BRPI0709258A2 - methods for improving plant yield characteristics, for producing a transgenic plant, for increasing plant yield over appropriate control plants, and for increasing abiotic stress resistance in plants relative to control plants, plant or part of the plant. same, construction, use of a construction, plant, plant part or plant cell, transgenic plant, harvestable parts of a plant, products, uses of a nucleic acid, isolated polypeptide, and isolated nucleic acid molecule - Google Patents

Abstract

METHODS FOR IMPROVING CHARACTERISTICS RELATED TO YIELD ON PLANTS, FOR THE PRODUCTION OF A TRANSGENIC PLANT, TO INCREASE PLANT YIELD IN RELATION TO SUITABLE CONTROL PLANTS AND TO INCREASE RESISTANCE TO ABIOTIC STRESS IN PLANTS IN PLANTATION OF PLANTS IN PLANTATION SAME, CONSTRUCTION, USE OF A CONSTRUCTION, PLANT, PART OF A PLANT OR PLANT CELL, TRANSGENIC PLANT, HARVESTING PARTS OF A PLANT, PRODUCTS, USES OF A NUCLEIC ACID, AND ISOLATED, ISOLATED, ISOLATED, ISOLATED, POLYPEDIC INSULATED CONSTRUCTION, AND ISOLATED ISOLATED, ISOLATED, AND ISOLATED, ISOLATED, AND ISOLATED. . The present invention generally relates to the field of molecular biology and relates to a method of improving characteristics related to plant yield, in particular to increase plant yield and / or early vigor, in relation to control plants. More specifically, the present invention relates to a method for improving performance-related characteristics comprising modifying the expression of a nucleic acid encoding a HAL3 polypeptide, MADS15 polypeptide, PLT transcription factor polypeptide, basic / helix-loop-helix transcription factor ( bHLH), or SPL15 transcription factor. The invention also provides constructs useful in the methods of the invention.

Description

'' METHODS FOR IMPROVING PLANT Yield CHARACTERISTICS, FOR PRODUCTION OF A TRANSGENIC PLANT, FOR INCREASING PLANT Yield FOR ADDITIONAL DECONTROL PLANTS AND FOR INCREASING RESISTANCE ABOUT PLANT, IN PLANT PLANTS, USES A CONSTRUCTION, PLANT, PART OF PLANT OR CELL, PLANT, TRANSGENIC PLANT, HARVESTING PARTS OF A PLANK, PRODUCTS, USE OF A NUCLEIC ACID, ISOLATED POLYPEPTIDE, AND "INSULATED ACID MOLECULE"

The present invention generally relates to the field of molecular biology and relates to a method of enhancing various plant growth characteristics by modulating expression in a plant of a nucleic acid encoding a growth related protein (GRP). The present invention also relates to plants having modulated expression of a GRP-encoding nucleic acid whose plants have improved growth characteristics relative to corresponding wild type plants or other control plants. The invention also provides constructs useful in the methods of the invention.

The ever-growing world population and the diminishing supply of arable land available for agriculture drives research to increase agriculture efficiency. Conventional means for cultural and horticultural breeding use selective breeding techniques to identify plants having desirable characteristics. However, such selective breeding techniques have several disadvantages, namely that these techniques are typically labor intensive and result in plants that often contain heterogeneous genetic components that may not always result in the desirable trait being passed on from parental plants. Advances in molecular biology have allowed humans to modify the germplasm of animals and plants. Plant genetic engineering requires the isolation and manipulation of genetic material (typically in the form of DNA or RNA) and the subsequent introduction of such genetic material into a plant. Such technology has the ability to generate crops or plants having various improved economic, agronomic or horticultural characteristics.

A feature of particular economic interest is increased production. Production is usually defined as the measurable production of a culture's economic value. This can be defined in terms of quantity and / or quality. Yield is directly dependent on several factors, for example, number and size of organs, plant architecture (eg number of branches), seed yield, leaf senescence and more. Root development, nutrient uptake, stress tolerance and early vigor can also be important factors in determining yield. Optimizing the factors mentioned above can thus contribute to increased crop production.

Seed production is a particularly important feature, since the seeds of many plants are important for human and animal nutrition. Crops such as corn, rice, wheat, canola and soya account for more than half of total human caloric intake, either through consumption of the seeds themselves or through the consumption of meat products raised from processed seeds. They are also a source of sugars, oils, and many types of metabolites used in industrial processes. Seeds contain an embryo (the source of new aerial parts and roots) and an endosperm (the nutrient source for embryo growth during germination and during early seedling growth. ). Seed development involves many genes, and requires the transfer of metabolites from the roots, leaves and stems to the developing seed. Oendosperm, in particular, assimilates the metabolic precursors of carbohydrates, oils and proteins and synthesizes them into storage macromolecules to fill the grain.

Another important feature for many cultures is early vigor. Improving early vigor is an important objective of modern rice breeding programs in both temperate and tropical rice cultivars. Long roots are important for proper ground anchorage in rice sown in water. Where rice is sown directly in flooded fields, and where plants must emerge rapidly through water, larger aerial parts are associated with vigor. Where furrow sowing is practiced, larger mesocotyls and coleoptils are important for good seedling emergence. The ability to manipulate early vigor in plantasseria of great importance in agriculture. For example, early vigor was a limitation for the introduction of maize hybrids (Zea mays L.) based on the European Atlantic Corn Belt germplasm.

An additional important feature is that of improved abiotic stress tolerance. Abiotic stress is a primary cause of crop loss around the world, reducing average yields for most major cultivars by more than 50% (Wang et al., Planta (2003) 218: 1-14). Abiotic stresses can be caused by drought, salinity, temperature extremes, chemical toxicity and oxidative stress.

The ability to improve plant tolerance to abiotic stress would be of great economic advantage to farmers around the world and would allow cultivation of crops during adverse conditions and in cropland may not be otherwise possible.

Harvest yield can therefore be increased by optimizing one of the factors mentioned above.

Depending on use, modification of certain production characteristics may be favored over others. For example for applications such as forage or wood production, or fuel resource, an increase in vegetative parts of a plant may be desirable, and for applications such as wheat, starch or oil production, an increase in seed parameters may be particularly desirable.

Even among seed parameters, some may be favored over others, depending on the application. Several mechanisms can contribute to increased seed production, whether in the form of increased seed size or increased seed numbers.

One approach to increasing yield (seed production and / or biomass) in plants may be by modifying the inherent growth mechanisms of a plant, such as the cell cycle or various signaling pathways involved in plant growth or defense mechanisms.

It has now been found that various growth traits can be improved in plants by modulating expression in a nucleic acid plant encoding a GRP (Growth Related Protein) in a plant. GRP can be one of the following: a MADS-box transcription factor (OsMADS15), a PLT transcription factor, a bHLH transcription factor, an SPL15 transcription factor, and a halotolerance protein (HAL3).

INTRODUCTION

Halotolerance Proteins

Many enzymatic processes in a cell require the development of Coenzyme A (CoA or CoASH). Coenzyme A (CoASH) is a highly polar molecule consisting of 3 'adenosine, 5'-diphosphate bound to 4-phosphopanthenic acid (Vitamin B5) and since then β-mercaptoethylamine. The free SH group may be esterified depending on the process in which CoA is involved. The CoA synthesis pathway was elucidated in bacteria, animals and plants. In plants, the conversion of the CoA4'-phosphopantothenyl cysteine (PPC) precursor to 4'-phosphopantethine is catalyzed by the flavoprotein 4'-phosphopantothenyl cysteine decarboxylase (PPCDC or HAL3, Kupke et al. J. Biol. Chem. 276, 19190 -19196, 2001). The gene coding for HAL3 proteins may be part of a small family of genes: the Arabidopsis genome comprises two isoforms (AtHAL3a and AtHAL3b; Espinoza-Ruiz et al. Plant J. 20, 529-539, 1999), in tobacco 3 HAL3 genes are present ( Yonamine et al., J. Exp. Bot. 55, 387-395, 2004), although emarroz HAL3 is a single copy gene. AtHAL3 and other HAL3 proteins are suggested to function as a trimer, with each monomer having a flavin mononucleotide (FMN) attached (Albert et al. Structure 8, 961-969,2000). It is postulated that the FMN cofactor plays a role in the reaction by redoxing 4'-phosphopantetein.

The molecular function of HAL3 proteins is not completely elucidated yet. HAL3 proteins show homology to the yeast SIS2 protein, which is involved in halotolerance (Ferrando et al., Mol. Cell.Biol. 15, 5470-5481). Ectopically expressing HALS plants or cells show improved salt, osmotic or lithium stress tolerance (Espinoza-Ruiz et al., 1999; Yonamine et al., 2004). Overexpression of HAL3a BY2 cell tobacco reportedly caused an increase in intracellular proline content, which may contribute to the salt tolerant phenotype (Yonamine et al., 2004). In addition, Arabidopsissuperexpressing AtHALSa plants had a faster growth rate than wild type plants (Espinoza-Ruiz et al., 1999). AtHAL3b protein has also been shown to interact with cell cycle protein CDKB1; 1 (International Patent 00/36124), so it has been postulated that AtHAL3b is useful for conferring salt stress tolerance on plants and for increasing plant growth rate under saline stress conditions. It has now been found that preferentially increasing expression of a nucleic acid encoding an expanding tissue HAL3 polypeptide generates plants having increased yield over control plants, whose yield increase is at least 10% compared to control plants.

Transcription Factors

Transcription factors are usually defined as proteins that bind to a cis-regulatory element (eg. An enhancer, a TATA box) and which, in association with RNA polymerase, are capable of reactivating and / or suppressing transcription. The Arabidopsis genome codes for at least 1533 transcriptional regulators, which account for -5.9% of their estimated total number of genes (Riechmann et al., 2000 (Science Vol. 290,2105-2109)).

MADS15

MADS-box genes are a large family of eukaryotic transcriptional regulator genes involved in various aspects of yeast, plant and animal development. MADS-box genes encode a strongly conserved MADS domain responsible for binding DNA to specific boxes in the regulatory region of their target genes. The gene family can be divided into two main strains, type I and type II. Type II genes are also called MIKC5 proteins referring to the four functional domains they possess (Figure 5, (Jack, Plant Mol.Biol. 46,515-520, 2001)):

MADS for DNA5 binding about 60 amino acids (highly conserved) located at the N-terminal end of the protein I for intervening (less conserved) domain involved in selective MADS dimer formation;

- K for keratin domain (well maintained) responsible for dimerization;

- C for C-terminal region (sequence variable and length) involved in transcriptional activation, or formation of a multimeric transcription factor complex.

More than 100 MADS-box genes have been identified in Arabidopsis, and have been phylogenetically classified into 12 strands, each having specific MADS consensus derivations (Thiessen et al. JMol. Evol. 43, 484-516, 1996). OsMADS15 belongs to the SQUA (paraSQUAMOSA, by Antirrhinum majus). SQUA genes are classified as function organ identity genes A with reference to the ABC floral organ identity specification model proposed by Coen and Meyerowitz in 1991 (Nature 353, 31-7). In addition to OsMADS15, OsMADS 14, OsMADS18, and OsMADS20 rice are also part of ciadoSQUA.

The SQUA ciate in dicotyledonous plants is subdivided into two subgroups, the subclades APl and FUL (in which OsMADS 15 is grouped). These subclasses differ essentially with respect to the specific amino acid motifs located at the C-terminus of their respective proteins (Litt and Irish, Genetics 165, 821-833, 2003). Due to the presence of a specific AP1 amino acid motif, dicotyledon AP1 cyst related proteins usually comprise a C-terminus farnesylation motif (this motif is CAAX, where C is cysteine, A is usually an aliphatic amino acid, and X is methionine, glutamine , serine, cysteine or alanine). In monocotyledonous plants, SQUA-chromium proteins are also subdivided into two major groups, which can be distinguished based on conserved C-terminal motifs located within the last 15 amino acids of the proteins: LPPWMLS (eg OsMADS 15, SEQ ID NO: 117 ) eLPPWMLR (e.g., OsMADS 18). In contrast to dedicated SQUA culled sequences, CQUI SQUA monocotyledon sequences lack a farnesylation motif at their terminus. C.MMADS 15 has been postulated to function in a complex with other proteins to control organ formation. However, even now no mutant with a visible phenotype has been identified; no experimental data has been presented regarding ectopic expression or diminished expression of OsMADSl 5 except for data presented in International Patent 01/14559 (European Patent 1209232, US Patent 6,995,302). In this description, transgenic plants of Fagopyrum esculentum expressing OsMADS15 towards direction showed increased branching, whereas transgenics expressing antisense construct had decreased growth and suppressed branching. Kalanchoèdaigremontiana transformed with the felt construction had a foliar development around the roots, which was adopted as an indication of increased branching. The authors determined that no changes were observed in terms of floral development and structure of these flowers. OsmADS 15 ortholog in Arabidopsis is APETALAI (AP1). Ectopic expression of APl results in decreased flowering time, and mutant APl exhibit delayed flowering and have abnormal flowers.

PLT

The AP2 (apetala2) / ERF (ethylene-responsive element binding factor) family comprises transcription factors with at least one highly conserved DNA binding domain, the AP2 domain. The AP2 domain was originally described in APETALA2 (AP2), an Arabidopsis protein involved in developmental programs such as meristem identity regulation and floral organ specification (Jofukuet al, (1994) Plant Cell 6, 1211-1225). AP2 / ERF proteins are divided into subfamilies based on whether they contain one (ERF subfamily) or two (AP2 subfamily) DNA binding domains (Figure 12). More than 140 AP2 / ERF genes have been identified in the Arabidopsis thaliana genome (Reichmann et al. (2000) Science 290: 2105-2110), of which up to 18 belong to the AP2 subfamily (Kim et al. (2006) Mol Bio Evol 23 (1): 107-120).

Two Arabidopsis transcription factor AP2 polypeptides encoded by the Plethora 1 (PLT1) and Plethora 2 (PLT2) genes, collectively referred to as the PLT genes, have been shown to be required for stem cell specification and maintenance specifically in the dereraiz meristem. In RT-PCR analysis, PLT transcripts were detected mainly in in roots, indicating that PLT expression is strongly associated with root identity. Ectopic expression of PLT in the Arabidopsis embryo induces homeotic transformation of apical domains into root, root, or hypocotyl stem cells (Aida et al (2004) Cell 119: 109-120).

United States Patent Application 2004/0045049 and International Patent Application W003 / 013227 provide the nucleic acid sequence encoding the Arabidopsis transcription factor PLT1 (referred to in applications as Gl 793). Overexpression of G1793 (using the CaMV 35S promoter) in Arabidopsis has been reported to produce changes in coltiledone morphology, a slight reduction in overall plant size and fine inflorescences (possibly with abnormal flowers) compared to wild type controls. G1793 superexpressors produce more deboning oil than control plants. Nucleic acid sequences encoding potential orthologs of Glycine max, Oryza sativa and Zea mays are provided.

International Patent Application WO 03/002751 establishes a Glycine max nucleic acid sequence encoding a PLT polypeptide, with sequence similarity to a maize nucleic acid sequence identified by (pormicroarrangement of DNA) profiling.

International Patent Application WO 05/075655 establishes sequences of Oryza sativa and Zea mays nucleic acids with sequence similarity to Arabidopsis PLT genes.

bHLH

Basic helix-loop-helix proteins (bHLH) are a superfamily of transcription factors that bind as dimers to specific DNA target sites and have been well characterized in nonvegetarian eukaryotes as important regulatory components in various biological processes. The distinguishing characteristics of the bHLH superfamily is a bipartite domain consisting of approximately 60 amino acids. This bipartite domain is comprised of a basic DNA binding region, which binds to an E-box consensus hexanucleotide and two α-helices separated by a variable circular region. The two α-helices promote dimerization, allowing the formation of homo and heterodimers between different family members. While the bHLH domain is evolutionarily conserved, there is little sequence similarity between strands beyond the domain.

Bailey et al, 2003 (The Plant Cell, Vol. 15, 2497-2501) report the total number of bHLH genes detected in Arabidopsis thalian as 162, making bHLH genes one of the largest families of transcription factors in Arabidopsis; rice genome according to reported contains 131 bHLH genes (Buck and Atchley, 2003 (J. Mol. Evol. 56: 742-750)). Heim et al., 2003 (Mol. Biol. Evol. 20 (5): 735-747) identified 12 subfamilies of Arabidopsis thaliana bHLH genes based on structural similarities.

Plant bHLH proteins that have been characterized have been reported to work on anthocyanin biosynthesis, defitochrome signaling, globulin expression, fruit dehiscence, skin and epidermal development (Buck and Atchley, 2003).

SPL

Squamosa promoter deletion protein (SPL) transcription factor polypeptides are structurally diverse proteins that share a highly conserved DNA binding domain (DBD) of about 80 amino acid residues in length (Klein etal. (1996) Mol Gen Genet 259 : 7-16; Cardon et al. (1999) Gene 237: 91-104) .The SPL transcription factor DNA consensus sequence binding site in the target gene promoter is 5'-TNCGTACAA-3 'where N represents any base . Within the SPL DBD are ten conserved cysteine (Cys) or histidine (His) residues (see Figure 23) of which eight are coordinating zinc residues linking two zinc ions necessary for the formation of SPL-specific zinc finger structure ( Yamasaki et al (2004) J MolBiol 337: 49-63). A second characteristic conserved within the SPD DBD is a bipartite nuclear localization signal. Outside the DBD, a microRNA (miR156) target motif (miRNA) is found in most nucleic acid sequences encoding SPL transcription factor polypeptides (either in the coding region, or in the 3 'UTR) along the plant doreino (Rhoades et al. (2002) Cell 110: 513-520). miRNAs control posttranscriptional SPL gene expression by directing mRNAsoding SPL for degradation or translational repression.

Riechmann et al. (Science 290: 2105-2109, 2000) report 16 SPL transcription factor polypeptides in Arabidopsis thaliana, compared with sequence similarity between them (in addition to the characteristics mentioned above), the size of the deduced SPL polypeptide ranging from131 to 927 amino acids. However, pairs of SPL transcription factor polypeptides sharing greater sequence homology were detected within the SPL family of this plant (Cardon et al. (1999)).

The transcription factor polypeptides SPL (found only in plants) characterized so far have been shown to function in plant development, particularly in flower development. Transgenic plants overexpressing a SPL3 transcription factor polypeptide have been shown to flourish earlier (Cardon et al. (1997) PlantJ 12: 367-377).

In European Patent Application EP1033405, nucleic acid sequences and deduced polypeptide of the SPL15 transcription factor polypeptide are disclosed.

In International Patent Application W003013227, the nucleic acid sequence (and deduced polypeptide sequence; internal reference G2346) is disclosed which encodes part of the transcription defector polypeptide SPL15 meanwhile 38 amino acids of the C-terminal transcription factor SPL15. Transgenic Arabidopsisthaliana plants constitutively overexpressing the modified SPL15 (or G2346) transcription factor polypeptide had slightly increased cotyledons. In late stages of development, the same plants showed no consistent difference from control plants.

SUMMARY OF THE INVENTION

Surprisingly, it has now been found that preferentially increasing expression of a nucleic acid encoding a HAL3 polypeptide in expanding tissues generates plants having improved yield characteristics, in particular increased early vigor and increased seed yield over control plants, whose increase in seed yield is at least 10% compared to control plants.

According to another embodiment of the present invention, a method is provided for improving yield characteristics, in particular for increasing early vigor of a plant and increasing seed production of a plant, relative to control plants, comprising preferably increasing expression of a plant. nucleic acid encoding an HAL3 polypeptide in expanding tissues of a plant.

Surprisingly, it has now been discovered that modulating expression of a nucleic acid encoding an MADS15 polypeptide generates plants having improved production characteristics, in particular increased production relative to control plants.

According to one embodiment, a method is provided for increasing plant yield over control plants, comprising increasing expression of a nucleic acid encoding an MADS15 polypeptide in a plant. Increased yield comprises increased vegetative biomass, but not increased seed yield.

According to another embodiment, the present invention provides methods for increasing plant production over control plants, comprising decreasing the level and / or activity of an endogenous MADS15 polypeptide. The increased yield comprised higher seed yield, but did not include increased vegetative biomass.

Surprisingly, it has now been found that increasing expression of a nucleic acid encoding a PLT transcription factor polypeptide generates plants having improved production characteristics, in particular increased production relative to control plants.

According to a further embodiment, a method is provided for increasing plant yield over control plants, comprising increasing expression in a plant of a nucleic acid by encoding a PLT transcription factor polypeptide.

Surprisingly, it has now been found that modular expression in a plant of a nucleic acid encoding a particular bHLH transcription factor class generates plants having improved production characteristics over control plants. The particular bHLH transcription defector class suitable for improving plant yield characteristics is described in detail below.

According to a further embodiment, the present invention provides a method for enhancing plant production characteristics relative to control plants, comprising modulating expression in a plant of a nucleic acid encoding a particular bHLH transcription defactor class.

Surprisingly, it has now been found that increasing a plant's expression of a nucleic acid encoding a transcription factor SPL15 polypeptide generates plants having improved production characteristics, in particular increased production relative to control plants.

According to a further embodiment, the invention provides a method for increasing plant production over control plants, comprising increasing expression in a plant of a nucleic acid encoding a transcription factor polypeptide SPL15.

Polypeptide (S) ZProtein (S)

The terms "polypeptide" and "protein" are used interchangeably herein and refer to amino acids in a polymeric form of any length.

Polynucleotide (s) / Nucleic acid (s) / Nucleic acid sequence (s) / Nucleotide sequence (s)

The terms "polynucleotide (s)", "nucleic acid sequence (s)", "nucleotide sequence (s)", "nucleic acid (s)" are used interchangeably herein and refer to nucleotides, or ribonucleotides or deoxyribonucleotides or a combination of both, in a polymeric shape of any length.

Control Plant (s)

The choice of suitable control plants is a dermal part of an experimental structure and may include corresponding wild-type plants or corresponding plants without the gene of interest. The control plant is typically of the same plant species or even the same variety as the plant to be verified. The control plant may also be a nullizigote of the plant to be verified. A "control plant" used here refers not only to whole plants, but also to plant parts, including seeds and parts of seeds.

Counterpart (s)

"Protein homologues" include peptides, oligopeptides, polypeptides, proteins and enzymes having amino acid substitutions, deletions and / or insertions with respect to the unmodified protein in question and having similar biological and functional activity to the unmodified protein from which they are derived.

A deletion refers to the removal of one or more amino acids from a protein.

An insert refers to one or more amino acid residues being introduced at a predetermined site in a protein. Insertions may comprise N-terminal and / or C-terminal fusions as well as single or multiple intra-sequence amino acid insertions. Generally, insertions within the amino acid sequence will be smaller than N- or C-terminal fusions of the order of about 1 to 10 residues. Examples of N- or C-terminal fusion proteins or peptides include the deletion domain or activation domain of a transcriptional activator as used in the two yeast hybrid systems, phage coat proteins, (histidine) -o label, glutathione label. S-transferase, protein A, maltose binding protein, dihydrofolate reductase, Tag · 100 epitope, c-myc epitope, FLAG® epitope, IacZ, CMP (calmodulin binding peptide), HA epitope, protein C epitope and VSV epitope.

A substitution refers to the substitution of protein amino acids for other amino acids having similar properties (such as hydrophobicity, hydrophilicity, antigenicity, propensity to form or break α-helical structures or similar β-leaf structures). Amino acid substitutions are typically single residues, but may be grouped depending on functional constraints placed on the polypeptide, insertions will usually be on the order of about 1 to 10 amino acid residues. Amino acid substitutions are preferably conservative amino acid substitutions. Conservative Substitution Tables are well known in the art (see, for example, Creighton (1984) Proteins. W.H. Freeman and Company and Table 1 below).

Table 1: Examples of conserved amino acid substitutions

<table> table see original document page 17 </column> </row> <table>

Amino acid substitutions, deletions and / or insertions may readily be made using synthetic peptide techniques well known in the art, such as solid phase peptide synthesis and the like, or by recombinant DNA manipulation. Methods for manipulating DNA sequences to produce substitution, insertion, or deletion variants of a protein are well known in the art. For example, techniques for making substitution mutations at predetermined DNA sites are well known to those of skill in the art and include T13-Gen mutagenesis (T7-Gen in vitro (USB, Cleveland, OH), QuickChange asitium directed mutagenesis (Stratagene, San Diego) , CA), site-directed mutagenesis by PCR or other site-directed mutagenesis protocols.

Derivatives "Derivatives" include peptides, oligopeptides, polypeptides which may, compared to the protein's naturally occurring amino acid sequence, such as the protein of interest, comprise amino acid substitutions with non-naturally occurring amino acid residues, or additions of non-naturally occurring amino acid residues. . "Derivatives" of a protein also encompass peptides, oligopeptides, polypeptides comprising naturally occurring altered deamino acid residues (glycosylated, acylated, prenylated, phosphorylated, myristoylated, sulfated etc.) or unnaturally altered deamino acid residues in a naturally occurring manner. of the polypeptide. A derivative may also comprise one or more non-amino acid substituents or additions compared to the amino acid sequence from which it is derived, for example a reporter molecule or other linker, covalently or not covalently linked to the amino acid sequence, such as a reporter molecule that is linked to. facilitate their detection by non-naturally occurring amino acid residues relative to the amino acid sequence of a non-naturally occurring protein.

Orthologist (s) / Paralog (s)

Orthologs and paralogs encompass evolutionary concepts used to describe ancestral gene relationships. Paralogues are genes within a species that originated by duplicating an ancestral gene and orthologs are genes from different organisms that originated through species.

Domain

The term "domain" refers to a set of amino acids conserved at specific positions along an alignment of evolutionarily related protein offsets. While amino acids at other positions may vary from homologous, amino acids that are highly conserved at specific positions indicate amino acids that are likely to be essential in the structure, stability or function of a protein. Identified by their high degree of conservation and aligned sequences from a family of protein homologs, they can be used as identifiers to determine if any polypeptide in question belongs to a previously identified polypeptide family.

Reason / Consensus Sequence / Signature

The term "motive" or "consensus sequence" or "signature" refers to a short conserved region in the evolutionarily related protein sequence. Motives are often highly conserved parts of domains, but may also include only part of the domain, or are located outside the conserved domain (if all motif amino acids fall outside a defined domain).

Hybridization

The term "hybridization" as defined herein is a process characterized by the fact that substantially homologous complementary nucleotide sequences anneal each other. The hybridization process may occur entirely in solution, i.e. both complementary nucleic acids are in solution. The hybridization process may also occur with one of the complementary nucleic acids immobilized to a matrix such as magnetic microspheres, Sepharose microspheres or any other resin. The hybridization process may further occur with one of the complementary nucleic acids immobilized to a solid support such as a denitrocellulose or nylon membrane or immobilized by eg photolithography to, for example, a silent glass support (the latter known as nucleic acid or microarray arrangements). as integrated nucleic acid circuits). In order to allow hybridization to occur, nucleic acid molecules are generally thermally or chemically denatured to merge a double strand into two single strands and / or to remove clamping structures or other secondary strands of single stranded nucleic acids.

The term "stringency" refers to the conditions under which hybridization may occur. Hybridization stringency is influenced by conditions such as temperature, salt concentration, ionic strength, and hybridization buffer composition. Generally, low stringency conditions are selected to be about 30 ° C lower than the thermal fusion point (Tm) for the specific sequence at a defined ionic strength and pH. Medium stringency conditions are when the temperature is 20 ° C below Tm, and high stringency conditions are when the temperature is 10 ° C below Tm. High stringency hybridization conditions are typically used to isolate hybridization sequences that have high sequence similarity to the target nucleic acid sequence. However, nucleic acids may shift in sequence and further encode a substantially identical polypeptide due to the degeneracy of the genetic code. Therefore medium stringency hybridization conditions may sometimes be necessary to identify such nucleic acid molecules.

Tm is the temperature at defined ionic strength and pH at which 50% of the target sequence hybridizes to a perfectly matched probe. ATm is dependent on the solution conditions and the basic composition and length of the probe. For example, larger sequences hybridize specifically at higher temperatures. The maximum hybridization rate is obtained from about 16 ° C to 32 ° C below Tm. The presence of monovalent cations in the hybridization solution reduces electrostatic repulsion between two nucleic acid strands thereby promoting hybrid formation, this effect is visible at sodium concentrations up to 0.4 M (higher concentrations, this effect can be ignored). Formamide reduces the fusion temperature of DNA-DNA and DNA-RNA duplexes by 0.6 to 0.7 ° C for each percent formamide, and addition of 50% formamide allows hybridization to be performed at 30 to 45 ° C, although the rate of hybridization is decreased. Incorrect base pairing reduces the hybridization rate and thermal stability of the duplexes. On average and for large probes, Tm decreases about 1 ° C by% mismatch. Tm can be calculated using the following equations, depending on the types of hybrids:

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

Tm = 81.5 ° C + 16.6xlog10 [Na +] at + 0.41x% [G / Cb] - SOOxtLc] -1 -0.6 lx% formamide

2) DNA-RNA or RNA-RNA hybrids:

Tm = 79.8 + 18.5 (log10 [Na +] a) + 0.58 (% G / Cb) +11.8 (% G / Cb) 2

- 820 / Lc

3) Oligo-DNA or oligo-RNAd hybrids:

For <20 nucleotides: Tm = 2 (In)

For 20-35 nucleotides: Tm = 22 + 1.46 (In)

to or to another monovalent cation, but only accurate within the range 0.01-0.4 M.

b only accurate for% GC in the range of 30% to 75%.

cL = duplex length in base pairs.

d Oligo, oligonucleotide; ln, effective primer length = 2x (G / C no.) + (A / T no.).

Nonspecific binding can be controlled using any of several known techniques such as, for example, blocking the membrane with protein-containing solutions, RNA, DNA, and SDS heterologous additions to the hybridization buffer, and Rnase treatment. For non-homologous probes, a series of hybridizations may be performed by varying either (i) progressively decreasing annealing temperature (e.g. from 68 ° C to 42 ° C) or (ii) progressively decreasing formamide concentration (eg 50% to 0 ° C). %). The artisan is aware of several parameters that can be changed during hybridization and that will either maintain or change stringency conditions.

In addition to hybridization conditions, specificity of hybridization typically also depends on the function of post hybridization washes. To remove background resulting from non-specific hybridization, samples are washed with dilute saline. Critical factors of such washes include the ionic strength and temperature of the final wash solution: the lower the salt concentration and the higher the wash temperature, the greater the stringency of the wash. Washing conditions are typically performed at or below hybridization stringency. Positive hybridization generates a signal that is at least twice that of the foregoing. Generally, stringent conditions suitable for nucleic acid hybridization assays or gene amplification detection procedures are as set forth above. More or less stringent conditions can also be selected. The skilled artisan is aware of various parameters which may be changed during washing and which will maintain or change stringency conditions.

For example, high stringency hybridization conditions typical for DNA hybrids greater than 50 nucleotides include hybridization at 65 ° C in Ix SSC or 42 ° C in Ix SSC and 50% formamide, followed by washing at 65 ° C in 0.3x SSC . Examples of medium stringency hybridization conditions for DNA hybrids greater than 50 nucleotides include hybridization at 50 ° C in 4x SSC or at 40 ° C in 6x SSC and 50% formamide, followed by washing at 50 ° C in 2x SSC. The hybrid length is the anticipated length for the hybridizing nucleic acid.

When nucleic acids of known sequence are hybridized, hybrid length can be determined by aligning the sequences and identifying the conserved regions described herein. IxSSC is 0.15M NaCl and 15mM sodium citrate; Hybridization solution and wash solutions may additionally include 5 χ Denhardt reagent, 0.5-1.0% SDS, 100 μg / ml denatured shredded salmon sperm DNA, 0.5% sodium pyrophosphate.

For the purposes of defining the stringency level, reference may be made to Sambrook et al. (2001) Molecular Cloning: Laboratory Manual, 3rd Edition Cold Spring Harbor Laboratory Press, CSH, New York or Current Protocols in Molecular Biology, John Wiley & Sons, N.Y. (1989 and annual updates).

Processing Variant

The term "processing variant" as used herein encompasses variants of a nucleic acid sequence in which selected introns and / or exons have been cut, substituted, displaced or added, or in which introns have been shortened or lengthened. Such variants will be one in which the biological activity of the protein is substantially maintained, this can be achieved by selectively maintaining functional protein segments. Such union variants can be found in nature or can be made by man. Methods for predicting and isolating such union variants are well known in the art (see for example Foissac and Schiex, BMC Bioinformatics. 2005; 6:25).

Allelic variant

Alleles or allelic variants are alternative forms of a given gene, located at the same chromosomal position. Allelic variants embrace Single Nucleotide Polymorphisms (SNPs) as well as Small Insert / Deletion Polymorphisms (INDELs). The size of INDELs is typically less than 100 bp. SNPs and INDELs form the largest set of sequence variants in naturally occurring polymorphic strains of most organisms.

Gene Shuffling / Directed EvolutionGene Shuffling or Directed Evolution consists of DNA shuffling repetitions followed by appropriate screening and / or selection to generate nucleic acid variants or protein-decoding portions having a modified biological activity (Castle et al., (2004) Science 304 (5674) ): 1151-4; U.S. Patents 5,811,238 and 6,395,547).

Regulator / Control Sequence / Promoter

The terms "regulatory element", "control sequence" and "promoter" are all interchangeably used herein and should be adopted in a broad context to refer to nucleic acid regulatory sequences capable of expressing the sequences to which they are linked. The term "promoter" typically refers to a nucleic acid control sequence located prior to the transcriptional onset of umgene and which is involved in recognition and binding of RNA polymerase and other proteins, thereby directing transcription of a operably linked nucleic acid. Covered by the above-mentioned terms are transcriptional regulatory sequences derived from a classic genomic and eukaryotic gene (including the TATA box that is required for accurate transcription initiation, with or without a CCAAT box sequence) and additional regulatory elements (ie sequences above the gene enhancers, and silencers). ) that alter degenerate expression in response to development and / or external stimuli, or a specific tissue way. Also included within the term is a transcriptional regulatory sequence of a classic prokaryotic gene, in which case it may include a -35 box sequence and / or -10 additional box regulatory sequences. The term "regulatory element" also encompasses a synthetic or derivative fusion molecule that confers, activates, or enhances expression of a nucleic acid molecule in a cell, tissue, or organ.

A "plant promoter" comprises regulatory elements, which mediate the expression of a coding sequence segment in plant cells. Therefore, a plant promoter need not be of plant origin, but may originate from viruses or microorganisms, for example viruses that attack plant cells. The "plant promoter" may also originate from a plant cell, e.g. from the plant which is transformed with the nucleic acid sequence to be expressed in the inventive process described herein. This also applies to other "vegetable" regulatory signals, such as "vegetable" terminators. Promoters prior to nucleotide sequencing useful in the methods of the present invention may be modified by one or more nucleotide substitution (s), insertion (s) and / or deletion (s) without interfering with the functionality or activity of any promoter, the open reading frame (ORF) or the 3 'regulatory region such as terminators or other 3' regulatory regions that are located outside the ORR. In addition, it is possible that the activity of the promoters is increased by modification of their sequence, or that they are completely replaced by more active promoters, even those of heterologous organisms. For plant expression, the nucleic acid molecule should, as described above, be operably linked to or comprise a suitable promoter expressing the right gene and with the required spatial expression pattern.

For the identification of functionally equivalent promoters, the promoter strength and / or expression pattern of a candidate promoter may be analyzed for example by operatively linking the promoter to a reporter gene and assaying the level and pattern of dogene reporter expression in various plant tissues. Suitable well known reporter genes include for example beta-glucuronidase or beta galactosidase. Promoter activity is assayed by measuring the enzymatic activity of beta-glucuronidase or beta-galactosidase. The promoter strength and / or expression pattern may be compared to those of a reference promoter (such as that used in the methods of the present invention). Alternatively, promoter strength can be assayed by quantifying mRNA levels or comparing mRNA levels of the nucleic acid used in the methods of the present invention, with gene mRNA levels responsible for maintaining cell metabolism such as 18S rRNA, using methods known in the art, such as Northern blotting. with densitometric analysis of autoradiograms, real-time quantitative PCR or RT-PCR (Heid et al., 1996 Genome Methods 6: 986-994). Generally by weak "promoter" is meant a promoter that directs expression of a decoding sequence at a low level. By "low level" is meant at levels of about 1 / 10,000 transcripts to about 1 / 100,000 transcripts at about 1 / 500,0000 transcripts per cell. Conversely, a "strong promoter" directs expression of a high level coding sequence, or in about 1/10 transcripts to about 1/100 transcripts to about 1 / 1,000 transcripts per cell.

Operationally linked

The term "operably linked" as used herein refers to a functional link between the promoter sequence and the gene of interest, such that the promoter sequence is capable of initiating transcription of the gene of interest.

Constitutive promoter

A "constitutive promoter" refers to a promoter that is transcriptionally active during most, but not necessarily all, growth and developmental phases and under most environmental conditions in at least one cell, tissue or organ. Table 2 below provides examples of constitutive promoters.Table 2: Examples of constitutive promoters

<table> table see original document page 27 </column> </row> <table>

Ubiquitous Promoter

An ubiquitous promoter is active in substantially all tissues or cells of an organism.

Development Regulated Promoter

A developmentally regulated promoter is active during certain stages of development or in parts of the plant undergoing evolutionary changes.

Inducible Promoter

An inducible promoter has onset of induced transcription or increased in response to a chemical stimulus (for a review see Gatz1997, Annu. Rev. Plant Physiol. Plant Mol. Biol., 48: 89-108), or may be "inducible" stress ", ie activated when a plant is exposed to various stress conditions, or a" pathogen-inducible "reactivated when a plant is exposed to exposure to multiple pathogens.

Promoter Specific Organ / Specific Tissue A specific organ or specific tissue promoter is one that is preferably capable of initiating transcription into certain organs or tissues, such as leaves, roots, seed tissue, etc. For example, a "specific promoter" is a promoter that is transcriptionally active predominantly in plant roots, substantially to the exclusion of any other parts of a plant, while still allowing any evasive expression in these other plant parts. Promoters capable of initiating transcription in certain cells are referred to herein only as "cell specific".

A specific green tissue promoter as defined herein is a promoter that is transcriptionally active predominantly green tissue, substantially to the exclusion of any other parts of a plant, while still allowing any evasive expression in these other plant parts.

Examples of specific green tissue promoters that may be used to perform the methods of the invention are shown in Table 3 below.

Table 3: Examples of specific green tissue promoters

<table> table see original document page 28 </column> </row> <table>

Another example of a tissue specific promoter is a meristem specific promoter, which is transcriptionally active predominantly in meristematic tissue, substantially to the exclusion of any other parts of a plant, while still allowing any elusive expression in these other plant parts.

The term "terminator" encompasses a control sequence which is a DNA sequence at the end of a transcriptional unit that signals 3 'processing and polyadenylation of a transcript-terminating primary transcript. The terminator may be derived from the natural gene, a variety of other plant genes, or T-DNA. The terminator to be added may be derived, for example, from the nopaline synthase or octopin synthase genes, or alternatively from another plant gene, or less preferably from any other eukaryotic gene.

