gly Thr Ser Gly Gly Gly Cys Gly Asp Asp Leu Asp Ile Thr

100 105 110

Leu Asp Arg Ser Ser Ser Arg Gly Thr Ser Asp Glu Glu Glu Asp Tyr

115 120 125

Gly Gly Glu Thr Cys Arg Lys Lys Leu Arg Leu Ser Lys Asp Gln Ser

130 135 140

Ala Val Leu Glu Asp Thr Phe Lys Glu His Asn Thr Leu Asn Pro Lys

145 150 155 160

Gln Lys Leu Ala Leu Ala Lys Lys Leu Gly Leu Thr Ala Arg Gln Val

165 170 175

Glu Val Trp Phe Gln Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr

180 185 190

Glu Val Asp Cys Glu Tyr Leu Lys Arg Cys Val Glu Lys Leu Thr Glu

195 200 205

Glu Asn Arg Arg Leu Glu Lys Glu Ala Ala Glu Leu Arg Ala Leu Lys

210 215 220

Leu Ser Pro Arg Leu Tyr Gly Gln Met Ser Pro Pro Thr Thr Leu Leu

225 230 235 240

Met Cys Pro Ser Cys Glu Arg Val Ala Gly Pro Ser Ser Ser Asn His

245 250 255

Asn Gln Arg Ser Val Ser Leu Ser Pro Trp Leu Gln Met Ala His Gly

260 265 270

Ser Thr Phe Asp Val Met Arg Pro Arg Ser

275 280

<210>257

<211>1284

<212>DNA

＜213〉Arabidopis thaliana

<400>257

aataatgatg gattgtaatt agccacctga caaaatctct catcattcaa agtactatat 60

taatcccctc tcttcttcat taccatctca catctctctc tatttctctt ccacaaagag 120

tcctaacttc gagttgaaac aaacaccatt tctcatctct atctcagaaa gaacaaacca 180

tttcgtgttc tttctttctc tattctcata aggaaatata attcctgaaa ctgttgagtt 240

cttgtgaaag gaaataaaaa acatgatgat gggcaaagaa gatctaggtt tgagcctaag 300

cttagggttt tcacaaaatc acaatcctct tcagatgaat ctgaatccta actcttcatt 360

atcaaacaat ctccagagac tcccatggaa ccaaacattc gatcctacat cagatcttcg 420

caagatagac gtgaacagtt ttccatcaac ggttaactgc gaggaagaca caggagtttc 480

gtcaccaaac agtacgatct caagcaccat tagcgggaag agaagtgaga gagaaggaat 540

ctccggaacc ggcgttggct ccggcgacga tcacgacgag atcactccgg atcgagggta 600

ctcacgtgga acctcagatg aagaagaaga cgggggcgaa acgtcgagga agaagctcag 660

gttatcaaaa gatcagtctg cttttctcga agagactttc aaagaacaca acactctcaa 720

tcccaaacag aagctagctt tggctaagaa gctgaacttg acggcaagac aagtggaagt 780

gtggttccaa aacagaagag ctagaaccaa gttaaagcaa acggaggtag attgcgaata 840

cttgaaacgg tgcgtagaga agctaacgga agagaaccgg agacttcaga aagaggctat 900

ggagcttcga actctcaagc tgtctccaca attctacggt cagatgactc caccaactac 960

actcatcatg tgtccttcgt gcgagcgtgt gggtggccca tcatcatcga accatcacca 1020

caatcacagg cccgtttcta tcaatccgtg ggttgcttgt gctggtcagg tggctcatgg 1080

gctgaatttt gaagccttgc gtccacgatc gtgatttttt attttagtgg tgggaaaagg 1140

gtgttttggt atttttcgtt atcgttatat agtctatctg tgtggggtca ttgtaatttt 1200

ggatgattgg ccttctcatg aactagtcct atgtatgatg caaccttaaa aagatttaaa 1260

ttagcaaaaa ttagttacaa actt 1284

<210>258

<211>283

<212>PRT

＜213〉Arabidopis thaliana

<400>258

Met Met Met Gly Lys Glu Asp Leu Gly Leu Ser Leu Ser Leu Gly Phe

1 5 10 15

Ser Gln Asn His Asn Pro Leu Gln Met Asn Leu Asn Pro Asn Ser Ser

20 25 30

Leu Ser Asn Asn Leu Gln Arg Leu Pro Trp Asn Gln Thr Phe Asp Pro

35 40 45

Thr Ser Asp Leu Arg Lys Ile Asp Val Asn Ser Phe Pro Ser Thr Val

50 55 60

Asn Cys Glu Glu Asp Thr Gly Val Ser Ser Pro Asn Ser Thr Ile Ser

65 70 75 80

Ser Thr Ile Ser Gly Lys Arg Ser Glu Arg Glu Gly Ile Ser Gly Thr

85 90 95

Gly Val Gly Ser Gly Asp Asp His Asp Glu Ile Thr Pro Asp Arg Gly

100 105 110

Tyr Ser Arg Gly Thr Ser Asp Glu Glu Glu Asp Gly Gly Glu Thr Ser

115 120 125

Arg Lys Lys Leu Arg Leu Ser Lys Asp Gln Ser Ala Phe Leu Glu Glu

130 135 140

Thr Phe Lys Glu His Asn Thr Leu Asn Pro Lys Gln Lys Leu Ala Leu

145 150 155 160

Ala Lys Lys Leu Asn Leu Thr Ala Arg Gln Val Glu Val Trp Phe Gln

165 170 175

Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu

180 185 190

Tyr Leu Lys Arg Cys Val Glu Lys Leu Thr Glu Glu Asn Arg Arg Leu

195 200 205

Gln Lys Glu Ala Met Glu Leu Arg Thr Leu Lys Leu Ser Pro Gln Phe

210 215 220

Tyr Gly Gln Met Thr Pro Pro Thr Thr Leu Ile Met Cys Pro Ser Cys

225 230 235 240

Glu Arg Val Gly Gly Pro Ser Ser Ser Asn His His His Asn His Arg

245 250 255

Pro Val Ser Ile Asn Pro Trp Val Ala Cys Ala Gly Gln Val Ala His

260 265 270

Gly Leu Asn Phe Glu Ala Leu Arg Pro Arg Ser

275 280

<210>259

<211>957

<212>DNA

＜213〉Arabidopis thaliana

<400>259

atgggggaaa gagatgatgg gttgggtttg agtctaagct tgggaaatag tcaacaaaaa 60

gaaccatctc tgaggttgaa tcttatgccg ttgacaactt cttcttcttc ttcttcgttt 120

caacacatgc acaatcagaa taacaatagc catccccaga agattcataa catctcttgg 180

actcatctgt ttcaatcttc tgggattaaa cgtacaactg cagagagaaa ctccgacgcc 240

gggtcatttc taagaggttt caacgtgaac agagctcagt cttcggtggc ggtagtggac 300

ttggaagaag aagccgccgt cgtctcgtct ccaaacagcg ccgtttcgag tctgagtgga 360

aataaaaggg atcttgcggt ggcgagagga ggagatgaaa acgaggcgga gagagcttct 420

tgctcacgcg gagggggaag cggtggtagc gacgatgaag acggcggaaa cggcgacgga 480

tcaaggaaga aactacggtt atcgaaggat caagctcttg ttctcgagga gacttttaaa 540

gaacatagca ctcttaatcc gaagcaaaag ctggctctag caaaacagtt gaatctaagg 600

gcaagacaag ttgaagtgtg gtttcagaac cgtagggcaa ggacgaagct gaaacaaacg 660

gaggttgatt gtgagtattt aaagagatgt tgcgataatc tgaccgagga gaatcgacgg 720

ct g cagaaag aagtgtcgga gctgagggcg ttgaagttgt ctccacatct ctacatgcac 780

atgactcctc ctactactct caccatgtgc ccttcttgcg aacgtgtctc ctcctctgcc 840

gccactgtga ccgctgctcc ttccactact actactccta cggtggtggg gcggccaagt 900

ccacagcgat taactccttg gactgctatt tctctccagc aaaaatcagg tcgctag 957

<210>260

<211>318

<212>PRT

＜213〉Arabidopis thaliana

<400>260

Met Gly Glu Arg Asp Asp Gly Leu Gly Leu Ser Leu Ser Leu Gly Asn

1 5 10 15

Ser Gln Gln Lys Glu Pro Ser Leu Arg Leu Asn Leu Met Pro Leu Thr

20 25 30

Thr Ser Ser Ser Ser Ser Ser Phe Gln His Met His Asn Gln Asn Asn

35 40 45

Asn Ser His Pro Gln Lys Ile His Asn Ile Ser Trp Thr His Leu Phe

50 55 60

Gln Ser Ser Gly Ile Lys Arg Thr Thr Ala Glu Arg Asn Ser Asp Ala

65 70 75 80

Gly Ser Phe Leu Arg Gly Phe Asn Val Asn Arg Ala Gln Ser Ser Val

85 90 95

Ala Val Val Asp Leu Glu Glu Glu Ala Ala Val Val Ser Ser Pro Asn

100 105 110

Ser Ala Val Ser Ser Leu Ser Gly Asn Lys Arg Asp Leu Ala Val Ala

115 120 125

Arg Gly Gly Asp Glu Asn Glu Ala Glu Arg Ala Ser Cys Ser Arg Gly

130 135 140

Gly Gly Ser Gly Gly Ser Asp Asp Glu Asp Gly Gly Asn Gly Asp Gly

145 150 155 160

Ser Arg Lys Lys Leu Arg Leu Ser Lys Asp Gln Ala Leu Val Leu Glu

165 170 175

Glu Thr Phe Lys Glu His Ser Thr Leu Asn Pro Lys Gln Lys Leu Ala

180 185 190

Leu Ala Lys Gln Leu Asn Leu Arg Ala Arg Gln Val Glu Val Trp Phe

195 200 205

Gln Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys

210 215 220

Glu Tyr Leu Lys Arg Cys Cys Asp Asn Leu Thr Glu Glu Asn Arg Arg

225 230 235 240

Leu Gln Lys Glu Val Ser Glu Leu Arg Ala Leu Lys Leu Ser Pro His

245 250 255

Leu Tyr Met His Met Thr Pro Pro Thr Thr Leu Thr Met Cys Pro Ser

260 265 270

Cys Glu Arg Val Ser Ser Ser Ala Ala Thr Val Thr Ala Ala Pro Ser

275 280 285

Thr Thr Thr Thr Pro Thr Val Val Gly Arg Pro Ser Pro Gln Arg Leu

290 295 300

Thr Pro Trp Thr Ala Ile Ser Leu Gln Gln Lys Ser Gly Arg

305 310 315

<210>261

<211>1357

<212>DNA

＜213〉Arabidopis thaliana

<400>261

ccacaacccc atatctcttt gtctgaaaaa actcttttgc atatataaat aaatataagc 60

catatctttt gaaactgtac tgctgcacaa gtgtagaggc agtggcacaa catctttaag 120

aaagagagag agaaagagtc atttattcat ttcctcgctt aaaaatcttg agcacccaga 180

cagaattctt ctttcttctt tctggggttg agaaaaatga gtgaaagaga tgatggattg 240

gggctaagtt tgagcttgag tttaggtttt aatcaaaagg acccgtcttc gaggttaaat 300

ccaatgcctc tggcttctta tgcatcttca tcacacatgc agcatatgca gcagagcaat 360

tataaccatc ctcaaaagat tcagaacact tggattaaca tgtttcagtc atcagagaga 420

aactcggaca tgagatcgtt tctccgggga atagacgtga acagagctcc atcgacggtg 480

gtggttgacg tggaggatga aggcgccgga gtttcgtctc cgaacagcac cgtctcaagc 540

gtgatgagcg ggaagaagag cgagcgagag ctaatggctg cggcaggtgc agttggagga 600

ggtagagtag aagataatga gattgagaga gcttcttgct cgctcggcgg tggtagcgac 660

gatgaagacg gtagcgggaa cggagatgac agttcgagga agaaactccg attgtctaaa 720

gaacaagctt tggttcttga agaaactttt aaagaacata gtacactcaa tccgaagcaa 780

aagatggctt tggctaagca attgaatctg aggacgagac aagttgaagt gtggttccaa 840

aaccgaaggg caaggacgaa gctgaagcaa acggaagtag actgtgaata tcttaagaga 900

tgttgcgaga atctaacgga tgagaatcgg agattgcaaa aggaagtgag tgagcttagg 960

gctttaaagc tttctccaca cttatacatg cacatgaaac ctcccactac tctcacaatg 1020

tgtccttctt gcgagcgagt cgctgttacg tcatcttcgt catcggtggc tcctcctgtg 1080

atgaattcat cgtctccgat gggtccgatg agtccgtggg ctgccatgcc tctacggcaa 1140

cgacctgctg ctggttctca ttagagttta attagtctaa agataatagg tttggtgatt 1200

tgtttttatt atattgttga acggacttgg aagttctttt caagagttgg tcccatgctc 1260

ttctttcctt tgttttattc agtttctatt tatagctgaa ttctggataa tggaaaatct 1320

actaatatta ttgtccaatt attagtttgg ggaaaga 1357

<210>262

<211>315

<212>PRT

＜213〉Arabidopis thaliana

<400>262

Met Ser Glu Arg Asp Asp Gly Leu Gly Leu Ser Leu Ser Leu Ser Leu

1 5 10 15

Gly Phe Asn Gln Lys Asp Pro Ser Ser Arg Leu Asn Pro Met Pro Leu

20 25 30

Ala Ser Tyr Ala Ser Ser Ser His Met Gln His Met Gln Gln Ser Asn

35 40 45

Tyr Asn His Pro Gln Lys Ile Gln Asn Thr Trp Ile Asn Met Phe Gln

50 55 60

Ser Ser Glu Arg Asn Ser Asp Met Arg Ser Phe Leu Arg Gly Ile Asp

65 70 75 80

Val Asn Arg Ala Pro Ser Thr Val Val Val Asp Val Glu Asp Glu Gly

85 90 95

Ala Gly Val Ser Ser Pro Asn Ser Thr Val Ser Ser Val Met Ser Gly

100 105 110

Lys Lys Ser Glu Arg Glu Leu Met Ala Ala Ala Gly Ala Val Gly Gly

115 120 125

Gly Arg Val Glu Asp Asn Glu Ile Glu Arg Ala Ser Cys Ser Leu Gly

130 135 140

Gly Gly Ser Asp Asp Glu Asp Gly Ser Gly Asn Gly Asp Asp Ser Ser

145 150 155 160

Arg Lys Lys Leu Arg Leu Ser Lys Glu Gln Ala Leu Val Leu Glu Glu

165 170 175

Thr Phe Lys Glu His Ser Thr Leu Asn Pro Lys Gln Lys Met Ala Leu

180 185 190

Ala Lys Gln Leu Asn Leu Arg Thr Arg Gln Val Glu Val Trp Phe Gln

195 200 205

Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu

210 215 220

Tyr Leu Lys Arg Cys Cys Glu Asn Leu Thr Asp Glu Asn Arg Arg Leu

225 230 235 240

Gln Lys Glu Val Ser Glu Leu Arg Ala Leu Lys Leu Ser Pro His Leu

245 250 255

Tyr Met His Met Lys Pro Pro Thr Thr Leu Thr Met Cys Pro Ser Cys

260 265 270

Glu Arg Val Ala Val Thr Ser Ser Ser Ser Ser Val Ala Pro Pro Val

275 280 285

Met Asn Ser Ser Ser Pro Met Gly Pro Met Ser Pro Trp Ala Ala Met

290 295 300

Pro Leu Arg Gln Arg Pro Ala Ala Gly Ser His

305 310 315

<210>263

<211>744

<212>DNA

＜213〉rice

<400>263

atgatggaga gggccgagga cctgcgcctg agcctcagtc tcagctcgcc gcttattgct 60

cctcgtactc accatgtcgc catgctgttc cacgctcctc cagagaaaag attcctggag 120

atgccgctgc tccctgctgc gaagcggagc gaggtcgtcg cggcagaaga ggagcgcgcg 180

ggcctgcgcg gcggcggcgg cagcgacgag gaggacggtg gctgcggcat cgacggctca 240

cgcaagaagc tccggctttc caaggaccag tccgccgtgc tcgaggacag cttccgggag 300

caccccaccc tcaaccccag gcagaaggca actttggcgc agcagctcgg gcttcggcct 360

cggcaggtcg aggtgtggtt tcagaacaga cgcgcaagga cgaagctgaa gcagacggag 420

gtggactgcg agttcctgaa gcgctgctgc gagacgctca cggaggagaa ccggaggctg 480

cagaaggagg tgcaggagct gcgagcgctc aagctcgtct cgccgcacct ctacatgaac 540

atgtccccgc ccaccacgct caccatgtgc ccctcctgcg agcgcgtctc caacaccaat 600

aacaactcca gcgccgccgc cgccgccgac cgccgcggca tcaggactac tactgccgca 660

ggcggcggca gcgtcgtcga caccgccgcc gacgggggca tcctctgcca ccgcccgatc 720

gccgtccggc cgcagcagtc atga 744

<210>264

<211>247

<212>PRT

＜213〉rice

<400>264

Met Met Glu Arg Ala Glu Asp Leu Arg Leu Ser Leu Ser Leu Ser Ser

1 5 10 15

Pro Leu Ile Ala Pro Arg Thr His His Val Ala Met Leu Phe His Ala

20 25 30

Pro Pro Glu Lys Arg Phe Leu Glu Met Pro Leu Leu Pro Ala Ala Lys

35 40 45

Arg Ser Glu Val Val Ala Ala Glu Glu Glu Arg Ala Gly Leu Arg Gly

50 55 60

Gly Gly Gly Ser Asp Glu Glu Asp Gly Gly Cys Gly Ile Asp Gly Ser

65 70 75 80

Arg Lys Lys Leu Arg Leu Ser Lys Asp Gln Ser Ala Val Leu Glu Asp

85 90 95

Ser Phe Arg Glu His Pro Thr Leu Asn Pro Arg Gln Lys Ala Thr Leu

100 105 110

Ala Gln Gln Leu Gly Leu Arg Pro Arg Gln Val Glu Val Trp Phe Gln

115 120 125

Asn Arg Arg Ala Arg Thr Lys Leu Lys Gln Thr Glu Val Asp Cys Glu

130 135 140

Phe Leu Lys Arg Cys Cys Glu Thr Leu Thr Glu Glu Asn Arg Arg Leu

145 150 155 160

Gln Lys Glu Val Gln Glu Leu Arg Ala Leu Lys Leu Val Ser Pro His

165 170 175

Leu Tyr Met Asn Met Ser Pro Pro Thr Thr Leu Thr Met Cys Pro Ser

180 185 190

Cys Glu Arg Val Ser Asn Thr Asn Asn Asn Ser Ser Ala Ala Ala Ala

195 200 205

Ala Asp Arg Arg Gly Ile Arg Thr Thr Thr Ala Ala Gly Gly Gly Ser

210 215 220

Val Val Asp Thr Ala Ala Asp Gly Gly Ile Leu Cys His Arg Pro Ile

225 230 235 240

Ala Val Arg Pro Gln Gln Ser

245

<210>265

<211>846

<212>DNA

＜213〉rice

<400>265

atgaggtcgt acatggacgg cggcggcgcg gcggcgtacg aggaggagga ggaggaggtt 60

gaggacgacg acggcggcgg cggcggcggc ggcggcggcg gcggtggggg gctcggggag 120

aagaagcggc ggctggcggc ggagcaggtg cgggcgctgg agcggagctt cgaggcggac 180

aacaagctgg acccggagcg gaaggcccgg atcgcccgcg accttcgcct ccaccctcgc 240

caggtcgccg tctggttcca gaaccgccgc gcgaggtgga agaccaagca gatcgagcgc 300

gacttcgccg ccctccgctc ccgccacgac gccctccgcc tcgagtgcga cgccctccgc 360

cgcgacaagg acgccctcgc cgccgagatc gccgacctcc gggacagggt ggacggccag 420

atgtccgtca agctggaggc cgtggccgcg gacgaacacc agccgcctcc gccgccgccg 480

ccgccgccac tggcgtataa cagcaaggtg gtggacggct cgacggacag cgactcgagc 540

gcggtgttca acgaggaggc gtcgccgtac tccggcgcgg ccatcgacca ccaccaccac 600

caaactccgg cgagctacga cacggcgggg ttcacctcct tcttcgcgcc atccaccacg 660

ctcacctcgt ccctctcctt cccttccatg ttccacgcgt catcgcattt cgatggccac 720