Modulation

The term "modulation" means in relation to gene expression or expression, a process in which the expression level is changed by gene expression compared to the control plant, preferably the expression level is increased. Unmodulated original expression may be from any expression of a structural RNA (rRNA, tRNA) or mRNA with subsequent translation. The term "modulating activity" shall mean any change in expression of inventive nucleic acid sequences or encoded proteins, which leads to increased plant production and / or growth.

Enhanced Expression / Overexpression

The term "increased expression" or "overexpression" as used herein means any form of expression that is additional to the original wild-type expression level.

Methods for enhancing expression of degene genes or products are well documented in the art and include, for example, overexpression directed by appropriate promoters, the use of transcription enhancers or translation enhancers. Isolated nucleic acids serving as promoter or enhancer elements may be introduced into an appropriate (typically before) non-heterologous form of a polynucleotide in order to increase expression of a nucleic acid by encoding the polypeptide of interest. For example, endogenous promoters may be altered in vivo by mutation, deletion, and / or substitution (see, Kmiec, U.S. Patent No. 5,565,350; Zarling et al., PCT / US93 / 03868), or isolated promoters. introduced into a plant cell at the appropriate orientation and distance of a gene of the present invention to control gene expression.

If polypeptide expression is desired, it is generally desirable to include a polyadenylation region at the 3 'end of a polynucleotide coding region. The polyadenylation region may be derived from the natural gene, a variety of other plant genes, or T-DNA. The 3 'end sequence to be added may be derived, for example, from the nopaline synthase or octopin synthase genes, or alternatively from another plant gene, or less preferably from any other eukaryotic gene.

An intron sequence can also be added to either the untranslated 5 'region (RTU) or the partial decoding sequence coding sequence to increase the amount of mature message accumulating in the cytosol. Inclusion of a processable intron in the transcription unit in both plant and animal constructs has been shown to increase gene expression at both mRNA and protein levels by up to 1000-fold (Buchman and Berg (1988) Mol. Cell biol. 8: 4395-4405; Callis et al. (1987) Genes Dev 1: 1183-1200). Such genetically expressing intron stimulation is enhanced when placed near the 5 'end of the transcription unit. Use of the Intron Adhl-S 1, 2, and 6 Corn Introns, the IntronBronze-I are known in the art. For general information see: The Maize Handbook, Chapter 116, Freeling and Walbot, Eds., Springer, N.Y. (1994).

Endogenous gene

Reference herein to an "endogenous" gene refers not only to the gene in question as found in a plant in its natural shape (ie, without any human intervention), but also refers to the same gene (or a substantially homologous nucleic acid / gene) in an isolated form subsequently (re) introduced into a plant (umtransgene). For example, a transgenic plant containing such a transgene may experience substantial reduction of transgene expression and / or substantial reduction of endogenous gene expression.

Diminished expression

Reference herein to "substantial reduction or elimination" is taken to mean a decrease in endogenous gene expression and / or polypeptide levels and / or polypeptide activity relative to control plants. Substantial reduction or elimination is in ascending order preferably at least 10%, 20%, 30%, 40% or 50%, 60%, 70%, 80%, 85%, 90%, or 95%, 96%, 97 %, 98%, 99% or more reduced compared to those control plants.

For the substantial reduction or elimination of endogenous umgene expression in a plant, a sufficient length of substantially contiguous nucleotides of a nucleic acid sequence is required. In order to perform gene silencing, this may be as little as 20, 19, 18, 17, 16, 15, 14, 13, 12, 11, 10 or fewer nucleotides, alternatively this may be as much as the entire gene (including 5 'and / or3' UTR, or in whole or in part). The substantially contiguous nucleotide stretch may be derived from the nucleic acid encoding the protein of interest (target gene), or from any nucleic acid capable of encoding a protein homolog, paralog or homologue of the protein of interest. Preferably, the substantially contiguous nucleotide stretch is capable of forming hydrogen bonds with the target gene (or sense or antisense strand), more preferably, the substantially contiguous nucleotide stretch is preferably 50%, 60%, 70%, 80%. %, 85%, 90%, 95%, 96%, 97%, 98%, 99%, 100% sequence identity with the target gene (either ribbons or antisense). The nucleic acid sequence encoding a (functional) polypeptide is not a requirement for the various methods discussed herein for substantially reducing or eliminating expression of an endogenous gene.

This substantial reduction or elimination of expression can be achieved using routine tools and techniques. A preferred method for substantially reducing or eliminating endogenous gene expression is to introduce and express in a plant a genetic construct in which the nucleic acid (in this case a substantially contiguous nucleotide portion derived from the gene of interest, or any nucleic acid capable of decoding an ortholog, homologue or homologue of any of the proteins of interest) is cloned as an inverted repeat (in part or in full), separated by a spacer (non-coding DNA).

In such a preferred method, expression of the endogenous gene is reduced or substantially eliminated by RNA-mediated silencing using an inverted repeat of a nucleic acid or part thereof (in this case a substantially contiguous nucleotide stretch derived from the gene of interest, or any capable nucleic acid). decoding a protein ortholog, paralogue or homologue of interest), preferably capable of forming a staple structure. Inverted repetition is cloned an expression vector comprising control sequences. A non-coding DNA nucleic acid sequence (a spacer, for example a matrix coupling region (MAR) fragment, an intron, a polylinker, etc.) is located between the two inverted nucleic acids forming the inverted repeat. After transcription of the inverted repeat, a chimeric RNA with a self-complementary structure is formed (partial or complete). This double stranded RNA structure is referred to as staple RNA (hpRNA). HpRNA is processed by the plant into siRNAs that are incorporated into an RNA-induced silencing complex (RISC). RISC additionally cleaves mRNA transcripts, thereby substantially reducing the number of mRNA transcripts to be translated into empolipeptides. For additional general details see for example, Grierson etal. (1998) International Patent 98/53083; Waterhouse et al. (1999) International Patent 99/53050).

Performance of the methods of the invention is not intended to introduce into a plant a genetic construct in which the nucleic acid is cloned as an inverted repeat, but any one or more of the well-known "gene silencing" methods may be used to achieve the same effects.

One such method for reducing endogenous gene expression is RNA-mediated gene expression silencing (formanegative regulation). Silencing in this case is triggered in a plant by a double stranded RNA (dsRNA) sequence that is substantially similar to the target gene endogenous. This dsRNA is further processed by the plant from about 20 to about 26 nucleotides called short interfering RNAs (siRNAs). SiRNAs are incorporated into an RNA-induced deletion complex (RISC) that cleaves the endogenous target dogene mRNA transcript, thereby substantially reducing the number of mRNA transcripts to be translated into a polypeptide. Preferably, the double stranded RNA sequence corresponds to a target gene.

Another example of an RNA silencing method involves the introduction of nucleic acid sequences or parts thereof (in this case a substantially contiguous stretch of nucleotides derived from the gene of interest, or any nucleic acid capable of encoding a protein homolog, paralog or homologue of the protein of interest). ) in an orientation felt in a plant. "Direction orientation" refers to a DNA sequence that is homologous to an mRNA transcript thereof. A plant should therefore be introduced so at least one copy of the nucleic acid sequence. The additional nucleic acid sequence will reduce endogenous dogene expression, generating a phenomenon known as co-suppression. Gene expression reduction will be more pronounced if multiple additional copies of a nucleic acid sequence are introduced into the plant, as there is a positive correlation between high transcript levels and co-suppression triggering.

Another example of an RNA silencing method involves the use of antisense nucleic acid sequences. An "antisense" nucleic acid sequence comprises a nucleotide sequence that is complementary to a "sense" nucleic acid sequence encoding a protein, i.e. complementary to the coding strand of a double stranded cDNA molecule or complementary to a transcribed demRNA sequence. The antisense nucleic acid sequence preferably is complementary to the endogenous gene to be silenced. The complementarity may be located in the "coding region" and / or the "non-coding region" of a gene. The term "coding region" refers to a nucleotide sequence region comprising codons that are translated into amino acid residues. The term "non-coding region" refers to the 5 'and 3' sequences flanking the coding region that are untranslated amino acid transcripts (also referred to as 5 'and 3' untranslated regions).

Antisense nucleic acid sequences can be construed according to the Watson and Crick base pairing rules. Sequence of antisense nucleic acids may be complementary to the entire nucleic acid sequence (in this case a substantially contiguous nucleotide sequence derived from the gene of interest, or any nucleic acid capable of encoding a protein ortholog, paralogue or homologue of interest), but may also be a oligonucleotide which is provided for only part of the nucleic acid sequence (including 5 'and 3' mRNA UTRs). For example, the antisense oligonucleotide sequence may be complementary to the region surrounding the transcription start site of an mRNA transcript encoding a polypeptide. The length of a suitable antisense oligonucleotide sequence is known in the art and can start from about 50, 45, 40, 35, 30, 25, 20, 15 or 10 nucleotides in length or less. An antisense nucleic acid sequence according to the invention may be constructed using chemical synthesis and enzymatic binding reactions using methods known in the art. For example, an antisense nucleic acid sequence (eg, an antisense oligonucleotide sequence) may be chemically synthesized using naturally occurring nucleotides or variously modified nucleotides constructed to increase the biological stability of molecules or to increase the physical stability of the duplex formed between acid sequences. antisense and sense nucleicides, eg, phosphorothioate derivatives and acridine-substituted nucleotides may be used. Examples of modified nucleotides that can be used to harness antisense nucleic acid sequences are well known in the art. Known nucleotide modifications include methylation, cyclization and caps and replacement of one or more of the naturally occurring nucleotides with an analog such as inosine. Other modifications of nucleotides are well known in the art.

The antisense nucleic acid sequence can be biologically produced using an expression vector in which a nucleic acid sequence has been subcloned in an antisense orientation (ie, RNA transcribed from the inserted nucleic acid will be from an antisense orientation to a target nucleic acid of interest). Preferably, production of antisense nucleic acid sequences in plants occurs by means of a stably integrated nucleic acid construct comprising a promoter, an operably linked antisense oligonucleotide, and a terminator.

Nucleic acid molecules used for silencing the methods of the invention (whether introduced into a plant or generated in situ) hybridize to or bind to mRNA transcripts and / or genomic DNA by encoding a polypeptide to thereby inhibit protein expression, eg, by inhibiting transcription. and / or translation. Hybridization may be by conventional nucleotide complementarity to form a stable duplex, or, for example, in the case of an antisense nucleic acid sequence that binds to DNA duplexes, through specific interactions in the main double strand slit. Antisense nucleic acid sequences can be introduced into a plant by transformation or direct injection into a specific tissue site. Alternatively, antisense nucleic acid sequences may be modified to target selected cells and then systemically administered. For example, for systemic administration, antisense nucleic acid sequences may be modified so that they specifically bind to receptors or antigens expressed on a selected cell surface, eg, by binding antisense nucleic acid sequence to peptides or antibodies that select for receptors or antigens. cell surface. Antisense nucleic acid sequences can also be released into cells using the vectors described herein.

In a further aspect, the antisense nucleic acid sequence is an α-anomeric nucleic acid sequence. A sequence of α-anomeric nucleic acids forms complementary strand-specific double strand hybrids in which, unlike bus units, strands run parallel to each other (Gaultier et al. (1987) Nucl AcRes 15: 6625-6641). The antisense nucleic acid sequence may also comprise a 2-o-methylribonucleotide (Inoue et al. (1987) Nucl AcRes 15, 6131-6148) or a chimeric RNA-DNA analog (Inoue et al. (1987) FEBS Lett. 215 , 327-330).

Substantial reduction or elimination of gene endogenous expression may also be performed using ribozymes. Ribozymes are ribonuclease activity-catalyzed RNA molecules that are capable of cleaving a single stranded nucleic acid sequence, such as an mRNA, to which they have a complementary region. Thus ribozymes (eg, hammerhead ribozymes (described in Haselhoff and Gerlach (1988) Nature 334, 585-591) can be used to catalytically cleave mRNA transcripts encoding a polypeptide, thereby substantially reducing the number of mRNA transcripts to translated into a polypeptide.

A ribozyme having specificity for a nucleic acid sequence can be constructed (see for example: Cech et al. U.S. Patent No. 4,987,071; and Cech et al. U.S. Patent No. 5,116,742).

Alternatively, mRNA transcripts corresponding to a nucleic acid sequence may be used to select a catalytic RNA for ribonuclease-specific activity from a set of RNA demolecules (Bartel and Szostak (1993) Science 261, 1411-1418). Use of ribozymes for gene silencing in plants is known in the art (eg, Atkins et al. (1994) International Patent 94/00012; Lenne et al. (1995) International Patent 95/03404; Lutziger et al. (2000) Patent International 00/00619, Prinsen et al (1997) International Patent 97/13865 and Scott et al (1997) International Patent 97/38116).

Gene silencing can also be achieved by insertion mutagenesis (eg, T-DNA insertion or detransposon insertion) or strategies as described by, among others, Angell and Baulcombe ((1999) Plant J 20 (3): 357-62), (Amplicon VIGS International Patent 98/36083), or Baulcombe (International Patent 99/15682). Gene silencing can also occur if there is a mutation in an endogenous gene and / or a mutation in a gene / nucleic acid subsequently introduced into a plant. Substantial reduction or deletion may be caused by a non-functional polypeptide.For example, a polypeptide may be linked to various interacting proteins, one or more mutation (s) and / or truncation (s) may therefore provide a polypeptide that still It is capable of binding to interacting proteins (such as receptor proteins) but which cannot exhibit their normal function (such as signaling ligand).

An additional approach to gene silencing is to target nucleic acid sequences complementary to the gene regulatory region (eg, the promoter and / or enhancers) to form triple helix structures that prevent gene transcription in target cells. See Helene, C., Anticancer Drug Res. 6, 569-84, 1991; Helene et al., Ann.N.Y. Acad. Know. 660, 27-36 1992; and Maher, L.J. Bioassays 14, 807-15, 1992.

Other methods, such as the use of antibodies directed to an endogenous polypeptide to inhibit its plant function, or signaling interference in which a polypeptide is involved, will be well known to one of skill. In particular, it may be contemplated that manufactured molecules may be useful for inhibiting the biological function of a target polypeptide, or for interfering with the signaling pathway in which the target polypeptide is involved.

Alternatively, a screening program may be adjusted to identify in a plant population natural variants of a gene whose variants encode reduced activity polypeptides. Such natural variants may also be used for example to perform homologous recombination.

Artificial and / or natural microRNAs (miRNAs) may be used to knock out gene expression and / or mRNA translation. Endogenous miRNAs are small single stranded RNAs typically 19-24 nucleotides in length. They function primarily for regular gene expression and / or mRNA translation. Most plant microRNAs (miRNAs) have perfect or near perfect complementarity with their target sequences. However, there are natural targets with up to five mismatches. They are processed from larger non-coding RNAs with reverse structures characteristic by Dicer family specific double stranded RNases. Upon processing, they are incorporated into the RNA-induced silencing complex (RISC) by binding to its major component, an Argonaut protein. MiRNAs serve as the specificity components of RISC, since they match target nucleic acid bases, primarily mRNAs, in the cytoplasm. Subsequent regulatory events include target mRNA cleavage and destruction and / or translational inhibition. Effects of miRNA overexpression are thus often reflected in decreased mRNA levels of target genes.

Artificial microRNAs (amiRNAs), which are typically 21 nucleotides in length, can be genetically engineered specifically to negatively regulate gene expression of one or multiple genes of interest. Targeting determinants of microRNAvegetal are well known in the art. Empirical parameters for target recognition have been defined and can be used to aid the design of specific amiRNAs, Schwab R, 2005. Convenient tools for the design and generation of amiRNAs and their precursors are also available to the public, Schwab et al., 2006.

For optimal performance, gene silencing techniques used to reduce expression in a plant of an endogenous gene require the use of monocotyledon nucleic acid sequences for monocotyledonous plants and dicotyledonous plants for dicotyledonous plants. Preferably, the nucleic acid sequence of any given plant species is introduced into that same species. For example, a rice nucleic acid sequence is transformed into a rice plant. However, it is not an absolute requirement that the nucleic acid sequence to be introduced originates from the same plant species as the plant into which it will be introduced. It is sufficient that substantial homology exists between the target endogenous gene and the nucleic acid to be introduced.

Examples of various methods for reducing or substantially eliminating expression in a plant of a gene endogenous are described above. One of ordinary skill in the art should readily be able to adapt the abovementioned methods for silencing to achieve reduction of expression of an endogenous gene in an entire plant or parts thereof through the use of an appropriate promoter, for example.

Selectable marker (geneVGene reporter

"Selectable marker", "selectable marker gene" or "reporter gene" includes any gene that confers a phenotype in a cell which is expressed to facilitate the identification and / or selection of cells transfected or transformed with an invention nucleic acid construct . These marker genes enable the identification of successful transfer of nucleic acid molecules through a number of different principles. Suitable markers can be selected from markers that confer antibiotic or herbicide resistance, introduce a new metabolic trait, or allow visual selection. Examples of selectable marker genes include genes conferring antibiotic resistance (such as npt11 which phosphorylates neomycin ecanamycin, or hpt, phosphorylating hygromycin, or genes conferring resistance, for example bleomycin, streptomycin, tetracycline, chloramphenicol, ampicillin, gentamicin, geneticin (G418), spectinomycin or blasticidine), herbicides (eg, bar providing resistance to Basta; aroA or gox providing resistance to glyphosate, or genes conferring resistance to, for example, imidazolinone, phosphinothricin or sulfonylurea), or genes that provide a metabolic trait (such as as manA which allows plants to use mannose as the sole source of carbon or xylose isomerase for xylose autilization, or antinutritive markers such as 2-deoxyglucose resistance). Expression of visual marker genes results in the formation of color (eg β-glucuronidase, GUS or β-galactosidase with their colored substrates, for example X-Gal), luminescence (such as luciferin / luceferase system) or fluorescence (Green Fluorescent Protein, GFP , derived from this). This list represents only a small number of possible markers. The skilled worker is familiar with such markers. Different markers are preferred depending on the organism and method of selection.

It is known that by stable or transient integration of nucleic acids into plant cells, only a minority of cells absorb the exogenous DNA and, if desired, integrate it into their genome, depending on the expression vector used and the transfection technique used.

To identify and select these integrants, a gene encoding a selectable marker (such as those described above) is typically introduced into host cells along with the gene of interest. These markers may be used on mutants in which these genes are non-functional, for example by deletion by conventional methods. In addition, nucleic acid molecules encoding a selectable marker may be introduced into a host cell in the same vector comprising the sequence encoding the polypeptides of the invention or used in the methods of the invention, or in a separate vector. Cells that have been stably transfected with the introduced nucleic acid can be identified by selection (for example, cells that integrate the selectable marker survive while the other cells die). Since marker genes, particularly antibiotic and herbicide resistance genes, do not are most required or desired in the transgenic host cell once nucleic acids have been successfully introduced, the process according to the invention to introduce nucleic acids advantageously employs techniques that enable the removal or excision of these marker genes. One such method is what is known as co-transformation. The co-transformation method employs two vectors simultaneously for transformation, vector loading the nucleic acid according to the invention and one second loading the marker gene (s). A large proportion of transformants receive or, in the case of plants, comprise (up to 40% or more of the transformants) both vectors. In case of transformation with Agrobacteria, the transformants usually receive only a portion of the vector, i.e. the T-DNA flanked sequence, which normally represents the expression drawer. The marker genes can subsequently be removed from the transformed plant by breeding. In another method, marker genes integrated into a transposon are used for transformation along with desired nucleic acid (known as Ac / Ds technology). Transformants may be cross-linked to a transposase source or transformants are transformed with a nucleic acid construct conferring expression of a transposase, either transiently or stably. In some cases (approx. 10%), the transposon skips outside the host cell name once transformation has been successful and is lost. In several additional cases, the transposon jumps to a different location. In these cases the marker gene should be eliminated by performing crosses. In microbiology, techniques have been developed that make possible, or facilitate, the detection of such events. An additional advantageous method relies on what is known as combining systems; whose advantage is that cross elimination can be missed. The best known system of this type is what is known as the Cre / lox system. Crel is a recombinase that removes sequences located between IoxP sequences. If the marker gene is integrated between the IoxP sequences, it is removed once transformation has been successful by recombinase expression. Additional recombination systems are the HIN / HIX, FLP / FRT and REP / STB systems (Tribble et al., J.Biol. Chem., 275, 2000: 22255-22267; Velmurugan et al., J. Cell Biol., 149 , 2000: 553-566). Site-specific integration into the plant genome of nucleic acid sequences according to the invention is possible. Of course, these methods can also be applied to microorganisms such as yeast, fungi or bacteria.

Transgenic / Transgene / Recombinant

For the purposes of the invention, "transgenic", "transgene" or "recombinant" means with reference to, for example, a nucleic acid sequence, expression drawer, gene construct or a vector comprising the nucleic acid sequence or an organism transformed with the nucleic acid sequences, listener expression cassettes according to the invention, all of these constructs caused by recombinant methods in which or

(a) the nucleic acid sequences encoding proteins useful in the methods of the invention, or

(b) genetic control sequence (s) that are operably linked with the nucleic acid sequence according to the invention, for example a promoter, or

(c) a) and b) are not located in their natural genetic environment or have been modified by recombinant methods, and it is possible for amodification to take the form of, for example, a substitution, addition, deletion, inversion or insertion of one or more nucleotide residues . The natural genetic environment is understood to mean the natural genomic or chromosomal locus in the original plant or the presence in a library. In the case of a genomic library, the natural genetic environment of the nucleic acid sequence is preferably maintained, at least in part. The environment flanks the nucleic acid sequence at least on one side and has a sequence length of at least 50 bp, preferably at least 500 bp, especially preferably at least 1000 bp, more preferably at least 5000 bp. A naturally occurring expression drawer - for example, the naturally occurring combination of the natural nucleic acid sequence promoter with the corresponding nucleic acid sequence encoding a polypeptide in the methods of the present invention, as defined above - becomes an expression drawer when this expression drawer is used. modified by unnatural, synthetic ("artificial") methods such as, for example, mutagenic treatment. Suitable methods are described, for example, in US Patent 5,565,350 or International Patent 00/15815.

A transgenic plant for the purposes of the invention is believed to mean, as above, that the nucleic acids used in the method of the invention are not in their natural locus in the genome of said plant, and it is possible for the nucleic acids to be expressed homologously or heterologously. However, as mentioned, transgenic also means that while the nucleic acids according to the invention used in the inventive method are in their natural position in the genome of a plant, the sequence has been modified with respect to the natural sequence, and / or which regulatory sequences of the sequences. have been modified.

Transgenic is preferably understood to mean expression of nucleic acids according to the invention at an unnatural locus in the genome, i.e. homologous or preferably heterologous expression of nucleic acids occurs. Preferred transgenic plants are mentioned here.

The term "introduction" or "transformation" as referred to herein includes the transfer of an exogenous polynucleotide to a host cell, regardless of the method used for transfer. Plant tissue capable of subsequent clonal propagation, either by organogenesis or embryogenesis, can be transformed with a genetic construct of the present invention and an entire plant regenerated therefrom. The particular tissue chosen will vary depending on the clonal propagation systems available for, and best suited to, the particular species being transformed. Exemplary tissue targets include leaf discs, pollen, embryos, cotyledons, hypocotyls, megagametophytes, callus tissue, existing tissue (eg, apical meristem, axillary buds, and root meristems), and induced meristem tissue (eg, cotyledon meristem in system). hypocotyl). The polynucleotide may be transiently or stably introduced into a host cell and may be kept unintegrated, for example, as a plasmid. Alternatively, it may be integrated into the host genome. The resulting transformed plant cell can then be used to regenerate a transformed plant in a manner known to those skilled in the art.

The transfer of exogenous genes to the genome of a plant is called transformation. Transformation of plant species is now a fairly routine technique. Advantageously, any of several transformation methods may be used to introduce the gene of interest into a suitable ancestral cell. The methods described for transformation and regeneration of plants from plant tissues or plant cells may be used for transient or stable transformation. Transformation methods include the use of liposomes, electroporation, chemicals that increase free DNA uptake, DNA injection directly into the plant, particle gun bombardment, virus or pollen transformation, and microprojection. Methods can be selected from the calcium / polyethylene glycol method for protoplasts (Krens, FA et al., (1982) Nature 296, 72-74; Negrutiu I et al. (1987) Plant Mol Biol 8: 363-373); electroporation protoplasts (Shillito RD et al. (1985) Bio / Technol 3, 1099-1102); microinjection in plant material (Crossway A et al., (1986) Mol. GenGenet 202: 179-185); DNA-coated particle bombardment or RNA (Klein TM et al., (1987) Nature 327: 70) infection with (non-integrative) viruses and the like. Transgenic plants, including transgenic crop plants, are preferably produced by Agrobacterium-mediated transformation. An advantageous transformation method is plant transformation. For this purpose, it is possible, for example, to allow agrobacteria to act on plant grains or to inoculate the plant system with agrobacteria. Particularly suitable in accordance with the invention has been shown to allow a suspension of transformed aggrobacteria to act on the intact plant or at least the floral primordia. The plant is subsequently grown until treated plant seeds are obtained (Clough and Bent, Plant J. (1998) 16, 735-743). Methods for Agrobacterium dearroz mediated transformation include well known methods for rice transformation, such as those described in any of the following: European Patent Application EP1198985 Al, Aldemita and Hodges (Plant 199: 612-617, 1996); Chan et al. (Plant Mol Biol 22 (3): 491-506, 1993), Hiei et al. (Plant J 6 (2): 271-282,1994), the descriptions of which are incorporated by reference herein as fully presented. In the case of maize transformation, the preferred method is as described or in Ishida et al. (Nat. Biotechnol 14 (6): 745-50,1996) or Frame et al. (Plant Physiol 129 (1): 13-22, 2002), the disclosures of which are incorporated by reference herein as shown in their entirety. Such methods are further described as an example in B. Jenes et al., Techniques for Gene Transfer, in: Transgenic Plants, Vol. 1, Engineering and Utilization, eds. S.D. Kung and R. Wu5Academic Press (1993) 128-143 and Potrykus Annu. Rev. Plant Physiol.Plant Molec. Biol. 42 (1991) 205-225). Nucleic acids or the expressed construct are preferably cloned into a vector, which is suitable for transforming Agrobacterium tumefaciens, for example pBin19 (Bevan et al., Nucl. Acids Res. 12 (1984) 8711). Talvector-transformed agrobacteria can be used in a manner known for plant transformation, such as plants used as a model, such as Arabidopsis (Arabidopsis thaliana is within the scope of the present invention not considered as a crop plant), or crop plants such as as, by way of example, tobacco plants, for example by immersing tarnished leaves or chopped leaves in an agrobacterial solution and then growing in a suitable medium. Plant transformation by Agrobacterium tumefaciens is described, for example, by Höfgen and Willmitzerem Nucl. Acid Res. (1988) 16, 9877 or is known among other ways from F.F. White, Vectors for Gene Transfer in Higher Plants; in Transgenic Plants, Vol. 1, Engineering and Utilization, eds. S.D. Kung and R. Wu, Academic Press, 1993, p. 15-38.

In addition to the transformation of somatic cells, which then have to be regenerated into intact plants, it is also possible to transform cells of plant meristems and in particular those cells that develop into gametes. In this case, transformed gametes follow natural plant development, generating transgenic plants. Thus, for example, Arabidopsis seeds are treated with agrobacteria and seeds obtained from growing plants from which a certain proportion is transformed and thus transgenic [Feldman, KA and Marks MD (1987). Mol Gen Genet 208: 274-289; Feldmann K (1992). In: C. Koncz, N-HChua and J Shell, eds, Methods in Arabidopsis Research. Word Scientific, Singapore, pp. 274-289]. Alternative methods are based on repeated removal of inflorescences and incubation of the excision site in the daroseta center with transformed agrobacteria, whereby seedlings can similarly be obtained at a later time (Chang (1994). Plant J. 5: 551-558; Katavic (1994), Mol Gen Genet, 245: 363-370). However, an especially effective method is the vacuum infiltration method with its modifications such as the "flower bud" method. In the case of vacuum infiltration of Arabidopsis, intact plants under reduced pressure are treated with an agrobacterial suspension [Bechthold, N (1993). C R Acad Sci Paris Life Sci, 316: 1194-1199], while in the case of the "flower bud" method the developing floral tissue is briefly incubated with a surfactant-treated agrobacterial suspension [Clough, SJ und Bent, AF (1998). The Plant J. 16, 735-743]. A certain proportion of transgenic seeds are collected in both cases, and thus can be distinguished from non-transgenic seeds by cultivating in the selective conditions described above. In addition, stable transformation of plastids is advantageous in that plastids are inherited maternally reducing or eliminating the risk of transgene flow through most crop pollen. Transformation of the chloroplast genome is usually achieved by a process that has been schematically presented in Klaus etal., 2004 [Nature Biotechnology 22 (2), 225-229]. Briefly the sequences to be transformed are cloned together with a selectable marker gene between flanking sequences homologous to the dechloroplast genome. These homologous flanking sequences direct specific site integration in the plastoma. Plastid transformation has been described for many different plant species and an overview is given in Bock (2001) Transgenic plastids in basic research and plant biotechnology. J Mol Biol.2001 Sep 21; 312 (3): 425-38 or Maliga, P (2003) Progress towards commercialization of plastid transformation technology. Trends Biotechnol.21, 20-28. Further biotechnological progress has recently been reported in the form of marker free plastid transformants, which may be produced by a transient co-integrated marker gene (Klaus et al., 2004, Nature Biotechnology 22 (2), 225-229).

T-DNA activation identification

Identification by T-DNA activation (Hayashi et al. Science (1992) 1350-1353), involves T-DNA insertion, usually containing a promoter (may also be a translation enhancer or an intron), genomic naregion of the gene of interest. or 10 kb before or after the decoding region of a gene in a configuration such that the promoter directs expression of the directed gene. Typically, expression regulation of the genide directed by its natural promoter is disrupted and the gene falls under the newly introduced promoter control. The promoter is typically embedded in a T-DNA. This T-DNA is randomly inserted into the plant genome, for example through Agrobacterium infection and leads to modified gene expression near the inserted T-DNA. The resulting transgenic plants show dominant phenotypes due to the modified expression of degenes near the introduced promoter.

TILLING

TILLING (Locally Targeted Induced Lesions in Genomes) is a mutagenesis technology useful for generating and / or identifying nucleic acids encoding proteins with modified expression and / or activity. TILLING also allows selection of plants carrying such mutant variants. These mutant variants may exhibit modified expression, either in force or in localization or in time (if mutations affect the promoter for example). These mutant variants may exhibit greater activity than that exhibited by the gene in its natural form. TILLING combines high density mutagenesis with rapid screening methods. The steps typically followed in TLLING are: (a) EMS mutagenesis (Redei GP and Koncz C (1992) InMethods in Arabidopsis Research, Koncz C, Chua NH, Schell J, eds.Singapore, World Scientific Publishing Co, pp. 16-82; Feldmann et al., (1994) In Meyerowitz EM, Somerville CR, eds, Arabidopsis, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, pp 137-172; Lightner Jand Caspar T. (1998) In J Martinez-Zapater, J Salinas , eds, Methods on Molecular Biology, Vol. 82. Humana Press, Totowa, NJ, pp 91-104); (b) DNA preparation and combination of individuals; (c) PCR amplification of a region of interest; (d) denaturation and annealing to allow heteroduplex formation; (e) DHPLC, where the presence of a heteroduplex in a combination is detected as an extra nocromatogram peak; (f) identification of the individual mutant; and (g) sequencing of the mutant PCR product. TILLING methods are well known in the art (McCallum et al., (2000) Nat Biotechnol 18: 455-457; reviewed by Stemple (2004) Nat Rev Genet 5 (2): 145-50).

Year Recombination

Homologous recombination allows introduction into a genome of a selected nucleic acid at a defined selected position. Homologous recombination is a standard technology routinely used in biological sciences for lower organisms such as yeast or mossPhyscomitrella. Methods for performing homologous recombination on plants have been described not only for model plants (Offringa et al. (1990) EMBO J 9 (10): 3077-84) but also for crop plants, eg rice (Terada et al. (2002) Nat. Biotech 20 (10): 1030-4; Iida and Terada (2004) Curr Opin Biotech 15 (2): 132-8).

Production

The term "production" generally means a measurable production of economic value, typically related to a specific culture, area, and time period. Individual plant parts contribute directly to production based on their number, size and / or weight, or current production is production per acre for one crop and year, which is determined by dividing total production (includes both harvested and evaluated yields) by planted acres.

Early vigor

Early vigor (healthy well-balanced growth active especially during early stages of plant growth) can result from increased plant fitness due to, for example, plants being better adapted to their environment (i.e. optimizing the use of energy resources and partitioning between shoots and roots). Early vigorous plants also show increased seedling survival and improved crop establishment, which often results in highly uniform fields (with the crop growing evenly, ie with most plants reaching the various stages of development substantially at the same time), and often better and bigger production. Therefore, early vigor can be determined by measuring various factors such as thousand seed weight, germination percentage, emergence percentage, seedling growth, seedling height, root length, root and shoot biomass and more.

Increase / Improve / Accentuation

The terms "increase", "improvement" or "accentuation" are interchangeable and shall mean in the sense of the request at least 5%, 6%, 7%, 8%, 9% or 10%, preferably at least 15% or 20%. more preferably 25%, 30%, 35% or 40% more production and / or growth compared to control plants as defined herein.

Seed production

Increased seed production may be manifested as one or more of the following: (a) an increase in seed biomass (total seed weight) which may be on an individual seed basis and / or per plant and / or per hectare or acre; b) increased number of flowers per plant; c) increased number of seeds (floods); (d) increased grain filling rate (which is expressed as the ratio of the number of full grains divided by the total number of grains); (e) increased harvest index, which is expressed as a proportion of the production of cut-off parts, such as seeds, divided by total biomass; and f) Increased Thousand Seed Weight (TKW), which is extrapolated from the number of full grains counted and its total weight. An increased TKW may result from increased seed size and / or seed weight, and may also result from increased embryo size and / or endosperm.

An increase in seed yield can also be manifested as an increase in seed size and / or deboning volume. In addition, an increase in seed yield may also be manifested as an increase in seed area and / or demented length and / or seed width and / or seed perimeter. Increased production may also result in modified architecture, or may occur because of modified architecture.

Plant

The term "plant" as used herein encompasses plants, ancestors and progeny of plants and plant parts, including seeds, shoots, stems, leaves, roots (including tubers), flowers, tissues and organs, characterized in that each of the mentioned above comprises the gene / nucleic acid of interest. The term "plant" also encompasses plant cells, suspension cultures, callus tissue, embryos, meristematic regions, gametophytes, sporophytes, emicrospore pollen, again characterized by the fact that each of the above comprises the gene / nucleic acid of interest. .

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocotyledonous and dicotyledonous plants including fodder or forage vegetables, ornamental plants, food crops, trees or shrubs selected from the list comprising Acer spp., Actinidiaspp. Abelmoschus spp., Agave sisalana, Agropyron spp., Agrostis stolonifera, Allium spp., Amaranthus spp., Ammophila arenaria, Ananas eomosus, Annona spp., Apium graveolens, Araehis spp. (e.g. Avena sativa, Avena fatua, Avena byzantina, Avena fatua var. sativa, Avena hybrida), Averrhoa starfruit, Bambusa sp., Benineasa hispida, Bertholletia exeelsea, Beta vulgaris, Brassica spp. (egBrassica napus Brassiea rapa ssp. [canola oilseed rape turnip rape]), Cadaba farinosa, Camellia sinensis, Canna indica, Cannabis sativa, Capsicum spp. Carex elata, Carica papctya, Carissa macrocarpa, Caryaspp. Carthamus tinetorius , Castanea spp., Ceiba pentandra, Cichoriumendivia, Cinnamomum spp., Citrullus lanatus, Citrus spp., Cocos spp., Coffeaspp. Colocasia eseulenta, Cola spp., Corchorus sp. Croeus sativus, Cueurbita spp., Cueumis spp., Cynara spp., Daucus earota, Desmodium spp., Dimocarpus longan, Dioseorea spp., Diospyros spp. sp., Eriobotryajaponiea, Eucalyptus sp., Eugenia uniflora, Fagopyrum spp., Fagus spp., Festuea arundinaeea, Fieus cariea, Fortunella spp., Fragaria spp., Ginkgobiloba, Glyeine spp. (e.g. Glycine max, Soya hispida or Soy max), Gossypiumhirsutum, Helianthus spp. (e.g. Helianthus annuus), Hemerocallis fulva, Hibiseus spp., Hordeum spp. (e.g. Hordeum vulgare), Ipomoea potatoes, Juglans spp., Laetuea sativa, Lathyrus spp., Lens eulinaris, Linumusitatissimum, Litehi ehinensis, Lotus spp., Luffa aeutangula, Lupinus spp., Luzula sylvatiea, Lyeopersie spp. (eg Lyeopersieon eseulentum, Lycopersicon lyeopersicum, Lyeopersieon pyriform), Maerotyloma spp., Malus spp., Malpighia emarginata, Mammea americana, Maniif spp. spp., Morus nigra, Musa spp., Nicotiana spp., Olea spp., Opuntia spp., Ornithopus spp., Oryza spp. (eg Oryza sativa, Oryza latifolia), Panicum miliaceum, Panicum virgatum, Passiflora edulis, Pastinaea sativa, Pennisetum sp., Persea spp., Petroselinum erispum, Phalaris arundinaeea, Phaseolus spp. . Pinus spp., Pistaeia vera, Pisum spp., Poa spp., Populus spp., Prosopis spp., Prunus spp., Psidium granatum, Pyrus eommunis, Quereus spp. ., Ricinus eommunis, Rubus spp., Saecharum spp., Salix sp., Sambueus spp., Seeale cereale, Sesamum spp., Sinapis sp., Solanum spp. (e.g. Solanum tuberosum, Solanum integrifolium or Solanum lycopersicum), Sorghum bieolor, Spinaeia spp., Syzygium spp., Tagetes spp., Tamarindus indica, Theobroma caeao, Trifolium spp. (eg Tritieum aestivum, Tritieum durum, Tritieum turgidum, Tritieum hybernum, Triticum maeha, Tritieum sativum or Tritieum vulgare), Tropaeolum minus, Tropaeolum majus, Vaeeinium spp., Vicia spp. , Zizania palustris, Ziziphus spp., Among others.

DETAILED DESCRIPTION OF THE INVENTION

Detailed Description for HAL3

According to a first embodiment of the present invention, a method is provided for increasing plant yield over control plants, preferably comprising increasing expression of a nucleic acid encoding an HAL3 polypeptide in expanding tissues of a plant.

Any reference hereinafter to a "protein useful in the methods of the invention" is taken to mean a HAL3 polypeptide as defined herein. Any reference hereinafter to a "nucleic acid useful in the methods of the invention" is taken to mean the nucleic acid capable of encoding such an HAL3 polypeptide. The nucleic acid to be introduced into a plant (and therefore useful for carrying out the methods of the invention) is any nucleic acid encoding the type of protein that will now be described, hereinafter also called the "HAL3 nucleic acid" or "HALT gene."

A "reference", "reference plant", "control", "control plant", "wild type" or "wild type plant" is in particular a cell, tissue, organ, plant, or part thereof which was not produced according to the method of the invention. Accordingly, the terms "wild type", "control" or "reference" are interchangeable and may be a cell or part of the plant such as an organelle or tissue, or a plant, which has not been modified or treated according to the descriptive method. according to the invention. Accordingly, the cell or plant part such as an organelle or wild-type plant, control or reference corresponds to the cell, plant or part thereof as much as possible and is in any other property but in the result of the invention process so identical to the subject. of the invention as possible. Thus, the wild type, control, or reference is treated identically or as identically as possible, claiming that only conditions or properties may be different that do not influence the quality of the property tested. This means in other words that wild type means (1) a plant, which carries the unchanged or unmodified form of a gene or allele, or (2) the initial material / plant from which plants produced by the process or method of the invention are derived.

Preferably, any comparison between wild type plants and plants produced by the method of the invention is made under analogous conditions. The term "analogous conditions" means that all conditions such as, for example, growing or growing conditions, assay conditions (such as buffer composition, temperature, substrates, pathogen strain, concentrations and the like) are maintained identical between experiments. to be compared.

The "reference", "control", or "wild type" is preferably a subject, eg an organelle, a cell, a tissue, a plant, which has not been modulated, modified or treated in accordance with the process described herein and is in any other property is as similar to the subject of the invention as possible. Reference, control, wild type is in its genome, transcriptome, proteome or metabolism as similar as possible to the subject of the present invention. Preferably, the term "reference" "control" or "wild type" organelle, cell, tissue or plant refers to an organelle, cell, tissue or plant which is almost identical to the organelle, cell, tissue or plant of the present invention or a part thereof is preferably 95%, more preferred is 98%, even more preferred is 99.00%, in particular 99.10%, 99.30%, 99.50%, 99.70%, 99.90%, 99 , 99%, 99, 999% or more. Preferably the "reference", "control", or "wild type" is preferably a subject, eg an organelle, a cell, a tissue, a plant, which is genetically identical to the cell organelle used according to the method of the invention except that molecules of nucleic acid or the gene product encoded by them are engineered, modulated or modified according to the inventive method.

In case a control, reference or wild type other than the subject of the present invention not only subject to the method of the invention cannot be provided, a control, reference or wild type may be a plant in which the cause for modulation of activity conferring the increased metabolites as described in examples.

The reported increase in activity of the polypeptide amounts in a cell, tissue, organelle, organ or organism or a part thereof is preferably to at least 5%, preferably to at least 10% or to at least 15%, especially preferably to at least at least 20%, 25%, 30% or more, most especially preferably are for at least 40%, 50% or 60%, more preferably for at least 70% or more in comparison as a control, reference or wild type.

The term "increased yield" is adopted to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground parts and / or below-ground parts. Preferably, the increase in yield is at least 10% over the yield of corresponding wild type plants.

In particular, such cut-off parts are seeds, and performance of the methods of the invention results in plants having increased seed production relative to control plant seed production.

The term "expression" or "gene expression" means the appearance of a phenotypic trait as a consequence of the transcription of a specific gene or specific genes. The term "expression" or "gene expression" in particular means the transcription of a gene orgenes into structural RNA (rRNA, tRNA) or mRNA with subsequent translation of the latter into a protein. The process includes DNA transcription, processing of the resulting mRNA product and its translation into an active protein.

Performance of the methods of the invention generates improved yield trait plants. In particular performance of the methods of the invention generates plants having increased yield, especially increased seed yield over control plants. The terms "production" and "seed production" are described in more detail in the "definitions" section here.

Reference in this place to improved yield characteristics is adopted to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground (cut-off) and / or below-cut (cut-off) parts. ground. In particular, such cut-off parts are seeds, and performance of the methods of the invention results in plants having increased seed yield over seed production from suitable control plants.

Using corn as an example, an increase in yield can be manifested as one or more of the following: increase in the number of plants per hectare or acre, an increase in the number of ears per plant, an increase in the number of rows, number of seeds per row, weight. dementing, weight of one thousand seeds, ear length / diameter, increased grain filling rate (which is the number of full grains divided by the total number of grains and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary panicles), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in milliseed weight, among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having increased seed production relative to control plants. Therefore according to the present invention there is provided a method for increasing plant seed production over control plant seed production, the method comprising preferably increasing expression of a nucleic acid encoding an HAL3 polypeptide in partisan tissues, preferably in expanding tissues. of a vegetable shoot.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit an increased growth rate (for at least part of their due cycle), relative to the growth rate of control plants in a matched maturity. your life cycle.

The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the entire plant. Plants having an increased rate of decrease may have a shorter life cycle. The proper cycle of a plant can be adopted to mean the time required to grow from dry mature seed to the stage where the plant produced dry mature seeds, similar to the initial material. This life cycle can be influenced by factors such as early vigor, growth rate, green index, flowering time and seed ripening speed. The increase in growth rate may occur at one or more stages in a plant's life cycle or during substantially the entire plant life cycle. Increased growth rate during the early stages of a plant's life cycle may allow for improved vigor. Increasing growth rate can alter the harvest cycle of a plant by allowing plants to be sown later and / or harvested earlier than otherwise possible (a similar effect can be obtained with early flowering time). If the growth rate is sufficiently increased, it may allow additional seed seeding of the same plant species (eg sowing and harvesting rice plants followed by sowing and harvesting additional rice plants within a conventional growing period).

Similarly, if the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing and harvesting of soybean, potato or any other suitable plant) . Additional harvest times of the same rhizome in the case of some crop plants may also be possible. Changing the harvesting cycle of a plant can lead to an increase in annual biomass production per acre (due to an increase in the number of times (say in a year) that any particular plant can be grown and harvested. An increase in growth rate may also allow the cultivation of plant crops in a wider geographical area than their wild counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or planting name (early cycle). or at the time of harvest (late cycle).

Such adverse conditions can be avoided if the harvest cycle is shortened.

The growth rate can be determined by deriving several growth curve parameters, such parameters can be: T-Mid (time used by plants to reach 50% of their maximum size) and T-90 (time used by plants to reach 90% of their maximum size). maximum size), among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having an increased growth rate relative to control plants. Therefore, in accordance with the present invention, there is provided a method for increasing plant growth rate, which method comprises modulating expression, preferably enhancing expression, in a plant of a nucleic acid encoding an HAL3 polypeptide as defined herein.

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general.

Mild stress on the other hand is defined here as any stress to which a plant is exposed that does not result in general plant growth arrest without the ability to resume growth. Mild stress in the sense of the invention leads to a reduction in growth of stressed plants. less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the plant. control non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "no stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location.

Performance of the methods of the invention generates plants grown under non-stress conditions or under conditions of mild drought increased yield over suitable control plants grown under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under non-stress conditions or under mild dry conditions, which method comprises increasing expression in a plant of a nucleic acid by encoding a HAL3 polypeptide.

Performance of the methods of the invention generates plants grown under nutrient deficiency conditions, particularly under nitrogen-deficient conditions, increased yield over control-grown plants under comparable conditions. Therefore, according to the present invention there is provided a method for increasing production in plants grown under nutrient deficient conditions, the method of which is to increase expression in a plant of a nucleic acid by encoding a HAL3 polypeptide. Nutrient deficiencies can result from nutrient deficiencies or excesses such as nitrogen, phosphates and other compounds containing phosphorus, potassium, calcium, cadmium, magnesium, manganese, iron and boron, among others.

According to a second preferred feature of the invention, performance of the methods of the invention generates plants having increased plant vigor relative to control plants, particularly during the early stages of plant development (typically three weeks after germination in the case of rice and maize, but this will vary from species to species) leading to early vigor. Therefore, according to the present invention, there is provided a method for increasing early vigor seedlings, the method of which comprises modulating, preferably increasing, a plant expression of a nucleic acid encoding a HAL3 polypeptide. Preferably the effective seedling increase is achieved by expressing the nucleic acid encoding the HAL3 polypeptide under the control of a shoot-specific promoter. Also provided is a method for producing plants having early vigor over control plants, the method of which comprises modulating, preferably enhancing, expression in a plant of a nucleic acid encoding a HAL3 polypeptide.

Early vigor can also result from increased vegetative fitness due, for example, to plants being better adapted to their environment (i.e. optimizing the use of energy resources and partitioning between shoots and roots). Plants having early vigor also show increased seedling survival and better crop establishment, which often results in highly uniform fields (with the crop growing evenly, ie with most plants reaching various stages of development at substantially the same time), consequently better and bigger production. Therefore, early vigor can be determined by measuring various factors such as 1,000 seed weight, germination percentage, emergence percentage, seedling growth, seedling height, root length, root and aerial biomass, and more.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Still preferably, the plant is a monocot plant. Examples of monocotyledonous plants include sugar cane. Most preferably aplanta is a cereal. Examples of cereals include rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats.

Other advantageous plants are selected from the Asteraceae group consisting of the genera Helianthus, Tagetes eg, Helianthus annus [sunflower], Tagetes lucida, Tagetes erecta orTagetes tenuifolia [Carnation], Brassicaceae such as Brassica, Arabadopsis and Brassica napus. , Brassica rapa ssp. [canola, rapeseed, turnip] or Arabidopsis thaliana. Fabaceae such as the genera Glycinee.g. Glycine max, Soy hispida or Soy max [soy] species. Linaceae such as Linum genera e.g. Linum usitatissimum, [flax, flax seed]; Poaceae such as the genera Hordeum, Secale, Avena, Sorghum, Oryza, Zea, Triticum e.g. Hordeum vulgare [barley] species; Secalecereale [rye], Avena sativa, Avena fatua, Avena byzantina, Avena fatuavar. sativa, Avena hybrida [oats], Sorghum bicolor [Sorghum, millet], Oryzasativa, Oryza latifolia [rice], Zea mays [corn, plus] Triticum aestivum, Tritieum durum, Tritieum turgidum, Triticum hychanum, Tritieum schaum or Tritieum sativum vulgare [wheat, bread-grade wheat, trigocum]; Solanaceae such as the genera Solanum, Lycopersicon e.g. the species Solanum tuberosum [potato], Lyeopersieon esculentum, Lycopersiconlyeopersieum, Lyeopersieon pyriform, Solanum integrifolium or Solanumlyeopersieum [tomato].

The term "HAL3 polypeptide" as defined herein refers to a protein belonging to the HFCD superfamily (Cys Descarboxylases containing Homo-oligomeric Flavine; Kupke J. Biol.Chem. 276, 27597-27604, 2001). These proteins share a flavin binding motif, conserved active site residues and are trimeric or dodecameric enzymes. HAL3 proteins preferably comprise of terminus At terminus C (i) a substrate binding helix, (ii) a His insertion motif, (iii) a PXMNXXMW motif and (iv) a substrate recognition clamp (Figure 1). These four domains are predicted to be involved in substrate binding, the insertion His motif comprises a conserved His residue that is involved in the active site so that the sequence in the substrate recognition clamp can be somewhat variable in sequence and length (Blaesse et al. , EMBOJ 19, 6299-6310, 2000). The structural features (i) to (iv) are also described in Kupke et al. (2001), the description of which is incorporated herein by reference. Typically, HAL3 polypeptides are capable of binding to FMN decoders.

Typically the substrate binding helix is comprised of sequence 1 motive with the following consensus sequence (SEQ ID NO: 6): (K / R) PR (V / I) (I / L) LAA (S / T) GSVA (A / S) (I / MA ^) KF (G / E / A / S) (N / S / I) L (C / V / A) (H / R7G) (C / S / I) (F / L ) (T / S / C) (E / D / Q) WA (E / D) V (RZK) AV (V / A / S).

The His insertion motif, PXMNXXMW motif, and substrate recognition scheme are part of sequence motif 2 with the following consensus sequence (SEQ ID NO: 7): VLHIELR (R / K / Q) WAD (V / I / A) (LM) (V / I) IAPLSANTL (G / A) KJAGG (L / M) CDNLLTC (W) (W) RAWD (Y / F) (T / S / N / D / K) KP (L / F / M / I) F (V / Y) APAMNT (L / F) MW (N / S / T) NPFT (E / S / Q / Y) (R / K) H (L / F / I) F (M ) (D / N / S) (E / L / Q) (L / M) G (W / L) (T / S / Y / I) L (W) PP (W / T) (K7T / S) K (R / T / K) LACGD (Y / H) G (N / T) GAM (A / S) E

wherein X1 may be any amino acid, preferably X1 is one of L, V, Ε, H, D, Μ, I or Q and characterized in that X2 may be any amino acid, preferably X2 is one of S, L5 T, A, or V.

These motifs form part of a larger conserved region in the protein, as an example, the conserved HAL3a region of Arabidopsis is given as SEQ ID NO: 8. Useful HAL3 polypeptides in the methods of the present invention are preferably in increasing order of at least 65%, 70%, 75%, 80%, 85%, 90% or 95% of sequence identity with the conserved region represented by SEQ ID NO: 8.

The HAL3 protein conserved region, as exemplified in SEQ ID NO: 8 and comprising the characteristics described above (i) to (iv), encompasses a Flavoprotein domain that can be identified using specialized data banks eg SMART (Schultz et al. (1998 ) Proc Natl.Acad Sci USA 95, 5857-5864; Letunic et al. (2002) Nucleic Acids Res 30,242-244), InterPro (Mulder et al. (2003) Nucl Acids Res 31, 315 -318), Prosite (Bucher and Bairoch (1994), A generalized profile syntax forbiomolecular sequence motifs and its funetion in automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology. Altman R., Brutlag D Karp P., Lathrop R., Searls D., Eds., Pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137 (2004)) or Pfam ( Bateman et al., NucleicAcids Research 30 (1): 276-280 (2002)). This FMN binding domain (Pfam entry PF02441, InterPro entry IPR003382) is typically found in flavoprotein enzymes. The terms "domain" and "reason" are defined in the definitions section above.

The conserved region in SEQ ID NO: 2, as represented by SEQ ED NO: 8, can also be identified in other HAL3 proteins using sequence alignment methods for comparison as described above. In some instances, the default parameters may be adjusted to modify the search stringency. For example using BLAST, the threshold of statistical significance (called the expected value ") to report pairings against database sequences can be increased to show less stringent pairings. In this way, virtually exact shortlines can be identified, including retained reasons 1 and 2 (SEQ ID). NO: 6 and 7), or pairings with one or more conservative changes in any position.

HAL3 polypeptides (at least in their native form) and their homologs typically catalyze the decarboxylation of 4'-phosphopantothenylcysteine to 4'-phosphopantetein, a step in decoenzyme A biosynthesis whose reaction can be tested in a biochemical assay; be tested by a complementation test with a mutant E. coli dfp strain (Kupke et al 2001; Yonamine et al 2004). The enzyme is also capable of decarboxylarpantothenyl cysteine to pantothenyl cysteamine. In addition, protein is involved in giving saline and osmotic stress tolerance to plants, which feature may be useful in a HAL3 bioassay.

SEQ ID NO: 2 (encoded by SEQ ID NO: 1) is an example of an HAL3 polypeptide comprising N-terminus C (i) a substrate binding helix, (ii) a His insertion motif, (iii) a motif PXMNXXMW and (iv) a substrate recognition clamp; and preferably in increasing order of at least 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with the conserved region represented by SEQ ID NO: 8. Additional examples of HAL3 polypeptides comprising characteristics ( i) through (iv) are given in Table A in the examples section below:

Homologs of an HAL3 polypeptide may also be used to carry out the methods of invention. Homologs (or protein homologs) can be readily identified using routinely techniques known in the art, such as by sequence alignment. Sequence alignment methods for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA, and TFASTA. GAP uses the Needleman and Wunsch algorithm ((1970) J Mol Biol 48: 443-453) to find alignment. of two complete sequences that maximizes the number of pairings and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The computer application for performing BLAST analysis is publicly available through the NationalCentre for Biotechnology Information. Counterparts can be readily identified using, for example, the ClustalW multi-sequence alignment algorithm (version 1.83), with standard paired alignment parameters, and a percentage scoring method. Global percentages of similarity and identity can also be determined using one of the methods available in the MatGAT computational application package (Campanella et al., BMC Bioinformatics. 2003 Jul 10; 4:29. MatGAT: anapplication that generates similarity / identity matrices using protein or DNAsequences. ). Secondary manual editing can be performed to optimize alignment between conserved motifs, as would be apparent to an art-versed person.

Sequence identity values can be determined along the entire conserved domain (as indicated above) or along the full length nucleic acid or amino acid sequence using the programs mentioned above using the default parameters.

Counterparts also include orthologs and paralogues. Orthologogalogues can be easily found by performing a so-called reciprocal blast search. This can be done by first BLAST wrapping a query string (for example, SEQ ID NO: 1 or SEQ IDNO: 2) to a BLAST search against any sequence database, such as the publicly available NCBI database. . BLASTN orTBLASTX (using default predetermined values) can be used when starting from a nucleotide sequence and BLASTP or TBLASTN (using default predetermined values) can be used when starting from a protein sequence. BLAST results can be optionally filtered. Full length sequences or filtered results or unfiltered results are then subjected to a second BLAST search against sequences from the organism from which the query sequence is derived (where the query sequence is SEQ IDNO: 1 or SEQ ID NO: 2 The second BLAST must therefore be Arabidopsis counter sequences. The results of the first and second BLAST search are then compared. A paralog is identified if a high score hit of the first BLAST is of the same species from which the query sequence is derived; An ortholog is identified if a high score hit is not the same species from which the query sequence is derived. Preferred orthologs are orthologs of AtHAL3a (SEQ ID NO: 2), AtHAL3b (SEQ ID NO: 10) or the large form of AtHAL3b (SEQ ID NO: 40). High score hits are those with a low E value. The smaller the E value, the more significant the score (or in other words the lower chance that the hit was found at random). Computation of the value E is well known in the art. For large families, ClustalW can be used, followed by a neighboring cluster tree, to help visualize related gene clustering and to identify pathologists and paralogues. In addition to E values, comparisons are also counted by percent identity. Percentage identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid sequences (or polypeptides) compared over a particular length. Preferably, HAL3 polypeptide homologs have in ascending order preferably at least 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more identity or similarity. with an unmodified HAL3 polypeptide as represented by SEQ ID NO: 2. Preferably, HAL3 polypeptide homologs are those represented by the sequences referred to in TableA.

The HAL3 polypeptide useful in the methods of the present invention may be a derivative of SEQ ID NO: 2. "Derivatives" include peptides, oligopeptides, polypeptides which may, compared to the amino acid sequence of the naturally occurring form of the protein, as shown in SEQ ID NO. : 2, comprising amino acid substitutions by non-naturally occurring amino acid residues, or non-naturally occurring amino acid residue additions. Protein derivatives as represented by the sequences listed in Table A are additional examples that may be suitable for use in the inventive methods.

Examples of HAL3 polypeptide encoding nucleic acids include but are not limited to those represented by the sequences listed in Table A. Variants of HAL3 polypeptide encoding nucleic acids may be suitable for use in the methods of the invention. Suitable variants include portions of nucleic acids encoding HAL3 polypeptides and / or nucleic acids capable of hybridizing to nucleic acids / genes encoding HAL3 polypeptides. Additional variants include splice variants and allelic variants of nucleic acids encoding HAL3 polypeptides.

The term "moiety" as defined herein refers to a DNA fragment encoding a polypeptide comprising N-terminus C-terminus characteristics (i) a de-substrate binding helix, (ii) an insertion His motif, (iii) a PXMNXXMW motif and (iv) a substrate recognition clamp; and in ascending order of preference at least 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with the conserved region represented by SEQ ID NO: 8.

A portion may be prepared, for example, by making one or more deletions to a nucleic acid encoding an HAL3 polypeptide. Portions may be used in isolation or they may be fused to other coding sequences (or non-coding) to, for example, produce a protein that combines various activities. When fused to other coding sequences, the resulting polypeptide produced by translation may be larger than predicted for the HAL3 moiety. Preferably, the moiety encodes a polypeptide having substantially the same biological activity as the HAL3 polypeptide of SEQ ID NO: 2. The moiety is typically at least 50, 100, 150 or 200 nucleotides in length, preferably at least 250, 300, 350 or 400. nucleotides in length, more preferably at least 450, 500,550, 600 or 650 nucleotides in length.

Preferably, the portion is a portion of a nucleic acid as represented by the sequences listed in Table A. More preferably, the portion is a portion of a nucleic acid represented by SEQ ID NO: 1.

The terms "fragment", "fragment of a sequence" or "part of a sequence" "portion" or "portion thereof" mean a truncated sequence of the original sequence referred to. The truncated sequence (nucleic acid or protein sequence) may vary widely in length; the minimum size being a sequence of sufficient size to provide a sequence with at least one comparable function and / or activity of the original sequence referred to or to hybridize to the nucleic acid molecule of the invention or used in the process of the invention under stringent conditions, while the maximum size is not critical. . In some applications, the maximum size is usually not substantially larger than that required to provide the desired activity (s) and / or functions (s) of the original sequence. A comparable function means at least 40%, 45% or 50%, preferably at least 60%, 70%, 80% or 90% or more of the original frequency.

Another variant of a HAL3 polypeptide-encoding nucleic acid useful in the methods of the present invention is a nucleic acid capable of hybridizing under reduced stringency conditions, preferably under stringent conditions, with an acid-derived probe as defined above, whose hybridization sequence encodes a polypeptide. comprising N-terminus C-terminus characteristics (i) a substrate binding helix, (ii) an insertion His motif, (iii) a PXMNXXMW motif and (iv) a substrate recognition clamp; and preferably has an increasing order of at least 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with the conserved region represented by SEQ ID NO: 8.

Preferably, the hybridization sequence is one that is capable of hybridizing to a nucleic acid as represented by (or probes derived from) the sequences listed in Table A, or a portion of either sequence (the target sequence). More preferably the hybridization sequence is capable of hybridizing to SEQ ID NO: 1 (or probes derived therefrom). Probes are generally less than 700 bp or 600 bp in length, preferably less than 500, 400 bp, 300 bp200 bp or 100 bp in length. Commonly, probe lengths for DNA-DNA hybridization such as Southern blotting range from 100 to 500 bp, while the hybridizing region in DNA-DNA hybridization probes such as PCR amplification are generally less than 50 but larger than 10 nucleotides. preferably they are 15.20, 25, 30, 35, 40, 45 or 50 bp in length.

The HAL3 polypeptide may be encoded by a processing variant. The term "processing variant" as used herein encompasses variants of a nucleic acid sequence in which selected introns and / or exons have been cut, substituted, displaced or added, or in which introns have been shortened or lengthened. Such variants will be one in which the substantial biological activity of the protein is maintained, which can be achieved by selectively retaining functional segments of the protein. Such union variants can be found in nature or can be made by man. Methods for making such joint variants are well known in the art.

Preferred splice variants are nucleic acid splice variants encoding an HAL3 polypeptide comprising from N-terminus to C (i) a substrate binding helix, (ii) a His-insertion motif, (iii) a PXMNXXMW motif, and (iv) a staple. substrate recognition; and having in ascending order of preference at least 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with the conserved region represented by SEQ ID NO: 8.

Further preferred splice variants of HAL3 polypeptide-encoding nucleic acids comprising characteristics as defined above are splice variants of a nucleic acid represented by any of the sequences listed in Table A. Most preferred is a processing variant of a nucleic acid sequence as represented by SEQ ID NO. : 1.

The HAL3 polypeptide may also be encoded by an allelic variant of a nucleic acid encoding a N-terminus C-terminating polypeptide (i) a desubstrate binding helix, (ii) an insertion His motif, (iii) a PXMNXXMW motif, and (iv ) a substrate recognition clamp; and in ascending order of preference at least 65%, 70%, 75%, 80%, 85%, 90% or 95% sequence identity with the conserved region represented by SEQ EDNO: 8.

Preferred allelic variants of HAL3-encoding polypeptide nucleic acids comprising characteristics as defined above are allelic variants of a nucleic acid as represented by any of the sequences listed in Table A. Most preferred is an allelic variant of a nucleic acid sequence as represented by SEQ ID NO: 1.

Directed evolution (or gene shuffling) can also be used to generate nucleic acid variants encoding HAL3 polypeptides. Site directed mutagenesis can be used to generate nucleic acid variants encoding HAL3 polypeptides. Several methods are available to achieve site-directed mutagenesis, the most common of which are PCR-based methods (Current Protocols in Molecular Biology. Wiley).

Eds.).

Nucleic acids encoding HAL3 polypeptides may be derived from any natural or artificial source. Nucleic acid may be modified from its native form in composition and / or environmental genetics by deliberate human manipulation. Preferably the nucleic acid HAL3 is from a plant, still preferably from a planted plant, more preferably from the Brassicaceae family, most preferably the nucleic acid is from Arabidopsis thaliana.

Increased expression of a nucleic acid encoding a HAL3 polypeptide, preferably in aerial parts of a plant, can be accomplished by introducing a genetic modification (preferably nolocus of a HAL3 gene). The locus of a gene as defined herein is taken to mean a genomic region, which includes the gene of interest e1OKB before or after the coding region.

Genetic modification may be introduced, for example, by any one or more of the following methods: T-DNA activation, TILLING, and homologous recombination (as described in the definitions section) or by introducing and enhancing expression of a nucleic acid encoding a HAL3 polypeptide preferably in tissues. Following introduction of genetic modification, an optional step of deselecting for enhanced expression of a nucleic acid encoding a HAL3 factor polypeptide is preferentially in expanding tissues, whose increased expression generates plants having increased yield.

Identification by T-DNA activation results in plant-transgenics that show dominant phenotypes due to modified expression of genes near the introduced promoter. The promoter to be introduced can be any promoter capable of enhancing nucleic acid expression encoding a preferably expanding plant HAL3 polypeptide of a plant.

A genetic modification can also be introduced into a gene encoding a HAL3 polypeptide using the technique of TLLrNG (Locally Induced Targeted Lesions in Genomes). Plants bearing such mutant variants have increased expression of a nucleic acid encoding an HAL3 polypeptide preferably in expanding vegetable tissues.

T-DNA activation and TILLING are examples of technologies that enable the generation of genetic modifications comprising preferentially enhancing expression of a nucleic acid encoding a HAL3 polypeptide in expanding plant tissues.

The effects of the invention may also be reproduced using homologous recombination. Preferably the nucleic acid to be targeted is the region controlling the natural expression of a nucleic acid encoding an HAL3 polypeptide in a plant. A specific promoterfraction for expression in aerial parts is introduced into this region, replacing it partially or substantially all of it.

A preferred method for introducing a genetic modification (which in this case need not be at the locus of an HAL3 gene) is to preferentially introduce and express in the aerial part of a plant a nucleic acid encoding an HAL3 polypeptide as defined above. Nucleic acid to be introduced into a plant may be a full-length nucleic acid or may be a hybridizing portion or sequence or other nucleic acid variant as defined above.

The methods of the invention rely on preferentially increased expression of a nucleic acid encoding an HAL3 polypeptide in the shoot tissue of a plant, preferably in the cell expansion zone of vegetative shoots.

The invention also provides genetic constructs and vectors for facilitating introduction and / or preferential expression of nucleic acid sequences useful in the methods according to the invention, in aerial parts, preferably in expanding tissues of a shoots plant.

Therefore, a gene construct is provided comprising:

(i) a nucleic acid encoding an HAL3 polypeptide as defined above;

(ii) one or more control sequences, of which at least one is an aerial-specific promoter operably linked to the nucleic acid of (i).

Constructs useful in the methods according to the present invention may be constructed using recombinant DNA technology well known to those skilled in the art. The gene constructs may be inserted into commercially available vectors suitable for transformation into plants and suitable for dogene expression of interest in the transformed cells. The invention therefore provides us with a gene construct as defined above in the methods of the invention.

Plants are transformed with a vector comprising the sequence of interest (i.e., a nucleic acid encoding a HAL3 polypeptide). The skilled artisan is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells containing the sequence of interest. The sequence of interest is operatively linked to one or more control sequences (at least one promoter).

Suitable promoters, which are functional in plants, are generally known. They may take the form of constitutive or inducible promoters. Suitable promoters may enable developmental and / or tissue specific expression in multicellular eukaryotes; thus, specific leaf, root, flower, seed, stoma, tuber or fruit promoters may be advantageously used in plants.

Different plant promoters usable in plants are such as, for example, USP, LegB4, DC3 promoter or parsley ubiquitin opromotor.

For plant expression, the nucleic acid molecule must, as described above, be operably linked to or comprise a suitable promoter expressing the gene at the right time and a specific manner or cell. Usable promoters are constitutive promoters (Benfey et al., EMBO J. 8 (1989) 2195-2202), such as those originating from plant viruses such as 35S CAMV (Franck et al., Cell 21 (1980) 285-294). , 19S CaMV (see also U.S. Patent 5,352,605 and International Patent 84/02913), 34S FMV (Sanger et al., Plant.Mol. Biol., 14, 1990: 433-443), the parsley ubiquitin promoter, or promoters. such as the Rubiscis small subunit promoter described in U.S. Patent 4,962,028 or the PRP1 plant promoters [Ward et al., Plant. Mol. Biol. 22 (1993)], SSU, PGEL1, OCS [Leisner (1988) Proc Natl Acad Sci USA 85 (5): 2553-2557], lib4, usp, but [Cornai (1990) Plant Mol Biol 15 (3): 373 -381], STLS1, ScBV (Schenk (1999) PlantMol Biol 39 (6): 1221-1230), B33, SAD1 or SAD2 (flax promoters, Jainet al., Crop Science, 39 (6), 1999: 1696- 1701) or us [Shaw et al. (1984) Nucleic Acids Res. 12 (20): 7831-7846]. Additional examples of plant constituent promoters are the sugar beet V-ATPase promoters (International Patent 01/14572). Examples of constitutive synthetic promoters are the Super promoter (International Patent 95/14098) and G-box derived engines (International Patent 94/12015). If appropriate, chemically inducible promoters may also be used, compare European Patent A 388186, European Patent A 335528, International Patent 97/06268. Stable constitutive expression of proteins according to the invention a plant may be advantageous. However, inducible expression of the polypeptide of the invention is advantageous if late expression before harvesting is advantageous as metabolic manipulation may lead to plant growth retardation.

Plant gene expression can also be facilitated through a chemically inducible promoter (for a review, see Gatz1997, Annu. Rev. Plant Physiol. Plant Mol. Biol., 48: 89-108). Chemically inducible promoters are particularly suitable when it is desired to express the gene in a specific time manner. Examples of such promoters are a salicylic acid inducible promoter (International Patent 95/19443), and abscisic acid inducible promoter (European Patent 335 528), a tetracycline inducible promoter (Gatz et al. (1992) Plant J. 2,397-404 ), a cyclohexanol or ethanol inducible promoter (International Patent 93/21334) or others as described herein.

Other suitable promoters are those that react to biotic or abiotic stress conditions, for example the pathogen-induced genePRP1 promoter (Ward et al., Plant. Mol. Biol. 22 (1993) 361-366), the heat-inducible hsp80 promoter. tomato paste (U.S. Patent 5,187,267), the potato cold-inducible alpha-amylase promoter (International Patent 96/12814) or the injury-inducible pinll promoter (European Patent AO 375,091) or others as described herein .

Preferred promoters are in particular those which induce gene expression in tissues and organs, in seed cells, such as endosperm cells and developing embryo cells. Suitable promoters are the rapeseed napin gene promoter (U.S. Patent 5,608,152), the USP Vicia faba promoter (Baeumlein et al., MolGen Genet, 1991, 225 (3): 459-67), the oleosin promoter. Arabidopsis (International Patent 98/45461), the Phaseolusvulgaris phaseolin promoter (U.S. Patent 5,504,200), the Bce4 promoter of (International Patent 91/13980), the bean arc5 promoter, the carrot DcG3 promoter, or the promoter. Legumina B4 (LeB4; Baeumlein et al., 1992, Plant Journal, 2 (2): 233-9), and promoters that cause expressively specific monocotyledonous plants such as corn, barley, wheat, rye, rice and the like. . Advantageous specific seed promoters are the sucrose binding protein promoter (International Patent 00/26388), the phaseolin promoter and the napin promoter. Suitable promoters that should be considered are the barley genelpt2 or Iptl promoter (International Patent 95/15389 and International Patent 95/23230), and the promoters described in International Patent 99/16890 (barley hordein gene, rice glutelin gene, rice orizine dogene, rice prolamine gene, gliadinade gene wheat, the wheat glutelin gene, the corn zein gene, the oat deglutelin gene, the sorghum casirin gene, and the decent secalin gene). Additional suitable promoters are Amy32b, Amy 6-6 and Aleurain [U.S. Patent 5,677,474], Bce4 (rapeseed) [U.S. Patent 5,530,149], glycine (soy) [European Patent 571,741], phosphoenolpyruvate carboxylase (soybean) ) [Japanese Patent 06/62870], ADR12-2 (soybean) [International Patent 98/08962], isocitrate lyase (rapeseed) [US Patent 5,689,040] or α-amylase (barley) [European Patent 781,849]. Other enhancers that are available for gene expression in plants are specific leaf motors such as those described in DE-A 19644478 or light-regulated promoters such as, for example, the petE-depregnant promoter.

Additional suitable plant promoters are the cytosolic FBPase promoter or the potato ST-LSI promoter (Stockhaus et al., EMBO J.8, 1989, 2445), the Glycine max phosphoribosyl pyrophosphate amidotransferase promoter (GenBank Accession No. U87999) or the specific node promoter described in European Patent AO 249,676.

In a preferred embodiment, the sequence of nucleic acids encoding an HAL3 polypeptide is operably linked to a specific aerial part. An aerial specific promoter suits any promoter capable of directing expression of the gene of interest in plant aerial vegetative tissues. Preferably, the specific air promoterpart is capable of preferentially directing expression of the genus of interest in vegetative airway expanding tissues, i.e. cell expanding nazone of vegetative airways. Reference herein to "preferentially" directing expression in the shoot is adopted to mean directing expression of any sequence operably linked to this in the vegetative aerial cell expansion zone substantially for the exclusion of directed expression elsewhere in the plant, in addition to any residual expression due to promoter evasive expression. The specific aerial promoter may be either a natural or a synthetic promoter.

Preferably, the specific shoot promoter is an isolated promoter of a beta-expansin gene, such as a rice beta-expansin EXBP9 promoter (International Patent 2004/070039), as represented by SEQ ID NO: 5 or a similar strength promoter and / or a promoter with a similar expression pattern as the rice betaexpansine promoter (ie a functionally equivalent promoter).

It should be clear that the applicability of the present invention is not restricted to nucleic acid encoding an HAL3 polypeptide represented by SEQ ID NO: 1, nor is the applicability of the invention restricted to expression of a nucleic acid encoding an HAL3 polypeptide when directed by a beta expansin promoter.

Optionally, one or more terminator sequences (also a control sequence) may be used in the construction introduced in a plant. Additional regulatory elements may include transcriptional as well as translational enhancers. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Such sequences must be known or readily obtainable by a person skilled in the art.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl. Other control sequences (in addition to promoter, enhancer, silencer, intron sequences, 3'UTR and / or 5'UTR regions) may be protein and / or RNA stabilizing agents.

For the detection and / or selection of successful transfer of nucleic acid sequences as described in the sequence protocol used in the process of the invention, it is advantageous to use marker genes (= reporter genes). These marker genes enable the identification of a successful transfer of nucleic acid molecules through a number of different principles, for example, by visual identification with the aid of fluorescence, luminescence or the wavelength variation of light that is discernible by the human eye, by resistance to herbicides or antibiotics, through which they are known as nutritional markers (auxotrophism markers) or antinutritive markers, through enzymatic assays or through phytohormones. Examples of such markers that may be mentioned are GFP (= fluorescent green protein); the luciferin / luciferase system, β-galactosidase with its colored substrates, for example, X-Gal, herbicide resistance to, for example, imidazolinone, glyphosate, phosphinothricin or sulfonylurea, and aantibiotic resistance to, for example, bleomycin, hygromycin, streptomycin, kanamycin, tetracycline, chloramphenicol, ampicillin, gentamicin, geneticin (G418), spectinomycin or blasticidine, to name but a few, nutritional markers such as the use of mannose or xylose, or antinutritive markers such as 2-deoxyglucose resistance. This list is as small a number of bullets as possible. The skilled worker is very familiar with such markers. Different markers are preferred depending on the organism and the selection method.

The present invention also encompasses viable plants by the methods according to the present invention. The present invention therefore provides such viable plants, plant parts or plant cells by the method according to the present invention, the plants or parts of which herein comprise a nucleic acid transgene encoding an HAL3 polypeptide under the control of an aerial-specific promoter.

The invention also provides a method for producing transgenic plants having increased production relative to control plants, comprising introducing and preferentially expressing a nucleic acid encoding a HAL3 polypeptide in the aerial part of a plant.

Host plants for nucleic acids or the vector used in the method according to the invention, the expression drawer or listener construct are in principle advantageously all plants which are capable of synthesizing the polypeptides used in the inventive method.

More specifically, the present invention provides a method for the production of transgenic plants having increased yield which method comprises:

(i) introducing and preferably expressing a nucleic acid encoding an HAL3 polypeptide in the aerial part of a plant; and

(ii) cultivate the plant cell under conditions that promote plant growth and development.

Nucleic acid can be introduced directly into a plant cell or into the plant itself (including introduction into a tissue, organ or any other part of a plant). According to a preferred feature of the present invention, nucleic acid is preferably introduced into a plant by transformation.

The transfer of exogenous genes to the genome of a plant is called transformation. In doing so the methods described for plant transformation and regeneration from plant tissue or plant cells are used for transient or stable transformation. For selecting transformed plants, the plant material obtained in the transformation is, as a rule, subject to selective conditions so that transformed plants can be distinguished from unprocessed plants. For example, seeds obtained in the manner described above may be planted and, after an initial period of growth, subjected to appropriate spray selection. An additional possibility is to grow the seeds, if appropriate after sterilization, on agar plates using a suitable deselecting agent so that only transformed seeds can grow to plants. Additional advantageous transformation methods, particularly for plants, are known to the skilled worker and are described here below.

Generally after transformation, plant cells or cell clumps are selected for the presence of one or more markers that are encoded by expressible genes in plants co-transferred with the gene of interest, and the transformed material is regenerated into an entire plant.

As mentioned Agrobacteria transformed with an expression vector according to the invention may also be used in a manner known per se for the transformation of plants such as experimental plants such as Arabidopsis or crop plants such as cereals, maize, oats, rye, barley. , wheat, soybean, rice, cotton, sugar beet, canola, sunflower, flax, hemp, potato, tobacco, tomato, carrot, bell pepper, rapeseed, tapioca, cassava, arrowroot, alfalfa, lettuce and various tree species, nuts, and vines, in particular oil-containing crop plants such as soybean, peanut, castor, sunflower, corn, cotton, flax, rapeseed, coconut, palm oil, saffron (<Carthamus tinctorius) or cocoa, for example, by bathing scarified leaves or leaf segments in an agrobacterial solution and subsequently cultivating them in suitable medium.