caagaactcc tcgtcggcgg cggcggcgcc ggcgcagtgg ccgacgccga cctcggaggc 780

gccggattct tcgccggcga cgagcacgcc ggcggcctct cctggtacgg cgccgagggt 840

tggtag 846

<210>266

<211>281

<212>PRT

＜213〉rice

<400>266

Met Arg Ser Tyr Met Asp Gly Gly Gly Ala Ala Ala Tyr Glu Glu Glu

1 5 10 15

Glu Glu Glu Val Glu Asp Asp Asp Gly Gly Gly Gly Gly Gly Gly Gly

20 25 30

Gly Gly Gly Gly Gly Leu Gly Glu Lys Lys Arg Arg Leu Ala Ala Glu

35 40 45

Gln Val Arg Ala Leu Glu Arg Ser Phe Glu Ala Asp Asn Lys Leu Asp

50 55 60

Pro Glu Arg Lys Ala Arg Ile Ala Arg Asp Leu Arg Leu His Pro Arg

65 70 75 80

Gln Val Ala Val Trp Phe Gln Asn Arg Arg Ala Arg Trp Lys Thr Lys

85 90 95

Gln Ile Glu Arg Asp Phe Ala Ala Leu Arg Ser Arg His Asp Ala Leu

100 105 110

Arg Leu Glu Cys Asp Ala Leu Arg Arg Asp Lys Asp Ala Leu Ala Ala

115 120 125

Glu Ile Ala Asp Leu Arg Asp Arg Val Asp Gly Gln Met Ser Val Lys

130 135 140

Leu Glu Ala Val Ala Ala Asp Glu His Gln Pro Pro Pro Pro Pro Pro

145 150 155 160

Pro Pro Pro Leu Ala Tyr Asn Ser Lys Val Val Asp Gly Ser Thr Asp

165 170 175

Ser Asp Ser Ser Ala Val Phe Asn Glu Glu Ala Ser Pro Tyr Ser Gly

180 185 190

Ala Ala Ile Asp His His His His Gln Thr Pro Ala Ser Tyr Asp Thr

195 200 205

Ala Gly Phe Thr Ser Phe Phe Ala Pro Ser Thr Thr Leu Thr Ser Ser

210 215 220

Leu Ser Phe Pro Ser Met Phe His Ala Ser Ser His Phe Asp Gly His

225 230 235 240

Gln Glu Leu Leu Val Gly Gly Gly Gly Ala Gly Ala Val Ala Asp Ala

245 250 255

Asp Leu Gly Gly Ala Gly Phe Phe Ala Gly Asp Glu His Ala Gly Gly

260 265 270

Leu Ser Trp Tyr Gly Ala Glu Gly Trp

275 280

<210>267

<211>918

<212>DNA

＜213〉rice

<400>267

atgaagaggc ccagctgcag aggctcctcc atggccatca tccatgacac ctctgatcaa 60

caagaggaca acatgaggtc gtacatggac ggcggcggcg cggcggcgta cgaggaggag 120

gaggaggagg ttgaggacga cgacggcggc ggcggcggcg gcggcggcgg cggcggtggg 180

gggctcgggg agaagaagcg gcggctggcg gcggagcagg tgcgggcgct ggagcggagc 240

ttcgaggcgg acaacaagct ggacccggag cggaaggccc ggatcgcccg cgaccttcgc 300

ctccaccctc gccaggtcgc cgtctggttc cagaaccgcc gcgcgaggtg gaagaccaag 360

cagatcgagc gcgacttcgc cgccctccgc tcccgccacg acgccctccg cctcgagtgc 420

gacgccctcc gccgcgacaa ggacgccctc gccgccgaga tcgccgacct ccgggacagg 480

gtggacggcc agatgtccgt caagctggag gccgtggccg cggacgaaca ccagccgcct 540

ccgccgccgc cgccgccgcc actggcgtat aacagcaagg tggtggacgg ctcgacggac 600

agcgactcga gcgcggtgtt caacgaggag gcgtcgccgt actccggcgc ggccatcgac 660

caccaccacc accaaactcc ggcgagctac gacacggcgg ggttcacctc cttcttcgcg 720

ccatccacca cgctcacctc gtccctctcc ttcccttcca tgttccacgc gtcatcgcat 780

ttcgatggcc accaagaact cctcgtcggc ggcggcggcg ccggcgcagt ggccgacgcc 840

gacctcggag gcgccggatt cttcgccggc gacgagcacg ccggcggcct ctcctggtac 900

ggcgccgagg gttggtag 918

<210>268

<211>305

<212>PRT

＜213〉rice

<400>268

Met Lys Arg Pro Ser Cys Arg Gly Ser Ser Met Ala Ile Ile His Asp

1 5 10 15

Thr Ser Asp Gln Gln Glu Asp Asn Met Arg Ser Tyr Met Asp Gly Gly

20 25 30

Gly Ala Ala Ala Tyr Glu Glu Glu Glu Glu Glu Val Glu Asp Asp Asp

35 40 45

Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Gly Leu Gly Glu

50 55 60

Lys Lys Arg Arg Leu Ala Ala Glu Gln Val Arg Ala Leu Glu Arg Ser

65 70 75 80

Phe Glu Ala Asp Asn Lys Leu Asp Pro Glu Arg Lys Ala Arg Ile Ala

85 90 95

Arg Asp Leu Arg Leu His Pro Arg Gln Val Ala Val Trp Phe Gln Asn

100 105 110

Arg Arg Ala Arg Trp Lys Thr Lys Gln Ile Glu Arg Asp Phe Ala Ala

115 120 125

Leu Arg Ser Arg His Asp Ala Leu Arg Leu Glu Cys Asp Ala Leu Arg

130 135 140

Arg Asp Lys Asp Ala Leu Ala Ala Glu Ile Ala Asp Leu Arg Asp Arg

145 150 155 160

Val Asp Gly Gln Met Ser Val Lys Leu Glu Ala Val Ala Ala Asp Glu

165 170 175

His Gln Pro Pro Pro Pro Pro Pro Pro Pro Pro Leu Ala Tyr Asn Ser

180 185 190

Lys Val Val Asp Gly Ser Thr Asp Ser Asp Ser Ser Ala Val Phe Asn

195 200 205

Glu Glu Ala Ser Pro Tyr Ser Gly Ala Ala Ile Asp His His His His

210 215 220

Gln Thr Pro Ala Ser Tyr Asp Thr Ala Gly Phe Thr Ser Phe Phe Ala

225 230 235 240

Pro Ser Thr Thr Leu Thr Ser Ser Leu Ser Phe Pro Ser Met Phe His

245 250 255

Ala Ser Ser His Phe Asp Gly His Gln Glu Leu Leu Val Gly Gly Gly

260 265 270

Gly Ala Gly Ala Val Ala Asp Ala Asp Leu Gly Gly Ala Gly Phe Phe

275 280 285

Ala Gly Asp Glu His Ala Gly Gly Leu Ser Trp Tyr Gly Ala Glu Gly

290 295 300

Trp

305

<210>269

<211>936

<212>DNA

＜213〉rice

<400>269

atgaagaggc ccaccagcag cagccgaaaa tccaaaaaac aaggagagga cctggcgttc 60

tctgaggagg gcagcttgcc cgcggtgacc atggagcaga aagatgaagc cgagatggag 120

gaggtggacg aggaggagga ggaggaggtc gacgaagaca tggccggcgg gcacgcggcg 180

cagtcgccgt cgccgtcgtg cgggctgggc gagaagaagc ggcggctggc gctggagcag 240

gtgcgcgctc tggagcggag cttcgacacg gacaacaagc tggacccgga ccgcaaggcc 300

cgcatcgcgc gcgacctcgg cctgcagccg cgccaggtcg ccgtctggtt ccagaaccgc 360

cgcgcccggt ggaagacgaa gcagctcgag cgcgacttcg ccgccctccg cgcccgccac 420

gacgccctcc gcgccgactg cgacgccctg cgccgcgaca aggacgccct cgccgccgag 480

attcgggagc tgagggagaa gctgcccacc aagccggcgg acacggcggc atcggtgaag 540

gtagaagccg gcaatgacgc ggcggccggc gccgcggccg ccacggtgtg caaggacggc 600

tcgtcggacg acagcgactc cagcgtggtg ttcaacgacg aggcgtcgcc gtactccggc 660

gcggccttca ttggattcgg cccgtcgttc ttggtcgacg acgcgtcggc ggcaaccgtg 720

ggctgctcgt cgtcgctccc cgcgctcgag tccaaatggc acggtccgta ctccgacgac 780

tcgtgcaaag gcggcgtcta tggcttcacg gaggaatggc tcgctgcctg ctccggcgag 840

atggccggca acgacgccgc cggcttcttc tccgacgagc acgcctccaa cctcaacttc 900

ggttggtgcg cgagtggtaa cgagggttgg gaatga 936

<210>270

<211>311

<212>PRT

＜213〉rice

<400>270

Met Lys Arg Pro Thr Ser Ser Ser Arg Lys Ser Lys Lys Gln Gly Glu

1 5 10 15

Asp Leu Ala Phe Ser Glu Glu Gly Ser Leu Pro Ala Val Thr Met Glu

20 25 30

Gln Lys Asp Glu Ala Glu Met Glu Glu Val Asp Glu Glu Glu Glu Glu

35 40 45

Glu Val Asp Glu Asp Met Ala Gly Gly His Ala Ala Gln Ser Pro Ser

50 55 60

Pro Ser Cys Gly Leu Gly Glu Lys Lys Arg Arg Leu Ala Leu Glu Gln

65 70 75 80

Val Arg Ala Leu Glu Arg Ser Phe Asp Thr Asp Asn Lys Leu Asp Pro

85 90 95

Asp Arg Lys Ala Arg Ile Ala Arg Asp Leu Gly Leu Gln Pro Arg Gln

100 105 110

Val Ala Val Trp Phe Gln Asn Arg Arg Ala Arg Trp Lys Thr Lys Gln

115 120 125

Leu Glu Arg Asp Phe Ala Ala Leu Arg Ala Arg His Asp Ala Leu Arg

130 135 140

Ala Asp Cys Asp Ala Leu Arg Arg Asp Lys Asp Ala Leu Ala Ala Glu

145 150 155 160

Ile Arg Glu Leu Arg Glu Lys Leu Pro Thr Lys Pro Ala Asp Thr Ala

165 170 175

Ala Ser Val Lys Val Glu Ala Gly Asn Asp Ala Ala Ala Gly Ala Ala

180 185 190

Ala Ala Thr Val Cys Lys Asp Gly Ser Ser Asp Asp Ser Asp Ser Ser

195 200 205

Val Val Phe Asn Asp Glu Ala Ser Pro Tyr Ser Gly Ala Ala Phe Ile

210 215 220

Gly Phe Gly Pro Ser Phe Leu Val Asp Asp Ala Ser Ala Ala Thr Val

225 230 235 240

Gly Cys Ser Ser Ser Leu Pro Ala Leu Glu Ser Lys Trp His Gly Pro

245 250 255

Tyr Ser Asp Asp Ser Cys Lys Gly Gly Val Tyr Gly Phe Thr Glu Glu

260 265 270

Trp Leu Ala Ala Cys Ser Gly Glu Met Ala Gly Asn Asp Ala Ala Gly

275 280 285

Phe Phe Ser Asp Glu His Ala Ser Asn Leu Asn Phe Gly Trp Cys Ala

290 295 300

Ser Gly Asn Glu Gly Trp Glu

305 310

<210>271

<211>810

<212>DNA

＜213〉rice

<400>271

atgagaagtc cagcagcgct tctcccggtg gttgcagatg gtggtggtgg tgttggggtg 60

gaggaggaga tggatgtgga cgaggacatg gcgatgtgcg gcggccgcgg cggcggcggc 120

ggggagaaga agcggcggct gagcgtggag caggtgcgcg cgctggagcg gagcttcgag 180

acggagaaca agctggagcc ggagcggaag gcgcggctgg cgcgcgacct cgggctgcag 240

ccgcgccagg tcgccgtctg gttccagaac cgccgcgcgc ggtggaagac caagcagctc 300

gagcgcgact acgccgcgct ccgccaatcc tacgacgcgc tccgcgccga ccacgacgcg 360

cttcgccgcg acaaggacgc cctcctcgcc gagatcaagg agctgaaggg gaagctcggc 420

gacgaggacg ccgcggcgag cttctcgtcg gtgaaggagg aggaggaccc ggcggcgtcc 480

gacgccgacc ccccggccac cggcgcgccg cagggctcgt ccgagagcga ctcgagcgcg 540

gtgctgaacg acgcggagat ccttccacac aagccagcgc cggcggccgc cgccgacgcc 600

gcggcctcgg aggagacgga ggcggtggtg accggcgccg cgctgctcca ccacgccgag 660

gtgttcttcc acgggcagct tctcaaggtg gacgacgacg aggcggcgtt cctaggcgac 720

gacggcgcgg cgtgcggcgg cttcttcgcc gacgagcatc tcccgtcgct gccgtggtgg 780

gccgagccca ccgagcaatg gacgacctag 810

<210>272

<211>269

<212>PRT

＜213〉rice

<400>272

Met Arg Ser Pro Ala Ala Leu Leu Pro Val Val Ala Asp Gly Gly Gly

1 5 10 15

Gly Val Gly Val Glu Glu Glu Met Asp Val Asp Glu Asp Met Ala Met

20 25 30

Cys Gly Gly Arg Gly Gly Gly Gly Gly Glu Lys Lys Arg Arg Leu Ser

35 40 45

Val Glu Gln Val Arg Ala Leu Glu Arg Ser Phe Glu Thr Glu Asn Lys

50 55 60

Leu Glu Pro Glu Arg Lys Ala Arg Leu Ala Arg Asp Leu Gly Leu Gln

65 70 75 80

Pro Arg Gln Val Ala Val Trp Phe Gln Asn Arg Arg Ala Arg Trp Lys

85 90 95

Thr Lys Gln Leu Glu Arg Asp Tyr Ala Ala Leu Arg Gln Ser Tyr Asp

100 105 110

Ala Leu Arg Ala Asp His Asp Ala Leu Arg Arg Asp Lys Asp Ala Leu

115 120 125

Leu Ala Glu Ile Lys Glu Leu Lys Gly Lys Leu Gly Asp Glu Asp Ala

130 135 140

Ala Ala Ser Phe Ser Ser Val Lys Glu Glu Glu Asp Pro Ala Ala Ser

145 150 155 160

Asp Ala Asp Pro Pro Ala Thr Gly Ala Pro Gln Gly Ser Ser Glu Ser

165 170 175

Asp Ser Ser Ala Val Leu Asn Asp Ala Glu Ile Leu Pro His Lys Pro

180 185 190

Ala Pro Ala Ala Ala Ala Asp Ala Ala Ala Ser Glu Glu Thr Glu Ala

195 200 205

Val Val Thr Gly Ala Ala Leu Leu His His Ala Glu Val Phe Phe His

210 215 220

Gly Gln Leu Leu Lys Val Asp Asp Asp Glu Ala Ala Phe Leu Gly Asp

225 230 235 240

Asp Gly Ala Ala Cys Gly Gly Phe Phe Ala Asp Glu His Leu Pro Ser

245 250 255

Leu Pro Trp Trp Ala Glu Pro Thr Glu Gln Trp Thr Thr

260 265

<210>273

<211>834

<212>DNA

＜213〉rice

<400>273

atgaagcgac ccggcggtgc cggcggcggc ggaggcagcc catcgctcgt cacgatggct 60

aattctagtg atgatggata tggaggggtt gggatggagg cggaggggga cgtggaggag 120

gagatgatgg cgtgcggcgg cggcggggag aagaagcggc ggctgagcgt ggagcaggtt 180

cgcgcgctgg agcggagctt cgaggtggag aacaagcttg agcctgagcg gaaggcgcgg 240

ctggcgcgcg acctcggcct gcagccgcgc caggtcgccg tctggttcca gaaccgccgc 300

gcgcggtgga agaccaagca gctcgagcgc gactacgccg cgctccgcca ttcctacgac 360

tccctgcgcc tcgatcacga cgcgctccgc cgcgacaagg acgccctcct cgccgagatc 420

aaggagctga aggcgaagct cggggacgag gaggcggcgg cgagcttcac gtcggtgaag 480

gaggagccgg cggcctccga cgggccaccg gcggcgggat ttgggtcgtc cgacagcgac 540

tcaagcgcgg tgctgaacga cgtggacgcg gccggcgccg cgcccgcggc gacggacgcg 600

ctggctccgg aggcgtgcac gtttctcggc gcgccgcccg ccgcgggcgc gggcgcgggc 660

gcagcggcgg cggcgagcca cgaggaggtg ttcttccacg gcaatttcct caaggtggag 720

gaggacgaga cggggttcct cgacgacgac gagccgtgcg gcgggttctt cgccgacgat 780

cagcccccgc cgctgtcgtc gtggtgggcc gagcccacgg agcactggaa ctga 834

<210>274

<211>277

<212>PRT

＜213〉rice

<400>274

Met Lys Arg Pro Gly Gly Ala Gly Gly Gly Gly Gly Ser Pro Ser Leu

1 5 10 15

Val Thr Met Ala Asn Ser Ser Asp Asp Gly Tyr Gly Gly Val Gly Met

20 25 30

Glu Ala Glu Gly Asp Val Glu Glu Glu Met Met Ala Cys Gly Gly Gly

35 40 45

Gly Glu Lys Lys Arg Arg Leu Ser Val Glu Gln Val Arg Ala Leu Glu

50 55 60

Arg Ser Phe Glu Val Glu Asn Lys Leu Glu Pro Glu Arg Lys Ala Arg

65 70 75 80

Leu Ala Arg Asp Leu Gly Leu Gln Pro Arg Gln Val Ala Val Trp Phe

85 90 95

Gln Asn Arg Arg Ala Arg Trp Lys Thr Lys Gln Leu Glu Arg Asp Tyr

100 105 110

Ala Ala Leu Arg His Ser Tyr Asp Ser Leu Arg Leu Asp His Asp Ala

115 120 125

Leu Arg Arg Asp Lys Asp Ala Leu Leu Ala Glu Ile Lys Glu Leu Lys

130 135 140

Ala Lys Leu Gly Asp Glu Glu Ala Ala Ala Ser Phe Thr Ser Val Lys

145 150 155 160

Glu Glu Pro Ala Ala Ser Asp Gly Pro Pro Ala Ala Gly Phe Gly Ser

165 170 175

Ser Asp Ser Asp Ser Ser Ala Val Leu Asn Asp Val Asp Ala Ala Gly

180 185 190

Ala Ala Pro Ala Ala Thr Asp Ala Leu Ala Pro Glu Ala Cys Thr Phe

195 200 205

Leu Gly Ala Pro Pro Ala Ala Gly Ala Gly Ala Gly Ala Ala Ala Ala

210 215 220

Ala Ser His Glu Glu Val Phe Phe His Gly Asn Phe Leu Lys Val Glu

225 230 235 240

Glu Asp Glu Thr Gly Phe Leu Asp Asp Asp Glu Pro Cys Gly Gly Phe

245 250 255

Phe Ala Asp Asp Gln Pro Pro Pro Leu Ser Ser Trp Trp Ala Glu Pro

260 265 270

Thr Glu His Trp Asn

275

<210>275

<211>375

<212>DNA

＜213〉rice

<400>275

atgtcatcct ctctctttct tcttcctcct ctctttcttt ctctcttctc tccccttcgg 60

ctagccgcag ggagcacggg ctggggcggt agtgggcggg gacaagagag gcggagggag 120

ctgaagcggc gccggcggcg gcacacgctc tccagcctca gcggcaagcg tggtgcgcca 180

tctgccgctg ccgccgccgt cggcggcagc gacgacgagg actccgacga cggatcccgc 240

aagaagctcc gcctctccaa ggaccaagcc gccgtcctcg aggacacctt caacaagcac 300

aacaccctca accccaagca gaaggcggcg ctggcgaggc agctgaatct gaagccgcgg 360

caggtggagg tgtag 375

<210>276

<211>124

<212>PRT

＜213〉rice

<400>276

Met Ser Ser Ser Leu Phe Leu Leu Pro Pro Leu Phe Leu Ser Leu Phe

1 5 10 15

Ser Pro Leu Arg Leu Ala Ala Gly Ser Thr Gly Trp Gly Gly Ser Gly

20 25 30

Arg Gly Gln Glu Arg Arg Arg Glu Leu Lys Arg Arg Arg Arg Arg His

35 40 45

Thr Leu Ser Ser Leu Ser Gly Lys Arg Gly Ala Pro Ser Ala Ala Ala

50 55 60

Ala Ala Val Gly Gly Ser Asp Asp Glu Asp Ser Asp Asp Gly Ser Arg

65 70 75 80

Lys Lys Leu Arg Leu Ser Lys Asp Gln Ala Ala Val Leu Glu Asp Thr

85 90 95

Phe Asn Lys His Asn Thr Leu Asn Pro Lys Gln Lys Ala Ala Leu Ala

100 105 110

Arg Gln Leu Asn Leu Lys Pro Arg Gln Val Glu Val

115 120

<210>277

<211>1767

<212>DNA

＜213〉rice

<400>277

atgttttatt caagtccctt ttgttcatct cctttgcaga ttccatttct gacccaaatt 60