In addition to the transformation of somatic cells, which then have to be regenerated in intact plants, it is also possible to transform cells of plant meristems and in particular those cells that develop into gametes. In this case, transformed gametes follow natural plant development, generating transgenic plants. Thus, for example, Arabidopsis seeds are treated with agrobacteria and seeds are obtained from developing plants from which a certain proportion is transformed and thus transgenic [Feldman, KA and Marks MD (1987). Mol Gen Genet 208: 274-289; Feldmann K (1992). In: C. Koncz, N-H Chua and J Shell, eds, Methods in Arabidopsis Research. WordScientific, Singapore, pp. 274-289]. Alternative methods are based on repeated removal of inflorescences and incubation of the rosette's nocturnal excision site with transformed agrobacteria, whereby seedlings can similarly be obtained at a later time (Chang (1994). Plant J. 5: 551-558; Katavic (1994), Mol Gen Genet, 245: 363-370). However, an especially effective method is the vacuum infiltration method with its modifications such as the "flower bud" method. In the case of vacuum infiltration of Arabidopsis, intact plants under reduced pressure are treated with an agrobacterial suspension [Bechthold, N (1993). C R Acad Sci Paris Life Sci, 316: 1194-1199], while in the case of the "flower bud" method the developing floral tissue is briefly incubated with a surfactant-treated agrobacterial suspension [Clough, SJ und Bent, AF (1998). The Plant J. 16, 735-743]. A certain proportion of transgenic seeds are collected in both cases, and thus can be distinguished from non-transgenic seeds by cultivating in the selective conditions described above. In addition, stable transformation of plastids is advantageous in that plastids are inherited maternally reducing or eliminating the risk of transgene flow through most crop pollen. Transformation of the chloroplast genome is usually achieved by a process, which has been schematically presented in Klaus etal., 2004 [Nature Biotechnology 22 (2), 225-229]. Briefly the sequences to be transformed are cloned together with a selectable marker gene between flanking sequences homologous to the dechloroplast genome. These homologous flanking sequences direct specific site integration in the plastoma. Plastid transformation has been described for many different plant species and an overview can be taken from Bock (2001) Transgenic plastids in basic research and plant biotechnology. J MolBiol. 2001 Sep 21; 312 (3): 425-38 or Maliga, P (2003) Progress towards commercialization of plastid transformation technology. Trends Biotechnol.21, 20-28. Further biotechnological progress has recently been reported in the form of marker free plastid transformants, which may be produced by a transient cointegrated marker gene (Klaus et al., 2004, Nature Biotechnology 22 (2), 225-229).

Genetically modified plant cells can be regenerated by all methods with which the skilled worker is familiar. Suitable methods can be found in the publications mentioned above by S.D. Kung and R. Wu, Potrykus or Hofen and Willmitzer.

Following DNA transfer and regeneration, computationally transformed plants can be evaluated, for example using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, or quantitative PCR, all techniques well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first-generation (or IT) transformed plant may be self-fertilized to generate second-generation homozygous (or T2) transformants, and T2 plants further propagated by classical breeding techniques. Generated transformed organisms may adopt a variety of shapes. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); grafts of transformed and unprocessed tissues (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention further extends to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that it exhibits the same trait (s). ) genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also includes host cells containing an isolated nucleic acid encoding an HAL3 polypeptide. Preferred host cells according to the invention are plant cells.

The invention also extends to cut-off parts of a plant such as, but not limited to seeds, leaves, fruits, flowers, stems, rhizomes, tubers and bulbs. The invention furthermore relates to derived products, preferably directly derived, from a cut-off part of such a plant, such as pellets or dry powders, oil, fat, fatty acids, starch or proteins.

The present invention also encompasses use of nucleic acids encoding HAL3 polypeptides and use of HAL3 polypeptides to enhance plant production as defined above in the methods of the invention.

Nucleic acids encoding HAL3 polypeptides, or HAL3 polypeptides, may find use in breeding programs in which a DNA marker that can be genetically linked to a HAL3 gene is identified. Nucleic acids / genes, or HAL3 polypeptides may be used to define a molecular marker. This DNA or protein marker can then be used in breeding programs to select plants having increased yield as defined above in the methods of the invention.

Allelic variants of a HAL3 nucleic acid / gene may also find use in marker-aided breeding programs. Such breeding programs sometimes require reduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis; alternatively, the program may start with a collection of allelic variants of the so-called "natural" origin caused unintentionally. Identification of allelic variants then occurs, for example, by PCR. This is followed by a step to select higher allelic variances of the sequence in question and which generate increased production. Selection is typically performed by monitoring growth performance of plants containing different allelic variants of the sequence in question. Growth performance can be monitored in a home-grown house or in the field. Additional optional steps include crossing plants in which the superior allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

A nucleic acid encoding an HAL3 polypeptide may also be used as probes to genetically map the genes of which they are a part of, and as markers for, traits linked to such genes. Such information may be useful in plant breeding to develop strains with desired phenotypes. Such use of HÃL3 nucleic acids requires only one nucleic acid sequence of at least 15 nucleotides in length. HAL3 nucleic acids can be used as markers of restriction fragment length polymorphism (RFLP). Southern blots (Sambrook J5Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) of restriction digested plant genomic DNA can be probed with HAL3 nucleic acids. The resulting banding pattern can then be subjected to genetic analysis using computer programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, nucleic acids may be used to probe Southern blots containing endonuclease-treated genomic DNAs from a set of individuals representing ancestor and progeny of a defined genetic crossover. Segregation of DNA polymorphisms is observed and used to calculate the position of HAL3 nucleic acid in the mapagenetic previously obtained using this population (Botstein et al. (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4: 37-41. Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 cross-breeding populations, inbreeding populations, randomly crossed populations, nearby isogenic strains, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and cited references. in this one).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify DNA sequence differences between the ancestors of the cross-mapping region corresponding to the current sequence of nucleic acids. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having increased yield as described above. This increased production may also be combined with other economically advantageous features, such as additional production enhancing features, tolerance to other abiotic and biotic stresses, features modifying various architectural features and / or biochemical and / or physiological features.

Detailed Description for MADS15 above

Surprisingly, it has now been found that modular expression in a plant of a nucleic acid encoding a MADS15 polypeptide generates plants having improved production characteristics relative to control plants. The particular class of MADS15 polypeptides suitable for improving plant yield characteristics is described in detail below.

The present invention provides a method for enhancing plant production characteristics over control plants, comprising modulating expression in a plant of a nucleic acid encoding a MADS15 polypeptide.

Any reference hereinafter to a "protein useful in the methods of the invention" is taken to mean an MADS15 polypeptide as defined herein. Any reference hereinafter to a "useful nucleic acid in the methods of the invention" is taken to mean the nucleic acid capable of encoding such a MADS15 polypeptide.

A preferred method for modulating (preferably enhancing) expression of a nucleic acid encoding a protein useful in the methods of the invention is to introduce and express into a plant a nucleic acid encoding a protein useful in the methods of the invention as defined below.

The nucleic acid to be introduced into a plant (and therefore useful for carrying out the methods of the invention) is any nucleic acid encoding the type of protein that will now be described, hereinafter also called "MADS15 nucleic acid" or "MADS15 gene".

The term "MADS15 polypeptide" as defined herein refers to a protein falling into the FUL-like MADS box protein group as outlined by Adam et al. (J. Mol. Evol. 62, 15-31). FUL-like MADSbox proteins are part of the SQUAMOSA subfamily and have the typical IMKCc architecture: an N-terminated MADS (M) domain, followed by an intervening domain (I) involved in dimerization, a highly conserved dekeratin domain (K), and a variable domain (C ) at end C, which is responsible for protein binding in interaction or transcriptional activation (De Bodt et al. Trends in Plant Science 8, 475-483).

Plant MADS15 polypeptides can also be identified by the presence of certain conserved motifs. The presence of these conserved motifs can be identified using methods for sequence alignment for comparison as described above. In some instances, the default parameters can be adjusted to modify the search stringency. For example, using BLAST, the statistical significance threshold (called the "expected" value) for reporting matches against database sequences can be increased to show less stringent matches. In this way, practically exact short pairings can be identified. Upon identification of an MADS15 polypeptide by the presence of these motifs, a person skilled in the art can readily derive the corresponding nucleic acid encoding opolipeptide comprising the relevant motifs, and use sufficient length of contiguous nucleotides thereof to perform any or more of the gene silencing methods described above ( for the reduction or substantial elimination of expression of an endogenous MADS15 gene).

Typically, the presence of at least one of motifs 1-4 should be sufficient to identify any query sequence as a MADS15, however, the presence of at least motifs 1 and 2 is preferred. The consensus sequence provided is based on the monocot sequences given in the sequence listing. One skilled in the art will be well aware that the consensus sequence may vary in some way if additional or different sequences (eg, dicotyledonous plants) are used for comparison.

Reason 1, located in the MADS domain (SEQ ID NO: 48): L (L / V) KKA (H / 1S0EIS (VI) L (C / Y) DAE (V / I / L) (A / G) (L / A / V) (I / V) (W) FS (T / P / A / N) KGKLYE (Y / F) (Y / F) (T / S) (D / N / E) S (K / C / R / S) M (D / E) (N / I / R / K) IL (E / D) R.

Preferably, this conserved sequence 1 motif has the following:

LLKKAHEISVLCDAEVA (L / A / V) I (W) FS (T / P) KGKLYEYATDS (C / R) M (D / E) (R / K) ILER,

more preferably the conserved motif of sequence 1 theme sequence:

LLKKAHEISVLCDAEVAAIVFSPKGKLYEYATDSRMDKILER

Reason 2, located in domain K (SEQ ID NO: 49): KLK (AyS) (K / R) (V / I) AND (A / T / S) (L / I) (Q / N) (K / R / N) (S / C / R) (Q / H) (R / K) HLMGE

Preferably, this conserved sequence 2 motif has the following:

KLKAK (V / I) AND (A / T) (L / I) QK (S / C) (Q / H) (R / K) HLMGEM Preferably, this conserved sequence motif has 2 sequence:

KLKAKIETIQKCHKHLMGE

Optionally, the MADS15 polypeptide or homologue thereof may comprise in domain C motif 3 and / or motif 4: motif 3 (SEQ ID NO: 50): Q (P / Q / V / A) QTS (S / F) (S / F ) (Y / F) (Y / F) (Y / C / F) (Y / M) Reason 4 (SEQ ID NO: 51) :( G / A / V / L) (W / H) XWMX (S /H)

characterized in that the first residue X in motif 4 can be any amino acid, but preferably L, P or H; and characterized by the fact that the second residue X can be any amino acid, but preferably V or L.

Alternatively, the C domain (starting behind the keratin domain, ie at R175 in SEQ ID NO: 44) may be characterized by increased glutamine occurrence (usually 3.93%, here at least 8.77% with a maximum of 26.73%). ), in addition, alanine, proline, and / or serine content may also be increased (above 7.8%, 4.85% and 6.89% respectively).

Alternatively formulated, a preferred MADS15 protein useful in the methods of the present invention comprises at least one N-terminal MADS domain corresponding to the SMART SM00432 domain (PfamPF00319) followed by a Keratin domain corresponding to the K-box region defined in Pfam as PFO1486, more preferably the proteinMADS15 has in its MADS domain the conserved signature of SEQ IDNO: 48 and in its domain K the conserved signature of SEQ ID NO: 49, more preferably, the protein MADS15 has the sequence given in SEQ ID NO: 44.

Examples of proteins useful in the methods of the invention and nucleic acids encoding them are given below in Table D of Example 8.

Also useful in the methods of the invention are homologues of any of the amino acid sequences given in table D.

Also useful in the methods of the invention are derived from any of the polypeptides given in Table D or orthologs or paralogs of any of the above mentioned SEQ ID NOs. A preferred derivative is a derivative of SEQ ID NO: 44. Derivatives of the polypeptides given in Table D are additional examples which may be suitable for use in the methods of the invention. Derivatives useful in the methods of the present invention preferably have biological and functional activity similar to the unmodified protein from which they are derived.

The invention is illustrated by transforming plants with the Oryza sativa nucleic acid sequence represented by SEQ ID NO: 43, encoding the polypeptide sequence of SEQ ID NO: 44, however the performance of the invention is not restricted to these sequences. The methods of the invention may advantageously be performed using any nucleic acid encoding a protein useful in the methods of the invention as defined herein, including orthologs and paralogs, such as any nucleic acid sequences given in table D. The amino acid sequences given in table D may be considered as serortologists and paralogues of the MADS15 polypeptide represented by SEQ IDNO: 44.

Orthologs and paralogues can easily be found by performing a so-called reciprocal blast search. Typically, this involves a first BLAST involving performing BLAST with a query string (for example, using any of the sequences listed in table D) against any sequence database, such as NCBI's publicly available database. BLASTN or TBLASTX (using default predetermined values) are generally used when starting from a nucleotide sequence, and BLASTP or TBLASTN (using default predefined values) when starting from a protein sequence. BLAST results can be optionally filtered. The full-length sequences of either the filtered or unfiltered results are then BLASTed back (according to BLAST) against the organism sequences from which the query sequence is derived (where the query sequence is SEQ ID NO: 43 or SEQ ID NO: 44, the second BLAST must therefore be against Oryza sativa sequences). The results of the first BLAST second are then compared. A parallel is identified if a high score of the first BLAST is of the same species from which the query sequence is derived, a return BLAST then ideally results in the query sequence as the highest hit; an ortholog is identified if a high score hit in the first BLAST is not the same species from which the query sequence is derived, and preferably results by returning BLAST in the query sequence being among the highest hits.

High score hits are those with a low value E. The lower the E value, the more significant the score (or in other words the lower the chance that the hit was found at random). E-value computation is well known in the art. . In addition to E values, comparisons are also scored by percent identity. Percent Identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. For large families, ClustalW can be used, followed by a neighboring cluster tree, to help visualize related gene clustering and to identify orthologs and paralogs.

Table D gives examples of orthologs and paralogs of the MADS15 protein represented by SEQ ID NO 44. Additional orthologs and paralogs can be readily identified using the BLAST procedure described above.

The proteins of the invention are identifiable by the presence of the conserved MADS and / or keratin domain (s) (shown in Figure 5). There are expert databases for identifying domains, for example, SMART (Schultz et al. (1998) Proc Natl Acad Sci USA 95, 5857-5864 Letunic et al (2002) Nucleic Acids Res 30, 242-244, InterPro (Mulderet al., 2003) Nucl Acids Res. 31, 315-318, Prosite (Bucher and Bairoch (1994), A generalized profile syntax for biomolecular sequences motifs andits function in automatic sequence interpretation. (In) ISMB-94; Proceedings2nd International Conference on Intelligent Systems for Molecular Biology.Altman R. , Brutlag D., Karp P., Lathrop R., Searls D., Eds., Pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137, (2004), or Pfam (Bateman et al., Nucleic Acids Research 30 (1): 276-280 (2002). A toolkit for in silico analysis of protein sequences is available on the Ex proteomics server. PASY (hosted by the Swiss Institute of Bioinformatics (Gasteiger et al., ExPASy: the proteomicsserver for in-depth protein knowledge and analysis, Nucleic Acids Res.31: 3784-3788 (2003)).

Domains can also be identified using derothine techniques, such as by sequence alignment. Methods for sequence alignment for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA, and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the global (i.e. spanning complete sequences) alignment of two sequences that maximizes the number of pairings and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The computer application for performing BLAST analysis is publicly available through the NationalCentre for Biotechnology Information (NCBI). Counterparts can be readily identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the paired alignment standard parameters, and a percentage scoring method. Global percentages of similarity and identity can also be determined using one of the available methods. in the MatGAT computer application package (Campanella et al., BMC Bioinformatics. 2003 Jul10; 4:29. MatGAT: an application that generates similarity / identity matrices using protein or DNA sequences). Secondary manual editing can be performed to optimize alignment between retained subjects, as would be apparent to a person skilled in the art. In addition, instead of using full-length sequences for identifying homologs, specific domains (such as the MADS or keratin domain, or one of the motives defined above) may also be used. The sequence identity values, which are given below in Example 10 as a percentage, were determined over the entire sequence of nucleic acids or amino acids, and / or over selected domains or conserved motive (s) using the programs mentioned above. using the default parameters.

In addition, a MADS15 protein may also be identifiable by its ability or inability to bind to or interact with other proteins. DNA binding activity and protein-protein interactions can be readily determined in vitro or in vivo using techniques well known in the art. Examples of in vitro assays for DNA binding activity include: gel retardation analysis using known MADS-box DNA binding domains (West et al. (1998) NuclAcid Res 26 (23): 5277-87), or assays of a yeast hybrid. An example of an in vitro assay for protein-protein interactions is hybrid yeast analysis (Fields and Song (1989) Nature 340: 245-6). Proteins known to interact with OsMADS 15 include other MADS proteins (such as those involved in the complex). (signaling signaling), receptor-like kinases such as Clavatal and Clavata2, Erecta, BRI1 or RSK. Additional details are provided in Example 13.

Nucleic acids encoding proteins useful in the methods of the invention need not be full length nucleic acids, since performance of the methods of the invention is not relied upon to use full length nucleic acid sequences. Examples of nucleic acids suitable for use in carrying out the methods of the invention include the nucleic acid sequences given in Table D, but are not limited to those sequences. Nucleic acid variants may also be useful for practicing the methods of the invention. Examples of such nucleic acid variants include portions of nucleic acids encoding a protein useful in the methods of the invention, nucleic acids hybridizing nucleic acids encoding a protein useful in the methods of the invention, nucleic acid splice variants encoding a protein useful in the methods of the invention, allelic variants of nucleic acids encoding a protein useful in the methods of the invention and nucleic acid variants encoding a protein useful in the methods of the invention that are obtained by gene shuffling. The terms portion, hybridization sequence, processing variant, allelic variant, and gene shuffling will now be described.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing into a plant a portion of any of the nucleic acid sequences given in table D, or a portion of a nucleic acid encoding an ortholog, paralog or homologous to any of the amino acid sequences given in table D.

Portions useful in the methods of the invention encode a polypeptide falling within the definition of a nucleic acid encoding a protein useful in the methods of the invention as defined herein and having substantially the same biological activity as the amino acid sequences given in Table D. Preferably, the portion is a portion. of any of the nucleic acids given in Table D. The portion is typically at least 400 consecutive nucleotides in length, preferably at least 600 consecutive nucleotides in length, more preferably at least 700 consecutive length nucleotides, and more preferably at least 800 consecutive nucleotides in length, the nucleotides being at least 400 consecutive nucleotides in length. consecutive nucleotides being any of the nucleic acid sequences given in Table D. More preferably the portion is a portion of the nucleic acid of SEQ ID NO: 43.

Preferably, the portion encodes an amino acid sequence comprising (any or more of) the MADS domain and the keratin as defined herein.

A portion of a nucleic acid encoding a MADS15 protein as defined herein may be prepared, for example, by making one or more nucleic acid deletions. The portions may be used in isolation or they may be fused to other decoding (or non-coding) sequences to, for example, produce a protein that combines various activities. When fused to other coding sequences, the resulting polypeptide produced by translation may be larger than predicted for the MADS15 protein moiety.

Another variant of nucleic acid useful in the methods of the invention is a nucleic acid capable of hybridizing under reduced stringency conditions, preferably under stringent conditions, to a nucleic acid encoding a MADS15 protein as defined herein, or with a portion as defined herein.

Hybridization sequences useful in the methods of the invention encode a polypeptide having a MADS and / or keratin domain (see alignment of Figure 6) and having substantially the same biological activity as the MADS15 protein represented by any of the amino acid sequences given in table D. A The hybridization sequence is typically at least 400 consecutive nucleotides in length, preferably at least 600 consecutive nucleotides in length, more preferably at least 700 consecutive length nucleotides, and more preferably at least 800 consecutive nucleotides in length, the consecutive nucleotides being any one of the acid sequences. Preferably, the hybridization sequence is one which is capable of hybridizing to any of the nucleic acids given in table D, or to a portion of any of these sequences, a portion being as defined above. More preferably, the hybridization sequence is capable of hybridizing to a nucleic acid as represented by SEQ ID NO: 43 or to a portion of this. Preferably, the hybridization sequence encodes an amino acid sequence comprising any or more of the motifs or domains as defined herein.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing into a plant a nucleic acid capable of hybridizing to any of the nucleic acids given in table D, or comprising introducing and expressing in a plant a nucleic acid. capable of hybridizing to a nucleic acid encoding an ortholog, paralog or homologue of any of the nucleic acid sequences given in table D.

Another nucleic acid variant useful in the methods of the invention is a processing variant encoding an MADS15 protein as defined above.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing into a plant a processing variant of any of the nucleic acid sequences given in table D, or a processing variant of a nucleic acid encoding a ortholog, paralog or homologue of any of the amino acid sequences given in table D.

Preferred splice variants are nucleic acid splice variants represented by SEQ ID NO: 43 or a nucleic acid-processing variant encoding a SEQ ID NO: 44 ortholog or paralog. Preferably, the processing-encoded amino acid sequence comprises any one. or more of the reasons or domains as defined here.

Another useful nucleic acid variant for carrying out the methods of the invention is an allelic variant of a nucleic acid encoding a MADS15 protein as defined above. Allelic variants useful in the methods of the present invention have substantially the same biological activity as the MADS15 protein of SEQ ID NO: 44.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing in an plant an allelic variant of any of the nucleic acids given in table D, or comprising introducing and expressing a plant an allelic variant of an acid. nucleic acid encoding aortologist, paralog or homologue of any of the amino acid sequences given in table D.

Preferably, the allelic variant is an allelic variant of SEQ ID NO: 43 or an allelic variant of a nucleic acid encoding aortologist or paralog of SEQ ID NO: 44. Preferably, the amino acid sequence encoded by the allelic variant comprises any one or more of the motifs or domains. as defined in this place.

An additional nucleic acid variant useful in the methods of the invention is a nucleic acid variant obtained by scrambling. Gene shuffling or directed evolution can also be used to generate nucleic acid variants encoding MADS15 proteins as defined above.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing into a plant a variant of any of the nucleic acid sequences given in Table D, or comprising introducing and expressing in a plant a variant of an acid. nucleicocoding an ortholog, paralog or homologue of any of the amino acid sequences given in table D, whose variant nucleic acid is obtained by gene shuffling.

Preferably, the gene shuffling variant nucleic acid encodes an amino acid sequence comprising any one or more of the motifs or domains as defined herein.

In addition, nucleic acid variants can also be obtained by site-directed mutagenesis. Several methods are available to achieve site-directed mutagenesis, the most common being PCR-based methods (Current Protocols in Molecular Biology. Wiley Eds.).

Nucleic acids encoding MADS15 proteins may be derived from any natural or artificial source. Nucleic acid may be modified from its native form in composition and / or environmental genetics by deliberate human manipulation. Preferably the nucleic acid encoding MADS15 is from a plant, still preferably from a monocot plant, more preferably from the Poaceae family, more preferably the nucleic acid is from Oryza sativa.

Any reference herein to a MADS15 protein is therefore adopted to mean a MADS15 protein as defined above. Any nucleic acid encoding such a MADS15 protein is suitable for use to perform the methods of the invention.

The present invention also encompasses plants or parts thereof (including seeds) viable by the methods according to the present invention. Plants or parts thereof comprise a nucleic acid transgene encoding a MADS15 protein as defined above. The invention also provides genetic constructs and vectors for facilitating introduction and / or expression of nucleic acid sequences in the methods according to the invention in a plant. Genetic constructs may be inserted into vectors, which may be commercially available, suitable for transformation into plants, and suitable for expression of the gene of interest in transformed cells. The invention also provides use of a gene construct as defined herein in the methods of the invention.

More specifically, the present invention provides a construction comprising

(a) nucleic acid encoding MADS15 protein as defined above;

(b) one or more control sequences capable of directing the nucleic acid sequence expression of (a); and optionally

(c) a transcription termination sequence.

Plants are transformed with a vector comprising the sequence of interest (ie, a nucleic acid encoding a MADS15 polypeptide as defined herein. The skilled artisan is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells containing The sequence of interest The sequence of interest is operatively linked to one or more control sequences (at least one promoter).

Advantageously, any type of promoter may be used to direct expression of the nucleic acid sequence.

The promoter may be a constitutive promoter, alternatively the promoter may be an inducible promoter, ie having transcription initiation induced or increased in response to a chemically inducible or stress-inducible promoter, or a pathogen-induced promoter. Additionally or alternatively, the promoter may be be a specific organ or tissue specific promoter, or the promoter may be a ubiquitous promoter, or the promoter may be developmentally regulated. In addition, the promoter may be specific organ or specific tissue or cell.

Preferably, the nucleic acid MADS15 or variant thereof is operably linked to a constitutive promoter. A preferred constructive promoter is one that is also substantially expressively. Still preferably the promoter is derived from a plant, more preferably a monocot plant. Most preferred is use of a GOS2 promoter (e.g. rice, SEQ ID NO: 47 or SEQ ID NO: 108). It should be clear that the applicability of the present invention is not restricted to the MADS15 nucleic acid represented by SEQ ID NO: 43, nor is the applicability of the invention restricted to the expression of a MADS15 nucleic acid when directed by a GOS2 promoter. Examples of other constitutive promoters or functionally equivalent promoters that may also be used to direct expression of a MADS15 nucleic acid are shown in the definitions section.

Optionally, one or more terminator sequences may be used in the construction introduced into a plant. Additional regulatory elements may include transcriptional enhancers as well as traditional. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Such sequences must be known or readily obtainable by one of ordinary skill in the art.

An intron sequence can also be added to the untranslated 5 'region (RTU) or coding sequence to increase the amount of mature message that accumulates in the cytosol.

Other control sequences (in addition to promoter, enhancer, silencer, intron sequences, 3'UTR and / or 5'UTR regions) may be protein and / or RNA stabilizing agents. Such sequences must be known or readily obtainable by a person skilled in the art.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl.

For detection of successful transfer of nucleic acid sequences as used in the methods of the invention and / or selection of transgenic plants comprising these nucleic acids, it is advantageous to use marker genes (or reporter genes). Therefore, the genetic construct may optionally comprise a selectable marker gene.

The invention also provides a method for producing transgenic plants having improved production characteristics relative to control plants, comprising introducing and expressing in a plant any nucleic acid encoding a MADS15 protein as defined above.

More specifically, the present invention provides a method for the production of transgenic plants having increased yield, which method comprises:

(i) introducing and expressing in a plant or plant cell an MADS15 nucleic acid or variant thereof; and

(ii) cultivate the plant cell under conditions that promote plant growth and development.

Nucleic acid can be introduced directly into a plant cell or into the plant itself (including introduction into a tissue, organ or any other part of a plant). According to a preferred feature of the present invention, nucleic acid is preferably introduced into a plant by transformation.

Genetically modified plant cells can be regenerated by all methods with which the skilled worker is familiar. Suitable methods can be found in the publications mentioned above by S.D. Kung and R. Wu, Potrykus or Hofen and Willmitzer.

Generally after transformation, plant cells or cell clusters are selected for the presence of one or more markers that are encoded by expressible genes in plants co-transferred with the gene of interest, following which the transformed material is regenerated into an entire plant. To select transformed plants, the plant material obtained in the transformation is, as a rule, subject to selective conditions so that transformed plants can be distinguished from unprocessed plants. For example, seeds obtained in the manner described above may be planted and, after an initial growth period, subjected to appropriate spray selection.

An additional possibility is to cultivate the seeds, if appropriate after sterilization, on agar plates using a suitable sorting agent so that only transformed seeds can grow to plants. Alternatively, transformed plants are screened for a selectable marker such as those described above.

Following DNA transfer and regeneration, computationally transformed plants can also be evaluated, for example by Southern analysis, for the presence of the gene of interest, number of copies, and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, both techniques well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first-generation (or IT) transformed plant may be self-fertilized and second-generation homozygous (or T2) transformants selected, and T2 plants may then be further propagated by classical breeding techniques.

The generated transformed organisms can adopt a variety of forms. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); transformed and unprocessed detained grafts (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention extends further to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that the same feature (s) exhibit the same genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also includes host cells containing an isolated nucleic acid encoding a MADS15 protein as defined above. Preferred host cells according to the invention are plant cells.

Host plants for nucleic acids or the vector used in the method according to the invention, the expression drawer or listener construct are in principle advantageously all plants which are capable of synthesizing the polypeptides used in the inventive method.

The invention also extends to cut-off parts of a plant such as, but not limited to seeds, leaves, fruits, flowers, stems, rhizomes, roots, tubers and bulbs. The invention furthermore relates to derived products, preferably directly derived, from a cut-off part of such a plant, such as pellets or dry powders, oil, fat, fatty acids, starch or proteins.

According to a preferred feature of the invention, modulated expression is increased expression.

As mentioned above, a preferred method for modulating (preferably enhancing) expression of a nucleic acid encoding a MADS15 protein is to introduce and express in a plant a nucleic acid encoding a MADS15 protein; however the effects of performing the method, i.e. improving production characteristics can also be achieved using other well known techniques. A description of some of these techniques will now follow.

One such technique is identification by T-DNA activation. The resulting transgenic plants show dominant phenotypes due to the modified expression of genes near the introduced promoter.

The effects of the invention may also be reproduced using the TILLING (Genome-Targeted Local Injury) technique, or by homologous recombination.

Reference in this place to improved yield characteristics is adopted to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground (cut-off) and / or below-cut (cut-off) parts. ground.

By taking maize as an example, an increase in yield may be manifested as one or more of the following: increase in the number of plants established per hectare or acre, an increase in the number of ears per plant, an increase in the number of rows, number of seeds per row, weight seed weight, one thousand seed weight, ear length / diameter, increase in grain fill rate (which is the number of kernels divided by the total number of kernels and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary seedlings), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in weight of one thousand seeds, among others.

In a particular embodiment, such cut-off parts are roots, and performance of the methods of the invention results in plantarising increased root growth relative to the root growth of suitable control plants. Root development is an essential determinant of plant growth and crop production since root is the main channel for extracting nutrients from the environment.

Increased root yield may for example be manifested as one of more of the following: a) increased amount of root mass (whereby a discrimination between thick roots and thin roots can be made by setting a certain threshold), b) increased average root diameter, c ) increased total root biomass, and d) increased root biomass / increased shoot biomass. For the purpose of this invention, it should be understood that the term 'root growth' encompasses all aspects of growth of the different parts that constitute the root system at different stages of its development, both in monocotyledon and dicotyledonous plants. It should be understood that increased root growth may result from marked growth of one or more of its parts including the primary root, lateral roots, adventitious roots, etc. all of which fall within the scope of this invention.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit an increased growth rate (for at least part of their due cycle), relative to the growth rate of control plants in a matched maturity. your life cycle. The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the entire plant. Plants having an increased growth rate may have a shorter life cycle. The proper cycle of a plant can be adopted to mean the time required to grow from dry mature seed to the stage where the plant produced dry mature seeds, similar to the initial material. This life cycle can be influenced by factors such as early vigor, growth rate, green index, flowering time and seed ripening speed. The increase in growth rate may occur at one or more stages in a plant's life cycle or during substantially the entire plant life cycle. Increased growth rate during the early stages of a plant's life cycle may allow for improved vigor. Increasing growth rate can alter the harvest cycle of a plant by allowing plants to be sown later and / or harvested earlier than otherwise possible (a similar effect can be obtained with early flowering time). If the growth rate is sufficiently increased, it may allow additional seed sowing of the same plant species (for example sowing and harvesting rice plants followed by sowing and harvesting additional rice plants all within a conventional growing period). If the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing and harvesting of soybeans, potatoes or any other suitable plant). Additional harvest times of the same rhizome in the case of some crop plants may also be possible. Changing the harvesting cycle of a plant can lead to an increase in annual biomass production per acre (due to an increase in the number of times (say in a year) that any particular plant can be grown and harvested. An increase in growth rate may also allow the cultivation of transgenic plants in a wider geographical area than their wild counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or planting time (early cycle) or at harvesting time (late cycle). Such adverse conditions can be avoided if the harvesting cycle is shortened. The growth rate can be determined by deriving several growth curve parameters, such parameters can be: T-Mid (the time used by plants for reach 50% of their maximum size) and T-90 (time used by plants to reach 90% of their maximum size), among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having an increased growth rate relative to control plants. Therefore, according to the present invention, there is provided a method for increasing plant growth rate, which method comprises modulating expression, preferably enhancing expression, in a plant of a nucleic acid encoding a MADS15 protein as defined herein.

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general.

Mild stress, on the other hand, is defined here as any stress to which a plant is exposed that does not result in stopping overall plant growth without the ability to resume growth.

Mild stress in the sense of the invention leads to a reduction in growth of stressed plants of less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the control plant under non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "no stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location.

Performance of the methods of the invention generates plants grown under non-stress conditions or under mild drought conditions increased yields compared to control plants grown under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under non-stress conditions or under mild dry conditions, which method comprises increasing expression in a plant of a nucleic acid encoding a MADS15 polypeptide.

Performance of the methods of the invention generates plants grown under nutrient deficiency conditions, particularly under nitrogen deficiency conditions, increased yield over control-grown plants under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under nutrient deficiency conditions, the method of which is to increase expression in a plant of a nucleic acid by encoding a MADS15 polypeptide. Nutrient deficiency can result from nutrient deficiency or excess such as nitrogen, phosphates and other compounds containing phosphorus, potassium, calcium, cadmium, magnesium, manganese, iron and boron, among others.

In a preferred embodiment of the invention, the increase in yield and / or growth rate occurs according to the methods of the present invention under non-stress conditions.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Still preferably, the plant is a monocot plant. Examples of monocot plants include sugar cane. Most preferably aplanta is a cereal. Examples of cereals include rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats.

The present invention also encompasses use of nucleic acids encoding the MADS15 protein described herein and use of these MADS15 proteins to improve plant yield characteristics.

Nucleic acids encoding the MADS15 protein described herein, or the MADS15 proteins themselves, may find use in enhancement programs in which a DNA marker is identified that can be genetically linked to a gene encoding MADS15. Nucleic acids / genes, or the MADS15 proteins themselves can be used to define a molecular marker. This DNA or protein marker can then be used in breeding programs to select plants for enhanced production traits as defined above in the invention methods.

Allelic variants of a MADS15 gene / nucleic acid encoding protein may also find use in marker-assisted enhancement programs. Such breeding programs sometimes require introduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis, alternatively, the program may start with a collection of so-called "natural" allelic variants caused unintentionally. Identification of allelic variants then occurs, for example by PCR. This is followed by a step for selecting higher allelic variants of the sequence in question and generating increased production. Selection is typically performed by monitoring plant growth performance containing different allelic variants of the sequence in question. Growth performance can be monitored in a greenhouse or in the field. Additional optional steps include crossing plants in which the upper allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

Nucleic acids encoding MADS15 proteins can also be used as probes to genetically and physically map the genes of which they are a part of, and as markers for traits linked to such genes. Such information may be useful in plant breeding in order to develop strains with desired phenotypes. Such use of MADS15 protein-encoding nucleic acids requires only one nucleic acid sequence of at least 15 nucleotides in length. Nucleic acids encoding MADS15 protein can be used as restriction fragment length polymorphism (RFLP) markers. Southernblots (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, ALaboratory Manual) of restriction digested plant genomic DNA can be probed with MADS15 protein-encoding nucleic acids. The resulting banding pattern can then be subjected to genetic analysis using computer programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, nucleic acids can be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs from a set of individuals representing the ancestor and progeny of a defined genetic cross. Segregation of DNA polymorphisms is observed and used to calculate the position of nucleic acid encoding MADS15 protein in the previously obtained mapagenetic using this population (Botstein et al. (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4: 37-41. Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 cross-breeding populations, inbreeding populations, randomly crossed populations, nearby isogenic strains, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and references cited in this).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify differences in DNA sequences between the ancestors of the mapping region in the region corresponding to the current nucleic acid sequence. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having improved production characteristics as described above. These characteristics may also be combined with other economically advantageous characteristics, such as additional production enhancing characteristics, tolerance to other abiotic and biotic stresses, characteristics modifying various characteristics of production. architecture and / or biochemical and / or physiological characteristics.

Detailed Description of MADS15 Below

It has now surprisingly been found that decreasing the level and / or activity of an endogenous MADS15 polypeptide generates plants with improved yield traits, in particular increased yield relative to corresponding wild type plants. The present invention therefore provides methods for stimulating yield characteristics, particularly for increasing yield of a plant over control plants, comprising decreasing the level and / or activity of an endogenous MADS15 polypeptide. Reference herein to "control plants" is adopted to mean corresponding wild-type plants in which there is no reduction in activity of the endogenous MADS15 polypeptide.

Advantageously, performance of the methods according to the present invention results in plants having increased yield, particularly increased seed yield and / or increased biomass, relative to corresponding wild type plants.

The term "increased yield" as defined herein is taken to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground parts and / or below-cut parts. from soil.

In particular, such cut-off parts include biomass / fats and / or seeds, and performance of the methods of the invention results in plants having increased yield (in vegetative and / or seed biomass) relative to the production of control plants.

Using corn as an example, an increase in yield can be manifested as one or more of the following: increase in the number of plants per hectare or acre, an increase in the number of ears per plant, an increase in the number of rows, number of seeds per row, weight. dementing, weight of one thousand seeds, ear length / diameter, increased grain filling rate (which is the number of full grains divided by the total number of grains and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary panicles), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in milliseed weight, among others.

The increase in seed yield can also be manifested as an increase in seed size and / or deboning volume. This may increase the amount, or change the composition of, substances in the seed, such as oils, proteins and carbohydrates.

According to a preferred feature, performance of the methods of the invention results in plants having increased yield, particularly biomass and / or increased seed yield. Therefore, in accordance with the present invention, there is provided a method for enhancing plant production, which method comprises decreasing the level of activity of a MADS15 polypeptide or homologue thereof, preferably by decreasing expression of a MADS15 gene or homologue thereof.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit increased growth rate (for at least part of their due cycle) relative to the growth rate of corresponding wild-type plants at a corresponding stage. in your life cycle. The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the plant. Plants having an increased growth rate may have a shorter life cycle. The life cycle of a plant can be adopted to mean the time required to grow from dry mature seed to the stage where the plant produced dry mature seeds, similar to the initial material. This life cycle can be influenced by factors such as early vigor, growth rate, flowering time and seed ripening speed. An increase in growth rate may occur at one or more stages in a plant's life cycle or substantially throughout the entire plant life cycle. Increased growth rate during the early stages of a plant's life cycle may allow for improved vigor. Increasing the growth rate may alter the crop cycle of a plant by allowing plants to be sown and / or harvested earlier than otherwise possible (a similar effect can be obtained with early flowering time). If the growth rate is sufficiently increased, it may allow additional sowing of seeds of the same plant species (eg sowing and harvesting rice plants followed by sowing and harvesting additional rice plants all within a conventional growing period). Similarly, if the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing, harvesting of soybean, potato or any other suitable plant) . Additional harvest times for mesmorizoma in the case of some crop plants may also be possible.