ttggctattc ctcatgatga gtttgtctca aactggtgct ctgttaatct gccagtgata 120

gaagaagacg ctaatcttga ttatgatcca tttggtgcag ctgagctggc actggcagct 180

gctggtaata agttaactga agctaaagca aactattctt gcccttttcg ccctatcagt 240

atgccttcta tagcatatgc gcagacaaga acatcatgtg tggtgaaaat aatagcaaat 300

ttgcatgttt ttgtcccaaa catatgcgaa gagcaagaaa gagacctttt tcttcagaaa 360

tttcagaagt acttggtatc agggaatcct agatcatcag ttgatcatcc agcatcagct 420

gatctcaagg ccactacagt ttgcagaaac ttgggatctt tgtctgagta tgctagatcg 480

ttaattccta ataacttgtt aaacgaggaa gatgtgcaat tgttaagtga atttgcttat 540

aagttacaaa cttggtgtaa atcacatgtt ggacagagta catcccaggc agtaaagatt 600

gatccgtcat cagaaagcaa ggaagacttt aagccactgc agcatccctt gataccaagt 660

actgttgttc cagattccag tataaataac cttccgaaga acatggaaga gcctacacca 720

acaaacatgg aagagcctgc accaacaccc tcaacaaagc aagagggaaa tgccagggat 780

gagactccta gaagcactgt cgctttaaat ggtgggttcc tgcagaattc agtcggccag 840

gacttagtcc atcttggtgt ggcgagaact agttcaggct ttctaggggg aggtaccagt 900

acaagtacag gatccctgcg ctgcaaaatg gatcttgatc ctgcatccag cagtatggac 960

catttcaaaa caccagatag aaaagaaagt ggtcttcagg atgatgagaa aggagacact 1020

cacatgtatg atgagagaca acctaagaga aggaagcgaa ccattatgaa tgataggcaa 1080

ataaacgaaa tcgagaaggc tcttattgat gagcctgaga tgcataagaa tgctgcttta 1140

ctgcaggcat ggtcagagaa gttaagtggg cagggctcgg agattacgtc atctcagcta 1200

aagaattggc tgaacaacag aaaagctaag cttgctcgta ttgcgaaaga aagaggagta 1260

ctatctgagg gtgagaacgc agataagcca tctacgccag ctactcccca ccactgtgac 1320

tcctcagaaa gtgctggcga ggagagctac ttgccacctg cgagggtcat gagtgctcta 1380

ggcatatcca agggcagcag atttgtgagc ccagatggca atgagacaac atcacaggca 1440

gaattcaatc aaaatatcat gcttagccgc cctttcacaa gatcattctc atttgaacct 1500

ggccgtcttg tttcgctcat tgataatgat gggaaggagg ttggcagggg aaagatcttc 1560

caagttgagg ggagactgca agggaaggcc ctgacagata ctcgcgtttg catcgttgat 1620

gttattgagc tcaagattga gaaatggcgg gagctacctc atccctcgga agcatcagga 1680

agaacattcc aagaggcaga atcaaggaac ggtggtgtga tgagggtcgc gtgggatgtc 1740

atcagactat ctccagtggt tcagtaa 1767

<210>278

<211>588

<212>PRT

＜213〉rice

<400>278

Met Phe Tyr Ser Ser Pro Phe Cys Ser Ser Pro Leu Gln Ile Pro Phe

1 5 10 15

Leu Thr Gln Ile Leu Ala Ile Pro His Asp Glu Phe Val Ser Asn Trp

20 25 30

Cys Ser Val Asn Leu Pro Val Ile Glu Glu Asp Ala Asn Leu Asp Tyr

35 40 45

Asp Pro Phe Gly Ala Ala Glu Leu Ala Leu Ala Ala Ala Gly Asn Lys

50 55 60

Leu Thr Glu Ala Lys Ala Asn Tyr Ser Cys Pro Phe Arg Pro Ile Ser

65 70 75 80

Met Pro Ser Ile Ala Tyr Ala Gln Thr Arg Thr Ser Cys Val Val Lys

85 90 95

Ile Ile Ala Asn Leu His Val Phe Val Pro Asn Ile Cys Glu Glu Gln

100 105 110

Glu Arg Asp Leu Phe Leu Gln Lys Phe Gln Lys Tyr Leu Val Ser Gly

115 120 125

Asn Pro Arg Ser Ser Val Asp His Pro Ala Ser Ala Asp Leu Lys Ala

130 135 140

Thr Thr Val Cys Arg Asn Leu Gly Ser Leu Ser Glu Tyr Ala Arg Ser

145 150 155 160

Leu Ile Pro Asn Asn Leu Leu Asn Glu Glu Asp Val Gln Leu Leu Ser

165 170 175

Glu Phe Ala Tyr Lys Leu Gln Thr Trp Cys Lys Ser His Val Gly Gln

180 185 190

Ser Thr Ser Gln Ala Val Lys Ile Asp Pro Ser Ser Glu Ser Lys Glu

195 200 205

Asp Phe Lys Pro Leu Gln His Pro Leu Ile Pro Ser Thr Val Val Pro

210 215 220

Asp Ser Ser Ile Asn Asn Leu Pro Lys Asn Met Glu Glu Pro Thr Pro

225 230 235 240

Thr Asn Met Glu Glu Pro Ala Pro Thr Pro Ser Thr Lys Gln Glu Gly

245 250 255

Asn Ala Arg Asp Glu Thr Pro Arg Ser Thr Val Ala Leu Asn Gly Gly

260 265 270

Phe Leu Gln Asn Ser Val Gly Gln Asp Leu Val His Leu Gly Val Ala

275 280 285

Arg Thr Ser Ser Gly Phe Leu Gly Gly Gly Thr Ser Thr Ser Thr Gly

290 295 300

Ser Leu Arg Cys Lys Met Asp Leu Asp Pro Ala Ser Ser Ser Met Asp

305 310 315 320

His Phe Lys Thr Pro Asp Arg Lys Glu Ser Gly Leu Gln Asp Asp Glu

325 330 335

Lys Gly Asp Thr His Met Tyr Asp Glu Arg Gln Pro Lys Arg Arg Lys

340 345 350

Arg Thr Ile Met Asn Asp Arg Gln Ile Asn Glu Ile Glu Lys Ala Leu

355 360 365

Ile Asp Glu Pro Glu Met His Lys Asn Ala Ala Leu Leu Gln Ala Trp

370 375 380

Ser Glu Lys Leu Ser Gly Gln Gly Ser Glu Ile Thr Ser Ser Gln Leu

385 390 395 400

Lys Asn Trp Leu Asn Asn Arg Lys Ala Lys Leu Ala Arg Ile Ala Lys

405 410 415

Glu Arg Gly Val Leu Ser Glu Gly Glu Asn Ala Asp Lys Pro Ser Thr

420 425 430

Pro Ala Thr Pro His His Cys Asp Ser Ser Glu Ser Ala Gly Glu Glu

435 440 445

Ser Tyr Leu Pro Pro Ala Arg Val Met Ser Ala Leu Gly Ile Ser Lys

450 455 460

Gly Ser Arg Phe Val Ser Pro Asp Gly Asn Glu Thr Thr Ser Gln Ala

465 470 475 480

Glu Phe Asn Gln Asn Ile Met Leu Ser Arg Pro Phe Thr Arg Ser Phe

485 490 495

Ser Phe Glu Pro Gly Arg Leu Val Ser Leu Ile Asp Asn Asp Gly Lys

500 505 510

Glu Val Gly Arg Gly Lys Ile Phe Gln Val Glu Gly Arg Leu Gln Gly

515 520 525

Lys Ala Leu Thr Asp Thr Arg Val Cys Ile Val Asp Val Ile Glu Leu

530 535 540

Lys Ile Glu Lys Trp Arg Glu Leu Pro His Pro Ser Glu Ala Ser Gly

545 550 555 560

Arg Thr Phe Gln Glu Ala Glu Ser Arg Asn Gly Gly Val Met Arg Val

565 570 575

Ala Trp Asp Val Ile Arg Leu Ser Pro Val Val Gln

580 585

<210>279

<211>6

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif I

<400>279

Arg Lys Lys Leu Arg Leu

1 5

<210>280

<211>24

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif II

<400>280

Thr Lys Leu Lys Gln Thr Gl u Val Asp Cys Glu Phe Leu Arg Arg Cys

1 5 10 15

Cys Glu Asn Leu Thr Glu Glu Asn

20

<210>281

<211>10

<212>PRT

＜213〉artificial sequence

<220>

＜223〉motif III

<400>281

Thr Leu Thr Met Cys Pro Ser Cys Glu Arg

1 5 10

<210>282

<211>2193

<212>DNA

＜213〉rice

<400>282

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140

cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200

tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260

gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320

ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380

gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440

aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500

ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560

atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620

acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680

cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740

ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800

gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860

tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920

attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980

tgtcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040

cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100

agctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160

tggtgtagct tgccactttc accagcaaag ttc 2193

<210>283

<211>58

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: p01294

<400>283

ggggacaagt ttgtacaaaa aagcaggctt cacaatgatg ttcgagaaag acgatctg 58

<210>284

<211>52

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: p00402

<400>284

ggggaccact ttgtacaaga aagctgggtt taggacctag gacgaagagc gt 52

<210>285

<211>622

<212>DNA

＜213〉Arabidopis thaliana

<400>285

atgagcagac ccggagattg gaactgcagg tcatgcagcc atctcaactt ccagcgccgt 60

gactcttgcc agcgatgcgg tgactctcgt tccggccccg gtggagttgg tggcttagac 120

tttggtaatt tcggtggcag agccatgtct gctttcggat tcaccaccgg ctccgacgtt 180

cgtcccggtg attggtactg caccgtggga aactgcggga cacacaactt cgccagtcgc 240

tccacctgct tcaaatgcgg cactttcaag gacgagaccg gcgctggagg cggaggtggt 300

ggcatcggcg gtccggccat gtttgacgcc gacattatgc ggtctagagt ccccggtaac 360

ggtggtcgct ctagctggaa atccggcgac tggatttgca ctaggattgg ttgcaatgag 420

cataactttg caagcagaat ggaatgcttc aggtgcaatg caccaaggga cttcagcaac 480

agaacctctt tctaagttat acaaactgct tttgaagtag ccttgtctac ccagctttct 540

tgtacaaagt tggcattata agaaagcatt gcttatcaat ttgttgcaac gaacaggtca 600

ctatcagtca aaataaaatc at 622

<210>286

<211>164

<212>PRT

＜213〉Arabidopis thaliana

<400>286

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Ser His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Ser Arg Ser Gly

20 25 30

Pro Gly Gly Val Gly Gly Leu Asp Phe Gly Asn Phe Gly Gly Arg Ala

35 40 45

Met Ser Ala Phe Gly Phe Thr Thr Gly Ser Asp Val Arg Pro Gly Asp

50 55 60

Trp Tyr Cys Thr Val Gly Asn Cys Gly Thr His Asn Phe Ala Ser Arg

65 70 75 80

Ser Thr Cys Phe Lys Cys Gly Thr Phe Lys Asp Glu Thr Gly Ala Gly

85 90 95

Gly Gly Gly Gly Gly Ile Gly Gly Pro Ala Met Phe Asp Ala Asp Ile

100 105 110

Met Arg Ser Arg Val Pro Gly Asn Gly Gly Arg Ser Ser Trp Lys Ser

115 120 125

Gly Asp Trp Ile Cys Thr Arg Ile Gly Cys Asn Glu His Asn Phe Ala

130 135 140

Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg Asp Phe Ser Asn

145 150 155 160

Arg Thr Ser Phe

<210>287

<211>717

<212>DNA

＜213〉upland cotton

<400>287

atgagcaggc caggagattg gaactgcagg tcatgccaac acctcaactt ccaaaggagg 60

gacaactgcc aacgttgcgg tgaatctcga tacggtgtta gagtcggctc gacattcggg 120

tttaccgctg gctcggacgt tcgacctggt gactggtatt gcacggctgg aaactgcggc 180

acccacaatt tcgccagccg gtctacttgt ttcaattgcg gcgcgttcaa ggacgagtcg 240

gctggaggtt tcgacttgga catgtctcga tcaagagggt tcggaggtaa ccgatccggc 300

tggaaatcag gggattggat atgtaccagg ttagggtgca atgaacataa ttttgctagc 360

agaatggaat gtttcagatg cagtgctcca agagaattca acaatagaac ttcatattaa 420

atcacatgca tgctactata tccatttttg ggtgttttga ggcaattaat aggagattat 480

aatgagagag tgttactggc tttgatgatg aacaccacaa gcattttgtc atgtttcttt 540

tataagattc atggcaatgt agtacttttt cttgtgatga ttatttatgg tttcaattct 600

atggttttat tgtcttttat tttagagagt tttatttata tttttgtaat ggtgctttgg 660

ctttattaaa agaagatgat tctcttctgt cttttcaaaa aaaaaaaaaa aaaaaaa 717

<210>288

<211>139

<212>PRT

＜213〉upland cotton

<400>288

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Asn Cys Gln Arg Cys Gly Glu Ser Arg Tyr Gly

20 25 30

Val Arg Val Gly Ser Thr Phe Gly Phe Thr Ala Gly Ser Asp Val Arg

35 40 45

Pro Gly Asp Trp Tyr Cys Thr Ala Gly Asn Cys Gly Thr His Asn Phe

50 55 60

Ala Ser Arg Ser Thr Cys Phe Asn Cys Gly Ala Phe Lys Asp Glu Ser

65 70 75 80

Ala Gly Gly Phe Asp Leu Asp Met Ser Arg Ser Arg Gly Phe Gly Gly

85 90 95

Asn Arg Ser Gly Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Leu Gly

100 105 110

Cys Asn Glu His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Ser

115 120 125

Ala Pro Arg Glu Phe Ash Asn Arg Thr Ser Tyr

130 135

<210>289

<211>937

<212>DNA

＜213〉Zea mays

<400>289

cgggcacgac cagcaacaca acagccgagc tttgcttagg caagatgaac aggaagccag 60

gagactggga ctgcagggcg tgccagcacc tcaacttcag ccgccgagac atatgccagc 120

gctgcagcga gccacgtgga gttgctgatc gtggcagtgg cggcggcgga ggaggcgact 180

acgccagctt cggtggccgc ggtggctcct ccttcggcgg cggctttggc gctgctggct 240

ccgacgtccg ccctggtgac tggtactgct cctgcggcgc gcacaacttc gccagccgct 300

ccagctgctt caagtgctcc gcctacaagg aggaggccgc tgtgaacagt ggcgctggcg 360

gctttgatgg cgacatgtca cgctcacggg gctacggctt cggcagcggt gctgctgctg 420

ctgctggtgc tggcgctgcc cgtactacca accgccccgg ttggaagtcc ggagactgga 480

tctgcaccag atccggatgc aacgagcaca acttcgccag caggatggag tgcttcaggt 540

gcaacgcacc gcgggactct ggcactgagg tgtaggatcg agcaagttaa aaagtctgca 600

gcgccgaaga aagcgacgac aagaggagtc ctcatcacgt cgtaacgtaa gagagagagt 660

agtggatttg caacaaagaa aaaaaaaaga cggccgcgat gctctgttac tagctagcta 720

gttttgttca accacccatg ccgtctcttc tcttttatta gatttggttt ggttctcata 780

gccctttaat taccatttgg gacctatgtt tgctgttctg tttcccgttc gtctcctccg 840

catgtttgcg cttggatcga gtcttgtgat gtaacccccc aaaaaacgct tgcttaacta 900

gtactgttgc ttcttaataa aaaaaaaaaa aaaaaaa 937

<210>290

<211>176

<212>PRT

＜213〉Zea mays

<400>290

Met Asn Arg Lys Pro Gly Asp Trp Asp Cys Arg Ala Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Ile Cys Gln Arg Cys Ser Glu Pro Arg Gly

20 25 30

Val Ala Asp Arg Gly Ser Gly Gly Gly Gly Gly Gly Asp Tyr Ala Ser

35 40 45

Phe Gly Gly Arg Gly Gly Ser Ser Phe Gly Gly Gly Phe Gly Ala Ala

50 55 60

Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Ser Cys Gly Ala His

65 70 75 80

Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys Cys Ser Ala Tyr Lys Glu

85 90 95

Glu Ala Ala Val Asn Ser Gly Ala Gly Gly Phe Asp Gly Asp Met Ser

100 105 110

Arg Ser Arg Gly Tyr Gly Phe Gly Ser Gly Ala Ala Ala Ala Ala Gly

115 120 125

Ala Gly Ala Ala Arg Thr Thr Asn Arg Pro Gly Trp Lys Ser Gly Asp

130 135 140

Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu His Asn Phe Ala Ser Arg

145 150 155 160

Met Glu Cys Phe Arg Cys Asn Ala Pro Arg Asp Ser Gly Thr Glu Val

165 170 175

<210>291

<211>501

<212>DNA

＜213〉rice

<400>291

atgaacagga agccaggaga ctgggactgc agggcgtgcc agcacctcaa cttcagccgc 60

cgggacctat gccagcgctg cggcgagccg cgtggcgccg ctgatcgcgg cagcggtggt 120

ggcggtgact acgccaactt cggcggccgt ggtggttcct ccttcggtgg aggctttggc 180

actggctctg atgtccgccc aggtgactgg tactgcaact gcggcgcgca caacttcgcc 240

agccgctcca gctgcttcaa gtgcgctgct ttcaaggacg atgctgccgt caacagtggc 300

ggcgctggtg cctttgatgg tggggacatg tcgcgctcgc ggggctacgg cttcggcagc 360

ggcgccgtcc gcgccagccg ccctggctgg aagtctggcg actggatttg caccaggtct 420

ggatgcaatg agcacaactt cgccagcagg atggagtgct tcaggtgcaa cgcaccgcgg 480

gactccggca ctgaggtgta a 501

<210>292

<211>166

<212>PRT

＜213〉rice

<400>292

Met Asn Arg Lys Pro Gly Asp Trp Asp Cys Arg Ala Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Leu Cys Gln Arg Cys Gly Glu Pro Arg Gly

20 25 30

Ala Ala Asp Arg Gly Ser Gly Gly Gly Gly Asp Tyr Ala Asn Phe Gly

35 40 45

Gly Arg Gly Gly Ser Ser Phe Gly Gly Gly Phe Gly Thr Gly Ser Asp

50 55 60

Val Arg Pro Gly Asp Trp Tyr Cys Asn Cys Gly Ala His Asn Phe Ala

65 70 75 80

Ser Arg Ser Ser Cys Phe Lys Cys Ala Ala Phe Lys Asp Asp Ala Ala

85 90 95

Val Asn Ser Gly Gly Ala Gly Ala Phe Asp Gly Gly Asp Met Ser Arg

100 105 110

Ser Arg Gly Tyr Gly Phe Gly Ser Gly Ala Val Arg Ala Ser Arg Pro

115 120 125

Gly Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu

130 135 140

His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg

145 150 155 160

Asp Ser Gly Thr Glu Val

165

<210>293

<211>859

<212>DNA

＜213〉Arabidopis thaliana

<400>293

ataagctctt acactcattt caactctctt tttcgtttcc attaccctca gaagaagatg 60

aataggccgg gagattggaa ctgcagattg tgtagccacc tcaacttcca gaggagggat 120

tcatgccaac gttgtagaga gcctagaccg ggcgggatca gtaccgattt actcagcggt 180

tttggtggcc gtccggttag tagctccttc ggtttcaaca ccgggcccga tgtgcgaccc 240

ggggattggt attgcaacct tggggattgt gggacacata attttgccaa taggtccagt 300

tgtttcaagt gtggtgccgc aaaagatgag ttttcatgct caagtgctgc tgcaacaacc 360

gggtttatgg acatgaatgt tggtccgaga cgtggccttt ttggttttgg cggcagcagt 420

agtggtggtg gtggtacggg ccgttctcct tggaaatctg gagattggat ttgcccaagg 480

tcaggctgta acgaacataa cttcgcaagc aggtcagagt gtttcaggtg taacgcacca 540

aaggaacttg ccaccgaacc accctattag tcatttagtc ctcctacctt cttcatcatt 600

tccaatactc tggaagcatt agaggagagc agaaaggtga agaatcgaag caagataccg 660

accgccctta atctcttgat ttgtttaatt tcttagattt gactcgtttt atatctcgta 720

gtagtcagac ctatgttaga atgtaagcat gtcgagagtt ttccgaagca ttttgttctg 780

atatcggtct cctagctagt atgtaagttt gtttgaatgt attcttattt ctgataaagt 840

tgtttccttc tgattccgc 859

<210>294

<211>170

<212>PRT

＜213〉Arabidopis thaliana

<400>294

Met Asn Arg Pro Gly Asp Trp Asn Cys Arg Leu Cys Ser His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Arg Glu Pro Arg Pro Gly

20 25 30

Gly Ile Ser Thr Asp Leu Leu Ser Gly Phe Gly Gly Arg Pro Val Ser

35 40 45

Ser Ser Phe Gly Phe Asn Thr Gly Pro Asp Val Arg Pro Gly Asp Trp

50 55 60

Tyr Cys Asn Leu Gly Asp Cys Gly Thr His Asn Phe Ala Asn Arg Ser

65 70 75 80

Ser Cys Phe Lys Cys Gly Ala Ala Lys Asp Glu Phe Ser Cys Ser Ser

85 90 95

Ala Ala Ala Thr Thr Gly Phe Met Asp Met Asn Val Gly Pro Arg Arg

100 105 110

Gly Leu Phe Gly Phe Gly Gly Ser Ser Ser Gly Gly Gly Gly Thr Gly

115 120 125

Arg Ser Pro Trp Lys Ser Gly Asp Trp Ile Cys Pro Arg Ser Gly Cys

130 135 140

Asn Glu His Asn Phe Ala Ser Arg Ser Glu Cys Phe Arg Cys Asn Ala

145 150 155 160

Pro Lys Glu Leu Ala Thr Glu Pro Pro Tyr

165 170

<210>295

<211>834

<212>DNA

＜213〉rice

<400>295

cctagtacta agaacagcaa ccacctctga aagctttaca agttcgcagg ggcttaactg 60

cgaatgaaca tccagaggaa gccaggagac tggaactgca aatcgtgcca gcatctcaac 120

ttcagccgcc gggactactg ccagcgctgc cataccccac gccaggacct gccgcttggc 180

gatggttatg tcccaggtgg tgtgctgtcc tccctggaca ttcgcccggg cgactggtac 240

tgcaactgcg gctatcacaa ctttgctagc cgagcaagct gcttcaaatg tggcgccatt 300

gtgaaggacc ttccagcagg ccaaggtggt ggtgttgcca acggtgactt tgcccgtgcc 360

ctcgacagca gcgcagttcg tgctgggtgg aaggcgggtg actggatttg cacaaggcct 420

ggttgcaacg tccacaactt tgcaagtagg attgagtgct ataggtgcaa tgcacctagg 480

gaagcaggta atgtgaagta agaaaagact gacgcgacca aagatcgtga gacgtgacga 540

gctggccgag gatgaataaa gtgactgctg tcagttgtca cattcacagg ctgcgatcca 600

gaaactatgg atgggactat gtagtaacgt gctgatatat tattgctaag tgttactact 660

gcatggttgc aagggtggta gaagtacctt agcttccaag atcgttgtaa tgtgtgagtt 720

taattttggc gttttaagta ataagtctag tgattgcagg attgtacggg gtaatgtact 780

tgcattatcc attataatct taagcatcca atataattat gcaaaaaaaa aaaa 834

<210>296

<211>145

<212>PRT

＜213〉rice

<400>296

Met Asn Ile Gln Arg Lys Pro Gly Asp Trp Asn Cys Lys Ser Cys Gln

1 5 10 15

His Leu Asn Phe Ser Arg Arg Asp Tyr Cys Gln Arg Cys His Thr Pro

20 25 30

Arg Gln Asp Leu Pro Leu Gly Asp Gly Tyr Val Pro Gly Gly Val Leu

35 40 45

Ser Ser Leu Asp Ile Arg Pro Gly Asp Trp Tyr Cys Asn Cys Gly Tyr

50 55 60

His Asn Phe Ala Ser Arg Ala Ser Cys Phe Lys Cys Gly Ala Ile Val

65 70 75 80

Lys Asp Leu Pro Ala Gly Gln Gly Gly Gly Val Ala Asn Gly Asp Phe

85 90 95

Ala Arg Ala Leu Asp Ser Ser Ala Val Arg Ala Gly Trp Lys Ala Gly

l00 105 110

Asp Trp Ile Cys Thr Arg Pro Gly Cys Asn Val His Asn Phe Ala Ser

115 120 125

Arg Ile Glu Cys Tyr Arg Cys Asn Ala Pro Arg Glu Ala Gly Asn Val

130 135 140

Lys

145

<210>297

<211>958

<212>DNA

＜213〉rice

<400>297

aagcctccat ccatccatcc attcatcaga gctcaagctc aagcaagcaa gcttgctagc 60

tagctgctga gcagcattcc agtctgctcc agctagctca gctctctctg ctctgctctt 120

tgctcgataa cgaccatcat cttcttcttc ttcttcttct tcttcttctt cttcttcttg 180

tacattctat ttgctcgata gatagataga tagatagata ggtagataga gatggagacg 240

aaggcggcgg cgatggcgat gaggaagccg ggggactgga gctgcaggtc gtgccagtac 300

gtgaacttct gcaagaggga ggcgtgccag cggtgcgggg aggcgaagct cggggtggag 360

cggacggact acgccgccat gggcggcggg tgggaggtga agcccggcga ctggtgctgc 420

cgctgctgcg ccgtcaacaa ctacgccagc cgcggcagct gcttcaagtg cggcgccgcc 480

aagaacgact ccgccgccgc cgtcgcccag ggctggggct tctccgtcgc ctcccaggcc 540

ggctggaaga acggcgactg gatctgcccc agaatggaat gcaacgtgca gaactacgct 600

aacagaaccg agtgcttccg gtgcaatttc cccagatact acgttgattg atctgacata 660

tatgatatat gccttgcact gagaagaaat gaaagggaaa taaaatttac gagatcgaag 720

atcgacggac aagattgatc ggccaccggt catgcccttc agggttttct tttttctatt 780

tttttttaga ataaccctcg ataacctgag ggtttttttt catttgtaag agcggtaacg 840

gtactaaaaa aacacaaaaa cggcacaaaa ttgtggctga tgactgatca gtcttcctct 900

tttttttccc aataaaaggc agataattaa gaaagcaaaa ttattaaaaa aaaaaaaa 958

<210>298

<211>139

<212>PRT

＜213〉rice

<400>298

Met Glu Thr Lys Ala Ala Ala Met Ala Met Arg Lys Pro Gly Asp Trp

1 5 10 15

Ser Cys Arg Ser Cys Gln Tyr Val Asn Phe Cys Lys Arg Glu Ala Cys

20 25 30

Gln Arg Cys Gly Glu Ala Lys Leu Gly Val Glu Arg Thr Asp Tyr Ala

35 40 45

Ala Met Gly Gly Gly Trp Glu Val Lys Pro Gly Asp Trp Cys Cys Arg

50 55 60

Cys Cys Ala Val Asn Asn Tyr Ala Ser Arg Gly Ser Cys Phe Lys Cys

65 70 75 80

Gly Ala Ala Lys Asn Asp Ser Ala Ala Ala Val Ala Gln Gly Trp Gly

85 90 95

Phe Ser Val Ala Ser Gln Ala Gly Trp Lys Asn Gly Asp Trp Ile Cys

100 105 110

Pro Arg Met Glu Cys Asn Val Gln Asn Tyr Ala Asn Arg Thr Glu Cys

115 120 125

Phe Arg Cys Asn Phe Pro Arg Tyr Tyr Val Asp

130 135

<210>299

<211>519

<212>DNA

＜213〉beet (Beta vulgaris)

<400>299

atgagtaggc caggtgattg gaattgtagg tcatgcagcc acttgaactt ccaaaggagg 60

gactcctgcc agcgctgtgg ggacgtacgt cctgacggcc gaggcggcgg agggggagga 120

ggagactttg gcagtagctt tggagggagg tcaggtgggt ccccctttgg tgggggtttt 180

gcagggcccg atgttaggcc cggtgattgg tattgtagca ttggcaactg tggggcccac 240

aactttgcca gcaggtctag ctgcttcaag tgtggggcct acaaagagga agctggctgt 300

ggtgatagca tgggccgttc acgtggagga ttttcctttg gtggcatcgg cggtggcggt 360

agcggtgccg ctaccggccg ctcaggctgg aaatccggtg actggatttg cactaggtcg 420

ggttgcaacg agcataactt cgctagtagg actgagtgct tcagatgcag ggaaccaagg 480

gactccggta atgcaatgct aaaggaagta ccatgctga 519

<210>300

<211>172

<212>PRT

＜213〉beet

<400>300

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Ser His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Val Arg Pro Asp