Changing the harvest cycle of a plant can lead to an increase in annual biomass production per acre (due to an increase in the number (say a year) that any particular plant can be grown and harvested). An increase in growth rate may also allow the cultivation of transgenic plants in a wider geographical area than their wild type counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or at the time of planting (early cycle) or harvest time (late cycle). Such adverse conditions can be prevented if the harvest cycle is shortened. Growth rate can be determined by deriving various parameters from growth curves, such parameters can be: T-Mid (time taken by plants to reach 50% of their maximum size) and T-90 (time taken by plants to reach 90% of their size) maximum), among others.

Performance of the methods of the invention generates plants having an increased growth rate. Therefore, according to the present invention, there is provided a method for increasing plant growth rate relative to control plants, which method preferably comprises reducing the level and / or expression activity of an endogenous MADS15 gene in a plant.

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general. Mild stress on the other hand is defined here as any stress to which a plant is exposed that does not result in stopping of general plant growth without the ability to resume growth. Mild stress in the sense of the invention leads to a reduction in growth of stressed plants of less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the control plant under non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "no stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location.

Performance of the methods of the invention generates plants grown under non-stress conditions or under mild drought conditions increased yields compared to control plants grown under comparable conditions. Therefore, according to the present invention there is provided a method for increasing production in plants grown under non-stress conditions or under mild dry conditions, which method comprises reducing expression of an endogenous MADS15 gene in a plant.

Performance of the methods of the invention generates plants grown under nutrient deficiency conditions, particularly under nitrogen deficiency conditions, increased yield over control-grown plants under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under nutrient deficiency conditions, the method of which decreases expression in a plant of a nucleic acid by encoding a MADS15 polypeptide. Nutrient deficiency can result from nutrient deficiency or excess such as nitrogen, phosphates and other compounds containing phosphorus, potassium, calcium, cadmium, magnesium, manganese, iron and boron, among others.

In a preferred embodiment of the invention, the increase in yield and / or growth rate occurs according to the methods of the present invention under non-stress conditions.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Still preferably, the plant is a monocot plant. Examples of monocotyledonous plants include sugar cane. Most preferably aplanta is a cereal. Examples of cereals include rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats.

Reference herein to a level "decrease" of an endogenous MADS15 protein in a plant is adopted to mean a reduction in protein concentration or substantial elimination of an endogenous MADS15 protein relative to endogenous MADS15 protein levels found in corresponding wild type plants. Substantial reduction or elimination may result in reduced or substantially abolished MADS15 protein activity in a plant.

Reference herein to a "decrease" in activity of an endogenous MADS15 protein in a plant is adopted to mean a reduction in MADS15 protein activity or substantial elimination of endogenous MADS15 protein relative to endogenous MADS15 protein reactivity levels found in wild type plants.

Preferably, the reduction in endogenous MADS15 protein level and / or activity is achieved by decreasing the expression of the endogenous MADS15 gene.

Reference herein to an "endogenous" MADS15 gene not only refers to MADS15 genes as found in a plant in its natural form (i.e., without any human intervention), but also refers to isolated MADS15 genes subsequently introduced into a plant. For example, a transgenic plant containing a MADS15 transgene may face a substantial reduction or deletion of the MADS15 transgene expression and / or a reduced or substantial deletion of an endogenous MADS15 gene expression.

This reduction (or substantial elimination) of endogenous MADS15 gene expression can be achieved using any one or more of several well known gene silencing methods. "Silencing" or "decreased" expression, as used herein, refers to a substantial reduction or elimination of MADS15 gene expression and / or MADS15 polypeptide levels and / or MADS15 polypeptide activity.

One such method for substantially reducing or eliminating endogenous MADS15 gene expression is RNA-mediated gene expression reduction (RNA silencing). Silencing in this case is triggered in a plant by a double-stranded RNA (dsRNA) molecule that is substantially homologous to a target MADS15 gene. EstedsRNA is further processed by the plant at about 21 to about 26 nucleotides called short interfering RNAs (siRNAs). SiRNAs are incorporated into an RNA-induced silencing complex (RISC) that cleaves the mRNA of a target MADS15 gene, thereby substantially reducing the number of MADS15 mRNAs to be translated into a MADS15 protein.

An example of an RNA silencing method involves introducing coding sequences or parts thereof in a sense orientation into a plant. The additional gene, or part of it, will silence an endogenous MADS15 gene, generating a phenomenon known as co-suppression. Reduction of MADS15 gene expression will be more pronounced if several additional copies are introduced into the plant as there is a positive correlation between high transcript levels and co-suppression triggering. Another example of an RNA silencing method involves the use of nucleic acid sequences. MADS15 antisense.

Antisense nucleic acid can be produced microbiologically using an expression vector in which a nucleic acid is subcloned in an antisense orientation (ie, RNA transcribed from the inserted nucleic acid will be from an antisense orientation to a target nucleic acid of interest, further described in the following subsection). Preferably, antisense nucleic acid production in plants occurs by means of a stably integrated transgene comprising an operable engine for constitutive expression in plants, an antisense oligonucleotide, and a terminator.

A preferred method for substantially reducing or eliminating endogenous MADS15 gene expression by RNA silencing is to use an expression vector in which an MADS15 gene or fragment thereof is an inverted repeat (in part or completely) separated by a spacer (non-coding DNA). ).

In yet another embodiment, the substantial reduction or elimination of endogenous expression of MADS15 may be obtained using antibodies. For example, a Tetrahymena L-19 IVS RNA derivative can be constructed in which the nucleotide sequence of the active site is complementary to the nucleotide sequence to be cleaved into an mRNA encoding MADS15.

Gene silencing can also be achieved by insertion mutagenesis or if there is a mutation in the endogenous MADS15 gene and / or a mutation in the subsequently isolated MADS15 gene introduced in a plant. Substantial reduction or elimination of MADS15 protein activity may be caused by a non-functional MADS15. For example, MADS15 binds to various interacting proteins; one or more mutation (s) and / or truncation (s) within the MADS box of a MADS15 can therefore support a MADS15 protein that is still capable of binding to interaction proteins but which cannot exhibit its normal function as a transcription factor.

An additional approach for gene silencing is to target complementary nucleotide sequences to the MADS15 dogene regulatory region (e.g., the MADS15 promoter and / or enhancers) to form triple helix structures that prevent transcription of the MADS15 gene into target cells.

Yet another approach to gene silencing is described by Hiratsu et al. (Plant J. 34, 733-739, 2003). This method does not depend on sequence homology with the targeted gene but involves the use of a repression sequence domain in transcriptional gene fusions, and has been used to modify traits of agronomic interest (Fujita et al., Plant Cell 17,3470-3488, 2005 and Mitsuda at al., Plant Cell 17, 2993-3006, 2005). Typically, a chimeric nucleotide fusion is made between a genecoding a protein capable of positively influencing targeted dogene expression (such as a transcriptional activator), and a denucleotide fragment encoding a repression domain. By expression of chimeric gene fusion, expression of the targeted gene is suppressed, usually in a dominant negative manner. Repression domains are well known in the art, for example the EAR motif present in some Zinc finger transcription factor AP2. Depression domain-based methods are well suited for overcoming gene redundancy for genevisate in the plant species of choice.

Examples are described above of various methods of gene enhancement (for the substantial reduction or elimination of endogenous MADS15 gene expression. The methods of the invention rely on reducing the expression of an endogenous MADS15 gene in a plant. A person skilled in the art should readily be able to adapt the above mentioned methods for silencing to achieve gene silencing on an entire plant or parts thereof through the use of an appropriate engine, for example.

It should be noted that the essence of the present invention has surprising and advantageous results found with substantial reduction or deletion of endogenous MADS15 gene expression in a plant, and is not limited to any particular method for such substantial reduction or deletion of endogenous MADS15 protein activity. The activity of a MADS15 polypeptide may also be decreased or eliminated by introducing a genetic modification (preferably at the locus of a MADS15 gene). The locus of a gene as defined herein is taken to mean a genomic region, which includes the e10 kb gene of interest before or after the coding region.

Genetic modification may be introduced, for example, by any one or more of the following methods: T-DNA inactivation, TILLING, site-directed mutagenesis, directed evolution, homologous recombination. Following introduction of genetic modification, it follows a step of deselecting for decreased activity of an MADS15 polypeptide, whose activity decrease generates plants having increased yield.

Identification by T-DNA inactivation involves insertion of a T-DNA into the genomic region of the gene of interest or 10 kb before or after the coding region of a gene in such a configuration that T-DNA inhibits expression of the target gene. Typically, dogene expression regulation targeted by its natural promoter is disrupted. T-DNA is randomly inserted into the plant genome, for example through Agrobacterium infection and leads to decreased gene expression near the inserted T-DNA. The resulting transgenic plants show phenotypes due to inhibited gene expression near the introduced T-DNA.

A genetic modification can also be introduced into a MADS15 gene by using the TILLING technique.

Site-directed mutagenesis and random mutagenesis can be used to generate MADS1 nucleic acid variants. Several methods are available to achieve site-directed mutagenesis, the most common being current protocols in molecular biology.Wiley Eds.

Directed evolution can also be used to generate MADS15 nucleic acid variants. This consists of DNA scrambling repeats followed by appropriate screening and / or selection to generate MADS15 nucleic acid variants or variants encoding MADS15 polypeptides having a modified (decreased or aborted) biological activity (Castle et al., (2004) Science 304 (5674) : 1151-4; U.S. Patent Nos. 5,811,238 and 6,395,547).

T-DNA activation, TILLING, site-directed mutagenesis, and site-directed evolution are examples of technologies that enable the generation of new MADS1 alleles and variants.

The effects of the invention may also be reproduced using homologous recombination. The nucleic acid to be targeted may be allele encoding an inactive protein or a reduced activity protein used to replace the endogenous gene or may be introduced into the endogenous gene, and must be directed to the MADS15 locus.

Other methods, such as the use of antibodies to endogenous MADS15 to inhibit its in plant function, or interference with the signaling pathway in which MADS15 is involved, will be well known to one of ordinary skill. Alternatively, a screening program may be established to identify natural variants of a MADS15 gene whose variants have reduced MADS15 activity, or no MADS15 activity. Such natural variants may also be used in the methods of the present invention.

For optimal performance, gene silencing techniques used for the substantial reduction or elimination of endogenous MADS15 gene expression require the use of MADS15 nucleic acid sequences from monocotyledonous plants for transformation into monocotyledonous plants. Preferably, an MADS15 nucleic acid from any given plant species is introduced into that same species. For example, a rice MADS15 nucleic acid (either a full length MADS15 sequence or fragment) is transformed into a rice plant. MADS15 nucleic acid need not be introduced in the same plant variety.

Reference herein to a "MADS15 gene" or "MADS75 nucleic acid" is adopted to mean a polymeric form of a deoxyribonucleotide or ribonucleotide polymer of any length, or double or single strand, or analogs thereof, which have the essential characteristics of a ribonucleotide. natural in that they can hybridize to nucleic acids in a similar manner to naturally occurring cholinucleotides. A "MADS15 gene" or a "MADS15 nucleic acid" refers to a sufficient length of substantially contiguous nucleotides of a gene encoding MADS15 to perform gene deletion; this can be as little as 20 or fewer nucleotides. A gene encoding a (functional) protein is not a requirement for the various methods discussed above for substantially reducing or deleting expression of an endogenous MADS15 gene.

The methods of the invention may be performed using a sufficient length of substantially contiguous nucleotide nucleotides from an MADS15 nucleic acid, which may consist of 20 or fewer nucleotides, which may be from any part of the MADS1 5 gene / nucleic acid, such as the 5 'end of the region. coding that is well-conserved among the MADS1 5 gene family, or coding for one of the conserved motifs described below.

MADS15 genes are well known in the art and useful in the methods of the invention are substantially contiguous dogene / plant MADS15 nucleic acid nucleotides described in Moon et al. (Plant Physiol. 120, 1193-1204, 1999).

Other MADS15 genes / nucleic acid sequences may also be used in the methods of the invention, and may be readily identified by one of ordinary skill in the art. MADS15 polypeptides can be identified by the presence of one or more of several well-known characteristics (see below). Upon identification of a MADS15 polypeptide, one skilled in the art can readily derive, by routine techniques, the corresponding coding nucleic acid sequence and use a sufficient length of contiguous nucleotides thereof to perform any one or more of the silencing methods described above.

The term "MADS15 polypeptide or homologue thereof" as defined herein refers to a protein falling into the group of FUL-like MADS box proteins as delineated by Adam et al. (J. Mol. Evol. 62, 15-31). FUL-like MADS box proteins are part of the SQUAMOSA subfamily and have the typical MIKCc architecture: an N-terminated MADS domain (M), followed by a dimerization intervening domain (I), a highly conserved keratin domain (K), and a variable domain (C) notomer C, which is responsible for protein binding in transcriptional interaction or activation (De Bodt et al. Trends in Plant Science 8, 475-483).

Plant MADS15 polypeptides can also be identified by the presence of certain conserved motifs. The presence of these conserved motifs can be identified using methods for sequence alignment for comparison as described above. In some instances, the default parameters can be adjusted to modify the search stringency. For example, using BLAST, the statistically significant threshold (called "expected value") for reporting matches against database sequences can be increased to show less stringent matches. In this way, short, practically accurate matches can be identified. By identifying an MADS15 polypeptide by the presence of these reasons , a person skilled in the art can readily derive the corresponding nucleic acid encoding opolipeptide comprising the relevant motifs, and use a sufficient length of contiguous nucleotides thereof to perform any one or more of the gene silencing methods described above (for the reduction or substantial deletion of a gene expression). Endogenous MADS15).

Typically, the presence of at least one of motifs 1-4 should be sufficient to identify any query sequence as a MADS15, however the presence of at least motifs 1 and 2 is preferred. The consensus sequence provided is based on the monocot sequences given in the sequence listing. One of ordinary skill in the art will be well aware that the consensus sequence may vary in some way if additional or different sequences (for example, dedicated-sequence sequences) are used for comparison.

Reason 1, located in the MADS domain (SEQ ID NO: 114):

L (LA ^) KKA (H / N) EIS (VI) L (C / Y) DAE (V / I / L) (A / G) (L / AAA) (I / V) (I / V) FS (T / P / AY) KGKLYE (Y / F) (Y / S) (T / S) (D / N / E) S (K / C / R / S) M (D / E) (> / I / R / K) IL (E / D) R.

Preferably, this conserved sequence 1 motif has the following:

LLKKAHEISVLCDAEVA (L / AA ^) I (W) FS (T / P) KGKLYEYATDS (C / R) M (D / E) (R / K) ILER, more preferably conserved motif of sequence 1 theme sequence:

LLKKAHEISVLCDAEVAAIVFSPKGKLYEYATDSRMDKILER

Reason 2, located in domain K (SEQ ID NO: 115):

KLK (A / S) (K / R) (V / I) And (A / T / S) (L / I) (Q / N) (K / R / N) (S / C / R) (Q / H) (R / K) HLMGE

Preferably, this conserved sequence 2 motif has the following:

KLKAK (V / I) AND (A / T) (L / I) QK (S / C) (Q / H) (R / K) HLMGE

More preferably, this conserved sequence motif has 2 sequence:

KLK ^ KIETIQKCHKHLMGE

Optionally, the MADS15 polypeptide or homologue thereof may comprise in domain C motif 3 and / or motif 4:

reason 3 (SEQ ID NO: 116):

Q (P / Q / V / A) QTS (Y / F) (Y / F) (Y / F) (Y / F) (Y / C / F) (Y / M) reason 4 (SEQ ID NO: 117):

(G / A / V / L) (W / P) XWMX (S / H)

characterized in that the first residue X in motif 4 can be any amino acid, but preferably L, P or H; and characterized by the fact that the second residue X can be any amino acid, but preferably V or L.

Alternatively, the C domain (starting behind the keratin domain, ie at Rl 75 in SEQ ID NO: 110) may be characterized by increased glutamine occurrence (usually 3.93%, here at least 8.77% with a maximum of 26.73 %), in addition, alanine, proline, and / or serine content may also be increased (above 7.8%, 4.85% and 6.89% respectively). Alternatively formulated, a preferred MADS15 protein useful in the methods of The present invention comprises at least one N-terminal MADS domain corresponding to the SMART SM00432 domain (PfamPF00319) followed by a Keratin domain corresponding to the K-box region defined in Pfam as PFO1486, more preferably the MADS15 protein has the conserved signature of SEQ IDNO: 114 and in its K domain the conserved signature of SEQ ID NO: 115, more preferably, the MADS15 protein has the sequence given in SEQ IDNO: 110.

Homologs, as defined above, can be readily identified using routine techniques well known in the art, such as sequence alignment; OsMADS 15 homologs may have been differently named in various plant species, so degene / protein names should not be used to identify orthologs or paralogs. Methods for sequence alignment for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol48: 443-453) to find the alignment of two complete sequences that maximizes the number of pairings and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The computer application for performing BLAST analysis is publicly available through the NationalCentre for Biotechnology Information. Homologous sequences can be readily identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with the paired alignment standard parameters, and a percentage scoring method. Secondary manual editing can be performed to optimize conserved intermotif alignment (see below), as would be apparent to an art-savvy person.

The various structural domains in a MADS15 protein can be identified using specialized databases eg SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic etal. (2002) Nucleic Acids Res 30 , 242-244), InterPro (Mulder et al., (2003) Nucl. Acids Res. 31, 315-318;), Prosite (Bucher and Bairoeh (1994), Ageneralized profile syntax for biomolecular sequences motifs and its funetionin automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology.10 Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137 (2004)) or Pfam (Bateman et al., Nucleic Acids Research 30 (1): 276-280 (2002)).

In addition, a MADS15 protein may also be identifiable by its ability to bind to DNA and interact with other proteins. DNA binding activity and protein-protein interactions can be readily determined in vitro or in vivo using techniques well known in the art. Examples of in vitro assays for DNA binding activity include: gel retardation analysis using known DNAMADS-box binding domains (West et al. (1998) Nucl Acid Res 26 (23): 5277-87), or a yeast hybrid. An example of an in vitro assay for protein-protein interactions is the analysis of two yeast hybrids (Fieldsand Song (1989) Nature 340: 245-6). Proteins known to interact with OsMADS 15 include other MADS proteins (such as those involved in the flowering signal complex), receptor-like kinases such as Clavata and Clavata2, Erecta, BRI1 or RSK.

Therefore by identifying an MADS15 polypeptide using one or more of the features described above, one skilled in the art can readily derive the corresponding nucleic acid by encoding the polypeptide, and use a sufficient length of substantially contiguous nucleotides thereof to perform any one or more of the gene silencing methods described. above (for the substantial reduction or deletion of expression of an endogenous MADS15 gene).

Preferred for use in the methods of the invention is a sufficient length of substantially contiguous nucleotides of SEQID NO: 109 (OsMADS15), or the use of a sufficient length of substantially contiguous nucleotides of an OsMADS 15 ortholog or sequence coding (SEQ ID NO : 109). Examples of such OsMADS 15 orthologs and paralogues are provided in Table D below. Close homologues of OsMADS 15 are the proteins represented by SEQ ID NO: 147, 149, 151, 153, 155, 157, 159, 161, 163,165, 167, 169, 171 and 173.

Orthologists in, for example, monocotyledonous plant species can be easily found by performing a so-called reciprocal blast search. This can be done by first blasting developing BLAST with a query string (for example, SEQ ID NO: 109 or SEQ ID NO: 110) against any matching database, such as the publicly available NCBI.BLASTN database. or TBLASTX (using default predetermined values) can be used when starting from a nucleotide sequence and BLASTP or TBLASTN (using default predetermined values) can be used when starting from a protein sequence. BLAST results can be optionally filtered. Full length sequences or filtered results or unfiltered results are then BLASTed back (secondBLAST) against organism sequences from which the query sequence is derived (where the query sequence is SEQ ID NO: 109 or SEQ ID NO: IlOo second BLAST must therefore be against rice sequences). The first and second BLAST results are then compared. A analog is identified if a high score hit from the first BLAST is of the same species from which the query sequence is derived; An orthologist is identified if a high score hit is not of the same kind from which the query frequency is derived. High score hits are those with a lower E value. The lower the E value, the more significant the score (or in other words, the lower the chance that the hit was found by chance). Value computing E is well known in the art. In the case of large families, ClustalW can be used, followed by a neighboring tree to help visualize related gene grouping and to identify orthologs and paralogues.

The source of the substantially contiguous umgene / MADS15 nucleic acid contiguous nucleotides can be any plant or fonteartificial source. For optimal performance, gene silencing techniques used for the substantial reduction or elimination of endogenous MADS15 gene expression require the use of MADSl 5 sequences from monocotyledonous plants for transformation into monocotyledonous plants. Preferably, MADSl 5 sequences from the Poaceae family are transformed into plants in the family. Poaceae Still preferably, a rice MADS15 nucleic acid (be it a full length MADS15 sequence or fragment) is transformed into a rice plant. MADS15 nucleic acid need not be introduced in the same plant variety. More preferably, the rice MADS15 nucleic acid is of a sufficient length of substantially contiguous nucleotides of SEQ ID NO: 109 (OsMADS 15) or a sufficient length of substantially contiguous nucleotides of an OsMADS15 ortholog or sequence coding (SEQ ID NO: 110). As mentioned above, one of ordinary skill in the art should be aware of what would constitute a sufficient length of substantially contiguous nucleotides to carry out any of the gene deletion methods defined above, this may be as little as 20 or substantially substantially contiguous nucleotides in some cases.

The invention also provides genetic constructs and vectors for facilitating introduction and / or expression of nucleotide sequences useful in the methods according to the invention.

Therefore, a gene construct is provided comprising one or more control sequences capable of directing expression of a sense and / or antisense MADS15 nucleic acid sequence in a plant to silence an endogenous MADS15 gene in the plant; and optionally a transcription termination sequence. Preferably, the control sequence is a constitutive and ubiquitous promoter.

A preferred construct for gene silencing is one comprising an inverted repeat of a MADS15 or fragment gene, preferably capable of forming a staple structure, whose reverse repeat is under the control of a constitutive promoter.

Therefore, the invention provides a construction comprising:

(a) an MADS15 nucleic acid capable of forming a staple structure;

(b) one or more control sequences capable of directing the nucleic acid sequence expression of (a); and optionally

(c) a transcription termination sequence.

Useful constructs in the methods according to this

The invention can be created using recombinant DNA technology well known to those skilled in the art. The gene constructs may be inserted into commercially available vectors suitable for transformation into plants and suitable for expression of the gene of interest in transformed cells. The invention therefore provides use of a gene construct as defined above in the methods of the invention.

The sequence of interest is operably linked to one or more control sequences (at least one promoter) capable of increasing expression in a plant.

Advantageously, any type of promoter may be used to direct expression of the nucleic acid sequence. The promoter may be an inducible promoter, i.e. having induced or increased transcription initiation in response to an evolutionary, chemical, environmental or physical stimulus. Additionally or alternatively, the promoter may be a tissue-preferred or cell-preferred promoter, i.e. one which is preferably capable of initiating transcription in certain tissues, such as leaves, roots, seed tissue etc, or even specific cells. Promoters capable of initiating transcription in certain tissues or cells are only referred to herein as "tissue specific", respectively "cell specific".

Preferably, the MADS15 nucleic acid or functional variant thereof is operably linked to a constitutive promoter. Preferably the promoter is a ubiquitous promoter and is expressed predominantly throughout the plant. Preferably, the constructive promoter capable of preferentially expressing nucleic acid throughout the plant has a comparable expression profile to a GOS2 promoter. More preferably, the constitutive promoter has the same expression profile as the rice GOS2 promoter, more preferably, the promoter capable of preferentially expressing nucleic acid throughout the plant is the rice GOS2 promoter (SEQ ID NO: 113 or SEQ ID NO: 174) . It should be clear that the applicability of the present invention is not restricted to the MADS15 nucleic acid represented by SEQ ID NO: 109, nor is the applicability of the invention restricted to the expression of a MADS15 nucleic acid when directed by a GOS2 promoter. An alternative constitutive promoter that is useful in the methods of the present invention is the high mobility protein promoter group (PR00170, SEQ ID NO: 40 in International Patent 2004070039). Examples of other constitutive promoters that may also be used to direct expression of an acid MADS15 are shown in the definitions section.

Optionally, one or more terminator sequences may also be used in the construction introduced into a plant. Additional regulatory elements may include transcriptional as well as traditional transcriptional enhancers. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Such sequences must be known or readily obtainable by one of ordinary skill in the art.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl. The genetic construct may optionally comprise a selectable marker gene.

The present invention also encompasses plants including viable vegetable parts by the methods of the present invention having increased production relative to control plants and which have reduced or substantially deleted expression of an endogenous MADS15 gene.

The invention further provides a method for producing transgenic plants having increased production over control plants, whose transgenic plants have reduced or substantially deleted expression of an endogenous MADS15 gene.

More specifically, the present invention provides a method for producing transgenic plants having increased seed production whose method comprises: introducing and expressing in a plant, plant part or plant cell a gene construct comprising one or more control sequences capable of preferably directing expression of a sequence of inverse repeating MADS15 nucleic acids in a plant to silence an endogenous MADS15 gene in the plant; and

cultivate the plant, plant part or plant cell under conditions that promote plant growth and development.

Preferably, the construct introduced into a plant is one comprising an inverted (in part or complete) repeat of a MADS15 gene or fragment thereof, preferably capable of forming a staple structure.

According to a preferred feature of the present invention, the construct is introduced into a plant by transformation.

Generally after transformation, plant cells or cell clusters are selected for the presence of one or more markers that are encoded by expressible genes in plants co-transferred with the gene of interest, following which the transformed material is regenerated into an entire plant.

Following DNA transfer and regeneration, computationally transformed plants can be evaluated, for example using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, or quantitative PCR, all techniques well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first generation (or IT) transformed plant may be self-fertilized to generate second generation (or T2) homozygous transformants, and T2 plants further propagated by classical breeding techniques.

The generated transformed organisms can adopt a variety of forms. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); transformed and unprocessed detained grafts (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention extends further to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that the same feature (s) exhibit the same genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also extends to cropable parts of a plant such as seeds and derived products, preferably directly derived, from a cropable part of such a plant, such as dried pellets or powders, oil, fat, fatty acids, starch or proteins.

The present invention also encompasses use of MADS15 nucleic acids for the substantial reduction or elimination of endogenous MADS15 gene expression in a plant to increase seedling production as defined above.

Nucleic acids encoding the MADS15 protein described herein, or the MADS15 proteins themselves, may find use in enhancement programs in which a DNA marker is identified that can be genetically linked to a gene encoding MADS15. Nucleic acids / genes, or the MADS15 proteins themselves can be used to define a molecular marker. This DNA or protein marker can then be used in breeding programs to select plants for enhanced production traits as defined above in the invention methods.

Allelic variants of a MADS15 gene / nucleic acid encoding protein may also find use in marker-assisted enhancement programs. Such breeding programs sometimes require introduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis, alternatively, the program may start with a collection of so-called "natural" allelic variants caused unintentionally. Identification of allelic variants then occurs, for example by PCR. This is followed by a step for selecting higher allelic variants of the sequence in question and generating increased production. Selection is typically performed by monitoring plant growth performance containing different allelic variants of the sequence in question. Growth performance can be monitored in a greenhouse or in the field. Additional optional steps include crossing plants in which the upper allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

Nucleic acids encoding MADS15 proteins can also be used as probes to genetically and physically map the genes of which they are a part of, and as markers for traits linked to such genes. Such information may be useful in plant breeding in order to develop strains with desired phenotypes. Such use of MADS15 protein-encoding nucleic acids requires only one nucleic acid sequence of at least 15 nucleotides in length. Nucleic acids encoding MADS15 protein can be used as restriction fragment length polymorphism (RFLP) markers. Southernblots (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, ALaboratory Manual) of restriction digested plant genomic DNA can be probed with MADS15 protein-encoding nucleic acids. The resulting banding pattern can then be subjected to genetic analysis using computer programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, nucleic acids can be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs from a set of individuals representing the ancestor and progeny of a defined genetic cross. Segregation of DNA polymorphisms is observed and used to calculate the position of nucleic acid encoding MADS15 protein in the previously obtained mapagenetic using this population (Botstein et al. (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4: 37-41. Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 cross-breeding populations, inbreeding populations, randomly crossed populations, nearby isogenic strains, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and cited references. in this one).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify differences in DNA sequences between ancestors of the cross-mapping region corresponding to the current sequence of nucleic acids. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having improved production characteristics as described above.

These features may also be combined with other economically advantageous features, such as additional production enhancing features, tolerance to other abiotic and biotic stresses, features modifying various architectural features and / or biochemical and / or physiological features.

Detailed Description of PLT

Surprisingly, it has now been found that increasing expression of a nucleic acid encoding a PLT transcription factor polypeptide generates plants having improved production characteristics, in particular increased production relative to control plants.

In accordance with the present invention, a method is provided for increasing plant yield over control plants, comprising increasing expression in a plant of a nucleic acid by encoding a PLT transcription factor polypeptide.

The term "increased yield" as defined herein is taken to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground parts and / or below-cut parts. from soil.

In particular, such cut-off parts are seeds, and performance of the methods of the invention results in plants having increased seed production relative to control plant seed production.

Using corn as an example, an increase in yield can be manifested as one or more of the following: increase in the number of plants per hectare or acre, an increase in the number of ears per plant, an increase in the number of rows, number of seeds per row, weight. dementing, weight of one thousand seeds, ear length / diameter, increased grain filling rate (which is the number of full grains divided by the total number of grains and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary panicles), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in milliseed weight, among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having increased seed production relative to control plants. Therefore according to the present invention, there is provided a method for increasing plant seed production over control plant seed production, the method comprising increasing expression in a plant of a nucleic acid encoding a PLT transcription factor polypeptide.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit increased growth rate (for at least part of their due cycle) relative to the growth rate of corresponding wild-type plants at a corresponding stage. in your life cycle. The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the plant. A plant having an increased growth rate may still exhibit early flowering. The increase in growth rate may occur at one or more stages in a plant's life cycle or substantially throughout the entire plant life cycle. Increased growth rate during the early stages of a plant's life cycle may allow improved vigor (increased seedling emergence). Increasing growth rate can alter a plant's harvest cycle by allowing plants to be sown later and / or harvested earlier than otherwise possible. If the growth rate is sufficiently increased, it may allow additional seed sowing of the same plant species (for example sowing and harvesting rice plants followed by sowing and harvesting additional rice plants all within a conventional growing period). If the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing and harvesting of soybeans, potatoes or any other suitable plant). Additional harvest times of the same rhizome in the case of some crop plants may also be possible. Changing the harvesting cycle of a plant can lead to an increase in annual biomass production per acre (due to an increase in the number of times (say in a year) that any particular plant can be grown and harvested. An increase in growth rate may also allow the cultivation of transgenic plants in a wider geographical area than their wild counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or planting time (early cycle) or at harvesting time (late cycle). Such adverse conditions can be avoided if the harvesting cycle is shortened. The growth rate can be determined by deriving several growth curve parameters, such parameters can be: T-Mid (the time used by plants for reach 50% of their maximum size) and T-90 (time used by plants to reach 90% of their maximum size), among others.

Performance of the methods of the invention generates plants having an increased growth rate. Therefore, according to the present invention there is provided a method for increasing plant growth rate, which method comprises increasing expression in a plant of a nucleic acid by encoding a PLT transcription factor polypeptide.

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general. Mild stress on the other hand is defined here as any stress to which a plant is exposed that does not result in stopping of general plant growth without the ability to resume growth. Mild stress in the sense of the invention leads to a reduction in growth of stressed plants of less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the control plant under non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "no stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location.

Performance of the methods of the invention generates plants grown under non-stress conditions or under mild drought conditions improved yield characteristics compared to control plants grown under comparable conditions. Therefore, according to the present invention, there is provided a method for improving yield characteristics, in particular for increasing yield, in plants grown under non-stress or mild dry conditions, which method comprises increasing a plant expression of a nucleic acid encoding a transcription factor polypeptide PLT.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Examples of typical crop plants grown for oil production include soybean, sunflower, cotton, canola, peanut, or palm. Examples of typical crop plants grown for starch production include rice, wheat, barley, corn or potato. Still preferably, the plant is a monocot plant. Examples of monocotyledonous plants include sugar cane. More preferably the plant is a cereal. Examples of cereals include rice, corn, wheat, barley, millet, rye, triticale, sorghum and oats.

The term "PLT transcription factor polypeptide" as defined herein refers to any polypeptide preferably comprising an increasing order of at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98 % or 99% string identity with domain AP2 represented by SEQ ID NO: 191.

Preferably the PLT transcription factor polypeptide of choice is a PLT transcription factor polypeptide which in its natural environmental genetics is essentially expressed in the roots (below ground) of the plant.

Still preferably, the transcription factor polypeptide PLT comprises either a motif but preferably both motif 1 as represented by SEQ ID NO: 192 PK (V / L) (A / E) DFLG and motif 2 as represented by SEQ ID NO: 209: (V / L) FX (M / V) WN (D / E), characterized in that X can be any amino acid, preferably motif 2 has the sequence of SEQ ID NO: 193 (V / L) F (T / Y / N) (M / V) WN (D / E).

Examples of PLT transcription factor polypeptides comprising (i) in ascending order preferably at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% Sequences with the AP2 domain represented by SEQ ID NO: 191 are proteins represented by the sequences given in Table I in the examples section.

Preferred examples are represented by SEQ ID NO: 176 and SEQ ED NO: 178.

The AP2 domain in a PLT transcription factor polypeptide can be identified using specialized databases e.g. SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864;

Letunic et al. (2002) Nucleic Acids Res 30, 242-244), InterPro (Mulder et al., (2003) Nucl. Acids Res. 31,315-318), Prosite (Bucher and Bairoch (1994), Ageneralized profile syntax for biomolecular sequences motifs and its funetionin automatic sequence interpretation. (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology.

Altman R, Brutlag D., Karp P., Lathrop R, Searls D., Eds., Pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137,

(2004)) or Pfam (Bateman et al., Nucleic Acids Research 30 (1): 276-280 (2002)). The AP2 domain of a PLT transcription factor comprises two repeats R1 and R2, each about 68 amino acids and separated by a linker region (Figure 13).

The AP2 domain as represented by SEQ ID NO: 191 may be identified using methods for sequence alignment for comparison as described above. In some instances, the default parameters may be adjusted to modify the search stringency. For example using BLAST, the threshold of statistical significance (called expected values ") for reporting pairings against database sequences can be increased to show less stringent pairings. Thus, practically exact short pairings can be identified, including reason 1 as represented by SEQ ID NO : 192 and motif 2 as represented by SEQ ID NO: 209, preferably as represented by SEQ ID NO: 193.

Homologs of a PLT transcription factor polypeptide may also be used to carry out the methods of invention. Homologs (or homologous proteins) can be readily identified using routine techniques well known in the art, such as by aligning sequences. Methods for sequence alignment for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) J Mol Biol 48: 443-453) to find the alignment of two complete sequences that maximizes the number of pairings and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of the similarity between the two sequences. The computer application for performing BLAST analysis is publicly available through the NationalCentre for Biotechnology Information. Counterparts can be readily identified using, for example, the ClustalW multi-sequence alignment algorithm (version 1.83), with standard paired alignment parameters, and a percentage scoring method. Secondary manual editing can be performed to optimize alignment between conserved motifs, as would be apparent to a person skilled in the art.

Counterparts also include orthologs and paralogues. Orthologs can be easily found by performing a so-called BLAST perform with a query string (for example, SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177 or SEQ ID NO: 178) against any sequence database. , such as NCBI's publicly available database. BLASTN or TBLASTX (using default default values) can be used when starting from a nucleotide sequence and BLASTP or TBLASTN (using default default values) can be used when starting from a protein sequence. BLAST results can be optionally filtered. The full-length sequences of either the filtered or unfiltered results are then BLASTed back (according to BLAST) against the organism sequences from which the query sequence is derived (where the query sequence is SEQ ID NO: 175, SEQ ID NO: 176, SEQ ID NO: 177 or SEQ ID NO: 178, the second BLAST must therefore be against sequences from Arabidopsis thaliana). The results of the first and second BLAST are then compared. A paralog is identified if a first BLAST high score hit is of the same species from which the query sequence is derived; An ortholog is identified if a high score hit is not the same species from which the query sequence is derived. High score hits are those with a low E value. The lower the E value, the more significant the score (or in other words, the lower the chance that the hit was found at random). Value computing E is well known in the art. For large families, ClustalW can be used, followed by a neighboring cluster tree, to help visualize related gene clusters and to identify orthologs and paralogues.

In addition to E values, comparisons are also punctuated by percent identity. Percent identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid sequences (or polypeptides) compared over a particular length. PLT transcription factor polypeptide homologues are preferably in decreasing order of at least 25%, 30% , 40%, 50%, 60%, 70%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% or more sequence identity or similarity (functional identity) with a polypeptide of unmodified PLT transcription factor as represented by SEQ ID NO: 176 or SEQ ID NO: 178. Percentage identity between PLT transcriptional defactor polypeptide homologs outside of AP2 domain so they say low.Preferably5 PLT transcription factor polypeptide homologues comprise one domain AP2 with ascending order of preference at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity with the AP2 domain represented by SEQ ID NO: 191 Still preferably, PLT transcription factor polypeptide homologues are as represented by SEQ ID NO: 180, SEQ ID NO: 182, SEQ ID NO: 184, SEQ ID NO: 188, SEQ ID NO: 200e SEQ ID NO: 202.

The PLT polypeptide may be a derivative of SEQ ED NO: 176or SEQ ID NO: 178. "Derivatives" include peptides, oligopeptides, polypeptides which may, compared to the naturally occurring amino acid sequence of the protein, such as that set forth in SEQ ID NO. : 176 or SEQ ID NO: 178, comprising amino acid substitutions with non-naturally occurring amino acid residues, or additions of non-naturally occurring amino acid residues. Derived from SEQ ID NO: 180, SEQ ID NO: 182, SEQ ID NO: 184, SEQ ID NO: 186, SEQ ID NO: 188, SEQ ID NO: 200 and SEQ ID NO: 202 are additional examples that may be suitable for use. in the methods of the invention as long as they have the same or similar biological activity.

In addition, PLT transcription factor polypeptides (at least in their native form) typically have DNA binding activity and an activation domain. One of ordinary skill in the art can easily determine the presence of a DNA activation domain and binding activity using routine techniques and procedures. Proteins interacting with PLT transcription factor compolipeptides (as, for example, in transcriptional complexes) can be readily identified using standard techniques by one of ordinary skill in the art.

Examples of nucleic acids encoding PLT transcription defector polypeptides include but are not limited to those represented by any of: 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: 199 and SEQ ID NO: 201. Nucleic acid variants encoding PLT transcription factor polypeptides may be suitable for use in the methods of the invention as long as they have the same or less biological activity. similar. Suitable variants include portions of nucleic acids encoding PLT transcription factor polypeptides and / or nucleic acids capable of hybridizing to nucleic acids / genes encoding PLT transcription factor polypeptides. Additional variants include binding variants and allelic variants of nucleic acids encoding PLT transcription factor polypeptides.