20 25 30

Gly Arg Gly Gly Gly Gly Gly Gly Gly Asp Phe Gly Ser Ser Phe Gly

35 40 45

Gly Arg Ser Gly Gly Ser Pro Phe Gly Gly Gly Phe Ala Gly Pro Asp

50 55 60

Val Arg Pro Gly Asp Trp Tyr Cys Ser Ile Gly Asn Cys Gly Ala His

65 70 75 80

Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys Cys Gly Ala Tyr Lys Glu

85 90 95

Glu Ala Gly Cys Gly Asp Ser Met Gly Arg Ser Arg Gly Gly Phe Ser

100 105 110

Phe Gly Gly Ile Gly Gly Gly Gly Ser Gly Ala Ala Thr Gly Arg Ser

115 120 125

Gly Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu

130 135 140

His Asn Phe Ala Ser Arg Thr Glu Cys Phe Arg Cys Arg Glu Pro Arg

145 150 155 160

Asp Ser Gly Asn Ala Met Leu Lys Glu Val Pro Cys

165 170

<210>301

<211>807

<212>DNA

＜213〉Arabidopis thaliana

<400>301

atgggagacg gaagagaagg agactgggaa tgtttaggat gcagaaacag gaattatgcg 60

tttagatcat tctgtaacag atgcaagcag cctcgtctta tcatggataa taatacttct 120

ccaaactcta agtggcttcc tcgtatcggc gattggatct gcactggttg tactaacaac 180

aattatgcat cacgagaaaa gtgcaagaaa tgtgggcaat ctaaggaagt agcagcattg 240

tcagcacttg ctatccctgg agcttctctt caaactcatc tccattactt cacccgtgga 300

cctgagtcac atgatcaacc tggttcttta ctcgcattct ctaacgctac aaatcaagct 360

tcggttcata aagaatggag gagtggtgac tggatttgca gatgtggttt tcacaattat 420

tcctctcgta tacagtgcaa aaagtgcaat gaaatagctc cactagccct tggtacaaag 480

agattagcat cagaagcttt ggctcatgaa tgggatagca aaagactgaa tcaaggatat 540

acaagcatgc aaacacagtc agcgatatat gcatcttttc ctggtatgag cctaggaagg 600

gtctcaaatt ggcaacttcc tctcccgttt ctacaacaac actcaacacc tgctttactt 660

ggaatgggag tgaaacaatg gcgtgatggc gactggatgt gtacaaattg caagaatcac 720

aattatgcat cacgagcaga gtgcaatagg tgcaagacta cacgagatat ccttgatcag 780

gacataactc caacagaaca atcttga 807

<210>302

<211>268

<212>PRT

＜213〉Arabidopis thaliana

<400>302

Met Gly Asp Gly Arg Glu Gly Asp Trp Glu Cys Leu Gly Cys Arg Asn

1 5 10 15

Arg Asn Tyr Ala Phe Arg Ser Phe Cys Asn Arg Cys Lys Gln Pro Arg

20 25 30

Leu Ile Met Asp Asn Asn Thr Ser Pro Asn Ser Lys Trp Leu Pro Arg

35 40 45

Ile Gly Asp Trp Ile Cys Thr Gly Cys Thr Asn Asn Asn Tyr Ala Ser

50 55 60

Arg Glu Lys Cys Lys Lys Cys Gly Gln Ser Lys Glu Val Ala Ala Leu

65 70 75 80

Ser Ala Leu Ala Ile Pro Gly Ala Ser Leu Gln Thr His Leu His Tyr

85 90 95

Phe Thr Arg Gly Pro Glu Ser His Asp Gln Pro Gly Ser Leu Leu Ala

100 105 110

Phe Ser Asn Ala Thr Asn Gln Ala Ser Val His Lys Glu Trp Arg Ser

115 120 125

Gly Asp Trp Ile Cys Arg Cys Gly Phe His Asn Tyr Ser Ser Arg Ile

130 135 140

Gln Cys Lys Lys Cys Asn Glu Ile Ala Pro Leu Ala Leu Gly Thr Lys

145 150 155 160

Arg Leu Ala Ser Glu Ala Leu Ala His Glu Trp Asp Ser Lys Arg Leu

165 170 175

Asn Gln Gly Tyr Thr Ser Met Gln Thr Gln Ser Ala Ile Tyr Ala Ser

180 185 190

Phe Pro Gly Met Ser Leu Gly Arg Val Ser Asn Trp Gln Leu Pro Leu

195 200 205

Pro Phe Leu Glu Gln His Ser Thr Pro Ala Leu Leu Gly Met Gly Val

210 215 220

Lys Gln Trp Arg Asp Gly Asp Trp Met Cys Thr Asn Cys Lys Asn His

225 230 235 240

Asn Tyr Ala Ser Arg Ala Glu Cys Asn Arg Cys Lys Thr Thr Arg Asp

245 250 255

Ile Leu Asp Gln Asp Ile Thr Pro Thr Glu Gln Ser

260 265

<210>303

<211>1264

<212>DNA

＜213〉Arabidopis thaliana

<400>303

ctatttctcc taccgtcgaa ctaaatccta gggcacaggt gaaatcgccg agtgacgtct 60

agccaccgca ccgcctcctt ctccggttat ctcgctacta atcagactat tccacagcta 120

tgtcacaggt agacaacaga aattcatcag cagccaagcg tgctagaact gacggggggc 180

gtagagaaga tgattggatc tgcccaagtt gtggcaatgt caacttttca ttcaggacaa 240

cttgcaatat gcgtaattgc actcagccta gacctgcaga tcataatgga aagtctgctc 300

ccaaacctat gcaacatcaa caaggtttct catcacccgg ggcatactta ggatctgggg 360

gtccccctcc agtatatatg ggcgggtcac catatggatc tcctctcttt aatggatcat 420

ctatgcctcc ttatgacgtc ccattttctg ggggttcgcc ttaccatttt aactataata 480

gccgaatgcc tgccggagct cattacagac cattacatat gtctggacca ccaccatacc 540

atggcggatc tatgatggga agtggtggta tgtatggaat gcctccacca atagacaggt 600

atggccttgg tatggcaatg ggtcctggtt ctgccgctgc catgatgcca agaccaaggt 660

tttacccaga tgaaaaatca caaaagagag attcaactcg cgataatgat tggacatgtc 720

cgaattgtgg taatgtaaac ttctcattca gaactgtatg taacatgagg aagtgcaaca 780

ctccaaagcc tggttctcag cagggtggaa gctcagataa aatatccaaa caaaatgcac 840

cggaagggag ctggaagtgt gataactgtg gaaatataaa ctacccattc aggagcaaat 900

gcaacaggca aaactgtgga gctgataagc ctggggatcg gtcgaatgga tctccgtccc 960

gtgcaccaga agagaacgat cagtgagtcg tagacatgtt tgggggttga gaagggtgag 1020

tctaatccag cgggtgtcgt aatgtcagtc aatgtctcgt caggtcaggt cgtccatgtt 1080

gctgcgtcgt cagtcaagtt gcttctatgc atttgtctct ttacttccct tgcgtggttg 1140

tggattgtat taataatgca aaaattgttt atttactttg tcgtcctcgt ggatcttgtc 1200

ttgctataga atcctcctcc aggtccatac tgttttacat cctaatctta agtaagttgg 1260

cacc 1264

<210>304

<211>288

<212>PRT

＜213〉Arabidopis thaliana

<400>304

Met Ser Gln Val Asp Asn Arg Asn Ser Ser Ala Ala Lys Arg Ala Arg

1 5 10 15

Thr Asp Gly Gly Arg Arg Glu Asp Asp Trp Ile Cys Pro Ser Cys Gly

20 25 30

Asn Val Asn Phe Ser Phe Arg Thr Thr Cys Asn Met Arg Asn Cys Thr

35 40 45

Gln Pro Arg Pro Ala Asp His Asn Gly Lys Ser Ala Pro Lys Pro Met

50 55 60

Gln His Gln Gln Gly Phe Ser Ser Pro Gly Ala Tyr Leu Gly Ser Gly

65 70 75 80

Gly Pro Pro Pro Val Tyr Met Gly Gly Ser Pro Tyr Gly Ser Pro Leu

85 90 95

Phe Asn Gly Ser Ser Met Pro Pro Tyr Asp Val Pro Phe Ser Gly Gly

100 105 110

Ser Pro Tyr His Phe Asn Tyr Asn Ser Arg Met Pro Ala Gly Ala His

115 120 125

Tyr Arg Pro Leu His Met Ser Gly Pro Pro Pro Tyr His Gly Gly Ser

130 135 140

Met Met Gly Ser Gly Gly Met Tyr Gly Met Pro Pro Pro Ile Asp Arg

145 150 155 160

Tyr Gly Leu Gly Met Ala Met Gly Pro Gly Ser Ala Ala Ala Met Met

165 170 175

Iro Arg Pro Arg Phe Tyr Pro Asp Glu Lys Ser Gln Lys Arg Asp Ser

180 185 190

Thr Arg Asp Asn Asp Trp Thr Cys Pro Asn Cys Gly Asn Val Asn Phe

195 200 205

Ser Phe Arg Thr Val Cys Asn Met Arg Lys Cys Asn Thr Pro Lys Pro

210 215 220

Gly Ser Gln Gln Gly Gly Ser Ser Asp Lys Ile Ser Lys Gln Asn Ala

225 230 235 240

Pro Glu Gly Ser Trp Lys Cys Asp Asn Cys Gly Asn Ile Asn Tyr Pro

245 250 255

Phe Arg Ser Lys Cys Asn Arg Gln Asn Cys Gly Ala Asp Lys Pro Gly

260 265 270

Asp Arg Ser Asn Gly Ser Pro Ser Arg Ala Pro Glu Glu Asn Asp Gln

275 280 285

<210>305

<211>1853

<212>DNA

＜213〉rice

<400>305

cagtttccac tttcttccaa agccgaaaaa tcgagagaga aagagcaaaa ccctaaccgc 60

ggcgctccgc ccccttccgc cgccggattc ctcctcctcc ttcgtccgcc ttcgccggcg 120

ccgccctgct ccgaaggagc ccaggccttg tcgccgcgtc gcttgccccg ccgccggttt 180

cagcagcagc cacccccacc accatgtctt ctcaggtcga caaccgtagc caatctgctg 240

gaaagcgtgc ccgcaccgac ggtgggcggc gtgaggacga ctgggtttgc cccagctgcc 300

agaacgtcaa cttcgccttc cgcaccacct gcaacatgcg caattgcaac caatccaggc 360

ccaccgacta tacgaaggat atgcagaaac ctatgcagac accgccgccc catttcccca 420

tgtcaggggg atacatgagc ccagggacgc cgccatccat gtacctcggg ggtggtgctc 480

ccccttatgg cacctccctc tatggcggac ctgctttacc acgttatggt attgctcagt 540

tccctggggg ttctggatat ccatatggct atggtggccg cctgccaatg gggagcccct 600

atgggccgcc gatgcatatg gcagggcctc catattctgc tggatccatg atgggaccag 660

gtggaatgta tgggatgccc atggacagat atagcttggg cttacctgct ggtcctggtc 720

caatgggcgc gagggctggt tcatattccg aggaaggatc ccagaagaag cctgcaggag 780

ctgggcgtga taatgattgg aaatgcccta attgcaacaa cattaacttt gcattccgga 840

cagtctgtaa catgagaaag tgcaatacac caagacctga aaaccagggg tccaaacctg 900

atggtgcaag aggtccaaaa ccaaagatgc cagaaggtag ttggaagtgc gagaagtgca 960

ataacataaa ctatccattc cgcacaaaat gtaaccgtcc aagttgcgag gctgaaaagc 1020

catttcagac aaacaatgct aatgagtcat ctgctgatca ggacaatcag ttgttgtcat 1080

gtaatattgt gaagctttta tcaaagctgc aactttcaca tgactttcaa gacaataatg 1140

agcaaacaaa ggtggatcct cccggtggcc ctgctggagt accgacaagt tcgcacgccc 1200

tgcgaatggc agctctcagt gagttgcaga acagtgttaa gagctgatac ccttttcatt 1260

ttccaaacaa ggtgagaact ttgatccatg tggatgtgct tctatcagga gcagctgaca 1320

gtattgcatg cagtgatctt gctaaggaag ttggagatgg gaaacatcgt gtgatggcca 1380

gccttgatgt aagaatgcaa atcagtctta ctccgtacca tatctgtgtg gagtgtgcac 1440

accggaagca tagtcatttt ttgaatggtc gattgcttct tccacagtaa agtctataat 1500

accagtgtgc aggtgttttt ttccctttct ttcagtgtga aggtgttttt ttaccccttt 1560

ctttcgttca ttcattcttt ttctttttcc ttgttgagac ggtatcctgg agcactttgt 1620

aatactctaa tgtgcttcgg aaagtagaat ataagttgtg cttatgggcg ggccaaaatg 1680

ctttggccgt tagttgcatg acagcctttc caacatgctg ctgcctagcc gtagttctcc 1740

caaaaatgaa accatcttga gatccgtttg cggttgaact gaaattagac ggtatctctg 1800

gtttcaacat catcctttaa accctggatg aaagatttgc aatattcctg cgg 1853

<210>306

<211>347

<212>PRT

＜213〉rice

<400>306

Met Ser Ser Gln Val Asp Asn Arg Ser Gln Ser Ala Gly Lys Arg Ala

1 5 10 15

Arg Thr Asp Gly Gly Arg Arg Glu Asp Asp Trp Val Cys Pro Ser Cys

20 25 30

Gln Asn Val Asn Phe Ala Phe Arg Thr Thr Cys Asn Met Arg Asn Cys

35 40 45

Asn Gln Ser Arg Pro Thr Asp Tyr Thr Lys Asp Met Gln Lys Pro Met

50 55 60

Gln Thr Pro Pro Pro His Phe Pro Met Ser Gly Gly Tyr Met Ser Pro

65 70 75 80

Gly Thr Pro Pro Ser Met Tyr Leu Gly Gly Gly Ala Pro Pro Tyr Gly

85 90 95

Thr Ser Leu Tyr Gly Gly Pro Ala Leu Pro Arg Tyr Gly Ile Ala Gln

100 105 110

Phe Pro Gly Gly Ser Gly Tyr Pro Tyr Gly Tyr Gly Gly Arg Leu Pro

115 120 125

Met Gly Ser Pro Tyr Gly Pro Pro Met His Met Ala Gly Pro Pro Tyr

130 135 140

Ser Ala Gly Ser Met Met Gly Pro Gly Gly Met Tyr Gly Met Pro Met

145 150 155 160

Asp Arg Tyr Ser Leu Gly Leu Pro Ala Gly Pro Gly Pro Met Gly Ala

165 170 175

Arg Ala Gly Ser Tyr Ser Glu Glu Gly Ser Gln Lys Lys Pro Ala Gly

180 185 190

Ala Gly Arg Asp Asn Asp Trp Lys Cys Pro Asn Cys Asn Asn Ile Asn

195 200 205

Phe Ala Phe Arg Thr Val Cys Asn Met Arg Lys Cys Asn Thr Pro Arg

210 215 220

Pro Glu Asn Gln Gly Ser Lys Pro Asp Gly Ala Arg Gly Pro Lys Pro

225 230 235 240

Lys Met Pro Glu Gly Ser Trp Lys Cys Glu Lys Cys Asn Asn Ile Asn

245 250 255

Tyr Pro Phe Arg Thr Lys Cys Asn Arg Pro Ser Cys Glu Ala Glu Lys

260 265 270

Pro Phe Gln Thr Asn Asn Ala Asn Glu Ser Ser Ala Asp Gln Asp Asn

275 280 285

Gln Leu Leu Ser Cys Asn Ile Val Lys Leu Leu Ser Lys Leu Gln Leu

290 295 300

Ser His Asp Phe Gln Asp Asn Asn Glu Gln Thr Lys Val Asp Pro Pro

305 310 315 320

Gly Gly Pro Ala Gly Val Pro Thr Ser Ser His Ala Leu Arg Met Ala

325 330 335

Ala Leu Ser Glu Leu Gln Asn Ser Val Lys Ser

340 345

<210>307

<211>1078

<212>DNA

＜213〉rice

<400>307

ctggtctctc gctctcttcg ctcgcggcgg cttctccgcc tccgcctccg ccgccctctc 60

ctcctctcgc ctcgccgccg gcgccgccag ccaccgcgcg ggtcgggcgg gccgtatctt 120

cctttctgtt ccgatggctt ccgcgaaggt ggagaaccgc ggcggcggtg ggttcggttc 180

gaagaggtca cgcaacgacg tgtctgtaag ggagggggac tggacttgtc ctcagtgtgg 240

taatgtcaac ttcagtttta gaaatgtttg caaccgcgga gcctgtggtg caccccgtcc 300

atcaccgagt ctaagcccaa gagtgccacc tcctcctgct gctggatatg atcggccaca 360

tcttggatat gatcggccac atctgtttta tggtagtgct ggcaccccac ctcctattcc 420

tcttggatct ggtagctatg gtgcccccta tccacatctt ggcttgcggt atggatatgg 480

tccaccagta ggacctcctg cttcatatgg ccttttttct tcttatggtc aacctggacc 540

aatgggcagt ccgatgggag gcatgggcta tggccctgga cctgagctag gccgatatgg 600

ttatggtttt agaggatctc caatgccggt ttctagccca tggtctggtg gagcattagt 660

ggaaaataat gacagctctg cttcacgcaa gcgtcgtgga ggcccagatg gaatggctga 720

gaatgactgg atctgcccaa agtgtgagaa tgtcaacttt tccttcagaa acagttgcaa 780

tatgaagaaa tgtggagctc caaggccaag ccctggatct aatgctaccc catgtcgcaa 840

agacaaggac gctccggaag ggagctggac ctgcccggag tgcaacaacc tgaactaccc 900

tttccgcacg gcgtgcaatc ggaaaggctg cggaagcagc aggccggcag cggccacggc 960

gaactaggac cctactaact ttgcgccggc gattgggggc agagagagta ttgttgtatc 1020

ctagctatat acaagttaat ttaaactgta aacggctcaa atatattttc ttgtggcc 1078

<210>308

<211>277

<212>PRT

＜213〉rice

<400>308

Met Ala Ser Ala Lys Val Glu Asn Arg Gly Gly Gly Gly Phe Gly Ser

1 5 10 15

Lys Arg Ser Arg Asn Asp Val Ser Val Arg Glu Gly Asp Trp Thr Cys

20 25 30

Pro Gln Cys Gly Asn Val Asn Phe Ser Phe Arg Asn Val Cys Asn Arg

35 40 45

Gly Ala Cys Gly Ala Pro Arg Pro Ser Pro Ser Leu Ser Pro Arg Val

50 55 60

Pro Pro Pro Pro Ala Ala Gly Tyr Asp Arg Pro His Leu Gly Tyr Asp

65 70 75 80

Arg Pro His Leu Phe Tyr Gly Ser Ala Gly Thr Pro Pro Pro Ile Pro

85 90 95

Leu Gly Ser Gly Ser Tyr Gly Ala Pro Tyr Pro His Leu Gly Leu Arg

100 105 110

Tyr Gly Tyr Gly Pro Pro Val Gly Pro Pro Ala Ser Tyr Gly Leu Phe

115 120 125

Ser Ser Tyr Gly Gln Pro Gly Pro Met Gly Ser Pro Met Gly Gly Met

130 135 140

Gly Tyr Gly Pro Gly Pro Glu Leu Gly Arg Tyr Gly Tyr Gly Phe Arg

145 150 155 160

Gly Ser Pro Met Pro Val Ser Ser Pro Trp Ser Gly Gly Ala Leu Val

165 170 175

Glu Asn Asn Asp Ser Ser Ala Ser Arg Lys Arg Arg Gly Gly Pro Asp

180 185 190

Gly Met Ala Glu Asn Asp Trp Ile Cys Pro Lys Cys Glu Asn Val Asn

195 200 205

Phe Ser Phe Arg Asn Ser Cys Asn Met Lys Lys Cys Gly Ala Pro Arg

210 215 220

Pro Ser Pro Gly Ser Asn Ala Thr Pro Cys Arg Lys Asp Lys Asp Ala

225 230 235 240

Pro Glu Gly Ser Trp Thr Cys Pro Glu Cys Asn Asn Leu Asn Tyr Pro

245 250 255

Phe Arg Thr Ala Cys Asn Arg Lys Gly Cys Gly Ser Ser Arg Pro Ala

260 265 270

Ala Ala Thr Ala Asn

275

<210>309

<211>662

<212>DNA

＜213〉overgrown with weeds blue or green (Brassica rapa)

<400>309

ggtcttgttt tctttcaata aacacataat aagatagctt ccaagttcat caagaaataa 60

acgtaagcgc acgcatacaa taacctatcg aagatgagca gacccggaga ctggaactgt 120

agatcatgca cccacctcaa cttccagcgc cgtgattctt gccagcgatg cggtgactcc 180

cgtttgggtg caggtggagt cggtggctta gagtttggtg atttcggcgg cagaggtatg 240

tctgcttttg gattcaccac gggctccgac gttcgtccag gtgactggta ctgcacagtt 300

ggaaactgcg ggacacataa ctttgccagc cgctccacct gcttcaaatg cggcactttc 360

aaggacgaat ccctcggtgg gggcggcggc ggtggcgtag gcgtaaggcg gtccggtcat 420

gttgacgctg acgttatgcg gtctagagtc tccggcaacg gtggccgctc cagctggaaa 480

tccggtgatt ggatttgcac caggcttggt tgcaatgagc ataactttgc aagcagaatg 540

gagtgcttca gatgcaatgc accaagggac ttcagcatga gaacctcttt ctaagttaca 600

caataatgcc tttgaagtag ccttgtttcc aatctcttgg gcaccgtttc aatcaatgga 660

aa 662

<210>310

<211>166

<212>PRT

＜213〉overgrown with weeds blue or green

<400>310

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Thr His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Ser Arg Leu Gly

20 25 30

Ala Gly Gly Val Gly Gly Leu Glu Phe Gly Asp Phe Gly Gly Arg Gly

35 40 45

Met Ser Ala Phe Gly Phe Thr Thr Gly Ser Asp Val Arg Pro Gly Asp

50 55 60

Trp Tyr Cys Thr Val Gly Asn Cys Gly Thr His Asn Phe Ala Ser Arg

65 70 75 80

Ser Thr Cys Phe Lys Cys Gly Thr Phe Lys Asp Glu Ser Leu Gly Gly

85 90 95

Gly Gly Gly Gly Gly Val Gly Val Arg Arg Ser Gly His Val Asp Ala

100 105 110

Asp Val Met Arg Ser Arg Val Ser Gly Asn Gly Gly Arg Ser Ser Trp

115 120 125

Lys Ser Gly Asp Trp Ile Cys Thr Arg Leu Gly Cys Asn Glu His Asn

130 135 140

Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg Asp Phe

145 150 155 160

Ser Met Arg Thr Ser Phe

165

<210>311

<211>749

<212>DNA

＜213〉Euphorbia esula L

<400>311

attttccctt cttcacccaa atatctctca ccttttaaca agagaaaagc aaaaagaaga 60

tgagcagacc aggagattgg aactgcaggt cctgccagca tctcaacttc cagagaaggg 120

actcgtgcca gcgctgtggg gactccaggt ccgggactgg aggttcagga gctgactttg 180

gcgggtttgg aagccgggtt gggtcctcat tcgggttcag caccgggtct gatgttcgac 240

ccggtgactg gtactgtact gctggcaact gtggggccca caactttgcc agccgggcaa 300

gttgcttcaa atgtggagtt tataaggatg attctggggc ccctggcggg tttgattccg 360

atatcctccg ggcctctaga ggttttggta gtggcagcaa tcgctcttct tggaaatctg 420

gtgattggat ctgcactcgg tggggatgta atgaacacaa ctttgcaagc agaatggagt 480

gtttcaaatg cagtgcccct agggacctta gtaacagaac ttcatactag acattttgga 540

tggtgcagca gccaagaaag agaattaaga ttactacttt taaattttat tttattttta 600

ctatttgttt tgggtgttat tagagtggaa aacaaacatg gcttaaaaac ccatgtttca 660

tccctaattt aagctaagga agcaaacccc ttttggcatt ttgtaaggca gatttaggat 720

tgtagtactt aattaattag tgggttgtt 749

<210>312

<211>156

<212>PRT

＜213〉Euphorbia esula L

<400>312

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Ser Arg Ser Gly

20 25 30

Thr Gly Gly Ser Gly Ala Asp Phe Gly Gly Phe Gly Ser Arg Val Gly

35 40 45

Ser Ser Phe Gly Phe Ser Thr Gly Ser Asp Val Arg Pro Gly Asp Trp

50 55 60

Tyr Cys Thr Ala Gly Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ala

65 70 75 80

Ser Cys Phe Lys Cys Gly Val Tyr Lys Asp Asp Ser Gly Ala Pro Gly

85 90 95

Gly Phe Asp Ser Asp Ile Leu Arg Ala Ser Arg Gly Phe Gly Ser Gly

100 105 110

Ser Asn Arg Ser Ser Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Trp

115 120 125

Gly Cys Asn Glu His Asn Phe Ala Ser Arg Met Glu Cys Phe Lys Cys

130 135 140

Ser Ala Pro Arg Asp Leu Ser Asn Arg Thr Ser Tyr

145 150 155

<210>313

<211>646

<212>DNA

＜213〉upland cotton

<400>313

ctccattctc ttatttaact caaactcacc accttttcca tttttctaat taagaaaaga 60

aaagatgagc aggccaggag attggaactg caggtcgtgc caacacctaa acttccaaag 120

gagggactcc tgccaacgct gcggggaatt ccggtcgggt gatcacttcg gtagctacgg 180

tggtggcagg ggtggctcct cctttggatt cgccaccggc tccgacgtcc gacctggtga 240

ttggtactgc actgcgggaa actgcggtac ccacaatttc gccagtcgtt ccagctgctt 300

caaatgtggt gcattcaagg acgaccctgc cggaggtttc gacagcgacg ttccgcgttc 360

tagaggattt ggcggcggta atcgatccgg ctggaaatcc ggcgactgga tatgtaccag 420

gtcgggatgc aatgagcata actttgctag ccgaatggaa tgtttcagat gcagtgcccc 480

aagagactt caccgctagaa cttcatacta aatacaagct atccagtttt gggtgttgca 540

aaccaagaga gactcaaatg agagaaaaaa ttttacgggg ttagggttac ccgggtaaat 600

atatggcttg ggggggtgtg gcaccagggt tgaaaacacc agggtg 646

<210>314

<211>148

<212>PRT

＜213〉upland cotton

<400>314

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Glu Phe Arg Ser Gly

20 25 30

Asp His Phe Gly Ser Tyr Gly Gly Gly Arg Gly Gly Ser Ser Phe Gly

35 40 45

Phe Ala Thr Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Thr Ala

50 55 60

Gly Asn Cys Gly Thr His Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys

65 70 75 80

Cys Gly Ala Phe Lys Asp Asp Pro Ala Gly Gly Phe Asp Ser Asp Val

85 90 95

Pro Arg Ser Arg Gly Phe Gly Gly Gly Asn Arg Ser Gly Trp Lys Ser

100 105 110

Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu His Asn Phe Ala

115 120 125

Ser Arg Met Glu Cys Phe Arg Cys Ser Ala Pro Arg Asp Phe Thr Ala

130 135 140

Arg Thr Ser Tyr

145

<210>315

<211>613

<212>DNA

＜213〉grape

<400>315

ggtcttcttc catctctgca cagttatcct gatatttcct tggaaaacat gagcaggcca 60

ggagattgga actgcaggtc atgccagcac atgaacttcc aaaggcgcga ttcctgccaa 120

cgctgcggtg acccaaaatc gggtgggggt gactttggaa gctttggtgg gaggggtgga 180

tcctcctttg ggttcacggg ctcggatgtc cgcccagggg actggtactg caatgcaggc 240

aactgtggag ctcacaactt tgctagccgc tctaactgct tcaagtgtgg tgcattcaaa 300

gatgagtctg ctgggggcta cgattccgac atgtcacgct cccgaggttt cgggttcggc 360

ggtggcagcg gccggtctgg gtggaaatcc ggtgattgga tatgcagcag gtctggatgc 420

aatgagcaca actttgctag cagaatggaa tgtttcagat gcaatgcccc gagggacttg 480

agtaacaaaa cttcatacta gacatatatc ttccattttt gggtactgca gccagccaag 540

agagaccaaa tcatagaata tctattaatc cttgtgttgt tattaatttc tttgctttgg 600

gtgctaggtt ctt 613

<210>316

<211>150

<212>PRT

＜213〉grape

<400>316

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Met Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Pro Lys Ser Gly

20 25 30

Gly Gly Asp Phe Gly Ser Phe Gly Gly Arg Gly Gly Ser Ser Phe Gly

35 40 45

Phe Thr Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Asn Ala Gly

50 55 60

Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Asn Cys Phe Lys Cys

65 70 75 80

Gly Ala Phe Lys Asp Glu Ser Ala Gly Gly Tyr Asp Ser Asp Met Ser

85 90 95

Arg Ser Arg Gly Phe Gly Phe Gly Gly Gly Ser Gly Arg Ser Gly Trp

100 105 110

Lys Ser Gly Asp Trp Ile Cys Ser Arg Ser Gly Cys Asn Glu His Asn

115 120 125

Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg Asp Leu

130 135 140

Ser Asn Lys Thr Ser Tyr

145 150

<210>317

<211>844

<212>DNA

＜213〉comospore poplar (Populus trichocarpa)

<400>317

tcagaccatt tccctttcat cttctagcta gcatagcttc tctcagaggt ttttgaaacc 60

cctttttgcc attcttctct cacttagtat acttagtctt ctctaatcaa ttcataagca 120

aatatgaaca ggccaggaga ctggaactgc aggtcatgcc aacacctcaa tttccagagg 180

cgtgactctt gccaacgttg tggggacccc aggtccgcag gtgattttgg gggtttcggt 240

gggcggggtg gctcatcact tgggttcacc gggtcggatg ttcgtcccgg tgattggtac 300

tgcactgccg gaaactgcgg ggcccacaac tttgctagcc gttctagttg cttcaaatgt 360

ggagtgtaca aggaaatgga ctccgccggg ggcttcgatt ctgatttttc tcgaactaga 420

gggtttggtg ggagcactgg aggtggcaat cgatctggat ggaaatccgg agactggatt 480

tgcactaggt ggggatgcaa cgaacataac tttgctagca gaatggagtg cttcaagtgc 540

aatgccccaa gagatcttag caacagaact tcatactaga tacaatttct ccatttcctg 600

ggactgcacc agccaagaac aaagacaaca tgattaaaaa atatatatca ctacttttca 660

ttttcttctt gatttctttt tgtattagct agctagggtc ttgaggcgag acatggttta 720

agaaccatgt ttactgcttt gagctatata taacccttgg cattttgtca ggcttcctgt 780

aggaagtata tcgttgtctt tgtgggcttt tgatctatta tattcattat tgtaaccaag 840

ggag 844

<210>318

<211>151

<212>PRT

＜213〉comospore poplar

<400>318

Met Asn Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Pro Arg Ser Ala

20 25 30

Gly Asp Phe Gly Gly Phe Gly Gly Arg Gly Gly Ser Ser Leu Gly Phe

35 40 45

Thr Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Thr Ala Gly Asn

50 55 60

Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys Cys Gly

65 70 75 80

Val Tyr Lys Glu Met Asp Ser Ala Gly Gly Phe Asp Ser Asp Phe Ser

85 90 95

Arg Thr Arg Gly Phe Gly Gly Ser Thr Gly Gly Gly Asn Arg Ser Gly

100 105 110

Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Trp Gly Cys Asn Glu His

115 120 125

Asn Phe Ala Ser Arg Met Glu Cys Phe Lys Cys Asn Ala Pro Arg Asp

130 135 140

Leu Ser Asn Arg Thr Ser Tyr

145 150

<210>319

<211>785

<212>DNA

＜213〉upland cotton

<400>319

ccttgcgtcc gctttcttct tcattgttcc tgtctttgct tactcacccc ttatatatct 60

gctttctcaa gtaggggaga aaatatgagc aggccaggag actggaattg caggtcatgc 120

caacacctca acttccaaag gagggactca tgccagcgct gtggagaacc aagacctggt 180

ggtggtgaca gaggcggcga ctatggaagc tttggtggca ggggtggctc atctttcggg 240

tttactggac ccgatgttag gcctggtgac tggtattgca ctgtgggcaa ctgcggtgct 300

cacaacttcg ccagcaggtc gagctgcttc aaatgtggtg cggccaaaga tgaatcatcc 360

ggaggatttg aaagtgacat cccacgtatg aggggttatg gttttagcac tggcagctct 420

agtcgctcta actggaaatc tggagactgg atttgcacca ggtcgggttg caatgaacac 480

aacttcgcca gcaggatgga atgtttcaga tgcaatgcac caagggactc cacccacaaa 540

tcttcatact aattaataaa attttcattt ttgggtactg cagtcgcaag agagagatca 600

gacaaggcaa tcccagatct tgctttcttt ttcttttgtt tgtttcttta ttttagatct 660

ttagatgaat catgtttgaa agacttcagt tgaagtactt aagctaatat caaagtacat 720

agggcatgca tgtaattgat gtatggtatc agcaatgtta tatcagtgtc tttgtgccaa 780

aaaaa 785

<210>320

<211>155

<212>PRT

＜213〉upland cotton

<400>320

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Glu Pro Arg Pro Gly

20 25 30

Gly Gly Asp Arg Gly Gly Asp Tyr Gly Ser Phe Gly Gly Arg Gly Gly

35 40 45

Ser Ser Phe Gly Phe Thr Gly Pro Asp Val Arg Pro Gly Asp Trp Tyr

50 55 60

Cys Thr Val Gly Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser

65 70 75 80

Cys Phe Lys Cys Gly Ala Ala Lys Asp Glu Ser Ser Gly Gly Phe Glu

85 90 95

Ser Asp Ile Pro Arg Met Arg Gly Tyr Gly Phe Ser Thr Gly Ser Ser

100 105 110

Ser Arg Ser Asn Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly

115 120 125

Cys Asn Glu His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn

130 135 140

Ala Pro Arg Asp Ser Thr His Lys Ser Ser Tyr

145 150 155

<210>321

<211>760

<212>DNA

＜213〉fireweed

<400>321

gtattcggaa tcctcaaaga aacacacact tcctatccat cacaagaaag atgagcaggc 60

caggagattg gaactgcagg tcatgccagc acttgaactt ccagaggagg gactcttgcc 120

aaagatgtgg ggagacgagg tatggtggtg ggggtggtgt gtttggtggt agaggaagta 180

tcatcagccc ttcagcattt ggcttcacag gcccagatgt ccgaccgggt gattggtact 240

gcaatgttgg caactgcggg gctcacaact ttgctagccg ctcgagctgc ttcaagtgtg 300

gtgcgttcaa ggatgactta gcttgtagtg gtggtggtgg tggtggtggt ggcgtttttg 360

atggtgatat gtcacgtggc aggggtttcg ggtttggtgg aggaagtggt ggtggaggtg 420

gtggcagcag ccgttcaggg tggaagtccg gtgactggat atgcggcagg cctggttgca 480

atgagcacaa ctttgcaagc agaatggaat gttttaggtg caacgcacct cgggaatcgg 540

gtaacaagtc tccttattaa gcaggttgcg gcatttccag ttccgggtat catggacttg 600

ctatctacca aacaggaaga gagataagtg atcgatcaga cagaatcatg aagagcatcc 660

agttgatagg aaattctaaa ctgaccccct ttttgcccat atcataatat gcaatcctat 720

ctttgatttt ttttttcctt ttgttttatt tttttttttt 760

<210>322

<211>169

<212>PRT

＜213〉fireweed

<400>322

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Glu Thr Arg Tyr Gly

20 25 30

Gly Gly Gly Gly Val Phe Gly Gly Arg Gly Ser Ile Ile Ser Pro Ser

35 40 45

Ala Phe Gly Phe Thr Gly Pro Asp Val Arg Pro Gly Asp Trp Tyr Cys

50 55 60

Asn Val Gly ASn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys

65 70 75 80

Phe Lys Cys Gly Ala Phe Lys Asp Asp Leu Ala Cys Ser Gly Gly Gly

85 90 95

Gly Gly Gly Gly Gly Val Phe Asp Gly Asp Met Ser Arg Gly Arg Gly

100 105 110

Phe Gly Phe Gly Gly Gly Ser Gly Gly Gly Gly Gly Gly Ser Ser Arg

115 120 125

Ser Gly Trp Lys Ser Gly Asp Trp Ile Cys Gly Arg Pro Gly Cys Asn

130 135 140

Glu His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro

145 150 155 160

Arg Glu Ser Gly Asn Lys Ser Pro Tyr

165

<210>323

<211>517

<212>DNA

＜213〉soybean

<400>323

tttctgcatc tctaatactt caactcactg tattatcttg agtaattttg cagagaagta 60

aaatatgagc aggccaggag actggaattg caggtcgtgc cagcacctga actttcagag 120

gagagactca tgccagcgat gtggggactc aaaatatgga gatagagttg ttgattttgg 180

tggttttgga ggaagaggag ggtcctcatt tggtttaact ggctcagatg ttcgccccgg 240

cgactggtac tgtgctgctg ctaactgtgg tgcacacaac tttgctagcc gctcaagctg 300

cttcaagtgt ggtgctttca aggatgactt ggctggagga ggctataaca gttctgacat 360

cttgcgctcc agagcttttg gtggcagtgg aagacctgga tggaaatctg gtgattggat 420

atgcagcaga tcaggatgca atgagcacaa ctttgctagc agaatggaat gttttaaatg 480

cagtgctccc agggacacgt actagaaaaa atgagtt 517

<210>324

<211>146

<212>PRT

＜213〉soybean

<400>324

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Ser Lys Tyr Gly

20 25 30

Asp Arg Val Val Asp Phe Gly Gly Phe Gly Gly Arg Gly Gly Ser Ser

35 40 45

Phe Gly Leu Thr Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Ala

50 55 60

Ala Ala Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe

65 70 75 80

Lys Cys Gly Ala Phe Lys Asp Asp Leu Ala Gly Gly Gly Tyr Asn Ser

85 90 95

Ser Asp Ile Leu Arg Ser Arg Ala Phe Gly Gly Ser Gly Arg Pro Gly

100 105 110

Trp Lys Ser Gly Asp Trp Ile Cys Ser Arg Ser Gly Cys Asn Glu His

115 120 125

Asn Phe Ala Ser Arg Met Glu Cys Phe Lys Cys Ser Ala Pro Arg Asp

130 135 140

Thr Tyr

145

<210>325

<211>889

<212>DNA

＜213〉comospore poplar

<400>325

gacaagttcc ttaaacaatc tattgctctt tcaactacta gctaggttac acttcatttc 60

tgtacgtgtt aattctccta tctttctcct ttctgctttc tcgagatgag cagaccagga 120

gattggaatt gcaggtcatg ccagcacttg aactttcaaa ggagggactc atgccagcgt 180

tgcggtgatc caaggcccgg agagagagat cactatggaa gtttcggtgg aagatcatca 240

gggggctcat tcggatttac gggccctgat gttaggcctg gtgattggta ttgcacggct 300

ggcaattgtg gagctcacaa ctttgctagt cgttcaagct gcttcaagtg tggtgtgtcc 360

aaggatgaat cctctggtgg tggacttgat gctgatatgt cacggatgag aggttatggc 420

ttcggcggag gcggaggcgg aggcagtggc tctagccgta attggaaatc cggagactgg 480

atttgcacca ggtccggttg caacgagcac aactttgcta gcaggactga atgctataga 540

tgcaatgcac caagagaatc tagcagcaac aagtcttcgt attaatcatc gatatcgata 600

gcatttttgt gtcctgcagt gctgcaagga gggaattaac gaagaaggct catgagaaat 660

cttggattca tctttttgct cttaattact tctatttttg ttcttttgga tagctgtact 720

cttaagctga gaagctttag aaggatcatg ggagtgaaat taagttcaag gagagctata 780

tatatatcat tagccagtaa tttgtcaggt tccctaaata tagacatgta gttctgtact 840

tggtatcaat gtctttgtgt tttgagacgg gtttaagccc ttgtcgttc 889

<210>326

<211>159

<212>PRT

＜213〉comospore poplar

<400>326

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Pro Arg Pro Gly

20 25 30

Glu Arg Asp His Tyr Gly Ser Phe Gly Gly Arg Ser Ser Gly Gly Ser

35 40 45

Phe Gly Phe Thr Gly Pro Asp Val Arg Pro Gly Asp Trp Tyr Cys Thr

50 55 60

Ala Gly Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe

65 70 75 80

Lys Cys Gly Val Ser Lys Asp Glu Ser Ser Gly Gly Gly Leu Asp Ala

85 90 95

Asp Met Ser Arg Met Arg Gly Tyr Gly Phe Gly Gly Gly Gly Gly Gly

100 105 110

Gly Ser Gly Ser Ser Arg Asn Trp Lys Ser Gly Asp Trp Ile Cys Thr

115 120 125

Arg Ser Gly Cys Asn Glu His Asn Phe Ala Ser Arg Thr Glu Cys Tyr

130 135 140

Arg Cys Asn Ala Pro Arg Glu Ser Ser Ser Asn Lys Ser Ser Tyr

145 150 155

<210>327

<211>723

<212>DNA

<213>Populus fremontii x Populus angustifolia

<400>327

agggaaaaag aaagcctaac cctcctgcct gcctcaagct taccccatta tcgaggtcta 60

aaacaagtat aaaaacagcc ttagccccaa tacgtaaatc acaagttcct taaacaatct 120

attgctcttt caactactag gtttcccact tcatttctgt gcctactgat tctcctctct 180

ttcaactttc tgccttctca agctagctag agaagggaag atgagcagac caggagattg 240

gaattgcagg tcatgccaac acttgaactt ccagaggagg gactcgtgcc agcgttgtgg 300

ggacccaagg cccggagaga gagatcatta tggaagtttt ggtggaagat cagggggctc 360

gttcggattt acggggcctg atgttaggcc cggtgactgg tattgctcgg ttggcaactg 420

tggagctcac aactttgcta gtcgttcaag ctgcttcaag tgtggtatgt ccaaggatga 480

atcctctggt ggtgggcttg atgctgacat ttcatggatg agaggttatg gcttcggcgg 540

aggcagcgcc tctagccgct ctaattggaa atccggagac tggatttgca ccaggtcagg 600

ttgcaacgag cacaactttg ctagcaggac tgagtgttac agatgcaatg caccaagaga 660

atcaggcagc aacaagtctt cgtattaatc atcaacatcg atcgcatttt tgtgtactgc 720

agt 723

<210>328

<211>155

<212>PRT

<213>Populus fremontii x Populus angustifolia

<400>328

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu Asn

1 5 10 15

Phe Gln Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Pro Arg Pro Gly

20 25 30

Glu Arg Asp His Tyr Gly Ser Phe Gly Gly Arg Ser Gly Gly Ser Phe

35 40 45

Gly Phe Thr Gly Pro Asp Val Arg Pro Gly Asp Trp Tyr Cys Ser Val

50 55 60

Gly Asn Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys

65 70 75 80

Cys Gly Met Ser Lys Asp Glu Ser Ser Gly Gly Gly Leu Asp Ala Asp

85 90 95

Ile Ser Trp Met Arg Gly Tyr Gly Phe Gly Gl y Gly Ser Ala Ser Ser

100 105 110

Arg Ser Asn Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys

115 120 125

Asn Glu His Asn Phe Ala Ser Arg Thr Glu Cys Tyr Arg Cys Asn Ala

130 135 140

Pro Arg Glu Ser Gly Ser Asn Lys Ser Ser Tyr

145 150 155

<210>329

<211>920

<212>DNA

＜213〉rice

<400>329

cacctcacag cattttccac cttagagctc accaacacag cagctgatac ttgtttaggg 60

taagacaaga tgaacaggaa gccaggagac tgggactgca gggcgtgcca gcacctcaac 120

ttcagccgcc gggacctatg ccagcgctgc ggcgggccgc gtggcgccgc tgatcgcggc 180

agcggtggtg gcggtgacta cgccaacttc ggcggccgtg gtggttcctc cttcggtgga 240

ggctttggca ctggctctga tgtccgccca ggtgactggt actgcaactg cggcgcgcac 300

aacttcgcca gccgctccag ctgcttcaag tgcgctgctt tcaaggacga tgctgccgtc 360

aacagtggcg gcgctggtgc ctttgacggt ggggacatgt cgcgctcgcg gggctacggc 420

ttcggcagcg gcgccgtccg cgccagccgc cctggctgga agtctggcga ctggatttgc 480

accaggtctg gatgcaatga gcacaacttc gccagcagga tggagtgctt caggtgcaac 540

gcaccgcggg actccggcac tgaggtgtaa tttgccgtac gtgtccgatc gatctggatc 600

cgatgaggct tgcagcagtg acgacgagca gcagaagcag cgttaagagt tgtgatgtct 660

acataagaag aagaagaaag tagaatgcaa aagaaatctc cccatggttt tactagtttt 720

gtttcttccc gttttagatt tggttctgat tcccatttgg gaggacccgt cgacccctga 780

ttatctatgt tttacccgtt ttatttcctg tttctttcgg catgtttgct cttcgatcga 840

gtcgtgtaac ccgaaacgct tgcgcttgag aagtattatt attattaact agtatgttgc 900

ttcttaaaaa aaaaaaaaaa 920

<210>330

<211>166

<212>PRT

＜213〉rice

<400>330

Met Asn Arg Lys Pro Gly Asp Trp Asp Cys Arg Ala Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Leu Cys Gln Arg Cys Gly Gly Pro Arg Gly

20 25 30

Ala Ala Asp Arg Gly Ser Gly Gly Gly Gly Asp Tyr Ala Asn Phe Gly

35 40 45

Gly Arg Gly Gly Ser Ser Phe Gly Gly Gly Phe Gly Thr Gly Ser Asp

50 55 60

Val Arg Pro Gly Asp Trp Tyr Cys Asn Cys Gly Ala His Asn Phe Ala

65 70 75 80

Ser Arg Ser Ser Cys Phe Lys Cys Ala Ala Phe Lys Asp Asp Ala Ala

85 90 95

Val Asn Ser Gly Gly Ala Gly Ala Phe Asp Gly Gly Asp Met Ser Arg

100 105 110

Ser Arg Gly Tyr Gly Phe Gly Ser Gly Ala Val Arg Ala Ser Arg Pro

115 120 125

Gly Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu

130 135 140

His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg

145 150 155 160

Asp Ser Gly Thr Glu Val

165

<210>331

<211>703

<212>DNA

＜213〉switchgrass

<400>331

ccacgcgtcc gcagaactcc ttgtcctagg ccccagaaac acaaacgacc gctctttgtt 60

taggcaagat gaacaggaag cctggagact gggattgcag ggcttgccaa cacctcaact 120

tcagtcggcg ggacctatgc cagcgctgtg gtgagccacg tggagctgct gaccgtggca 180

gcggcggtgg aggtgactat gccaacttcg gtggccgtgg tggctcctcc ttcggcggtg 240

gctttggtgc tggctctgat gtccgccctg gtgactggtt atgttcctgc ggcgcgcaca 300

acttcgccag ccgctccaac tgcttcaagt gctctgcctt caaggaggag gctgctgtca 360

acagtggtgc tggtggcttt gatggtgaca tgtcacgctc gcgctacggc ttcggtggcg 420

gtgctgcccg caccaaccgc cctggttgga agtctggaga ctggatctgc accaggtccg 480

gatgcaacga gcacaacttt gccagcagga tggagtgttt caggtgcaac gcaccacggg 540

actccggcac tgaggtgtag gatcggagct ggtgcttcga acggaccccg acgagcagaa 600

gcagcaagaa acctgataag aagagtagta gatttgcaaa aggcatcccg gatgctctat 660

tactgttttg ttccacccct accgcttcct tttagaattt ggt 703

<210>332

<211>163

<212>PRT

＜213〉switchgrass

<400>332

Met Asn Arg Lys Pro Gly Asp Trp Asp Cys Arg Ala Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Leu Cys Gln Arg Cys Gly Glu Pro Arg Gly