The term "portion" as defined herein refers to a DNA fragment encoding a polypeptide preferably comprising in ascending order at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98 % or 99% sequence identity to the AP2 domain represented by SEQ ID NO: 191. The portion may additionally comprise or a motif but preferably either motif 1 as represented by SEQ ID NO: 192 and / or motif 2 as represented by SEQID NO: 209. ; preferably, motif 2 has the sequences as represented by SEQ ID NO: 193.

A portion may be prepared, for example, by making one or more deletions to a nucleic acid encoding a PLT transcription factor factor polypeptide. The portions may be used in isolation or may be fused to other (or non-coding) coding sequences to, for example, produce a protein that combines various activities. When fused to other coding sequences, the resulting opolipeptide produced by translation may be larger than that predicted for the PLT transcription factor portion. Preferably, the tag encodes a polypeptide with substantially the same biological activity as the PLT transcription factor polypeptides of SEQID NO: 176 and SEQ ID NO: 178.

Preferably, the portion is a portion of a nucleic acid as represented by any of the sequences listed in Table I of the Examples. More preferably the portion is a portion of a nucleic acid as represented by SEQ ID NO: 175 and SEQ ID NO: 177.

Another variant of a nucleic acid encoding a PLT5 transcription factor polypeptide useful in the methods of the present invention is a nucleic acid capable of hybridizing under reduced stringency conditions, preferably under stringent conditions, with a nucleic acid-derived probe as defined above, whose hybridization sequence encodes a polypeptide comprising in ascending order preferably at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% sequence identity to the AP2 domain represented by SEQID NO: 191. The hybridization sequence may further comprise or a motif but preferably both motif 1 as represented by SEQ ID NO: 192 and / or motif 2 as represented by SEQ ID NO: 209, preferably as represented by SEQ ID NO: 193.

Preferably, the hybridization sequence is one that is capable of hybridizing to a nucleic acid as represented by (or probes derived from) any of the sequences listed in Table I of the Examples, or a portion of any of the above sequences (the target sequence). More preferably the hybridization sequence is capable of hybridizing to SEQ ID NO: 175 or SEQ ID NO: 177 (or probes derived therefrom). Probes are generally less than 1000 bp in length, preferably less than 500 bp in length. Commonly, probe lengths for DNA-DNA hybridization such as Southern blotting range from 100 to 500 bp, while the hybridizing region in hybridization probes. DNA-DNA such as PCR amplification are generally smaller than 50 but larger than chenucleotides.

The PLT transcription factor polypeptide may be encoded by a processing variant. The term "processing variant" as used herein encompasses variants of a nucleic acid sequence in which selected introns and / or exons have been cut, substituted, displaced or added, or in which introns have been shortened or lengthened. Such variants will be ones in which the substantial biological activity of the protein is maintained, which may be selectively retained by functional segments of the protein. Such union variants may be found in nature or may be man-made. Methods for making such joint variants are well known in the art.

Preferred splice variants are nucleic acid splice variants encoding a PLT transcription factor polypeptide preferably comprising in ascending order at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98 % or 99% sequence identity to the AP2 domain represented by SEQ ID NO: 191. Linking variants may additionally comprise either a motif but preferably a motive 1 as represented by SEQ ID NO: 192 and / or motif 2 as represented by SEQ ID NO: 209, preferably as represented by

SEQ ID NO: 193.

Further preferred splice variants of nucleic acids encoding PLT transcription factor polypeptides comprising features as defined above are splice variants of a nucleic acid as represented by any of the sequences listed in Table I of the Examples. More preferred is a process-variant nucleic acid sequence as represented by SEQ ID NO: 175 and SEQ ID NO: 177. The PLT transcription factor polypeptide may also be encoded by an allelic variant of a nucleic acid encoding a polypeptide comprising in ascending order. preferably at least 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identity mismatch with the AP2 domain represented by SEQ ID NO: 191. further comprising or a motif but preferably as motif 1 as represented by SEQ ED NO: 192 and / or motif 2 as represented by SEQ ID NO: 209, preferably as represented by SEQ ID NO: 193.

Preferred allelic variants of PLT transcription factor polypeptide-encoding nucleic acids comprising characteristics as defined above are splice variants of a nucleic acid represented by any of the sequences listed in Table I of the Examples. Most preferred is an allelic variant of a nucleic acid sequence as represented by SEQ ID NO: 175 and SEQ ID NO: 177.

Increased expression of a nucleic acid encoding a PLT transcription factor polypeptide leads to increased levels of the corresponding polypeptide or mRNA, which may equal the increased activity of the PLT transcription factor polypeptide; or activity can also be increased when there is no change in polypeptide levels, or even when there is a reduction in polypeptide levels. This can occur when the intrinsic properties of the polypeptide are changed, for example by making mutant versions that are more active than the polypeptide. Wild type.

Directed evolution (or gene shuffling) can be used to generate nucleic acid variants encoding PLT transcription factor polypeptides. This consists of DNA scrambling repeats followed by appropriate screening and / or selection to generate nucleic acid variants or portions thereof encoding transcription factor FPL polypeptides or homologues or portions thereof having a biologically modified activity (Castle et al., (2004) Science 304 ( 5674): 1151-4; U.S. Patent Nos. 5,811,238 and 6,395,547).

Site-directed mutagenesis can be used to generate nucleic acid variants encoding PLT transcription factor polypeptides. Several methods are available to achieve site-directed mutagenesis, most commonly being PCR (current protocols inmolecular biology. Wiley Eds.) Based methods.

Directed evolution and site directed mutagenesis are examples of technologies that enable the generation of nucleic acid variants encoding modified activity PLT transcription factor polypeptides useful for carrying out the methods of the invention.

Nucleic acids encoding PLT transcription factor polypeptides may be derived from any natural or artificial source. Nucleic acid may be modified from its native form into genomic composition and / or environment through deliberate human manipulation. Preferably the PLT transcription factor nucleic acid is from a plant, preferably from a dicotyledonous plant, preferably from the Brassicaceae family, more preferably from the genus Arabidopsis, more preferably the nucleic acid is from Arabidopsisthaliana.

The methods of the invention rely on enhanced expression of a nucleic acid encoding a PLT transcription factor polypeptide in a plant. The nucleic acid may be a full length nucleic acid or may be a nucleic acid hybridization or otherwise portion or sequence as defined above.

The methods of the invention rely on enhanced expression of a nucleic acid encoding a PLT transcription factor polypeptide in a plant. The invention also provides genetic constructs and vectors for facilitating introduction and / or expression in a plant of nucleic acid sequences useful in methods according to the invention. the invention.

Therefore, a gene construct is provided comprising: (i) A nucleic acid encoding a PLT transcription factor polypeptide as defined above;

(ii) One or more control sequences, of which at least one is a constitutive half-force promoter.

Constructs useful in the methods according to the present invention may be constructed using recombinant DNA technology well known to those skilled in the art. The gene constructs may be inserted into commercially available vectors suitable for transformation into plants and suitable for dogene expression of interest in the transformed cells. The invention therefore provides use of a gene construct as defined above in the methods of the invention.

Plants are transformed with a vector comprising the sequence of interest (i.e., a nucleic acid encoding a PLT transcription factor polypeptide). The skilled artisan is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells containing the sequence of interest. The sequence of interest is operatively linked to one or more control sequences (at least one promoter).

Nucleic acid encoding a PLT transcription factor polypeptide or variant thereof is operably linked to a constructive promoter, preferably a constitutive half-force promoter. A constitutive engine is transcriptionally active during most, but not necessarily all, phases of its growth and development and is substantially ubiquitously expressed at moderate levels. Motor force and / or expression pattern can be analyzed as described in the definitions section. The promoter strength and / or expression pattern may for example be compared to those of a well-characterized preferred reference reference promoter, such as the Cab27 promoter (weak expression, GenBank AP004700) or the putative protochlorophidiductase promoter (strong expression, GenBank AL606456 ). Preferably the promoter used in the methods of the present invention is derived from a plant, still preferably a monocot plant. Most preferred is use of a GOS2 (rice) engine (SEQ ID NO: 194 or alternatively SEQ ID NO: 210). It should be clear that the applicability of the present invention is not restricted to the nucleic acid represented by SEQ ID NO: 175 or SEQ ID NO: 177, nor is the applicability of the invention restricted to expression of a nucleic acid encoding a PLT transcription factor polypeptide when directed by a GOS2 promoter. Examples of other constructive promoters that may also be used to direct expression of a nucleic acid encoding a PLT transcription factor polypeptide are shown in the definitions section.

Optionally, one or more terminator sequences (also a control sequence) may be used in the construction introduced in a plant. Additional regulatory elements may include transcriptional as well as translational enhancers. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Other control sequences (in addition to promoter, enhancer, silencer, intron sequences, 3'UTR and / or 5'UTR regions) may be protein and / or RNA stabilizing agents. Such sequences must be known or readily obtainable by a person. versed in art.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl. The genetic construct may optionally comprise a selectable marker gene.

The present invention also encompasses viable plants by the methods according to the present invention. The present invention therefore provides plants, plant parts or plant cells of these viable by the method according to the present invention, wherein the plants or parts thereof comprise a nucleic acid transgene encoding a PLT transcription factor polypeptide under the control of a constructive promoter, preferably a non-viral constitutive promoter.

The invention also provides a method for producing transgenic plants having increased production over control plants, comprising introducing and preferentially expressing a nucleic acid encoding a PLT transcription factor polypeptide in a plant.

More specifically, the present invention provides a method for the production of transgenic plants having increased yield which method comprises:

(i) introducing and expressing a nucleic acid encoding a PLT transcription factor polypeptide in a plant; and

(ii) cultivate the plant cell under conditions that promote plant growth and development.

Nucleic acid can be introduced directly into a plant cell or into the plant itself (including introduction into a tissue, organ or any other part of a plant). According to a preferred feature of the present invention, nucleic acid is preferably introduced into a plant by transformation. Generally after transformation, plant cells or cell clumps are selected for the presence of one or more markers that are encoded by expressible genes in co-transferred plants. with the gene of interest, following which the transformed material is regenerated into an entire plant. Following DNA transfer and regeneration, putatively transformed plants can be evaluated, for example using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, or quantitative PCR, all techniques being well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first generation (or IT) transformed plant may be self-fertilized to generate second generation (or T2) homozygous transformants, and T2 plants further propagated by classical breeding techniques.

The generated transformed organisms can adopt a variety of forms. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); transformed and unprocessed detained grafts (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention extends further to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that the same feature (s) exhibit the same genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also includes host cells containing an isolated nucleic acid encoding a PLT transcription factor polypeptide. Preferred host cells according to the invention are plant cells.

The invention further extends to cut-off parts of a plant such as, but not limited to seeds, leaves, fruits, flowers, stems, rhizomes, tubers and bulbs. The invention furthermore relates to derived products, preferably directly derived, from a cut-off part of such a plant, such as pellets or dry powders, oil, fat, fatty acids, starch or proteins.

The present invention also encompasses use of nucleic acids encoding PLT transcription factor polypeptides and use of PLT transcription factor polypeptides to increase plant yield as defined above in the methods of the invention.

Enhanced expression of a nucleic acid encoding a PLT transcription factor polypeptide can be accomplished by introducing genetic modification (preferably at the locus of a PLT transcription factor gene). The locus of a gene as defined herein is adopted to mean a genomic region, which includes the gene of interest and IOKBantes or after the coding region.

Genetic modification may be introduced by alternative methods such as that described above, for example by any of the following: T-DNA activation, TILLING, and homologous recombination. Following introduction of genetic modification, there is an optional step of selecting for increased expression of a nucleic acid encoding a PLT3 transcription factor polypeptide whose increased expression generates plants having increased yield.

Identification by T-DNA activation results in plant-transgenics that show dominant phenotypes due to modified expression of genes near the introduced promoter. The promoter to be introduced is a constitutive medium-strength promoter capable of enhancing nucleic acid expression encoding a PLT transcription factor polypeptide in a plant.

A genetic modification can also be introduced into a gene encoding a transcription factor polypeptide PLT using the TILLING (Locally Targeted Injuries into Genomes) technique.

The effects of the invention may also be reproduced using homologous recombination. The nucleic acid to be introduced (which may be a nucleic acid encoding a PLT transcription factor factor polypeptide or variant thereof as defined above) is directed to the olocus of a PLT gene. The nucleic acid to be targeted may be an improved enhancement used to replace the endogenous gene or may be introduced into the endogenous gene.

T-DNA activation, TILLING, and homologous recombination are examples of technologies that enable the generation of genetic modifications comprising increasing expression of a nucleic acid by encoding a PLT transcription factor polypeptide in plants.

Nucleic acids encoding PLT transcription factor polypeptides, or PLT transcription factor polypeptides, may find use in breeding programs in which a DNA marker that can be genetically linked to a PLT transcription factor gene is identified. Nucleic acids / genes, or PLT transcription factor polypeptides can be used to define a molecular marker. This DNA or protein marker can then be used in breeding programs to select plants having increased yield as defined above in the methods of the invention.

Allelic variants of a PLT transcription factor gene nucleic acid / gene may also find use in marker-aided enhancement programs. Such breeding programs sometimes require the introduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis, or alternatively, the program may start with a collection of unintentionally caused "natural" allelic variants.

Identification of allelic variants then occurs, for example, by PCR. This is followed by a step for selecting higher allelic variants of the sequence in question and generating increased production. Selection is typically performed by monitoring plant growth performance containing different allelic variants of the sequence in question. Decreasing performance can be monitored in a greenhouse or in the field.

Additional optional steps include crossing plants in which the upper allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

A nucleic acid encoding a PLT transcription factor polypeptide may also be used as probes to genetically and physically map the genes of which they are a part of, and as markers for traits linked to such genes. Such information may be useful in plant breeding in order to develop strains with desired phenotypes. Such use of PLT transcription factor nucleic acids requires only a nucleic acid sequence of at least 15 nucleotides of length. PLT transcription factor nucleic acids can be used as restriction-fragment length polymorphism (RFLP) markers. Southern blots (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) of restriction digested plant genomic DNA can be probed with PLT transcription factor nucleic acids. The resulting banding pattern can then be subjected to genetic analysis using computer programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, nucleic acids may be used to probe Southern blots containing endonuclease-treated genomic DNAs from a set of individuals representing ancestor and progeny of a defined genetic crossover. Segregation of DNA polymorphisms is observed and used to calculate the position of the PLT transcription factor nucleic acid in the genetic map previously obtained using this population (Botstein et al. (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4: 37-41. Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 cross-breeding populations, inbreeding populations, randomly crossed populations, nearby isogenic strains, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and cited references. in this one).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify differences in DNA sequences between ancestors of the cross-mapping region corresponding to the current sequence of nucleic acids. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having increased yield as described above. This increased production may also be combined with other economically advantageous features, such as additional production enhancing features, tolerance to other abiotic and biotic stresses, features modifying various architectural features and / or biochemical and / or physiological features.

Detailed Description of bHLHS Surprisingly, it has now been found that modulating expression in a plant of a nucleic acid encoding a particular bHLH transcription factor class generates plants having improved production characteristics relative to control plants. The particular bHLH transcription defector class suitable for improving plant yield characteristics is described in detail below.

The present invention provides a method for enhancing plant production characteristics over control plants, comprising modulating expression in a plant of a nucleic acid by encoding a particular class of bHLH transcription factor.

A preferred method for modulating (preferably enhancing) expression of a nucleic acid encoding a bHLH transcription factor is to introduce and express into a plant a nucleic acid encoding a particular class of bHLH transcription factor as further defined below.

The nucleic acid to be introduced into a plant (and therefore useful for carrying out the methods of the invention) is any nucleic acid encoding the type of bHLH transcription factor that will now be described. A bHLH transcription factor as defined herein refers to a polypeptide represented by any of SEQ ID NO 213, SEQ ID NO: 217, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301. The invention is illustrated by transforming plants with the Oryza sativa sequences represented by SEQ ID NO: 212, encoding the polypeptide of SEQ ID NO: 213. SEQ ID NO: 215 deOryza sativa (encoded by SEQ ID NO: 214) and Oryzasativa SEQ ID NO: 217 (encoded by SEQ ID NO: 216) are paralogs of the SEQ ID NO: 213 polypeptide. SEQ ID NO: 219 from Arabidopsis thaliana (encoded by SEQ ID NO: 218) and SEQ ID NO: 225 from Arabidopsis thaliana (encoded by SEQ ID NO: 224) are orthologs of the polypeptide of SEQ ID NO: 213. Arabidopsis thaliana SEQID NO: 221 (encoded by SEQ ID NO: 220) and Arabidopsis SEQID NO: 223 thaliana (encoded by SEQ ID NO: 222) are variants of SEQ ID NO: 218 and SEQ ID NO: 219.

Orthologs and paralogues can easily be found by performing a so-called reciprocal blast search. Typically this involves a first BLAST involving performing BLAST with a query string (e.g. SEQ ID NO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216, SEQ ID NO: 217, SEQ ID NO: 218, SEQ ID NO: 219, SEQ ID NO: 224 or SEQ ID NO: 225) against any sequence database, such as the publicly available NCBI database. BLASTN or TBLASTX (using default defaults) are generally used when starting from a nucleotide sequence, and BLASTP or TBLASTN (using default defaults) when starting from a protein sequence. BLAST results may be optionally filtered. The full length sequences of the filtered or unfiltered results are then BLASTed back (according to BLAST) against sequences from the organism from which the query sequence is derived (where the query sequence is SEQ IDNO: 212, SEQ ID NO: 213, SEQ ID NO: 214, SEQ ID NO: 215, SEQ ID NO: 216 or SEQ ID NO: 217, the second BLAST must therefore be Oryza's counterparts, where the query sequence is SEQ ID NO: 218, SEQID NO: 219, SEQ ID NO: 224 or SEQ ID NO: 225, the second BLAST should therefore be against Arabidopsis sequences). The results of the first BLAST second are then compared. A parallel is identified if a high score of the first BLAST is of the same species from which the query sequence is derived, a return BLAST then ideally results in the query sequence as the highest hit; an ortholog is identified if a high score hit in the first BLAST is not the same species from which the query sequence is derived, and preferably results by returning BLAST in the query sequence being among the highest hits.

High score hits are those with a low value E. The lower the E value, the more significant the score (or in other words the lower the chance that the hit was found at random). E-value computation is well known in the art. . In addition to E-values, comparisons are also scored by percentage identity. Percent Identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. Preferably the score is greater than 50, more preferably greater than 100; and preferably the value

E is smaller than e-5, more preferably less than e-6. An example detailing the identification of orthologs and paralogues is given in Example 31 and Example 30 respectively in this place. For large families, ClustalW can be used, followed by a neighboring cluster tree, to help visualize related gene clustering and to identify orthologs and paralogs.

Examples of orthologs obtained by the BLAST procedure mentioned above are SEQ ID NO: 227 from Medicago truncatula (coded by SEQ ID NO: 226) and SEQ ID NO: 229 from Hordeum vulgare (coded by SEQ ID NO: 228). Additional orthologs and paralogs can be readily identified using the BLAST procedure described above by following the procedure given in the examples section.

The proteins represented by SEQ ID NO: 297, 299 and 301 have been hitherto unknown. Therefore, the invention also provides hitherto unknown bHLH transcription factors and bHLH transcription factor encoding nucleic acids.

According to a further embodiment of the present invention, there is therefore provided an isolated nucleic acid molecule comprising:

(i) a nucleic acid represented by any one of SEQID NO: 296, SEQ ID NO: 298 and SEQ ID NO: 300;

(ii) the complement of a nucleic acid represented by any one of SEQ ID NO: 296, SEQ ID NO: 298 and SEQ ID NO: 300;

(iii) a nucleic acid encoding a bHLH transcription factor having (a) in ascending order preferably at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the amino acid sequence represented by any one of SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301, and (b) one bHLH domain.

According to a further embodiment of the present invention, there is also provided an isolated polypeptide comprising:

(i) an amino acid sequence represented by any one of SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301;

(ii) an amino acid sequence having (a) preferably in ascending order at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96% , 97%, 98%, 99% or more sequence identity with any of the amino acid sequences represented by SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301, and (b) a bHLH domain;

(iii) derived from any of the amino acid sequences given in (i) or (ii) above.

Polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 221, SEQ ID NO 223, SEQ ID NO 225, SEQ ID NO: 227, SEQ ID NO: 229, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, or orthologs or dialogues of any of the above mentioned SEQ ID NOs all comprise a bHLH domain.

BHLH domains are well known in the art and can readily be identified by those skilled in the art. The family is defined by a bHLH signature domain, which consists of 60 or more amino acids with two functionally distinct regions. A base region, located at the N-terminal end of the domain, is involved in DNA ligation and consists of 15 or more amino acids with a high number of basic residues. An HLH region at the C-terminal end functions as a dimerization domain and comprises mainly hydrophobic residues forming two aliphatic propellers separated by a variable loop and length loop region.

A bHLH domain can be identified using methods for sequence alignment for comparison. In some instances, default parameters may be adjusted to modify the stringent stringency. For example using BLAST, the threshold of statistical significance (called the E value) for reporting pairings against database sequences can be increased to show less stringent pairings. In this way, virtually exact short pairings can be identified.

Methods for sequence alignment for comparison are well known in the art, such methods include GAP, BESTFIT, BLAST, FASTA and TFASTA. GAP uses the algorithm of Needleman and Wunsch ((1970) JMol Biol 48: 443-453) to find the global alignment (along the entire sequence) of two sequences that maximizes the number of pairings and minimizes the number of gaps. The BLAST algorithm (Altschul et al. (1990) JMol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of similarity between the two sequences. The computational application for performing BLAST analysis is publicly available through the National Center for Biotechnology Information (NCBI). Homologists can be readily identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83), with standard paired alignment parameters, and a percentage count method. Secondary manual editing can be performed to optimize alignment between conserved motifs, as would be apparent to a person skilled in the art.

There are also expert databases for domain identification. The bHLH domain in a bHLH transcription factor can be identified using, for example, SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic et al. (2002) Nucleic Acids Res 30, 242-244), InterPro (Mulder et al., (2003) Nucl. Acids.Res. 31,315-318), Prosite (Bucher and Bairoch (1994), A generalized profilesyntax for biomolecular sequence motifs and its funetion in automaticsequence interpretation (In) ISMB-94; Proceedings 2nd International Conference on Intelligent Systems for Molecular Biology Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., Pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137 (2004)) or Pfam (Bateman etal., Nucleic Acids Research 30 (1): 276-280 (2002)). The HLH domain structure in the InterPro database is named IPROO1092 and IPRO1 1598; PFOOO10 in the Pfam database; SM00353 in the SMART ePS50888 database in the PROSITE database. In addition, the alignment shown in Figure 18 highlights the bHLH domain of bHLH transcription factor useful in the methods of the invention.

Polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 257, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 or Orthologs or paralogues of any of the SEQ ID NOs mentioned above typically display considerable sequence divergence outside the conserved bHLH domain.

Figure 19a is a matrix showing the similarities and general identities (in bold) of the bHLH-like proteins described above. Even though identities appear to be relatively low, polypeptides having sequence identities falling within the variations shown in the matrix may be adopted as orthologs or paralogs of any kind. from SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219 or SEQ LD NO 225; This can be confirmed by the reciprocal deblast procedure described above. Typically, bHLH transcription factor encoding nucleic acids useful in the methods of the invention are preferably in ascending order of at least 13%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55 %, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity with bHLH transcription factors represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219 or SEQ ID NO 225.

The matrix shown in Figure 19b shows similarities and identities (in bold) along the bHLH domain, where certainly the values are higher than when considering full length proteins. Typically, nucleic acids encoding bHLH transcription factors useful in the methods of the invention have bHLH domains preferably in ascending order of at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65. %, 70%, 75%, 80%, 85%, 90%, 95% or more sequence identity to the bHLH domain of any of the polypeptides represented by any one of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219 or SEQ ID NO 225.

An amino acid pattern, termed a 5-9-13 configuration, can be found at three positions within the basic bHLH domain region (see Fig. 4 of Heim et al., 2003 (Mol. Biol. Evol. 20 (5): 735-747) and Figure 18 where three upward-pointing arrows show the configuration). Polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 257, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 or Orthologs or paralogs of any of the above-mentioned SEQ ED NOs preferably comprise a K / R ER5 configuration, more preferably an RER5 configuration within the bHLH domain, typically within the basic region of the domain. The presence of this configuration is not a prerequisite for performing the methods of the invention, so some variation in the K / R ER configuration is acceptable. Arabidopsis bHLH polypeptides were grouped into twelve subfamilies according to structural similarities (see Fig. 4 of Heimet al., 2003). Group IX members constitute bHLH polypeptides having an RER configuration. In addition, a Group VI member comprises an RER configuration.

Polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 257, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 or Orthologs or analogues of any of the above-mentioned SEQ ID NOs may also comprise (in addition to a bHLH domain and optionally a K / R ER 5-9-13 motif, preferably an RER motif) a domain designated PFB26111 (see Pfam database). The domain is also indicated in Figure 18.

Typically, nucleic acids encoding bHLH transcription factors (as defined above) having a bHLH domain and having, in ascending order preferably, at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85 %, 90%, 95% or more sequence identity to domainPFB26111 (see Figure 18) of any of the polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219 SEQ ID NO: 225, SEQ ID NO: 297, SEQ ID NO: 299, or SEQ ID NO: 301 are useful in the methods of the invention.

Nucleic acids encoding the polypeptides represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQID NO : 301, or orthologs or paralogues of any of the above mentioned SEQ ID NOs need not be full length nucleic acids, since performance of the methods of the invention is not relied upon for full length nucleic acid sequences. Examples of nucleic acids suitable for use to carry out the methods of the invention include but are not limited to those represented by any of: SEQ ID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO : 220, SEQ ID NO: 222, SEQ ID NO: 224, SEQ ID NO: 226, SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, and SEQ ID NO: 300.Nucleic Acid Variants They may also be useful for practicing the methods of the invention. Examples of such nucleic acid variants include nucleic acid moieties encoding a bHLH transcription factor as defined herein, nucleic acid splice variants encoding a bHLH transcription factor as defined herein, allelic nucleic acid variants encoding a bHLH transcription factor as defined herein and nucleic acid variants encoding a bHLH transcription factor as defined herein which are obtained by gene shuffling. The terms portion, processing variant, allelic variant, and gene shuffling will now be described.

A portion of a nucleic acid encoding a bHLH transcription factor as defined herein may be prepared, for example, by making one or more nucleic acid deletions. The portions may be used in isolation or they may be fused to other (or non-coding) coding sequences to, for example, produce a protein that combines various activities. When fused to other coding sequences, the resulting polypeptide produced by translation may be greater than predicted for the bHLH transcription factor factor portion.

According to the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing into a plant a portion of a nucleic acid encoding a bHLH transcription factor represented by any of SEQID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, or a portion of a nucleic acid encoding orthologs, paralogs, or homologues of any of the SEQ ID NOs mentioned above.

Portions useful in the methods of the invention encode a polypeptide having a bHLH domain (as described above) and have substantially the same biological activity as the bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, or orthologs or paralogues of any of the SEQID NOs mentioned above. The portion is typically at least 150 consecutive nucleotides in length, preferably at least 300 consecutive nucleotides in length, more preferably at least 400 consecutive nucleotides in length, and more preferably at least 500 consecutive nucleotides in length. Preferably, the moiety is a portion of a nucleic acid as represented by any one of SEQ ID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 220, SEQ ID NO: 222 , SEQ ID NO: 224, SEQ ID NO: 226, SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, and SEQ ID NO: 300. More preferably the portion is a portion of a nucleic acid as represented by SEQ ID NO: 212.

Another variant of nucleic acid useful in the methods of the invention is a nucleic acid capable of hybridizing under reduced stringency conditions, preferably under stringent conditions, with a nucleic acid encoding a bHLH transcription factor as defined herein, or with a moiety as defined herein. Hybridization sequences useful in the methods of the invention encode a polypeptide having a bHLH domain (as described above) and having substantially the same biological activity as the bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 211, SEQ ID NO: 217 SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 or having substantially the same biological activity as orthologs or paralogues of any of the above SEQ ID NOs. The hybridization sequence is typically at least 150 consecutive nucleotides in length, preferably at least 300 consecutive nucleotides in length, more preferably at least 400 consecutive nucleotides in length, and more preferably at least 500 consecutive nucleotides in length. Preferably, the hybridization sequence is one which is capable of hybridizing to any nucleic acid represented by SEQ ID NO: 212, SEQ ID NO: 214, SEQID NO: 216, SEQ ID NO: 218, SEQ ID NO: 220, SEQ ID NO: 222, SEQ ID NO: 224, SEQ ID NO: 226, SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, and SEQ ID NO: 300 or with a portion of any of the above sequences, a portion being as defined above. Most preferably the hybridization sequence is capable of hybridizing to a nucleic acid as represented by SEQ ID NO: 212, or portions thereof.

In accordance with the present invention, there is provided a method for enhancing plant production characteristics, comprising introducing and expressing into a plant a nucleic acid capable of hybridizing to a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, or comprising introducing and expressing in a plant a nucleic acid capable of hybridizing with a nucleic acid encoding aortologist, paralogue or homologue of any of the above mentioned SEQ ID NOs.

Another nucleic acid variant useful in the methods of the invention is a processing variant encoding a bHLH transcription factor as defined above.

In accordance with the present invention, there is provided a method for improving plant yield characteristics, comprising introducing and expressing in a plant a nucleic acid processing variant encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 225, SEQ ID NO: 297, SEQ ID NO: 299, and SEQ ID NO: 301, or a processing variant of a nucleic acid encoding an ortholog, analog or homolog of any of the SEQ IDNOs mentioned above.

Preferred splice variants are splice variants of a nucleic acid encoding bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297 , SEQ ID NO: 299, and SEQ ID NO: 301, or coupling variants encoding orthologs or dialogues of any of the above mentioned SEQ ID NOs. Further preferred are nucleic acid binding variants represented by any of SEQID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 220, SEQ ID NO: 222, SEQ ID NO. : 224, SEQ ID NO: 226, SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, and SEQ ID NO: 300. Most preferred is a processing variant of a nucleic acid represented by SEQ ID NO: 212 .

Another nucleic acid variant useful for carrying out the methods of the invention is an allelic variant of a nucleic acid encoding a bHLH transcription factor as defined above. According to the present invention, there is provided a method for improving plant yield characteristics, comprising introducing and expressing in a plant is an allelic variant of a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 297, SEQ ID NO: 299, and SEQ ID NO: 301, or comprising introducing and expressing in a plant an allelic variant of a nucleic acid encoding an ortholog, paralog or homologue of any of the above mentioned SEQ ID NOs.

The allelic variant may be an allelic variant of a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO. : 297, SEQ ID NO: 299, and SEQ ID NO: 301, or an allelic variant of a nucleic acid encoding orthologs or paralogues of any of the above mentioned SEQ ID NOs. Further preferred are allelic nucleic acid variants represented by any of SEQID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 220, SEQ ID NO: 222, SEQ ID NO: 224, SEQ ID NO: 226 and SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, SEQ ID NO: 300. Most preferred is an allelic variant of a nucleic acid as represented by SEQ ID NO: 212.

An additional nucleic acid variant useful in the methods of the invention is a nucleic acid variant obtained by scrambling. Gene shuffling or directed evolution can also be used to generate nucleic acid variants encoding bHLH transcription factors as defined above.

In accordance with the present invention, a method is provided for improving plant yield characteristics, comprising introducing and expressing in a plant a variant of a nucleic acid encoding a bHLH transcription factor represented by any of SEQID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ and ID NO: 301, or comprising introducing and expressing in a variant of a nucleic acid encoding an ortholog, paralog or homolog. of any of the above mentioned SEQ IDNOs, whose nucleic acid is obtained by scrambling.

In addition, nucleic acid variants can also be obtained by site-directed mutagenesis. Several methods are available to achieve site-directed mutagenesis, the most common being PCR-based methods (current protocols in molecular biology. Wiley Eds.).

Also useful in the methods of the invention are nucleic acids encoding homologues of any of the amino acids represented by SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 orortologists or paralogues of any of the SEQ ID NOs mentioned above.

Also useful in the methods of the invention are nucleic acids encoding derivatives of any of the amino acids represented by SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 orortologists or paralogues of any of the SEQ ID NOs mentioned above. Derived from SEQ ID NO: 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO: 221, SEQ ID NO: 223, SEQ ID NO: 225, SEQ ID NO: 227 and SEQ ID NO: 229, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 are additional examples that may be suitable for use in the inventive methods.

In addition, bHLH transcription factors (at least in their native form) typically have DNA binding activity and a reactive domain. A person skilled in the art can easily determine the presence of a domain of DNA activation and binding activity using tools and routine techniques.

Nucleic acids encoding bHLH transcription factors may be derived from any natural or artificial source. Nucleic acid may be modified from its native form in genomic composition and / or environment through deliberate human manipulation.

Preferably the nucleic acid encoding bHLH transcription factor is from a plant, still preferably from a monocot, more preferably from the Poaceae family, more preferably Oryza sativa nucleic acid.

Any reference in this place to a bHLH transcription factor is therefore adopted to mean a bHLH transcription factor as defined above. Any nucleic acid encoding such a bHLH transcription factor is suitable for use to perform the methods of the invention.

The present invention also encompasses plants or parts thereof (including plant cells) viable by the methods according to the present invention. Plants or parts thereof comprise a nucleic acid transgene encoding a bHLH transcription factor as defined above.

The invention also provides genetic constructs and vectors for facilitating introduction and / or expression of nucleic acid sequences in the methods according to the invention in a plant.

Therefore, a gene construct is provided comprising:

(i) Any nucleic acid encoding a bHLH-type discription factor as defined above;

(ii) One or more control sequences operably linked to the nucleic acid of (i).

Constructs useful in the methods according to the present invention may be constructed using recombinant DNA technology well known to those skilled in the art. The gene constructs may be inserted into commercially available vectors suitable for transformation into plants and suitable for dogene expression of interest in the transformed cells. The invention therefore provides us with a gene construct as defined above in the methods of the invention.

Plants are transformed with a vector comprising the sequence of interest (i.e., a nucleic acid encoding a bHLH-like transcription factor). The skilled artisan is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells containing the sequence of interest. The sequence of interest is operatively linked to one or more control sequences (at least one promoter).

Advantageously, any type of promoter, whether natural or synthetic, may be used to direct expression of the nucleic acid sequence. The promoter may be an inducible promoter, i.e. having induced or increased transcriptional initiation in response to an evolutionary, chemical, environmental or physical stimulus. The promoter may be a tissue specific promoter, i.e. one which is preferably capable of initiating transcription in certain tissues, such as leaves, roots, seed tissue, etc.

According to a preferred feature of the invention, the nucleic acid encoding a bHLH-like transcription factor is operatively linked to a constitutive promoter. A constructive promoter is transcriptionally active during most but not necessarily all phases of its growth and development and is substantially ubiquitously expressed. The constitutive promoter is preferably a GOS2 promoter, more preferably the constitutive promoter is a rice GOS2 promoter, yet preferably the constructive promoter is represented by the nucleic acid sequence substantially similar to SEQ ID NO: 230 or SEQ ID NO: 233, most preferably the constitutive promoter is as represented by SEQ IDNO: 233.

It should be clear that the applicability of the present invention is not restricted to the bHLH transcription factor encoding nucleic acid represented by SEQ ID NO: 212, nor is the applicability of the invention restricted to the expression of such a bHLH transcription factor encoding nucleic acid when directed by a GOS2 promoter. Examples of other constitutive promoters that may also be used to carry out the methods of the invention are shown in the definitions section.

Optionally, one or more terminator sequences (also a control sequence) may be used in the construction introduced in a plant. Additional regulatory elements may include transcriptional as well as translational enhancers. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Such sequences must be known or readily obtainable by a person skilled in the art.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl. Other control sequences (other than motor, enhancer, silencer, intron sequences, 3'UTR and / or 5'UTR regions) may be protein and / or RNA stabilizing agents. The genetic construct may optionally comprise a selectable marker gene.

The invention also provides a method for producing transgenic plants having improved production characteristics over control plants, comprising introducing and expressing in a plant any nucleic acid encoding a bHLH-like transcription factor factor polypeptide as defined above.

More specifically, the present invention provides a method for the production of transgenic plants having improved production characteristics, the method of which comprises:

(i) introducing and expressing a nucleic acid encoding a bHLH-like transcription factor (as defined herein) in a cellulose; and

(ii) cultivate the plant cell under conditions that promote plant growth and development.

Nucleic acid can be introduced directly into a plant cell or into the plant itself (including introduction into a tissue, organ or any other part of a plant). According to a preferred feature of the present invention, nucleic acid is preferably introduced into a plant by transformation.

Generally after transformation, plant cells or cell clusters are selected for the presence of one or more markers that are encoded by expressible genes in plants co-transferred with the gene of interest, following which the transformed material is regenerated into an entire plant.

Following DNA transfer and regeneration, computationally transformed plants can be evaluated, for example using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, or quantitative PCR, all techniques well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first generation (or IT) transformed plant may be self-fertilized to generate second generation (or T2) homozygous transformants, and T2 plants further propagated by classical breeding techniques.

The generated transformed organisms can adopt a variety of forms. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); transformed and unprocessed detained grafts (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention extends further to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that the same feature (s) exhibit the same genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also includes host cells containing an isolated nucleic acid encoding a bHLH transcription factor as defined above. Preferred host cells according to the invention are plant cells.

The invention also extends to cut-off parts of a plant such as, but not limited to seeds, leaves, fruits, flowers, stems, rhizomes, tubers and bulbs. The invention furthermore relates to derived products, preferably directly derived, from a cut-off part of such a plant, such as dried pellets or powders, oil, fat, fatty acids, starch or proteins. According to a preferred feature of the invention, modulated expression is increased expression.

As mentioned above, a preferred method for modulating (preferably enhancing) expression of a bHLH transcription factor encoding nucleic acid is to introduce and express in a plant a bHLH transcription factor encoding a nucleic acid; however the effects of performing the method, i.e. improved production characteristics can also be achieved using other well known techniques. A description of some of these techniques will now follow.

One such technique is identification by T-DNA activation. The resulting transgenic plants show dominant phenotypes due to modified expression of genes near the introduced promoter. The effects of the invention may also be reproduced using the TILLING technique. The effects of the invention may also be reproduced using homologous recombination.

Reference in this place to the term improved yield characteristics is adopted to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground (cut-off) and / or below-cut (cut-off) parts from soil.

In particular, such cut-off parts are seeds, and performance of the methods of the invention results in plants having increased seed production relative to control plant seed production.