20 25 30

Ala Ala Asp Arg Gly Ser Gly Gly Gly Gly Asp Tyr Ala Asn Phe Gly

35 40 45

Gly Arg Gly Gly Ser Ser Phe Gly Gly Gly Phe Gly Ala Gly Ser Asp

50 55 60

Val Arg Pro Gly Asp Trp Leu Cys Ser Cys Gly Ala His Asn Phe Ala

65 70 75 80

Ser Arg Ser Asn Cys Phe Lys Cys Ser Ala Phe Lys Glu Glu Ala Ala

85 90 95

Val Asn Ser Gly Ala Gly Gly Phe Asp Gly Asp Met Ser Arg Ser Arg

100 105 110

Tyr Gly Phe Gly Gly Gly Ala Ala Arg Thr Asn Arg Pro Gly Trp Lys

115 120 125

Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu His Asn Phe

130 135 140

Ala Ser Arg Met Glu Cys Phe Arg Cys Asn Ala Pro Arg Asp Ser Gly

145 150 155 160

Thr Glu Val

<210>333

<211>435

<212>DNA

＜213〉puncture vine clover

<400>333

atgaacagga aaatgagctg gtctggagga gattggatgt gtggtgcttg cgagcacata 60

aatttcaaga agagagaagc atgccaaaat tgtggatacc caaagtatgg aggccctgac 120

ccatcgacct atagatataa caggactgaa acgttggcag gggactggtt ttgcacttct 180

atgaactgtg gagctcacaa ctatgcaagc cgatcaaact gctatagatg tggtgcattt 240

aaagatcctt attcttctgg atatgggggt aacatggtgg gttctggagg atatggatca 300

gattgtagtt ctcccccagg atggaaaagt ggagactgga tttgccctag aattggctgt 360

ggaatccata attatgcaag caggacagag tgctacaaat gcaaaatgcc aagggattat 420

ggtggtgcag actga 435

<210>334

<211>144

<212>PRT

＜213〉puncture vine clover

<400>334

Met Asn Arg Lys Met Ser Trp Ser Gly Gly Asp Trp Met Cys Gly Ala

1 5 10 15

Cys Glu His Ile Asn Phe Lys Lys Arg Glu Ala Cys Gln Asn Cys Gly

20 25 30

Tyr Pro Lys Tyr Gly Gly Pro Asp Pro Ser Thr Tyr Arg Tyr Asn Arg

35 40 45

Thr Glu Thr Leu Ala Gly Asp Trp Phe Cys Thr Ser Met Asn Cys Gly

50 55 60

Ala His Asn Tyr Ala Ser Arg Ser Asn Cys Tyr Arg Cys Gly Ala Phe

65 70 75 80

Lys Asp Pro Tyr Ser Ser Gly Tyr Gly Gly Asn Met Val Gly Ser Gly

85 90 95

Gly Tyr Gly Ser Asp Cys Ser Ser Pro Pro Gly Trp Lys Ser Gly Asp

100 105 110

Trp Ile Cys Pro Arg Ile Gly Cys Gly Ile His Asn Tyr Ala Ser Arg

115 120 125

Thr Glu Cys Tyr Lys Cys Lys Met Pro Arg Asp Tyr Gly Gly Ala Asp

130 135 140

<210>335

<211>513

<212>DNA

＜213〉puncture vine clover

<400>335

atgagcagac caggagattg gaactgcagg acatgcaacc acctcaactt tcaaagaaga 60

gaatcttgcc aacgatgtgg ggagtcaaga atgacttctg gctgcggcgc cgttgatttt 120

ggtggctcct ttcttggtgg aagaggctct agctcccctt ttcctttcac caccggccct 180

gatgtccgtc ctggtgactg gtattgcact gttggaaact gtggagctca caactttgcc 240

agccgctcca gctgcttcaa atgtggtgcc cctaaggata ttgatacctt ctcctctgac 300

tcctcagaca tgccacgatt attgagatca ccatacggtt ttggagcagg cagcgctggt 360

ggcggtgcct ccactcgccc cggctggaaa tccggtgact ggatatgcac caggtctggg 420

tgtaacgagc ataacttcgc caatagaatg gaatgctacc gatgcaacgg tccaagggac 480

tctagtactg gaagatcttc ctatttatcg tga 513

<210>336

<211>170

<212>PRT

＜213〉puncture vine clover

<400>336

Met Ser Arg Pro Gly Asp Trp Asn Cys Arg Thr Cys Asn His Leu Asn

1 5 10 15

Phe Gln Arg Arg Glu Ser Cys Gln Arg Cys Gly Glu Ser Arg Met Thr

20 25 30

Ser Gly Cys Gly Ala Val Asp Phe Gly Gly Ser Phe Leu Gly Gly Arg

35 40 45

Gly Ser Ser Ser Pro Phe Pro Phe Thr Thr Gly Pro Asp Val Arg Pro

50 55 60

Gly Asp Trp Tyr Cys Thr Val Gly Asn Cys Gly Ala His Asn Phe Ala

65 70 75 80

Ser Arg Ser Ser Cys Phe Lys Cys Gly Ala Pro Lys Asp Ile Asp Thr

85 90 95

Phe Ser Ser Asp Ser Ser Asp Met Pro Arg Leu Leu Arg Ser Pro Tyr

100 105 110

Gly Phe Gly Ala Gly Ser Ala Gly Gly Gly Ala Ser Thr Arg Pro Gly

115 120 125

Trp Lys Ser Gly Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu His

130 135 140

Asn Phe Ala Asn Arg Met Glu Cys Tyr Arg Cys Asn Gly Pro Arg Asp

145 150 155 160

Ser Ser Thr Gly Arg Ser Ser Tyr Leu Ser

165 170

<210>337

<211>826

<212>DNA

＜213〉yucca (Yucca filamentosa)

<400>337

ggcacaagct ctcacgcaac cccttgtctc cattttactc ttccttcctc tttcaaggga 60

cttgagaaga tgaacaggaa gccaggagac tggaactgca ggtcatgcca gcaccttaat 120

ttcagccgca gggactcgtg ccagcgctgc ggcgaccccc ggtcgatcgg cagcgagcga 180

tcggactacc cgggcttcgt cgggggccgc ggggggtcct cattcgggtt cagcggctcg 240

gacgtcaggc ccggggactg gtactgccag tgcggggccc acaacttcgc cagccgctcg 300

agctgcttca agtgcaatgc tttcaaggat gagtccgccg ctagcggcgg cctcgacggc 360

ggcgacatgc tgagatccag ggggtttggc ttcggcggcg gcggcggcgc tcgcagtggc 420

tggaagtctg gtgactggat ttgcaacagg tctggctgca atgagcacaa cttcgctagc 480

aggatggaat gcttccgatg caatgcacct cgagattcgg gcactgaggt ttaaggagac 540

agccaagaga gaagtgacga gatgagatca gtgatgatga taccagtagt aatctcgagt 600

tattactagt tttgccacca gccccaactt tatctccttt tgatgttttt ttagagcccc 660

cttctaaacg gaggtgtctt ctctttttca tccgtttgtt tgcttttgaa agcttgcact 720

taccctttac acccccccgc ccccctccac cttgtagttt gtgttgagag tgtctttgta 780

agtctttgga gaagggctcc tttagcttct tgggggagtc ccttct 826

<210>338

<211>154

<212>PRT

＜213〉yucca

<400>338

Met Asn Arg Lys Pro Gly Asp Trp Asn Cys Arg Ser Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Ser Cys Gln Arg Cys Gly Asp Pro Arg Ser

20 25 30

Ile Gly Ser Glu Arg Ser Asp Tyr Pro Gly Phe Val Gly Gly Arg Gly

35 40 45

Gly Ser Ser Phe Gly Phe Ser Gly Ser Asp Val Arg Pro Gly Asp Trp

50 55 60

Tyr Cys Gln Cys Gly Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe

65 70 75 80

Lys Cys Asn Ala Phe Lys Asp Glu Ser Ala Ala Ser Gly Gly Leu Asp

85 90 95

Gly Gly Asp Met Leu Arg Ser Arg Gly Phe Gly Phe Gly Gly Gly Gly

100 105 110

Gly Ala Arg Ser Gly Trp Lys Ser Gly Asp Trp Ile Cys Asn Arg Ser

115 120 125

Gly Cys Asn Glu His Asn Phe Ala Ser Arg Met Glu Cys Phe Arg Cys

130 135 140

Asn Ala Pro Arg Asp Ser Gly Thr Glu Val

145 150

<210>339

<211>614

<212>DNA

＜213〉barley

<400>339

cggcacgagg cacttctcat cttggagctc agcagcacag catccaagct ttttcttcgg 60

caagatgaac aggaagccag gagactggga ctgcaggtcg tgccagcacc tcaacttcag 120

tcgccgggac ctatgccagc gctgcggtga gccacgtagt gccgctgacc gtggcagcgt 180

cggtggtgct cttggtggtg actacgccaa ctttggcggc cgtggcgggg gtggttcctc 240

atttggtgcc ggctttggtg ccggctctga cgtccgcccg ggtgactggt actgcacctg 300

tggagcgcac aacttcgcca gccgctccag ctgcttcaag tgtgctgctt tcaaggagga 360

agctgccgtc aatggtggcg ctggtggctt tgatggtgac atgtcacgct caaggggctt 420

tggctttggc gctgtcggtg gcatgggtgg cggcatggga gccggcgcag ccggtggtcg 480

tgccagtcgc cctggctgga agtctcgcga ctggatttgc accaggtctg gatgcaacga 540

gcacaacttc gccagcaggc aggagtgctt caggtgcaac gcgccgaggg actccggtag 600

cgccacacca tacg 614

<210>340

<211>183

<212>PRT

＜213〉barley

<400>340

Met Asn Arg Lys Pro Gly Asp Trp Asp Cys Arg Ser Cys Gln His Leu

1 5 10 15

Asn Phe Ser Arg Arg Asp Leu Cys Gln Arg Cys Gly Glu Pro Arg Ser

20 25 30

Ala Ala Asp Arg Gly Ser Val Gly Gly Ala Leu Gly Gly Asp Tyr Ala

35 40 45

Asn Phe Gly Gly Arg Gly Gly Gly Gly Ser Ser Phe Gly Ala Gly Phe

50 55 60

Gly Ala Gly Ser Asp Val Arg Pro Gly Asp Trp Tyr Cys Thr Cys Gly

65 70 75 80

Ala His Asn Phe Ala Ser Arg Ser Ser Cys Phe Lys Cys Ala Ala Phe

85 90 95

Lys Glu Glu Ala Ala Val Asn Gly Gly Ala Gly Gly Phe Asp Gly Asp

100 105 110

Met Ser Arg Ser Arg Gly Phe Gly Phe Gly Ala Val Gly Gly Met Gly

115 120 125

Gly Gly Met Gly Ala Gly Ala Ala Gly Gly Arg Ala Ser Arg Pro Gly

130 135 140

Trp Lys Ser Arg Asp Trp Ile Cys Thr Arg Ser Gly Cys Asn Glu His

145 150 155 160

Asn Phe Ala Ser Arg Gln Gl u Cys Phe Arg Cys Asn Ala Pro Arg Asp

165 170 175

Ser Gly Ser Ala Thr Pro Tyr

180

<210>341

<211>54

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm5539

<400>341

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgag cagacccgga gatt 54

<210>342

<211>51

<212>DNA

＜213〉artificial sequence

<220>

＜223〉primer: prm5540

<400>342

ggggaccact ttgtacaaga aagctgggta gacaaggcta cttcaaaagc a 51

<210>343

<211>2193

<212>DNA

＜213〉rice

<400>343

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120

catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180

tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240

tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300

aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360

atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420

ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480

ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540

gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600

tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660

tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720

aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780

aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840

acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900

tccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggag aggcaaagaa 960

aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020

ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080

cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140

cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200

tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260

gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320

ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380

gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440

aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500

ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560

atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620

acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680

cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740

ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800

gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860

tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920

attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980

tttcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040

cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100

agctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160

tggtgtagct tgccactttc accagcaaag ttc 2193

<210>344

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉the motif sequence 1

<220>

＜221〉variant

<222>(1)..(1)

＜223 〉/replace=" Arg "/replace=" Asp "/replace=" Asn "

<400>344

Gly Asp Trp

1

<210>345

<211>3

<212>PRT

＜213〉artificial sequence

<220>

＜223〉the motif sequence 2

<400>345

Gly Ser Trp

1

<210>346

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉the motif sequence 3

<220>

＜221〉variant

<222>(3)..(3)

＜223 〉/replace=" Cys "/replace=" Ser "

<220>

＜221〉variant

<222>(4)..(4)

＜223 〉/replace=" Lys "

<400>346

Asn Phe Gln Arg Arg

1 5

<210>347

<211>5

<212>PRT

＜213〉artificial sequence

<220>

＜223〉the motif sequence 4

<220>

＜221〉variant

<222>(2)..(2)

＜223 〉/replace=" Tyr "

<220>

＜221〉variant

<222>(3)..(3)

＜223 〉/replace=" Ser "/replace=" Pro "

<220>

＜221〉variant

<222>(4)..(4)

＜223 〉/replace=" Ser "/replace=" Phe "

<400>347

Asn Phe Ala Asn Arg

1 5

Claims (154)

1. increase the method for plant seed output with respect to control plant, it comprises the expression of nucleic acid in plant of regulating coding AZ polypeptide, randomly selection has the plant of the output of increase, and wherein said AZ polypeptide comprises at least one ankyrin to be repeated and at least one C3H1 Zinc finger domain.

2. the process of claim 1 wherein that described adjusting expression is by preferably introducing the genetic modification realization in the locus of coding AZ polypeptide or its homologue.

3. the method for claim 2, wherein said genetic modification is by a kind of realization in T-DNA activation, TILLING, site-directed mutagenesis or the orthogenesis.

4. increase the method for plant seed output with respect to the corresponding wild-type plant, it is included in introduces and expresses AZ nucleic acid or its variant in the plant, wherein said AZ nucleic acid encoding polypeptide, described polypeptide comprise at least one ankyrin to be repeated and at least one C3H1 Zinc finger domain.

5. the method for claim 4, the homologue of wherein said nucleic acid encoding AZ protein s EQ ID NO:2.

6. the method for claim 4, wherein said variant be AZ nucleic acid part or can with the sequence of AZ nucleic acid hybridization, described part or hybridization sequences coded polypeptide, described polypeptide comprise at least one ankyrin to be repeated and at least one C3H1 Zinc finger domain.

7. claim 5 or 6 method, wherein said AZ nucleic acid or its variant overexpression in plant.

8. each method in the claim 5 to 7, wherein said AZ nucleic acid or its variant are plant origins, preferably from dicotyledons, more preferably from Cruciferae, most preferably described nucleic acid is from Arabidopis thaliana.

9. each method in the claim 5 to 8, wherein said AZ nucleic acid or its variant effectively are connected with seed specific promoters.

10. the method for claim 9, wherein said seed specific promoters is the WS118 promotor.

11. each method in the claim 1 to 10, the seed production of wherein said increase comprises the thousand seed weight of increase.

12. plant by each method acquisition in the claim 1 to 11.

13. construct, it comprises:

(i) AZ nucleic acid or its variant;

(ii) with effective one or more regulating and controlling sequences that are connected of the nucleotide sequence of (i).

14. the construct of claim 13, wherein said regulating and controlling sequence are seed specific promoters or constitutive promoter.

15. the construct of claim 14, wherein said seed specific promoters are the WS118 promotors, preferably the WS118 promotor shown in SEQ ID NO:55 or SEQ ID NO:9.

16. the construct of claim 13, wherein said constitutive promoter are the GOS2 promotors, preferably the GOS2 promotor shown in SEQ ID NO:56 or SEQ ID NO:54.

17. plant, it is transformed by each construct in the claim 13 to 16.

18. be used to prepare the method for transgenic plant that has the seed production of increase with respect to the corresponding wild-type plant, described method comprises:

(i) in plant or vegetable cell, introduce and express AZ nucleic acid or its variant; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

19. have the transgenic plant of the seed production of increase, it produces because of introducing AZ nucleic acid or its variant in described plant.

20. each transgenic plant in the claim 12,17 or 19, wherein said plant is a monocotyledons, and as sugarcane, or wherein said plant is cereal, as rice, corn, wheat, barley, grain, rye, oat or Chinese sorghum.

21. the part gathered in the crops of each plant in the claim 12,17,19 or 20.

22. the part gathered in the crops of the plant of claim 21, wherein said gather in the crops the part be seed.

23. directly from the product of the part gathered in the crops of the plant of the plant of claim 20 and/or claim 21 or 22.

24.AZ the purposes of nucleic acid/gene or its variant, or the purposes of AZ polypeptide or its homologue, its be used for respect to corresponding wild-type plant increase seed production.

25. the purposes of claim 24, the seed production of wherein said increase comprises the thousand seed weight of increase.

26.AZ the purposes of nucleic acid/gene or its variant, or the purposes of AZ polypeptide or its homologue, it is as molecule marker.

27. the purposes of each construct in the claim 13 to 16, it is used for preparing the method for plant that has the seed production of increase with respect to control plant.

28. under abiotic stress, increase the method for plant biomass with respect to control plant, it comprises the expression of nucleotide sequence in plant of regulating coding synovial sarcoma transhipment (SYT) polypeptide, randomly selection has the plant of the output of increase, wherein said SYT polypeptide comprises from the N-terminal to the C-terminal: (i) according to the preferred sequence that increases, have at least 20% with SNH structural domain SEQ ID NO:58,25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, the SNH structural domain of 99% sequence identity; (ii) be rich in the Met structural domain; (iii) be rich in the QG structural domain.

29. the method for claim 28, wherein said SNH structural domain comprises the residue that shows with black in Fig. 6.

30. the method for claim 28, wherein said SNH structural domain is by shown in the SEQ ID NO:57.

31. each method in the claim 28 to 30, wherein said SYT polypeptide also comprise following one or more: (i) SEQ ID NO:146; (ii) SEQ ID NO:147; (iii) be positioned at and be rich in the Met structural domain before the SNH structural domain N-terminal.

32. each method in the claim 28 to 31, wherein said abiotic stress are one of following two kinds: salt stress or drought stress.

Coerce 33. each method in the claim 28 to 31, wherein said abiotic stress are the nutrition operabilities that reduces, the preferred nitrogen operability that reduces is coerced.

34. each method in the claim 28 to 33, the wherein said expression of being regulated realizes by following one or more: T-DNA activation, TILLING or homologous recombination.

35. each method in the claim 28 to 33, the wherein said expression of being regulated realizes by the nucleotide sequence of introducing in plant, plant part or vegetable cell and expression coding SYT polypeptide.

36. each method in the claim 28 to 35, the output of wherein said increase be following one or more: seed ultimate production, full seed number, the full rate of seed, TKW and harvest index.

37. the method for claim 35, wherein said nucleotide sequence are in the following SYT nucleic acid variant one or more:

(i) part of SYT nucleotide sequence;

(ii) can with the nucleotide sequence of SYT nucleic acid array hybridizing;

The (iii) splice variant of SYT nucleotide sequence;

The (iv) allelic variant of SYT nucleotide sequence;

(v) reorganize the SYT nucleotide sequence that obtains by gene;

(vi) by site-directed mutagenic obtained SYT nucleotide sequence.

38. the method for claim 35 or 37, any in the SYT polypeptide that provides in the wherein said nucleic acid sequence encoding table 6 or aforementioned any SEQ ID NO directly to homologue or collateral line homologue.

39. the method for claim 35 or 37, the SYT polypeptide shown in wherein said nucleic acid sequence encoding SEQ ID NO:60, SEQ ID NO:151 or the SEQ ID NO:153.

40. each method in the claim 28 to 39, nucleotide sequence overexpression in plant of wherein said coding SYT polypeptide.

41. each method in the claim 35,37 or 38, the nucleotide sequence of wherein said coding SYT polypeptide is a plant origin.

42. each method in the claim 35,37 to 41, the nucleotide sequence of wherein said coding SYT polypeptide effectively is connected with constitutive promoter.

43. the method for claim 42, wherein said constitutive promoter are from plant, preferably from monocotyledons.

44. the method for claim 42 or 43, wherein said constitutive promoter are the GOS2 promotors.

45. plant, plant part or vegetable cell, it is obtained by each method in the claim 28 to 44.

46. isolated nucleic acid sequences, it comprises and is selected from following nucleotide sequence:

(a) isolated nucleic acid sequences shown in SEQ ID NO:150 and the SEQ ID NO:152;

(b) isolated nucleic acid sequences of the polypeptide shown in coding SEQ ID NO:151 and the SEQ ID NO:153;

(c) isolated nucleic acid sequences, its sequence can derive out as the result of genetic code degeneracy from the polypeptide shown in SEQID NO:151 and the SEQ ID NO:153;

(d) isolated nucleic acid sequences, its encoded polypeptides has at least 70% identity with (a) the nucleotide sequence encoded polypeptide in (c);

(e) isolated nucleic acid sequences, homologue, derivative or the active fragments of the polypeptide of its coding shown in SEQ ID NO:151 and SEQ ID NO:153, described homologue, derivative or fragment are plant origin, and advantageously comprise from N-terminal to C-terminal:

(i) the SNH structural domain that has certain sequence identity with SNH structural domain SEQ ID NO:58, described sequence identity is at least 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% according to the preferred sequence that increases;

(ii) be rich in the Met structural domain;

(iii) be rich in the QG structural domain;

(f) isolated nucleic acid sequences or its complement, described isolated nucleic acid sequences can with above-mentioned (a) to (c) amplifying nucleic acid molecular hybridization, the plant protein in wherein said hybridization sequences or its complement coding (a) to (e);

Wherein, being regulated of nucleotide sequence is expressed in the output that increases plant under the abiotic stress with respect to control plant.

47. construct, it comprises:

(i) nucleotide sequence of coding as top defined SYT polypeptide;

(ii) one or more regulating and controlling sequences that can drive the expression of (i) amplifying nucleic acid sequence; Randomly

(iii) transcription termination sequence; Or

(iv) as the nucleotide sequence in the claim 46.

48. the construct of claim 47, wherein said regulating and controlling sequence is a constitutive promoter.

49. the construct of claim 48, wherein said constitutive promoter are the GOS2 promotors.

50. the construct of claim 48, wherein said GOS2 promotor is shown in SEQ ID NO:56 or SEQ ID NO:145.

51. plant, plant part or vegetable cell, it is transformed by each construct in the claim 47 to 50.

52. prepare the method for the transgenic plant of the output that has increase under abiotic stress, wherein said plant optimization is a monocotyledons, described method comprises:

(i) nucleotide sequence of introducing and expression coding SYT polypeptide in plant or vegetable cell;

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

53. the method for claim 52, it comprises that the plant that is (ii) obtained by described incubation step by hybridization produces plant and it comprises one or more offsprings of the part of seed.

54. have transgenic plant or its part of the output of increase under abiotic stress, it produces because of the nucleotide sequence of introducing coding SYT polypeptide in described plant or its part, the output of described increase is with respect to control plant.

55. each transgenic plant in the claim 45,51 or 54, wherein said plant is a monocotyledons, and as sugarcane, or wherein said plant is cereal, as rice, corn, wheat, barley, grain, rye, oat, triticale or Chinese sorghum.

56. the part gathered in the crops of each plant in the claim 45,51,54 or 55.

57. the part gathered in the crops of the plant of claim 56, wherein said gather in the crops the part be seed.

58. from, preferred directly from the product of the part gathered in the crops of the plant of the plant of claim 45 and/or claim 56 or 57.

59.SYT the purposes of nucleotide sequence, or the purposes of SYT polypeptide, or the purposes of each construct in the claim 47 to 50, it is used for increasing output with respect to control plant under abiotic stress.

60. the purposes of claim 59, the output of wherein said increase comprise following one or more: seed ultimate production, full seed number, the full rate of seed, TKW and harvest index.

61. the purposes of the nucleotide sequence of coding SYT polypeptide, or the purposes of SYT polypeptide, it is as molecule marker.

62. increase the method for plant biomass with respect to control plant, it comprises the expression of nucleotide sequence in plant shoot divides that increases coding cpFBPase polypeptide, randomly selection has the plant of the output of increase, and wherein said cpFBPase polypeptide is brought into play function and comprised in chloroplast(id): (i) at least one FBPase structural domain; (ii) fragment is inserted at least one redox regulation and control.

63. the method for claim 62, the expression of wherein said increase are by following one or more realizations: T-DNA activation, TILLING and homologous recombination.

64. the method for claim 62, the expression of wherein said increase realizes by the nucleotide sequence of introducing in plant shoot divides and expression coding cpFBPase polypeptide.

65. each method in the claim 62 to 64, wherein said nucleotide sequence are in the following cpFBPase nucleic acid variant one or more:

(i) part of cpFBPase nucleotide sequence;

(ii) can with the nucleotide sequence of cpFBPase nucleic acid array hybridizing;

The (iii) splice variant of cpFBPase nucleotide sequence;

The (iv) allelic variant of cpFBPase nucleotide sequence;

(v) reorganize the cpFBPase nucleotide sequence that obtains by gene;

(vi) by site-directed mutagenic obtained cpFBPase nucleotide sequence.

66. each method in the claim 62 to 65, the arbitrary cpFBPase polypeptide that provides among the wherein said nucleic acid sequence encoding table C or aforementioned any SEQ ID NO directly to homologue or collateral line homologue.

67. each method in the claim 62 to 66, wherein said nucleotide sequence is derived from photosynthetic cells, and preferred source more preferably is derived from green alga from terrestrial plant.

68. each method in the claim 62 to 67, the cpFBPase polypeptide shown in the wherein said nucleic acid sequence encoding SEQID NO:155.

69. each method in the claim 62 to 68, wherein said nucleotide sequence effectively are connected with can driving in plant shoot branch expression promoter.

70. the method for claim 69, wherein said promotor is a constitutive promoter, preferably the GOS2 promotor.

71. each method in the claim 62 to 70, the output of wherein said increase be selected from following one or more: the i) seed weight of Zeng Jiaing; The ii) full seed number of Zeng Jiaing; The iii) full rate of the seed of Zeng Jiaing; The iv) harvest index of Zeng Jiaing.

72. plant, from part that comprises seed and the cell of this type of plant, it can be obtained by each method in the claim 62 to 71, wherein said plant or its part comprise the nucleotide sequence that increases the coding cpFBPase polypeptide of expressing, and described nucleotide sequence effectively is connected with the expression promoter that drives in plant shoot divides.

73. construct, it comprises:

(i) nucleotide sequence of coding cpFBPase polypeptide;

The regulating and controlling sequence that (ii) one or more nucleotide sequences that can drive in (i) are expressed in plant shoot divides; Randomly

(iii) transcription termination sequence.

74. the construct of claim 73, wherein said nucleotide sequence is derived from green alga, and described regulating and controlling sequence is the GOS2 promotor.

75. the purposes of the construct of claim 73 or 74, it is used for increasing the output of plant.

76. plant, plant part or vegetable cell, its construct by claim 73 or 74 transforms.

77. preparation has the method for transgenic plant of the output of increase with respect to control plant, described method comprises:

(i) nucleotide sequence of introducing and expression coding cpFBPase polypeptide in plant shoot branch or vegetable cell; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

78. with respect to the transgenic plant that control plant has the output of increase, the output of described increase produces because of the expression of the increase of nucleotide sequence in plant shoot divides of coding cpFBPase polypeptide.

79. each plant in the claim 72,76 or 78, wherein said plant is crop plants or unifacial leaf or cereal, as rice, corn, wheat, barley, grain, rye, Chinese sorghum or oat.

80. the part gathered in the crops of each plant in the claim 72,76,78 or 79, wherein said gather in the crops the part be seed.

81. product from the part gathered in the crops of the plant of the plant of claim 79 and/or claim 80.

82. the purposes of the nucleotide sequence of coding cpFBPase polypeptide, it is used for increasing plant biomass with respect to control plant.

83. the purposes of claim 82, the plant biomass of wherein said increase be selected from following one or more: the i) seed production of Zeng Jiaing; The ii) full seed number of Zeng Jiaing; The iii) full rate of the seed of Zeng Jiaing; The iv) harvest index of Zeng Jiaing.

84. method with respect to multiple output correlated character in the control plant improvement plant, it comprises regulates little inducibility kinases (SIK) nucleic acid and/or the expression of SIK polypeptide in plant, wherein during the overexpression of the nucleic acid that comprises coding SIK polypeptide when the described expression of being regulated in plant, described output correlated character is the increase of every strain plant flowers number; And reduce or when removing the expression of endogenous SIK gene in plant substantially when the described expression of being regulated comprises, wherein said output correlated character is one or more in the full rate of the thousand seed weight (TKW) that increases, the harvest index (HI) that increases and increase.

85. the method for claim 84, wherein said minimizing or remove endogenous SIK expression of gene substantially and realize by the genetic expression of RNA mediation is reticent.

86. the method for claim 85, the silence of wherein said RNA mediation is by suppressing realization altogether.

87. the method for claim 85, the silence of wherein said RNA mediation realizes by using antisense SIK nucleotide sequence.

88. the method for claim 84 or 85, wherein said minimizing or remove endogenous SIK expression of gene substantially and realize that by the inverted repeats that uses SIK nucleic acid described inverted repeats preferably can form hairpin structure.

89. each method in the claim 84 to 88, wherein said endogenous SIK gene, or be used to reduce or the SIK nucleic acid of removing endogenous SIK genetic expression substantially is plant origin or artificial source.

90. the method for claim 89, wherein said SIK nucleic acid be from monocotyledons, and be used for transforming monocots; Or from dicotyledons, and be used to transform dicotyledons.

91. the method for claim 90, wherein said SIK nucleic acid be from Gramineae, and be used for transforming gramineous plant, more preferably wherein said SIK nucleic acid is from rice, and is used to transform rice plant.

92. each method among the claim 89-91, wherein said nucleic acid comprises among SEQ IDNO:209, SEQ ID NO:211 or the SEQ ID NO213-225 each or the Nucleotide of successive basically of the sufficient length of the arbitrary nucleic acid that provides in table 8 or the table 9.

93. the method for claim 84, wherein said overexpression is by introducing and express the nucleic acid realization of coding SIK polypeptide in plant.

94. the method for claim 84 or 93, wherein said SIK polypeptide is by SEQ ID NO:210 or it is directly to shown in homologue or the collateral line homologue.

95. claim 84,93 or 94 method, wherein said SIK polypeptide comprise following one or more, be preferably following whole:

(i) ATP-binding domain;

(ii) serine threonine kinases avtive spot mark;

(iii) be rich in the ser structure territory;

(iv) one or more myristoylations site;

(v) kinase activity.

96. plant, its plant part or vegetable cell, it is obtained by each method in the claim 84 to 92, wherein said plant, its plant part or vegetable cell be owing to using the SIK nucleotide sequence to have the endogenous SIK expression of gene of reduction, and have in the full rate of the thousand seed weight (TKW) of increase, the harvest index (HI) that increases and increase one or more with respect to control plant.

97. plant, its plant part or vegetable cell, it is obtained by each method in the claim 93 to 95, wherein said plant, its plant part or vegetable cell have the expression of the SIK coding nucleic acid of increase with respect to control plant, and have every strain plant flowers number of increase with respect to control plant.

98. construct, it comprises:

(a) the SIK polypeptide represented by SEQ ID NO:210 of coding, or it is directly to the nucleic acid of homologue or collateral line homologue;

(b) one or more regulating and controlling sequences that can drive the expression of (a) amplifying nucleic acid sequence; Randomly

(c) transcription termination sequence.

99. the construct of claim 98, wherein said regulating and controlling sequence comprises constitutive promoter, and preferably from the GOS2 promotor of rice, preferably the promotor with SEQ ID NO:56 or SEQ ID NO:226 has the promotor of suitable express spectra.

100. be used for reducing the construct that endogenous SIK gene is expressed plant, it comprises one or more regulating and controlling sequences, SIK nucleic acid and optional transcription termination sequence.

101. the construct of claim 100, wherein said SIK nucleic acid are the inverted repeats under constitutive promoter control, described inverted repeats preferably can form hairpin structure.

102. the construct of claim 101, wherein said constitutive promoter are the GOS2 promotors from rice, preferably the promotor with SEQ ID NO:56 or SEQ ID NO:226 has the promotor of suitable express spectra.

103. plant, plant part or vegetable cell, it is transformed by each construct in the claim 98 to 102.

104. preparation has the method for transgenic plant of every strain plant flowers number of increase with respect to control plant, described method comprises:

(i) in plant, introduce and express, or it is directly to the coding nucleic acid of homologue or collateral line homologue by the SIK polypeptide shown in the SEQ ID NO:210; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

105. preparation has one or more the method for transgenic plant in the output of full rate, increase of the thousand seed weight (TKW) of increase, the harvest index (HI) that increases and increase with respect to control plant, described method comprises:

(i) introducing and expression are used the SIK nucleotide sequence and reduce the genetic constructs that endogenous SIK gene is expressed in plants in plant, plant part or vegetable cell, and described genetic constructs comprises one or more regulating and controlling sequences; With

(ii) under the condition that promotes plant-growth and growth, cultivate plants, plant part or vegetable cell.

106. have the transgenic plant of every strain plant flowers number of increase with respect to control plant, it is characterized in that described plant has the SIK peptide coding expression of nucleic acids of increase with respect to control plant.

107. have the full rate of the thousand seed weight (TKW) of increase, the harvest index (HI) that increases and increase, one or more transgenic plant in the output with respect to control plant, it is characterized in that described plant has the endogenous SIK expression of gene of minimizing with respect to control plant.

108. each transgenic plant in the claim 96,97,103,106 or 107, wherein said plant is a monocotyledons, and as sugarcane, or wherein said plant is cereal, as rice, corn, wheat, barley, grain, rye, oat or Chinese sorghum.

109. the part gathered in the crops of the transgenic plant of claim 108.

110. the part gathered in the crops of claim 109, wherein said gather in the crops the part be seed.

111. product from the part gathered in the crops of the plant of the plant of claim 108 and/or claim 110.

112.SIK the purposes of coding nucleic acid, it is used for by the overexpression of SIK coding nucleic acid plant, increases every strain plant flowers number with respect to control plant.

113.SIK the purposes of nucleic acid, it is used for by reducing plant or remove endogenous SIK expression of gene substantially, increases in thousand seed weight (TKW), harvest index (HI) and full rate, the output one or more with respect to control plant.

114. be used for the method with respect to control plant improved seed stock content, it comprises the expression of coding nucleic acid in plant of regulating II class HD-Zip transcription factor.

115. the method for claim 114, wherein said II class HD-Zip transcription factor comprises following: (i) homeodomain box; (ii) leucine zipper; (iii) motif I, II and the III of random order.

Motif I:RKKLRL, or have the motif I that one or more conserved amino acids replace in any site, and/or have the motif I of one or two non-conservative change in site arbitrarily; With

Motif II:TKLKQTEVDCEFLRRCCENLTEEN or have motif II that one or more conserved amino acids replace in any site and/or have at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or the motif of higher sequence identity with the preferred sequence that increases with motif II; With

Motif III:TLTMCPSCER or have motif III that one or more conserved amino acids replace in any site and/or have one, the motif III of two or three non-conservative changes in site arbitrarily.

Arbitrary nucleotide sequence that 116. the method for claim 114 or 115, the nucleic acid of wherein said coding II class HD-Zip transcription factor are tables to be provided among the H or its part or can with the sequence of arbitrary nucleic acid hybridization of providing among the table H.

117. each method in the claim 114 to 116, the wherein said expression of being regulated activates in mark, TILLING or the homologous recombination any one or more by T-DNA and realizes.

118. each method in the claim 114 to 117, the wherein said expression of being regulated realizes by the nucleic acid of introducing in plant and expression coding II class HD-Zip transcription factor.

119. the method for any aforementioned claim, the stock content of wherein said improvement comprise the seed that has one or more content in lipid, oil, lipid acid, starch, sugar and the protein of improvement with respect to the control plant respective items.

120. the method for claim 119, the content of wherein said improvement are the oil-contgs with respect to the seed increase of control plant.

121. each method in the claim 118 to 120, wherein said nucleic acid and constitutive promoter preferably effectively are connected with the GOS2 promotor.

122. the method for any aforementioned claim, the nucleic acid of wherein said coding II class HD-Zip transcription factor is plant origin, preferably from dicotyledons, and also preferably from cress, more can be preferably from Arabidopis thaliana.

123. plant or it comprise the part of seed, it can obtain by the method for any aforementioned claim, and wherein said plant or its part comprise the nucleic acid of fgs encoder II class HD-Zip transcription factor.

124. construct, it comprises:

(a) the coding nucleic acid of II class HD-Zip transcription factor of definition as mentioned;

(b) one or more regulating and controlling sequences that effectively are connected with (i) amplifying nucleic acid.

125. the construct of claim 124, wherein said one or more regulating and controlling sequences are constitutive promoter at least, preferably the GOS2 promotor.

126. the construct of claim 124 or 125, the nucleic acid of wherein said coding II class HD-Zip transcription factor is the arbitrary nucleotide sequence that provides among the table H, or straight any nucleotide sequence to homologue, collateral line homologue or homologue of any peptide sequence that provides among the coding schedule H.

127. the purposes of each construct in the claim 124 to 126, it is used to prepare the plant that has the seed storage thing content of improvement with respect to control plant.

128. plant, plant part or vegetable cell, it is transformed by each construct in the claim 124 to 127.

129. preparation has the method for the transgenic plant of stock content in the seed of improvement with respect to control plant, described method comprises:

(i) nucleic acid of introducing and expression coding II class HD-Zip transcription factor in plant, plant part or vegetable cell, the nucleotide sequence that provides among the preferred table H, or be included in introduce in the plant and expression table H in the arbitrary amino acid sequence that provides directly to the nucleic acid sequence encoding of homologue, collateral line homologue or homologue; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth; With

(iii) from plant (ii), gather in the crops seed; Randomly

(iv) from seed (iii), extract in lipid, oil, lipid acid, starch, sugar and the protein any one or more.

130. have the transgenic plant of stock content in the seed of improvement with respect to control plant, the content of described improvement produces because of the expression of nucleic acid of the coding II class HD-Zip transcription factor that increases.

131. claim 123,128 or 130 transgenic plant, wherein said plant is to cultivate the crop plants that is used to produce oil, preferably Semen Brassicae campestris, canola oil dish, flax, soybean, Sunflower Receptacle, corn, oat, rye, barley, wheat, pepper, Flower of Aztec Marigold, cotton, oil palm, coconut, flax, castor-oil plant, peanut, olive, avocado, sesame, jatropha.

132. the part gathered in the crops of the plant of claim 131, wherein said gather in the crops the part be seed.

133. from the product of the part gathered in the crops of the plant of the plant of claim 131 and/or claim 132, preferred wherein said product is one or more in lipid, oil, lipid acid, starch, sugar and the protein.

134. the purposes of the nucleic acid of coding II class HD-Zip transcription factor, it is used for the content with respect to control plant improved seed stock.

135. the purposes of claim 134, wherein said II class HD-Zip transcription factor are the nucleotide sequences that provides among the table H, or the straight nucleic acid sequence encoding to homologue, collateral line homologue or homologue of the arbitrary amino acid sequence that provides among the coding schedule H.

136. be used for strengthening the method for plant output correlated character, it comprises regulates the expression of the coding proteinic nucleic acid of SYB1 in plant, the proteinic sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain.

137. the method for claim 136, the proteinic nucleic acid of wherein said coding SYB1 are arbitrary nucleic acid SEQ ID NO that table provides among the J, or its part, or can with the sequence of arbitrary nucleic acid SEQ ID NO hybridization of providing among the table J.

Any one or more is realized in mark, TILLING, site-directed mutagenesis, orthogenesis or the homologous recombination 138. the method for claim 136 or 137, the wherein said expression of being regulated activate by T-DNA.

139. the method for claim 136 or 137, the wherein said expression of being regulated is by introducing in plant and expressing the proteinic nucleic acid of coding SYB1 and realize, the proteinic sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain.

140. each method in the claim 136 to 139, the output correlated character of wherein said reinforcement are the seed productions that increases.

141. the method for claim 139 or 140, wherein said nucleic acid effectively is connected with constitutive promoter, preferably effectively is connected with the GOS2 promotor.

142. each method in the claim 136 to 141, the proteinic nucleic acid of wherein said coding SYB1 is plant origin, preferably from dicotyledons, also preferably from Cruciferae, more preferably from Arabidopsis, most preferably from Arabidopis thaliana.

143. plant or it comprise the part of seed, it is obtained by each method in the claim 136 to 142, and wherein said plant or its part comprise the proteinic recombinant nucleic acid of coding SYB1.

144. construct, it comprises:

(i) the proteinic nucleic acid of coding SYB1, the proteinic aminoacid sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain;

(ii) one or more regulating and controlling sequences that can drive the expression of (i) amplifying nucleic acid sequence; Randomly

(iii) transcription termination sequence.

145. the construct of claim 144, wherein said one or more regulating and controlling sequences are constitutive promoter at least, preferably the GOS2 promotor.

146. the purposes of the construct of claim 144 or 145, it is used to prepare the plant that has the output correlated character of reinforcement with respect to control plant, the especially plant of the seed production of Zeng Jiaing.

147. plant, plant part or vegetable cell, its construct by claim 144 or 145 transforms.

148. preparation has the method for the transgenic plant of enhanced yield correlated character with respect to control plant, described method comprises:

(i) in plant, introduce and express the proteinic nucleic acid of coding SYB1, the proteinic aminoacid sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain; With

(ii) culturing plants cell under the condition that promotes plant-growth and growth.

149. have the transgenic plant of the output of increase with respect to suitable control plant, or from the vegetable cells of described transgenic plant, the output of described increase is expressed because of the increase of the proteinic nucleic acid of coding SYB1 and is produced, the proteinic aminoacid sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain.

150. claim 143,147 or 149 transgenic plant, wherein said plant is crop plants or monocotyledons or cereal, as rice, corn, wheat, barley, grain, rye, Chinese sorghum or oat, or from the vegetable cells of described transgenic plant.

151. the part gathered in the crops of each plant in the claim 143,147,149 or 150, the wherein said preferably seed of part of gathering in the crops.

152. product from the part gathered in the crops of the plant of the plant of claim 150 and/or claim 151.

153. the purposes of the proteinic nucleic acid of coding SYB1, it is used for strengthening plant output correlated character, the proteinic aminoacid sequence of wherein said SYB1 comprises 3 RanBP type Zinc finger domains and randomly one or more low complex degree structural domain, but does not have and nonoverlapping other structural domains of described Zinc finger domain.

154. the purposes of claim 153, wherein said enhanced yield correlated character are the seed productions that increases with respect to control plant.

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