Using corn as an example, an increase in yield can be manifested as one or more of the following: increase in the number of plants established per hectare or acre, an increase in the number of spikes per plant, an increase in the number of rows, number of seeds per row, seed weight, thousand seed weight, ear length / diameter, increase in grain fill rate (which is the number of kernels divided by the total number of grains and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary seedlings), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in weight of one thousand seeds, among others.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit increased growth rate (for at least part of their due cycle) relative to the growth rate of corresponding wild-type plants at a corresponding stage. in your life cycle. The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the plant. A plant having an increased growth rate may still exhibit early flowering. The increase in growth rate may occur at one or more stages in a plant's life cycle or substantially throughout the entire plant life cycle. Increased growth rate during the early stages of a plant's life cycle may allow improved vigor (increased seedling emergence). Increasing growth rate can alter a plant's harvest cycle by allowing plants to be sown later and / or harvested earlier than otherwise possible. If the growth rate is sufficiently increased, it may allow additional seed sowing of the same plant species (for example sowing and harvesting rice plants followed by sowing and harvesting additional rice plants all within a conventional growing period). If the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing and harvesting of soybeans, potatoes or any other suitable plant). Additional harvest times of the same rhizome in the case of some crop plants may also be possible. Changing the harvesting cycle of a plant can lead to an increase in annual biomass production per acre (due to an increase in the number of times (say in a year) that any particular plant can be grown and harvested. An increase in growth rate may also allow the cultivation of transgenic plants in a wider geographical area than their wild counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or planting time (early cycle) or at harvesting time (late cycle). Such adverse conditions can be avoided if the harvesting cycle is shortened. The growth rate can be determined by deriving several growth curve parameters, such parameters can be: T-Mid (the time used by plants for reach 50% of their maximum size) and T-90 (time used by plants to reach 90% of their maximum size), among others.

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general. Mild stress on the other hand is defined here as any stress to which a plant is exposed that does not result in stopping of general plant growth without the ability to resume growth. Mild stress in the sense of the invention leads to a reduction in growth of stressed plants of less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the control plant under non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "no stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location.

Performance of the methods of the invention generates plants grown under non-stress conditions or under mild drought conditions increased yields compared to control plants grown under comparable conditions. Therefore, according to the present invention there is provided a method for increasing production in plants grown under non-stress conditions or under mild dry conditions, which method comprises increasing a plant's expression of a nucleic acid encoding a bHLH transcription factor.

Performance of the methods of the invention generates plants grown under nutrient deficiency conditions, particularly under nitrogen deficiency conditions, increased yield over control-grown plants under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under nutrient deficiency conditions, the method of which is to increase expression in a plant of a nucleic acid by encoding a MADS15 polypeptide. Nutrient deficiency can result from nutrient deficiency or excess such as nitrogen, phosphates and other compounds containing phosphorus, potassium, calcium, cadmium, magnesium, manganese, iron and boron, among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having an increased growth rate relative to control plants, particularly during the early stages of plant development (typically three weeks after germination in the case of rice and maize, but this will vary from species to species) leading to early vigor. Therefore, according to the present invention, there is provided a method for increasing the rate of plant growth, which method comprises modulating, preferably increasing, expression in a plant of a nucleic acid encoding a bHLH transcription factor. The present invention therefore also provides a method for obtaining plants having early vigor over control plants, the method of which comprises modulating, preferably increasing, expression in a plant of a nucleic acid encoding a bHLH transcription factor.

Early vigor can also result from or be manifested as increased plant suitability relative to control plants, for example, because plants are better adapted to their environment (i.e. being better able to cope with various abiotic or ubiotic stress factors). Plants having early vigor also show better crop establishment (with the crop growing evenly, i.e. most plants reaching the various stages of development substantially at the same time), and show better growth and often better yield. Early vigor can be determined by measuring various factors such as seedling growth rate, seed weight, germination percentage, emergence percentage, seedling height, root length and shoot biomass and more.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Still preferably, the plant is a monocot plant. Examples of monocotyledonous plants include sugar cane. Most preferably aplanta is a cereal. Examples of cereals include rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats. Plants in which early vigor is a particularly desirable trait include rice, corn, wheat, sunflower, sorghum.

The present invention also encompasses use of nucleic acids encoding bHLH transcription factors and use of bHLH transcription factor polypeptides to improve production characteristics.

Nucleic acids encoding bHLH transcription factor polypeptides, or bHLH transcription factors themselves, may find use in breeding programs in which a DNA marker is identified that can be genetically linked to a bHLH transcription factor encoding gene. Nucleic acids / genes, or bHLH transcription factors themselves can be used to define a molecular marker. This DNA or protein marker can then be used in breeding programs to select plants having increased yield as defined above in the methods of the invention.

Allelic variants of a nucleic acid / gene encoding bHLH transcription factor may also find use in marker-aided enhancement programs. Such breeding programs sometimes require introduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis, alternatively, the program may start with a collection of so-called "natural" allelic variants caused unintentionally. Identification of allelic variants then occurs, for example by PCR. This is followed by a step for selecting higher allelic variants of the sequence in question and generating increased production. Selection is typically performed by monitoring plant growth performance containing different allelic variants of the sequence in question. Growth performance can be monitored in a greenhouse or in the field. Additional optional steps include crossing plants in which the upper allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

A nucleic acid encoding a bHLH transcription factor can also be used as probes to genetically map the genes of which they are a part of, and as markers for, traits linked to such genes. Such information may be useful in plant breeding to develop strains with desired phenotypes. Such use of nucleic acids encoding bHLH transcription factor requires only a nucleic acid sequence of at least 15 nucleotides in length. Nucleic acids encoding bHLH transcription factor can be used as restriction fragment length polymorphism (RFLP) markers. Southern blots (Sambrook J, Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) of restriction digested plant genomic DNA can be probed with nucleic acids encoding bHLH transcription factor. The resulting decay pattern can then be subjected to genetic analysis using computational programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, nucleic acids can be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs from a set of individuals representing the ancestor and progeny of a defined genetic cross. Segregation of DNA polymorphisms is observed and used to calculate the position of nucleic acid encoding bHLH transcription factor in the genetic map previously obtained using this population (Botstein et al. (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (1986) Plant Mol. Biol. Reporter 4: 37-41. Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 cross-breeding populations, inbreeding populations, randomly crossed populations, nearby isogenic strains, and other sets of individuals may be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and cited references. in this one).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify differences in DNA sequences between ancestors of the cross-mapping region corresponding to the current sequence of nucleic acids. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having improved production characteristics as described above. These characteristics may also be combined with other economically advantageous characteristics, such as additional production enhancing characteristics, tolerance to other abiotic and biotic stresses, characteristics modifying various production characteristics. architecture and / or biochemical and / or physiological characteristics.

Detailed Description of SPL15

Surprisingly, it has now been found that increasing a plant's expression of a nucleic acid encoding a transcription factor SPL15 polypeptide generates plants having improved production characteristics, particularly increased production, relative to control plants. Therefore, the invention provides a method for stimulating particular production characteristics for increasing plant production relative to control plants, comprising increasing expression in a plant of a nucleic acid encoding a SPL15 transcription factor factor polypeptide.

A preferred method for enhancing expression of a nucleic acid encoding an SPL15 transcription factor polypeptide is to introduce and express in a plant a nucleic acid encoding an SPL15 transcription factor polypeptide.

The term "SPL15 transcription factor polypeptide" as defined herein refers to a polypeptide comprising from N-terminal to C-terminal: (i) Reason 1 as represented by SEQ ID NO: 276; and (ii) in ascending order of preferably at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% sequence identity with the SPL DBD represented by SEQ ID NO: 277; and (iii) Reason 2 as represented by SEQ ID NO: 278.

The most conserved amino acids within Reason 1 are XLXFGXXXYFX, and within Reason 2 DSXXALSLLSX (where X is a specified subset of differing amino acids for each position, as shown in SEQ ID NO: 276 and SEQ ID NO: 277). Within Reason 1 and Reason 2, one or more conservative changes in any position are permitted, and / or one, two or three non-conservative changes in any position.

Additionally, the transcription factor polypeptide SPL15 may comprise either or both of the following: (a) a G / S rich stretch preceding the SPL DBD; and (b) the W (S / T) L tripeptide at the C-terminal end of the polypeptide.

An example of an SPL15 transcription factor polypeptide as defined above comprising from N-terminal to C-terminal: (i) Reason 1 as represented by SEQ ID NO: 276; and (ii) in ascending order of preference at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% sequence identity with the SPL DBD represented by SEQ ID NO: 277; and ( iii) Reason 2 as represented by SEQ ID NO: 278; and further comprising: (a) a G / S-rich stretch preceding the SPL DBD; and (b) the W (S / T) L polypeptide at the C- end of the polypeptide is represented as SEQ ID NO: 235. Such additional examples are represented by any one of SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287 or orthologs or paralogues of any of the SEQED NOs mentioned above. The invention is illustrated by transforming plants with the Arabidopsis thaliana sequence represented by SEQ ID NO: 234, encoding the polypeptide of SEQ ID NO: 235. SEQ ID NO: 237 of Arabidopsis thaliana (encoded by SEQ ED NO: 236) is a dopolipeptide analog of SEQ ID NO: 235. SEQ ID NO: 239 (encoded by SEQ ID NO: 238, from Aquilegia formosa χ Aquilegia pubescens), SEQ ID NO: 241 (encoded by SEQ ID NO: 240, from Gossypium hirsutum), SEQ ID NO: 243 (encoded by SEQ ID NO: 242, from Ipomoea nil), SEQ ID NO: 245 (encoded by SEQ ID NO: 244, from Lactuca sativa), SEQ ID NO: 247 (encoded by SEQ ID NO: 246, from Malus SEQ ID NO: 249 (encoded by SEQ ID NO: 248 by Medieago truneatulà), SEQ ID NO: 251 (encoded by SEQ ID NO: 250 by Nicotiana bentamiana), SEQ ID NO: 253 (encoded by SEQ ID NO: 252, by Oryza sativa), SEQ ID NO: 255 (encoded by SEQ ID NO: 254, by Oryza sativa), SEQ ID NO: 257 (encoded by SEQ ID NO: 256, by Solanum tuberosum SEQ ID NO: 259 (encoded by SEQ ID NO: 258, by Vitis vinifera), SEQ ID NO: 261 (encoded by SEQ ID NO: 260, by Zea mays), SEQ ID NO: 263 (encoded by SEQ ID NO: 262, Zea mays), SEQ ID NO: 283 (encoded by SEQ ID NO: 282, Brassiea rapa), SEQ ID NO: 285 (encoded by SEQ ID NO: 284, Glyeine max), and SEQ ID NO: 287 (encoded by SEQ ID NO: 286, Populustremuloides) are orthologs of the polypeptide of SEQ ID NO: 235. Orthologs and paralogues can easily be found by performing a so-called reciprocal blast search. This can be done by a first BLAST involving performing BLAST with a query string (for example, SEQ ID NO: 234 or SEQ ID NO: 235) against any sequence database, such as the publicly available NCBI database. BLASTN or TBLASTX (using default default values) can be used when starting from a nucleotide sequence and BLASTP or TBLASTN (using default default values) can be used when starting from a polypeptide sequence. BLAST results can be optionally filtered. The full-length sequences of either the filtered results or the unfiltered results are then BLASTed back (according to BLAST) against organism sequences from which the query sequence is derived (where the query string is SEQ ID NO: 234 or SEQ ID NO: 235, the second BLAST should therefore be against Arabidopsis sequences). The results of the first BLAST second are then compared. A paralog is identified if a high score of the first BLAST is of the same species from which the query sequence is derived, a return BLAST then ideally results in the query sequence as the highest hit (other than itself); Aortologist is identified if a high score hit in the first BLAST is not of the same species from which the query sequence is derived and preferably results by returning BLAST in the query sequence among the highest hits. High score hits are those with a low E value. The lower the E value, the more significant the score (in other words, the lower the chance that the hit was found). Value computing E is well known in the art. In addition to E-values, comparisons are also scored by percent identity. Percent Identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. An example detailing the identification of orthologs and paralogues is given in Example 41. In the case of large families, ClustalW can be used, followed by a neighbor grouping tree, to help visualize related gene grouping and to identify orthologs and paralogues. In Figure 22, paralogosis orthologs of the transcription factor polypeptide SPL15 cluster together.

Polypeptides represented by any of SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO : 251, SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 263, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ IDNO: 287, or orthologs or paralogues of any of the above SEQ ID NOs, all comprise an SPL DBD preferably in ascending order of at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% string identity with SPL15 DBD represented by SEQID NO: 277.

SPL DBDs are well known in the art and can be readily identified by those skilled in the art. An SPL DBD can be identified using methods for sequence alignment for comparison. Sequence alignment methods for comparison include GAP, BESTFIT, BLAST, FASTA, and TFASTA. GAP uses the Needleman and Wunsch algorithm ((1970) J Mol Biol 48: 443-453) to find the alignment of two complete sequences that maximizes the number of matches and minimizes the number of gaps. The BLAST algorithm (Altschulet al. (1990) J Mol Biol 215: 403-10) calculates percent sequence identity and performs a statistical analysis of similarity between the two sequences. The computational application for performing BLAST analysis is publicly available through the National Center for BiotechnologyInformation. Counterparts can be readily identified using, for example, the ClustalW multiple sequence alignment algorithm (version 1.83) available from GenomeNet service at Kyoto University Bioinformatics Center, with standard paired alignment parameters, and a percentage scoring method. Secondary manual editing can be performed to optimize alignment between conserved subjects, as would be apparent to a person skilled in the art. The alignment shown in Figures 23 and 24 highlights SPL DBD in SPLl5 transcription factor polypeptides. Preferably, SPL15 transcription factor polypeptides useful in the methods of the invention comprise an SPL DBD having, in ascending order, preferably at least 65%, 70%, 80%, 85%, 90%, 95% or 98% identity. of sequence with the SPL DBD represented by SEQID NO: 277.

In some instances, default parameters may be adjusted to modify the search stringency. For example using BLAST, the threshold of statistical significance (called the expected value ") to report pairings against database sequences can be increased to show less stringent pairings. In this way, nearly exact shortlines can be identified. Reason 1 as represented by SEQ ID NO: 276 and Reason 2 as represented by SEQ IDNO: 278 both comprised of the SPPL15 transcription factor polypeptides useful in the methods of the invention can be identified in this manner (Figure 24.) Within Reason 1 and Reason 2, one or more conservative changes in any position are permitted. , and / or one, two or three non-conservative changes (s) at any position.The W (S / T) tripeptide L at the C-terminal end of the polypeptide can be similarly identified (Figure 24).

There are also special databases for domain identification. SPL DBD in an SPL15 transcription factor polypeptide can be identified using, for example, SMART (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95, 5857-5864; Letunic etal. (2002) Nucleie Aeids Res 30, 242-244; hosted by the EMBL at Heidelberg, Germany), InterPro (Mulder et al., (2003) Nucl. Acids Res. 31,315-318; hosted by the European Bioinformaties Institute (EBI) in the United Kingdom), Prosite (Bucher and Bairoeh (1994), A generalized profile syntax forbiomolecular sequences motifs and its fiinction in automatic sequence interpretation. (In) ISMB-94; Projections 2nd International Conference on Intelligent Systems for Molecular Biology. Altman R., Brutlag D., Karp P., Lathrop R., Searls D., Eds., Pp53-61, AAAIPress, Menlo Park; Hulo et al., Nucl. Acids Res. 32: D134-D137, (2004), The ExPASy server proteins is available as a service to the scientific. community (hosted by the SwissInstitute of Bioinformatics (SIB) in Switzerland) or Pfam (Bateman et al., NucleicAcids Research 30 (1): 276-280 (2002), hosted by the Sanger Institute in the United Kingdom). The SPL DBD in the InterPro database is designated IPR004333, PF03110 in the Pfam database, and PS51141 in the PROSITE database.

In addition, the presence of G / S-rich stretch preceding the SPL DBD can also be readily identified (Figure 24). Primary amino acid composition (in%) to determine if a polypeptide domain is rich in specific amino acids can be calculated using application programs. ExPASy server computers, in particular the ProtParam tool (Gasteiger E et al. (2003) ExPASy: the protein server for in-depth protein knowledge and analysis. Nucleic Acids Res31: 3784-3788). The protein composition of interest can then be compared with the average amino acid composition (in%) in the Swiss-Prot Protein Sequence database. Within this database, the content of Gly (G) and Ser (E) are both 6.9% (up to 13.8%). As an example, the G / S-rich stretch preceding the SPL DBD of SEQ ID NO: 235 contains 14.5% G and 25.5% S (up to 40%). As defined in this place, a G / S-rich stretch has a combined G and S content (in terms%) above that found in the average amino acid composition (in terms%) of the proteins in the Swiss-Prot Protein Sequence. Both G and S belong to the category of very small amino acids.

Nucleic acid encoding the polypeptides represented by any of SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 257, SEQ ID NO: 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287, or orthologs or analogues of any of the above SEQ ID NOs, do not need full length nucleic acids, since performance of the methods of the invention is unreliable in use. of full-length nucleic acid sequences. In addition, examples of suitable nucleic acids for use in carrying out the methods of the invention include but are not limited to those represented by any of: SEQ ID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ ID NO: 286. Nucleic acid variants may also be useful for practicing the methods of the invention. Examples of such variants include nucleic acid moieties, splice variants, allelic or naturally occurring variants or obtained by DNA manipulation.

A portion may be prepared, for example, by making one or more deletions to a nucleic acid encoding a SPL15 transcription factor polypeptide as defined above. The portions may be used in isolation or may be fused to other decoding (or non-coding) sequences to, for example, produce a protein that combines various activities. When fused to other coding sequences, the resulting polypeptide produced by translation may be greater than that predicted for the SPL15 transcription factor factor portion. Portions useful in the methods of the invention encode a SPL15 transcription factor polypeptide (as described above) and have substantially the same biological activity as the SPL15 transcription factor polypeptide represented by either SEQ ED NO: 235, SEQ ID NO: 237, SEQ ID NO : 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287, or orthologs or paralogues of any of the SEQ ID NO mentioned above . Examples of portions may include nucleotides encoding Reason 1 as represented by SEQ ID NO: 276, in ascending order of preferably at least 65%, 70%, 80%, 85%, 90%, 95% or 98% identity. sequence with the SPL DBD represented by SEQID NO: 277, and Reason 2 as represented by SEQ ID NO: 278. Portions may additionally include nucleotides encoding the G / I-rich stretch I am the W (S / T) L tripeptide (but not necessarily the same). nucleotide encoding amino acid sequences between any of these). The portion is typically at least 250 nucleotides in length, preferably at least 500 nucleotides in length, more preferably at least 750 nucleotides in length, and most preferably at least 1,000 nucleotides in length. Preferably, the portion is a portion of a nucleic acid as represented by any one of SEQ ID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284e SEQ ID NO: 286. More preferably the portion is a portion of a nucleic acid as represented by SEQ ID NO: 234.

Another nucleic acid variant useful in the methods of the invention is a nucleic acid capable of hybridizing under reduced stringency conditions, preferably under stringent conditions, with a nucleic acid encoding a transcription factor SPL15 polypeptide as defined above, or a portion as defined above. .

Hybridization sequences useful in the methods of the invention encode a polypeptide comprising from N-terminal to C-terminal: (i) Reason 1 as represented by SEQ ID NO: 276; and (ii) in ascending order preferably at least 65%, 70%, 75%, 80%, 85%, 90%, 95% or 98% sequence identity with the SPL DBD represented by SEQ ID NO: 277; iii) Reason 2 as represented by SEQ ID NO: 278, and having substantially the same biological activity as the SPL15 transcription factor polypeptides represented by SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 255, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287, orortologists or paralogues of any of the above SEQ ID NOs. The hybridization sequence is typically at least 250 nucleotides in length, preferably at least 500 nucleotides in length, more preferably at least 750 nucleotides in length, and more preferably at least 1000 nucleotides in length. Preferably, the hybridization sequence is one that is capable of hybridizing with any of the nucleic acids represented by SEQ ID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ IDNO: 286, or with a portion of any of the above-mentioned Sequences, a portion being as defined above. More preferably the hybridization sequence is capable of hybridizing to SEQ ID NO: 234, or portions thereof.

Another nucleic acid variant useful in the methods of the invention is a processing variant encoding a SPL15 transcription factor polypeptide as defined above.

Preferred splice variants are splice variants of a nucleic acid encoding SPL15 transcription factor polypeptide represented by any of SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, or SEQ ID NO: 287, union listeners encoding orthologs or dialogues of any of the above SEQ ID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO : 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ ID NO: 286. Most preferred is a variant of processing of a nucleic acid as represented by SEQID NO: 234.

Another nucleic acid variant useful for carrying out the methods of the invention is an allelic variant of a nucleic acid encoding a SPL15 transcription factor polypeptide as defined above.

The allelic variant may be an allelic variant of a nucleic acid encoding an SPL15 transcription factor polypeptide represented by any of SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 249, SEQ ID NO: 241, SEQ ED NO: 243 , SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO : 261, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, or SEQ ID NO: 287, or an allelic variant of a nucleic acid encoding orthologs or paralogs of any of the above mentioned SEQ ID NOs. Further preferred are allelic nucleic acid variants represented by any of SEQID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ ID NO: 286. Most preferred is an allelic variant of a nucleic acid as represented by SEQ ID NO: 234.

An additional nucleic acid variant useful in the methods of the invention is a nucleic acid variant obtained by scrambling. Gene shuffling or directed evolution may also be used to generate nucleic acid variants encoding SPL15 transcription factor polypeptides as defined above.

In addition, nucleic acid variants can also be obtained by site-directed mutagenesis. Several methods are available to achieve site-directed mutagenesis, the most common being PCR-based methods (Current Protocols in Molecular Biology. Wiley (Eds)).

Also useful in the methods of the invention are nucleic acids encoding homologues of any of the amino acids represented by SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 251, SEQ ID NO: 263, SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287, or orthologs or paralogues of any of the above SEQ ID NOs.

Also useful in the methods of the invention are nucleic acids encoding derivatives of any of the amino acids represented by SEQ ID NO: 235, SEQ ID NO: 237, SEQ ID NO: 239, SEQ ID NO: 241, SEQ ID NO: 243, SEQ ID NO: 245, SEQ ID NO: 247, SEQ ID NO: 249, SEQ ID NO: 251, SEQ ID NO: 253, SEQ ID NO: 257, SEQ ID NO: 259, SEQ ID NO: 261 SEQ ID NO: 283, SEQ ID NO: 285, SEQ ID NO: 287, or orthologs or paralogues of any of the SESE ID NOs mentioned above. "Derivatives" include peptides, oligopeptides, polypeptides which may, compared to the amino acid sequence of the naturally occurring protein form, such as that shown in SEQ ID NO: 235, comprise amino acid substitutions with non-naturally occurring amino acid residues, or amino acid residue additions not naturally occurring.

In addition, SPL15 transcription factor polypeptides (at least in their native form) typically have DNA binding activity and an activation domain. A person skilled in the art can easily determine the presence of a DNA activation domain and binding activity using routine tools and techniques.

Nucleic acids encoding SPL15 transcription factor polypeptides may be derived from any natural or artificial source.

Nucleic acid may be modified from its native form into genomic composition and / or environment through deliberate human manipulation. Preferably the nucleic acid encoding SPL15 transcription factor polypeptide is from a plant, still preferably from a dicotyledonous, more preferably from the Brassicacea family, most preferably the nucleic acid is from Arabidopsis thaliana.

Any reference herein to an SPL15 transcription factor polypeptide is therefore adopted to mean an SPL15 transcription factor factor peptide as defined above. Any nucleic acid encoding such an SPL15 transcription factor polypeptide is suitable for use in performing the methods of the invention.

The invention also provides genetic constructs and vectors for facilitating introduction and / or expression of nucleic acid sequences in the methods according to the invention in a plant.

Therefore, a gene construct is provided comprising:

(i) A nucleic acid encoding an SPL15 transcription factor polypeptide as defined above;

(ii) One or more control sequences operably linked to the nucleic acid of (i).

Constructs useful in the methods according to the present invention may be constructed using recombinant DNA technology well known to those skilled in the art. The gene constructs may be inserted into commercially available vectors suitable for transformation into plants and suitable for dogene expression of interest in the transformed cells. The invention therefore provides us with a gene construct as defined above in the methods of the invention.

Plants are transformed with a vector comprising the sequence of interest (i.e., a nucleic acid encoding a SPL1 transcription factor polypeptide). The skilled artisan is well aware of the genetic elements that must be present in the vector in order to successfully transform, select and propagate host cells containing the sequence of interest. The sequence of interest is operationally linked to one or more control sequences (at least one promoter).

Advantageously, any type of promoter, whether natural or synthetic, may be used to direct expression of the nucleic acid sequence. The promoter may be an inducible promoter, i.e. having induced or increased transcriptional initiation in response to an evolutionary, chemical, environmental or physical stimulus. The promoter may be a tissue specific promoter, i.e. one which is preferably capable of initiating transcription in certain tissues, such as leaves, roots, seed tissue, etc.

According to the invention, nucleic acid encoding a transcription factor SPL15 polypeptide is operably linked to a constitutive promoter. The constitutive promoter is preferably a HMGB (high mobility group B) promoter, more preferably the constitutive promoter is a rice HMGB promoter, yet preferably the constitutive promoter is represented by the nucleic acid sequence substantially similar to SEQ ID NO: 279, more preferably the constitutive promoter is as represented by SEQ ID NO: 279 or SEQ ID NO: 294.

It should be clear that the applicability of the present invention is not restricted to nucleic acid encoding a SP15 transcription factor polypeptide as represented by SEQ ID NO: 234, nor is the applicability of the invention restricted to the expression of such a nucleic acid encoding an SPL15 transcription factor polypeptide. when directed by a MHGB promoter. Examples of other constitutive promoters that may also be used to carry out the methods of the invention are shown in the definitions section.

Additional regulatory elements for enhancing expression of nucleic acids or genes, or gene products, may include transcriptional as well as translational enhancers. Those skilled in the art will be aware of terminator and enhancer sequences that may be suitable for use in carrying out the invention. Other control sequences (other than motor, enhancer, silencer, intron sequences, 3'UTR and / or 5'UTR regions) may be protein and / or RNA stabilizing agents. Such consequences must be known or readily obtainable by one of ordinary skill in the art. Optionally, one or more terminator sequences (also a control sequence) may be used in the construction introduced in a plant.

An intron sequence can also be added to either the untranslated 5 'region (RTU) or the partial decoding sequence coding sequence to increase the amount of mature message accumulating in the cytosol.

The genetic constructs of the invention may additionally include a replication source sequence that is required for maintenance and / or replication in a specific cell type. An example is when a genetic construct is required to be maintained in a bacterial cell as an episomal genetic element (e.g., plasmid or cosmid molecule). Preferred sources of replication include, but are not limited to, fl-ori and colEl.

The genetic construct may optionally comprise a selectable marker gene.

The invention also provides a method for producing transgenic plants having increased production relative to control plants, comprising introducing and expressing in a plant a nucleic acid encoding a SPL15 transcription factor polypeptide as defined above.

More specifically, the present invention provides a method for producing transgenic plants having increased production relative to control plants, the method of which comprises:

(i) introducing and expressing a nucleic acid encoding a SPL15 transcription factor polypeptide in a plant cell; and

(ii) cultivate the plant cell under conditions that promote plant growth and development.

Nucleic acid can be introduced directly into a plant cell or into the plant itself (including introduction into a tissue, organ or any other part of a plant). According to a preferred feature of the present invention, nucleic acid is preferably introduced into a plant by transformation.

Generally after transformation, plant cells or cell clusters are selected for the presence of one or more markers that are encoded by expressible genes in plants co-transferred with the gene of interest, following which the transformed material is regenerated into an entire plant.

Following DNA transfer and regeneration, computationally transformed plants can be evaluated, for example using Southern analysis, for the presence of the gene of interest, copy number and / or genomic organization. Alternatively or additionally, newly introduced DNA expression levels can be monitored using Northern and / or Western analysis, or quantitative PCR, all techniques well known to those of ordinary skill in the art.

The generated transformed plants can be propagated in a variety of ways, such as by clonal propagation or classical breeding techniques. For example, a first generation (or IT) transformed plant may be self-fertilized to generate second generation (or T2) homozygous transformants, and T2 plants further propagated by classical breeding techniques.

The generated transformed organisms can adopt a variety of forms. For example, they may be chimeras of transformed cells and unprocessed cells; clonal transformants (e.g., all cells transformed to contain the expression drawer); transformed and unprocessed detained grafts (e.g., in plants, a rhizomatransformed grafted to an unprocessed offspring).

The present invention clearly extends to any cell or plant produced by any of the methods described herein, and to all plant parts and propagules thereof. The present invention extends further to encompass the progeny of a transformed or transfected primary whole cell, tissue, organ or plant that has been produced by any of the methods mentioned above, the sole requirement being that the same feature (s) exhibit the same genotypic (s) and / or phenotypic (s) than those produced by the ancestor in the methods according to the invention.

The invention also includes host cells containing an isolated nucleic acid encoding a SP15 transcription factor polypeptide as defined above. Preferred host cells according to the invention are plant cells.

The invention also extends to cut-off parts of a plant such as, but not limited to seeds, leaves, fruits, flowers, stems, rhizomes, tubers and bulbs. The invention furthermore relates to derived products, preferably directly derived, from a cut-off part of such a plant, such as pellets or dry powders, oil, fat, fatty acids, starch or proteins.

As mentioned above, a preferred method for enhancing expression of a nucleic acid encoding an SPL15 transcription factor polypeptide is to introduce and express into a plant a nucleic acid encoding an SPL15 transcription factor polypeptide; however the effects of performing the method, i.e. increasing production, can also be achieved using other well known techniques. A description of some of these techniques will now follow.

One such technique is identification by T-DNA activation. The resulting transgenic plants show dominant phenotypes due to modified expression of genes near the introduced promoter.

The effects of the invention may also be reproduced using the TILLING technique.

The effects of the invention may also be reproduced using homologous recombination.

"Increased yield" as defined herein is meant to mean an increase in biomass (weight) of one or more parts of a plant, which may include above-ground (cut-off) and / or below-ground (cut-off) parts .

In particular, such cut-off parts include biomass / fats and / or seeds, and performance of the methods of the invention results in plants having increased yield (in vegetative and / or seed biomass) relative to the production of control plants.

Using corn as an example, an increase in yield can be manifested as one or more of the following: increase in the number of plants per hectare or acre, an increase in the number of ears per plant, an increase in the number of rows, number of seeds per row, weight. dementing, weight of one thousand seeds, ear length / diameter, increased grain filling rate (which is the number of full grains divided by the total number of grains and multiplied by 100), among others. Using rice as an example, an increase in yield can be manifested as an increase in one or more of the following: number of plants per hectare or acre, number of panicles per plant, number of spikelets per panicle, number of flowers (rapiers) per panicle (which is expressed as a ratio of the number of full grains to the number of primary panicles), increase in grain fill rate (which is the number of full grains divided by the total number of grains and multiplied by 100), increase in milliseed weight, among others.

Since transgenic plants according to the present invention have increased yields, these plants are likely to exhibit increased growth rate (for at least part of their due cycle) relative to the growth rate of corresponding wild-type plants at a corresponding stage. in your life cycle. The increased growth rate may be specific to one or more parts of a plant (including seeds), or may be substantially throughout the plant. A plant having an increased growth rate may still exhibit early flowering. The increase in growth rate may alter the crop cycle of a plant allowing plants to be sown later and / or harvested earlier than otherwise possible. If the growth rate is sufficiently increased, it can allow additional sowing of seeds of the same plant species (eg sowing and harvesting rice plants followed by sowing and harvesting additional rice plants all within a conventional growing period). Similarly, if the growth rate is sufficiently increased, it may allow additional sowing of seeds of different plant species (eg sowing and harvesting maize plants followed by, for example, optional sowing, harvesting of soybean, potato or any other suitable plant) . Additional harvesting times for mesmorizoma in the case of some crop plants may also be possible. Changing the harvest cycle of a plant may lead to an increase in annual biomass production per acre (due to an increase in the number of crops (say that in one crop). year) that any particular plant can be grown and harvested). An increase in growth rate may also allow the cultivation of transgenic plants in a wider geographical area than their wild type counterparts, as territorial limitations to cultivate a crop are often determined by adverse environmental conditions or at the time of planting (early cycle) or harvest time (late cycle). Such adverse conditions can be prevented if the harvest cycle is shortened. Growth rate can be determined by deriving various parameters from growth curves, such parameters can be: T-Mid (time taken by plants to reach 50% of their maximum size) and T-90 (time taken by plants to reach 90% of their size) maximum), among others.

According to a preferred feature of the present invention, performance of the methods of the invention generates plants having an increased growth rate relative to control plants. Therefore, in accordance with the present invention, a method for increasing plant growth rate is provided, the method of which comprises increasing expression in a plant of a nucleic acid encoding a SPL15 transcription factor polypeptide

An increase in yield and / or growth rate occurs when the plant is in non-stress conditions or whether the plant is exposed to various stresses compared to control plants. Plantastipically respond to more slowly growing stress exposure. Under severe stress conditions, the plant may even stop growing in general.

Mild stress, on the other hand, is defined here as any stress to which a plant is exposed that does not result in stopping overall plant growth without the ability to resume growth.

Mild stress in the sense of the invention leads to a reduction in growth of stressed plants of less than 40%, 35% or 30%, preferably less than 25%, 20% or 15%, more preferably less than 14%, 13%, 12%, 11% or 10% or less compared to the control plant under non-stress conditions. Due to advances in agricultural practices (irrigation, fertilization, pesticide treatments) severe stresses are not often encountered in cultivated crop plants. As a consequence, compromised growth induced by mild stress is often an undesirable feature for agriculture. Soft stresses are the day-to-day biotic and / or abiotic (environmental) stresses to which a plant is exposed. Abiotic stresses may be due to drought or excess water, anaerobic stress, saline stress, chemical toxicity, oxidative stress and hot, cold or freezing temperatures. Biotic stress can be osmotic stress caused by water stress (particularly due to drought), saline stress, oxidative stress, or ionic stress. Biotic stresses are typically those caused by pathogens such as bacteria, viruses, fungi and insects.

In particular, the methods of the present invention may be carried out under non-stress conditions or under mild dry conditions to increase plants yield over control plants. As reported in Wang et al. (Planta (2003) 218: 1-14), abiotic stress leads to a series of morphological, physiological, biochemical and emolecular changes that adversely affect plant growth and productivity. Dryness, salinity, extreme temperatures and oxidative stress are known to be interconnected and may induce injury. growth and cell growth through similar mechanisms. Rabbani et al. (Plant Physiol (2003) 133: 1755-1767) describes a particularly high degree of "interference" between drought stress and high salinity stress. For example, drought and / or salinity are manifested primarily as osmotic stress, resulting in disruption of homeostasis and ion distribution in the cell. Oxidative stress, which often accompanies high or low temperature, salinity or drought stress, can cause functional and structural protein denaturation. As a consequence, these various environmental stresses often activate cell signaling pathways and similar cellular responses, such as stress protein production, antioxidant enhancement, compatible solute accumulation, and decay suspension. The term "non-stress" conditions as used herein are those environmental conditions that allow for optimal plant growth. People skilled in the art will be aware of normal soil and climate conditions for a given location. Performance of the methods of the invention generates cultivated plants under no stress or under mild drought conditions increased production compared to control plants grown under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under non-stress conditions or under mild dry conditions, which method comprises increasing expression in a plant of a nucleic acid encoding a MADS15 polypeptide.

Performance of the methods of the invention generates plants grown under nutrient deficiency conditions, particularly under nitrogen deficiency conditions, increased yield over control-grown plants under comparable conditions. Therefore, according to the present invention, there is provided a method for increasing production in plants grown under nutrient deficiency conditions, the method of which is to increase expression in a plant of a nucleic acid by encoding a MADS15 polypeptide. Nutrient deficiency can result from nutrient deficiency or excess such as nitrogen, phosphates and other compounds containing phosphorus, potassium, calcium, cadmium, magnesium, manganese, iron and boron, among others.

The methods of the invention are advantageously applicable to any plant.

Plants which are particularly useful in the methods of the invention include all plants belonging to the Viridiplantae superfamily, in particular monocot and dicot plants including forage or forage vegetables, ornamental plants, food crops, trees or shrubs. According to a preferred embodiment of the present invention, the plant is a crop plant. Examples of crop plants include soybean, sunflower, canola, alfalfa, rapeseed, cotton, tomato, potato, and tobacco. Still preferably, the plant is a monocot plant. Examples of monocotyledonous plants include sugar cane. Most preferably aplanta is a cereal. Examples of cereals include rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats.

The present invention also encompasses viable plants by the methods according to the present invention. The present invention therefore provides plants, parts and cells of such plants viable by the methods according to the present invention, which plants or parts or cells comprise a nucleic acid transgene encoding an SPL15 transcription factor polypeptide as defined above.

The present invention also encompasses use of nucleic acids encoding SPL15 transcription factor polypeptides to increase production in a plant compared to production in a control plant.

One such use refers to increasing plant production, yields as defined above. Yield may include in particular one or more of the following: increased above ground biomass, increased number of flowers per panicle, increased seed production, increased total number of seeds, increased number of full seeds, increased Mill Seed Weight (TKW) and increased harvest index. .

Nucleic acids encoding SPL15 transcription factor polypeptides may find use in breeding programs in which a DNA marker that can be genetically linked to a gene encoding SPL15 transcription factor polypeptide is identified. Nucleic acids encoding SPL15 transcription factor polypeptides may be used to define a molecular marker. This marker can then be used in breeding programs to select plants having increased seed production. Nucleic acids encoding SPL15 transcription defector polypeptides may be, for example, a nucleic acid represented by any one of SEQ ID NO: 234, SEQ ID NO: 236, SEQID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 252, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ ID NO: 286.

Allelic variants of a nucleic acid encoding an SPL15 transcription factor polypeptide may also find use with marker-assisted breeding programs. Such breeding programs sometimes require the introduction of allelic variation by mutagenic treatment of plants, using for example EMS mutagenesis, alternatively, the program may start with a collection of so-called "natural" allelic variants caused unintentionally. Identification of allelic variants then occurs, for example by PCR. This is followed by a step to select higher allelic variants of the sequence in question and which generate increased seed yield. Selection is typically performed by monitoring plant growth performance containing different allelic variants of the sequence in question, for example, allelic variants different from any of SEQ ID NO: 234, SEQ ID NO: 236, SEQ ID NO: 238, SEQ ID NO: 240, SEQ ID NO: 242, SEQ ID NO: 244, SEQ ID NO: 246, SEQ ID NO: 248, SEQ ID NO: 250, SEQ ID NO: 254, SEQ ID NO: 254, SEQ ID NO: 256, SEQ ID NO: 258, SEQ ID NO: 260, SEQ ID NO: 262, SEQ ID NO: 282, SEQ ID NO: 284 and SEQ ID NO: 286. Growth performance can be monitored in a green house or in the field. Additional optional steps include crossing plants in which the superior allelic variant has been identified with another plant. This can be used, for example, to make a combination of interesting phenotypic characteristics.

Nucleic acids encoding SPL15 transcription factor polypeptides may also be used as probes to map genetically and physically the genes of which they are a part of, and as markers for traits linked to such genes. Such information may be useful in plant breeding in order to develop strains with desired phenotypes. Such use of nucleic acids encoding SPL15 transcription factor polypeptides requires only one nucleic acid sequence of at least 15 nucleotides in length. Nucleic acids encoding SPL15 transcription factor polypeptides can be used as restriction fragment length polymorphism (RFLP) markers. Southern blots (Sambrook J5 Fritsch EF and Maniatis T (1989) Molecular Cloning, A Laboratory Manual) of restriction digested plant genomic DNA can be probed with a nucleic acid encoding SPL15 transcription factor polypeptide. The resulting decay pattern can then be subjected to genetic analysis using computational programs such as MapMaker (Lander et al. (1987) Genomics 1: 174-181) in order to construct a genetic map. In addition, the nucleic acid can be used to probe Southern blots containing restriction endonuclease-treated genomic DNAs from a set of individuals representing the ancestor and progeny of a defined genetic cross. Segregation of DNA polymorphisms is observed and used to calculate the position of nucleic acid encoding SPL15 transcription factor polypeptide in the genetic map previously obtained using stapopulation (Botstein et al (1980) Am. J. Hum. Genet. 32: 314-331).

The production and use of probes derived from parause plant genes in genetic mapping is described in Bernatzky and Tanksley (GENETICS 112 (4): 887-898, 1986). Numerous publications describe genetic mapping of cDNA-specific clones using the methodology described above or variations thereof. For example, F2 crossbreeding populations, inbreeding populations, randomly crossed populations, nearby isogenic lineages (NIL), and other sets of individuals can be used for mapping. Such methodologies are well known to those skilled in the art.

Nucleic acid probes can also be used for physical mapping (ie, sequence allocation in physical maps; see Hoheisel et al. In: Non-mammalian Genomic Analysis: A Practical Guide, Academic press 1996, pp. 319-346, and cited references. in this one).

In another embodiment, nucleic acid probes may be used in direct mapping by fluorescence in situ hybridization (FISH) (Trask (1991) Trends Genet. 7: 149-154). Although current FISH mapping methods favor the use of large clones (several kb to several hundred kb; see Laan et al. (1995) Genome Res.5: 13-20), improvements in sensitivity may allow FISH mapping performance using smaller probes. .

Several methods based on nucleic acid amplification for genetic and physical mapping can be performed using nucleic acids. Examples include allele-specific amplification (Kazazian (1989) J. Lab. Clin. Med. 11: 95-96), PCR amplified fragment polymorphism (CAPS; Sheffield et al. (1993) Genomics 16: 325-332), allele-specific binding (Landegren et al. (1988) Science 241: 1077-1080), Nucleotide Extension Reactions (Sokolov (1990) Nucleic Acid Res.18: 3671), Radiation Hybrid Mapping (Walter et al. (1997) Nat. Genet. 7: 22-28) and Appropriate Mapping (Dear and Cook (1989) NucleicAcid Res. 17: 6795-6807). For these methods, a nucleic acid sequence is used to design and produce primer pairs for use in amplification or for primer extension reactions. The conception of such initiators is well known to those skilled in the art. In methods employing PCR-based genetic mapping, it may be necessary to identify differences in DNA sequences between ancestors of the cross-mapping region corresponding to the current sequence of nucleic acids. This, however, is generally not required for mapping methods.

The methods according to the present invention result in plants having increased yield as described above. These characteristics may also be combined with other economically advantageous characteristics, such as additional improved production characteristics, tolerance to other abiotic and biotic stresses, modifying and various architectural characteristics and / or biochemical and / or physiological characteristics.

Description of figures

The present invention will now be described with reference to the following figures in which:

Fig. 1 represents the schematic structure of the AtHAL3a polypeptide comprising from N-terminus to C-terminus (i) the desubstrate binding helix (single underline), (ii) the His insertion motif (double underline), (iii) the PXMNXXMW motif (dotted) and (iv) the substrate recognition clamp (ripple). The N-terminal and C-terminal ends of HAL3 proteins are not very conserved.

Fig. 2 shows a pEXP :: HAL3 binary vector for increased Oryza sativa expression of an Arabidopsisthaliana HAL3 nucleic acid under the control of a beta expansin promoter.

Fig. 3 shows a multiple alignment of several HAL3 sequences from Arabidopsis thaliana AtHAL3b (At_NP_973994), Sorghumbicolor (Sb), Arabidopsis thaliana AtHAL3a (Ath0218), Gossipium hirsutum (Cg), Hordeum vulgare (Hv), Oryza s) vinifera (Vv), Glycinemax (Gm), Solanum tuberosum (St), Zea mays (Zm), and Pinus sp (Pg).

Fig. 4 details examples of sequences useful for carrying out the methods according to the present invention: SEQ ID NO: 1 and SEQ ID NO: 2 represent the nucleic acid sequence and the protein sequence of AtHABa. SEQ ID NO: 3 and SEQ ID NO: 4 are the forward and reverse primer sequences used to isolate the AtHAL3 gene. SEQ ID NO: 5 is the beta expansin promoter sequence used in the methods of the present invention. SEQ ID NO: 9 to SEQ ED NO: 42 represent examples of full or partial length DNA / protein sequences useful in the methods of the invention or for isolating corresponding full length sequences. In some cases, sequences were grouped from EST sequences, with lower quality sequencing. As a consequence, few amino acid substitutions can be expected.

Fig. 5 (A) shows the domain structure of the MADS15 protein, (B) represents the sequence of SEQ ID NO: 44 with the bold MADS domain and the italic keratin domain. Between the MADS domain and the keratin domain, the intervening domain is located while domain C is located C-terminally to the keratin domain.

Fig. 6 shows a multiple alignment of various MADS15 proteins. Asterisks indicate identical amino acids, colonies represent highly conservative substitutions, dots represent less conservative substitutions.

Fig. 7 shows the binary vector for enhanced expression in Oryza sativa of a nucleic acid encoding Oryzasativa MADS15 protein under the control of a GOS2 promoter.

Fig. 8 details examples of sequences useful for carrying out the methods according to the present invention.

Fig. 9 is a schematic representation of a full length OsMADS 15 polypeptide. Typical domains (M, MADS; I, intervening domain; K, keratin K-box region; C, C-terminal variable region) are indicated, the lines above and below the diagram show the protein regions involved in DNA binding, dimerization and multimerization with interaction proteins.

Fig. 10 shows the binary vector pGOS :: MADS15hp MADS15 RNA parasilization in Oryza sativa using a staple construct under the control of a constitutive promoter (GOS2).

Fig. 11 details examples of sequences useful for carrying out the methods according to the present invention, or useful for isolating such sequences. Sequences may result from public EST associations, consequently of lower quality. As a consequence, few amino acid substitutions can be expected. Start (ATG) and stop codons delimit nucleic acid sequences when they encode full length MADS15 polypeptides. However both 5 'and 3' UTRs may also be used for carrying out the methods of the invention.

Fig. 12 shows the schematic classification of AP2 / ERF transcription factor polypeptides according to the domains present: the AP2 subfamily with two AP2 repeats and the ERP subfamily with only one AP2 repeating. The ERP subfamily is further subdivided into transcription factors comprising a B3 domain in addition to the AP2 repeat (called RAV), or not. PLT transcription factor polypeptides belong to the AP2 subfamily with two AP2 repeats.

Fig. 13 represents a schematic presentation of the PLT transcription factor polypeptide structure. The AP2 domain comprises the two (wrapped) AP2 repeats separated by a linker region. The approximate positions of the nuclear localization signal (NL S), motif 1PK (V / L) (A / E) DFLG and motivp 2 (V / L) FX (M / V) WN (D / E) are marked as boxes. .

Fig. 14 is an alignment of PLT transcription defector polypeptide sequences (from Table 4) compared to other AP2 domain transcription factor polypeptides (from Table 4 below). The nuclear delocalization (NLS) signal, motif 1 PK (V / L) (A / E) DFLG and motif 2 (V / L) FX (M / V) WN (D / E) are wrapped. Identical residues are blackened, conservative residues are grayed out.

Table 4: Domain Transcription Factor Polypeptides

AP2 aligned against the PLT transcription factor polypeptides used to carry out the methods of the invention.

<table> table see original document page 229 </column> </row> <table>

Fig. 15 is a photograph of 40 seeds of a control (left) plant compared to 40 seeds of a transgenic plant with increased expression of a PLT transcription factor polypeptide.

Fig. 16 shows a pGOS2 :: PLT binary vector for increased Oryza sativa expression of an Oryza sativa PLT transcription factor nucleic acid under the control of a GOS2 promoter.

Fig. 17 details examples of sequences useful for carrying out the methods according to the present invention (SEQ ID NO: 175 to SEQ ID NO: 188). Partial sequences (SEQ ID NO: 189 and SEQ ID NO: 190) useful for isolating corresponding full length sequences are also presented.

Fig. 18 shows an alignment of bHLH transcription factors as defined above. The sequences were aligned using AlignX software from Vector NTI suite (InforMax, Bethesda, MD). Multiple alignment was made with a gap opening penalty of 10 and a gap extension of 0.01. Secondary manual editing was also performed where necessary to better position some conserved regions. Domain bHLH is indicated inside the solid box and domain PFB26111 is indicated inside the hatched box. Also indicated by the three upward pointing arrows is the setting 5-9-13 (K / R ER). The single downward pointing arrow indicates the glutamic acid site for E-BOX recognition.

Fig. 19 shows a matrix of similarity and identity between bHLH transcription factors of various species. Percentage identity is shown in bold. Fig. 19a is a matrix over full length sequences and Fig. 19b is a matrix over the bHLH domain only. Further details are provided in Example 34.

Fig. 20 shows a binary vector pGOS2 :: bHLH, for increased Oryza sativa expression of a nucleic acid encoding Oryza sativa bHLH transcription factor under the control of a GOS2 promoter.

Fig. 21 details examples of sequences useful for carrying out the methods according to the present invention.

Fig. 22 Neighbor cluster tree result after multiple sequence alignment of all Arabidopsis thaliana SPL transcription factor polypeptides and SPL15 transcription factor polypeptide orthologs using CLUSTAL W (1.83) (in GenomeNet service at Kyoto University Bioinformatics Center ), and default values (Blosum 62 as a scoring matrix, small gap opening penalty of 10; gap extension penalty of 0.05). Arabidopsis thaliana SPL15 transcription factor polypeptide groups with the other SPL15 transcription factor polypeptide pathologists and paralogues, as shown in parentheses.

Fig. 23 shows an alignment of the DNA binding domain (DBD) of SP15 transcription factor polypeptide orthologs and paralogs. The sequences were aligned using AlignX software from VectorNTI suite (InforMax, Bethesda, MD). Multiple alignment was made with a gap opening penalty of 10 a gap length of 0.01.

Conserved Cys and His residues involved in zinc ion binding are wrapped. The bipartite nuclear localization signal (NLS) is underlined.

Fig. 24 is an alignment of SPL15 transcription factor polypeptide orthologs and paralogs. The sequences were aligned using the AlignX Vector NTI suite program (InforMax, Bethesda, MD). Multiple alignment was done with a gap opening penalty of a 0.01 gap length. Secondary manual editing has also been done where necessary to better position some conserved regions. The three main characterized domains, from N-terminal to C-terminal, are encased and identified as Reason 1, the SPL and Reason 2 DNA-binding domain. In addition, the G / Spreed-rich portion of the SPL DBD is labeled with Xs. and the W (S / T) L tripeptide at the C-terminal end of the also encased polypeptide.

Fig. 25 shows a binary vector pHMGB :: SPL15, for increased Oryza sativa expression of a nucleic acid encoding an Arabidopsis thaliana SPL15 transcription factor polypeptide under the control of an HMGB promoter.

Fig. 26 details examples of sequences useful for carrying out the methods according to the present invention.

Examples

The present invention will now be described with reference to the following examples, which are by way of illustration only. The following examples are not intended to fully or otherwise limit the scope of the invention.

DNA Manipulation: Unless otherwise determined, recombinant DNA techniques are performed according to standard protocols described in (Sambrook (2001) Molecular Cloning: Alaboratory Manual, 3rd Edition Cold Spring Harbor Laboratory Press, CSH, New York) or in Volumes 1 and 2 by Ausubel et al. (1994), Current Protocolsin Molecular Biology, Current Protocols. Standard materials and methods for plant molecular work are described in Plant Molecular BiologyLabfax (1993) by R.D.D. Croy, published by BIOS Scientific PublicationsLtd (UK) and Blackwell Scientific Publications (UK).

Example Section A: HAL3

Example 1: Identification of Sequences Related to Nucleic Acid Sequence Used in the Methods of the Invention

Sequences (full length cDNA, ESTs, or genomic) related to the nucleic acid sequence used in the methods of the present invention have been identified among those maintained in the Entrez Nucleotides database at the National Center for Biotechnology Information (NCBI) using database sequence search tools. , such as the Basic Local Alignment Tool (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997) Nucleic AcidsRes. 25: 3389-3402). The program is used to find local unequal regions between sequences by comparing nucleic acid or polypeptide sequences with sequence database and calculating the statistical significance of pairings. For example, the nucleic acid-encoded polypeptide of the present invention was used for the TBLASTN algorithm, with default settings and the low complexity sequence skip filter. The result of the analysis was paired comparison, and ranked according to the probability score (E value), where the score reflects the likelihood that a particular alignment will occur at random (the lower the E value, the more meaningful the hit). In addition to E values, comparisons were also scored by percent identity. Percentage identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid sequences (or polypeptides) compared over a particular length. In some instances, the default parameters may be adjusted to modify the search stringency. For example the Epode value may be increased to show less stringent pairings. In this way, practically exact short pairings can be identified.

Table A provides a list of nucleic acid sequences related to the nucleic acid sequence used in the methods of the present invention.

Table A: Examples of HAL3 Polypeptides:

<table> table see original document page 233 </column> </row> <table>

In some instances, related sequences have been experimentally grouped and publicly described by research institutions, such as The Institute for Genomic Research (TIGR). The Eukaryotic Gene Orthologs (EGO) database can be used to identify such related consequences, either by keyword searching or by using the BLAST algorithm with the nucleic acid or polypeptide sequence of interest.

Example 2: Cloning of Nucleic Acid Sequence in Methods of the Invention

The nucleic acid sequence used in the invention methods was PCR amplified using a custom Arabidopsis thaliana seedling decDNA library (in pCMVSport 6.0; Invitrogen, Paisley, UK) as a template. PCR was performed using Hifi Taq DNA Polymerase under standard conditions using 200 ng of template in a 50 μΐ PCR mix. The primers used were prm00957 (SEQ ID NO: 3; sense, bold start codon:

5 'aaaaagcaggctcacaatggagaatgggaaaagagac 3') and prm00958 (SEQ ID NO: 4; reverse, complementary: 5 'agaaagctgggttggttttaactagttccaccg 3'), which includes AttB sites for Gateway recombination. Amplified PCR fragment was purified also using standard method. The first step of the Gateway procedure, the BP reaction, was then performed during which the PCR fragment recombines in vivo with pDONR201 oplasmide to produce, according to Gateway terminology, an "input clone", pHAL3. Plasmid pDONR201 was purchased from Invitrogen as part of Gateway® technology.

Example 3: Expression Vector Construction The pHAL3 input clone was subsequently used in an LR reaction with pEXP, a destination vector used for Oryza sativa transformation. This vector contains as functional elements within the T-DNA limits: a selectable plant marker; a traceable marker expression drawer; and a Gateway drawer intended for in vivo LR combining with the nucleic acid sequence of interest already cloned into the input clone. A rice beta expansin promoter (SEQID NO: 5) for specific shoot expression was localized before the gateway.

After the LR recombination step, the resulting expression vector pEXP :: HAL3 (Figure 2) was transformed into Agrobacterium strain LBA4044 and subsequently to Oryza sativa plants.

Example 4: Culture Transformation

Transformed Agrobacterium containing the expression vectors were used independently to transform Oryzasativa plants. Dried ripe seeds of the Japanese rice cultivar Nipponbare have been husked. Sterilization was performed by incubating for one minute in 70% ethanol followed by 30 minutes in 0.2% HgCl 2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus-inducing medium). After incubation in the dark for four weeks, scutellus-derived callosembriogenes were excised and even propagated. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic decal pieces were subcultured in fresh medium 3 days before co-cultivation (to stimulate cell division activity).

Agrobacterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobacterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD60) of about 1. The suspension was then transferred to a Petri dish and the calluses immersed in the suspension for 15 minutes. Callused, they were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 25 ° C.

Co-cultured calli were cultured in medium containing 2,4-D for 4 weeks in the dark at 28 ° C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Primary transformants were transferred from a tissue culture chamber to a greenhouse. After quantitative PCR analysis to verify T-DNA insert copy number, only single copy transgenic plants exhibiting tolerance to the deletion agent were maintained for TI seed harvest. Seeds were then collected three to five months after transplantation. The method produced single locus transformants at a rate of up to 50% (Aldemita andHodges 1996, Chan et al. 1993, Hiei et al. 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural Acepa Al 88 (University of Minnesota) or A188 hybrids as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are collected from the corn plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm. Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and plants GMOs are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium containing the selection agent (eg imidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2- 3 weeks, or until aerial parts develop. The green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat processing

Wheat processing is performed with the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybeans are processed according to a modification of the method described in Texas A&M Patent 5,164,310. Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. Epicotyl and remnant cotyledon are further grown to develop axillary nodes.

These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / canola transformation

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188).

The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used. Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobaeterium (containing the expression vector) bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobaeterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

Alfalfa Transformation

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K2SO4, and 100 μm acetoseringone. Explantants are washed in half-strength Murashige-Skoog medium (Murashige andSkoog, 1962) and placed on the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobaeterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 5: Evaluation Procedure

5.1 Evaluation Configuration

Approximately 30 independent rice TO transformants were generated. Primary transformants were transferred from a tissue culture chamber to a greenhouse for TI seedlings. Seven events, of which progeny T1 secreted 3: 1 for presence / absence of transgene, were retained. For each of these events, approximately 10 Tl seedlings containing the transgene (hetero and homozygotes) and approximately 10 Tl seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression. Transgenic plants and corresponding nullizigotes were grown side by side at random positions. Greenhouse conditions were short days (12 hours of light), 28 ° C in the light and 22 ° C in the dark, and a relative humidity of 70%.

Four T1 events were additionally evaluated in generation T2 following the same evaluation procedure as for T1 generation but with more individuals per event. From the sowing stage to the maturity stage the plants were passed several times through a digital image cabinet. At each moment digital images (2048x1536 pixels, 16 million colors) were taken from each plant from at least 6 different angles.

5.2 Statistical analysis: t-test and F-test

A two-way ANOVA (analysis of variance) was used as a statistical model for the general assessment of plant phenotypic characteristics. An F test was performed on all measured parameters of all plants of all events transformed with the gene of the present invention. The F test was performed to verify for an effect of the gene over all transformation events and to verify for a general dogene effect, also known as a "global gene effect". The threshold for significance for a true overall gene effect has been adjusted to a 5% probability level for the F test. A significant Faponta test value for a gene effect means that it is not just the mere presence or position of the gene that is causing the abnormalities. differences in phenotype.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" are plants treated in the same manner as the transgenic plant, but from which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test is set at 10% probability level. Results for some events may be above or below this threshold. This is based on the hypothesis that a gene can only have an effect on certain positions in the genome, and that the occurrence of this position-dependent effect is not uncommon. This type of gene effect is also called in this place a "gene strain effect". The value of ρ is obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the hypothesis (ie, that there is no transgene effect). ) is correct.

Example 6: Evaluation Results

The mature primary panicles were collected, counted, bagged, barcoded and then dried for three days in an oven at 37 ° C. The panicles were then traced and all seeds were collected and counted. The full shells were separated from the empty ones using an air-blowing device. The empty shells were discarded and the remaining fraction was counted again. The full shells were weighed on an analytical balance. The number of full grains was determined by counting the number of full husks that remained after the desparing step. Total seed yield (total seed weight) was measured by weighing all full bark collected from a plant. Total number of seeds per plant was measured by counting the number of bark collected from a plant. The harvest index (HI) in the present invention is defined as the proportion between total seed yield and shoot (mm2) multiplied by a factor 106. The total number of flowers per panicle as defined in the present invention is the ratio of the total number of seeds to the number of mature primary panicles. The grain fill rate as defined in the present invention is the ratio (expressed as a%) of the number of full grains to the total number of seeds (or rapiers).

As shown in Table B, seed yield, full grain number, and harvest index are increased in plantastransgenic preferentially expressing a nucleic acid encoding a HAL3 polypeptide in the shoot compared to control plants.

Table B shows the average yield increase (total deboned weight), increase in number of full grains and increase of harvest percentage in percent, calculated from transgenic events compared to control plants, in the Tl generation.

Table B

<table> table see original document page 242 </column> </row> <table>

Example 7: Early Vigor Assessment Results

Early vigor was determined by counting the total number of plant shootings broken down from the bottom. This value was averaged for photos taken at the same time from different angles and converted to a physical surface value expressed in mm squared by calibration. The results described below are for plants three weeks after germination.

Early plant vigor (if determined by aerial area) was seen in six of seven IT generation events, with an overall increase in aerial area for transgenic seedlings of 27% compared with control plants. Four of these Tl events were additionally evaluated in T2 generation, and all These four events generated an increase in aerial area for transgenic seedlings compared to control plants, with an overall increase in aerial area for transgenic seedlings of 33% compared with control plants. The results also showed to be statistically significant with the ρ value of the F test being less than 0.0001 (T2 generation) indicating that the observed effect is probably deive to the transgene rather than gene position or strain effect, see table C.

Table C

<table> table see original document page 243 </column> </row> <table>

Example Section B: MADS15 Augmented

Example 8: Identification of Sequences Related to SEQID NO: 43 and SEQID NO: 44

Sequences (full length cDNA, EST or genomic) related to SEQ ID NO: 43 and / or protein sequences related to SEQ LD NO: 44 were identified among those maintained in the Entrez Nucleotides database at the National Center for BiotechnologyInformation (NCBI) using search engines. database sequences such as the Basic Local Alignment Tool (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997) Nucleic Acids Res. 25 : 3389-3402). The program is used to find regions of local similarity between sequences by comparing nucleic acid or polypeptide sequences with sequence database and calculating the statistical significance of identities. The polypeptide encoded by SEQ ID NO: 43 was used for the TBLASTN algorithm, with default settings and the filter to bypass low-complexity sequence counterweight. The result of the analysis was viewed by paired comparison, drawn according to the probability score (E value), where the score reflects the likelihood that a particular alignment will occur in the case (the lower the E value, the more significant the hit). In addition to E values, comparisons were also scored by percent identity. Percent identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. In some instances, the default parameters may be adjusted to modify the stringent stringency.

In addition to the publicly available nucleic acid sequences available from NCBI, particular database sequences are also searched following the same procedure as described above.

Table D provides a list of protein nucleic acid sequences related to the nucleic acid sequence represented by SEQ ID NO: 43 and the protein sequence represented by SEQ ID NO: 44. Table D: Nucleic acid sequence related nucleic acid sequences ( SEQ ID NO: 43) useful in the methods of the present invention, and the corresponding deduced polypeptides.

<table> table see original document page 245 </column> </row> <table>

Example 9: Alignment of Relevant Polypeptide Sequences

AlignX of the NTI Vector (Invitrogen) is based on the popular progressive alignment Clustal algorithm (Thompson et al. (1997) Nucleic Aeids Res 25: 4876-4882; Chenna et al. (2003). Nueleie Aeids Res31: 3497-3500). A phylogenetic tree can be constructed using a neighbor grouping algorithm. Default values are for gap gap penalty of 10, gap gap penalty of 0.1 and the selected weight matrix is Blosum 62 (if polypeptides are aligned).

The result of multiple alignment of sequences using relevant polypeptides to identify those useful for carrying out the methods of the invention is shown in Figure 6.

Example 10: Calculation of Global Percentage Identity Polypeptide Entrencies of Useful for Carrying Out the Methods of the Invention

Overall percentages of similarity and identity full-length polypeptide sequences useful for carrying out the methods of the invention were determined using one of the methods available in the art, the Global Matrix Alignment Tool (BMC Bioinformatics. 2003 4:29. MatGAT: an applicationthat generates similarity / identity matrices using protein or DNA sequences.Campanella JJ5 Bitincka L, Smalley J; hosted computer application by Ledion Bitincka). MatGAT computational application generates similarity / identity matrices for DNA or protein sequences without having to pre-align data. The program performs a series of paired alignments using the Myers and Miller global alignment algorithm (with a gap gap penalty of 12, and a narrow gap penalty of 2), calculates similarity and identity using for example Blosum62 (for polypeptides), and then put the results into a distance matrix. Sequence similarity is shown at the bottom half of the divider line and sequence identity is shown at the top half of the diagonal divider line. Parameters used in the comparison were:

Score Matrix: Blosum62

First Gap: 12

Extension Gap: 2

Results of computer application analysis are shown in Table E for similarity and overall identity in the complete length of polypeptide sequences (excluding partial polypeptide sequences). Percentage identity is given above dadiagonal and percentage similarity is given below diagonal.

The percent identity between polypeptide sequences useful for carrying out the methods of the invention may be as low as 40% amino acid identity compared to SEQ ID NO: 44.

Table E: MatGAT results for similarity and overall identity at full length of polypeptide sequences.

<table> table see original document page 247 </column> </row> <table> <table> table see original document page 248 </column> </row> <table>

Example 11: Identification of domains comprised of polypeptide sequences useful for carrying out the methods of the invention.

The Integrated Resource of Protein Families, Domains and Sites (InterPro) database is an integrated interface for signature data banks commonly used for text-based search queries. The InterPro database combines these databases, which use different methodologies and varying degrees of well-characterized overprotein biological information to derive protein signatures. Contributing banks include SWISS-PROT, PROSITE, TrEMBL5 PRINTS, ProDom and Pfam, Smart and TIGRFAMs. Interpro is hosted at the EuropeanBioinformatics Institute in the United Kingdom.

InterPro scan results of the polypeptide sequence as represented by SEQ ID NO: 44 are presented in Table F. Table F: Polypeptide sequence InterPro scan results as represented by SEQ ID NO: 44

Database Accession number Accession namePRINTS PR00404 MAD S DOMA INPFAM PF00319 SRF-TFSMART SM00432 MADSPROFILE PS50066 MADS BOX 2SUPERF AMILY SSF55455 SRF-IikePFAM PFO1486K-BOX1205456TXR19X45RTXX19PROXLE 19F MIX SIXPROFILT PROTEIN

Example 12: Topology prediction of polypeptide sequences useful for carrying out the methods of the invention (subcellular localization, transmembrane ...)

TargetP 1.1 predicts subcellular localization of proteinase and eukaryotes. The location designation is based on the predicted presence of any of the N-terminal presequences: chloroplast transit peptide (cTP), mitochondrial targeting peptide (mTP), or secretory signal peptide (SP). Scores on which the final forecast is based are not probabilities really, and they do not necessarily add to one. However, the location with the highest score is most likely to agree with TargetP, and the relationship between the scores (the confidence class) may be a indication of how accurate the forecast is. Confidence class (RC) ranges from 1 to 5, where 1 indicates the strongest forecast. TargetP is maintained on the server of the Technical University of Denmark.

For predicted sequences to contain an N-terminal pre-sequence a potential cleavage site may also be predicted.

Several parameters were selected, such as organism group (non-plant or vegetable), shortcut sets (none, pre-defined shortcut set, or user-specified shortcut set), and cleavage site prediction calculation (yes or no) .

TargetP 1.1 analysis results of the polypeptide sequence as represented by SEQ ID NO: 44 are presented in Table G. The "plant" organism group was selected, none defined, and the predicted length of the requested transit peptide. Subcellular localization of the polypeptide sequence as represented by SEQ ID NO: 44 may be the cytoplasm or nucleus, no transit peptide is predicted.

Table G: TargetP 1.1 analysis of polypeptide sequence as represented by SEQ ID NO: 44

<table> table see original document page 250 </column> </row> <table>

Many other algorithms can be used to perform such analyzes, including:

· ChloroP 1.1 hosted on TechnicalUniversityofDenmark server;

• Protein Prowler Subcellular Localization Predictor version 1.2 hosted on the Institute for Molecular Bioscience server, University of Queensland, Brisbane, Australia;

· PENCE Proteome Analyst PA-GOSUB 2.5 hosted at the University of Alberta Server, Edmonton, Alberta, Canada;

• TMHMM, hosted on Technical Universityof Denmark server

Example 13: Assay related to the polypeptide sequences useful for carrying out the methods of the invention Lim et al. (PMB 44, 513-527, 2000) describe two assays for measuring protein-protein interactions that can be applied to MADS15. In the assay of two yeast hybrids, a truncated form of MADS1 (comprising the complete MADS Box, the IeK domain but lacking C-terminal part of domain C) was used as bait to test interaction with MADS15. In the in vitro precipitation assay, GST and GST-linked MADS proteins were produced and immobilized on Sepharose4B glutathione. Resin bound GST / GST-MADS1 was mixed with S-labeled MADS15 proteins or fragments thereof. After washing, the interaction proteins were eluted and analyzed by SDS-PAGE. One of ordinary skill in the art will appreciate that the MADS15 protein may be used as bait in the screening of two hybrids or may be immobilized on deglutathione resin as well. In addition, a MADS15 protein, when used according to the methods of the present invention, will result in increased rice root yield, measured as an increased ratio of root to shoot biomass.

Example 14: Nucleic acid sequence cloning as represented by SEQID NO: 43

The Oryza sativa MADS15 gene was PCR amplified using a Oryza sativa seedling cDNA library (Invitrogen, Paisley, UK) as a model. After reverse transcription of seedling extracted RNA, cDNAs were cloned into pCMV Sport 6.0. Mean bank insert size was 1.5 kb and the original number of clones was 1.59 χ 10 cfu. Original titer was determined to be 9.6 χ 10 cfü / ml after the first amplification of 6 χ 1011 cfu / ml. After plasmid extraction, 200 ng of template was used in a 50 μΐ PCR mix. Prm06892 Initiators (SEQ ID NO: 45; sense, bold start codon, AttBl site in italics: S ^ ggggacaagtttgtacaaaaaagagaggcttaaaggggggggggggggggggggggggggggggggggggggggggg prm06893 (SEQ ID NO: 46; reverse, complementary, AttB2 site in italics: 5'-ggggaccactttgtacaagaaagctgggtttggccga.cga.cgacg3iC-3 '), which includes the ATB sites for Gateway recombination, were used for PCR amplification. PCR was performed using HiFi Taq DNA polymerase under standard conditions. A PCR fragment of about 915 bp was amplified and purified also using standard methods. The first step of the Gateway procedure, the BP reaction, was then performed, during which the PCR fragment would be serecombin in vivo with plasmid pDC) NR201 to produce, according to Gateway terminology, an "input clone", pMADS15. PlasmidopDC) NR201 was purchased from Invitrogen as part of the Gateway® technology.

Example 15: Construction of expression vector using nucleic acid sequence as represented by SEQID NO: 43

The input clone pMADS15 was subsequently used in an LR reaction with pGOS2, a targeting vector used for Oryza sativa transformation. This vector contains as functional elements within the limits of T-DNA: a selectable plant marker; a drawer traceable marker expression; and a Gateway drawer intended for in vivo LR combining with the nucleic acid sequence of interest already cloned into the input clone. A rice GOS2 promoter (SEQ ID NO: 108, alternatively, SEQ ID NO: 47 is equally useful) for constructive expression was located before this Gateway drawer.

Following the LR recombination step, the resulting expression vector pGOS2 :: MADS15 (Figure 7) was transformed into Agrobacterium strain LBA4044 according to methods well known in the art.

Example 16: Plant Transformation

Rice processing

Agrobacterium containing the expression vector was used to transform Oryza sativa plants. Dried ripe seeds of the Japanese Nipponbare rice cultivar were peeled. Sterilization was performed by incubating for one minute in 70% ethanol, followed by 30 minutes in 0.2% HgCl2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus induction medium). After incubation in the dark for four weeks, scutellus-derived embryogenic calli were excised and propagated in the same medium. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic callus pieces were subcultured in fresh medium 3 days prior to co-cultivation (to stimulate cell division activity).

Agrobacterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobacterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD60 °) of about 1. The suspension was then transferred to a petri dish and the calluses immersed in the suspension for 15 minutes. Callus were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 25 ° C. Co-cultured horses were grown in medium containing 2,4-D for 4 weeks in the dark at 25 ° C. 28 ° C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Primary transformants were transferred from a tissue culture chamber to a greenhouse. After quantitative PCR analysis to verify T-DNA insert copy number, only single copy transgenic plants exhibiting tolerance to the deletion agent were maintained for TI seed harvest. Seeds were then collected three to five months after transplantation. The method produced single locus transformants at a rate of up to 50% (Aldemita andHodges 1996, Chan et al. 1993, Hiei et al. 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural Alpa 88 (University of Minnesota) or Al 88 hybrids as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are collected from the corn plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm. Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and plants GMOs are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium containing the selection agent (eg imidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. The green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat processing

Wheat processing is performed with the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybeans are processed according to a modification of the method described in Texas A&M Patent 5,164,310. Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. The epicotyl and the remnant cotyledon are further cultivated to develop axillary nodes. These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / canola transformation

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188).

The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used. Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobacterium (containing the expression vector) bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobacterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

Alfalfa Transformation

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K2SO4, and 100 µg acetoseringone. Explantants are washed in half-strength Murashige-Skoog medium (Murashige andSkoog, 1962) and placed on the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobaeterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 17: Phenotypic Evaluation Procedure

17.1 Evaluation Configuration

Approximately 35 independent rice TO transformants were generated. Primary transformants were transferred from a tissue culture chamber to a greenhouse for TI seedlings. Six events, of which progeny T1 secreted 3: 1 for presence / absence of transgene, were retained. For each of these events, approximately 10 Tl seedlings containing the transgene (hetero and homozygotes) and approximately 10 Tl seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression.

Transgenic plants and corresponding nullizigotes were grown side by side in random positions. Greenhouse conditions were short days (12 hours of light), 28 ° C in the light and 22 ° C in the dark, and a relative humidity of 70%.

Four T1 events were additionally evaluated in generation T2 following the same evaluation procedure as for T1 generation but with more individuals per event. From the sowing stage to the maturity stage the plants were passed several times through a digital image cabinet. At each moment digital images (2048x1536 pixels, 16 million colors) were taken from each plant from at least 6 different angles.

17.2 Statistical analysis: F test

A two-way ANOVA (analysis of variance) was used as a statistical model for the general evaluation of plant phenotypic characteristics. An F test was performed on all measured parameters of all plants of all events transformed with the gene of the present invention. The F test was performed to check for a gene effect on all transformation events and to check for a general gene effect, also known as the overall gene effect. The threshold for significance for a true overall gene effect is set at a 5% probability level for the F test. A significant F test value points to a gene effect, meaning that it is not just the mere presence or apposition of the gene that is causing the differences. in phenotype.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" are plants treated in the same manner as the transgenic plant, but from which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test is set at 10% probability level. Results for some events may be above or below this threshold. This is based on the hypothesis that a gene can only have an effect on certain positions in the genome, and that the occurrence of this position-dependent effect is not uncommon. This type of gene effect is also called in this place a "gene strain effect". The value of ρ is obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the hypothesis (ie, that there is no transgene effect). ) is correct.

17.3 Measured Parameters

From the sowing stage to the maturity stage the plants were passed several times through a digital imaging cabinet. At each moment digital images (2048x1536 pixels, 16 million decors) were taken from each plant from at least 6 different angles.

The plant shoot (or leaf biomass) was determined by counting the total number of pixels in the digital images of plant partisans discriminated from the background. This value was averaged for at-the-moment photos of different angles and converted to a physical surface value expressed in mm squared by calibration. Experiments show that the plant shoot measured in this way correlates with the plant shoot biomass . The aerial part is the measured area at the moment the plant reached its maximum leaf biomass. Early vigor is the aerial plant area (seedling) three weeks after germination. Increase in root biomass is expressed as an increase in total root biomass (measured as the maximum observed root biomass during the life of a plant); or as an increase in the root / shoot index (measured as the ratio between root and shoot biomass in the active root and shoot growth period).

Example 18: Results of Phenotypic Evaluation of Plantastransgenics

Upon analysis of the plants as described above, the applicants found that plants transformed with the MADS15 gene construct had a higher root yield, expressed as index / shoot, compared to plants lacking the MADSl 5 transgene. The increase was 9.4% (value ρ 0.0207) in T1 and 25.7% (value of ρ 0.002) in T2. The values of ρ show that the increases were significant.

Example Section C: MADS15 Decreased

See also Examples 8 through 13 for identification and sequence characterization related to MADS15.

Example 19: Gene Cloning

The Oryza sativa MADS15 gene was PCR amplified using a Oryza sativa seedling cDNA library (Invitrogen, Paisley, UK) as a model. After reverse transcription of seedling extracted RNA, cDNAs were cloned into pCMV Sport 6.0. The average bank insert size was 1.5 kb and the original number of clones was of the order of 1.59 χ 10 7 cfii. Original titer was determined to be 9.6 x 105 cfu / ml after first amplification of 6 x 105 cfu / ml. After plasmid extraction, 200 ng of template was used in a 50 μΐ PCR mix. Prm06892 Initiators (SEQ ID NO: 111; sense, bold start codon, AttBl site in italics: '-gggacaagtttgtacaaaaaagcaggcttaaacagggggggg-eg prm06893 (SEQ ID NO: 112; reverse, complementary, italicAttB2 site:

5 '-gggaccactttgtacaagaaagctgggttíggccgacgacgacgac-3'), which includes AttB sites for Gateway recombination, were used for PCR amplification. PCR was performed using Hifi Taq DNA polymerase under standard conditions. A PCR fragment of about 915 bp was also purified and purified using standard methods. The deCRP fragment was subsequently used for the preparation of a staple construct using techniques known in the art. The first step of the Gateway procedure, the BP reaction, was then performed, during which the staple construct recombines in vivo with plasmid PDONR201 to produce, according to Gateway terminology, an "input cloning", pMADS15hp. Plasmid pDONR201 was purchased from Invitrogen as part of Gateway® technology.

Example 20: Vector Construction

The input clone pMADS15hp was subsequently used in an LR reaction with p01519, a destination vector for the inverted repeat construct. This vector contains as functional elements within all T-DNA boundaries: a selectable plant marker; a traceable marker expression drawer; and a Gateway drawer intended for LR in vivo matching so that the input clone sequence of interest is integrated as an inverted repeat. A GOS2 dearroz promoter (SEQ ID NO: 174, alternatively, SEQ ID NO: 113 is equally useful) for constitutive expression was located prior to this Gateway drawer.

After the LR recombination step, the resulting inverse repeat expression vector, Figure 10) was transformed into Agrobacterium strain and subsequently to Oryzasativa plants.

Example 21: Plant Transformation

Rice processing

Agrobacterium containing the expression vector was used to transform Oryza sativa plants. Dried ripe seeds of the Japanese Nipponbare rice cultivar were peeled. Sterilization was performed by incubating for one minute in 70% ethanol, followed by 30 minutes in 0.2% HgCl2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus induction medium). After incubation in the dark for four weeks, scutellus-derived embryogenic calli were excised and propagated in the same medium. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic callus pieces were subcultured in fresh medium 3 days prior to co-cultivation (to stimulate cell division activity).

Agrobacterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobacterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD60) of about 1. The suspension was then transferred to a Petri dish and the calluses immersed in the suspension for 15 minutes. Callus-laden were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 250 ° C. Co-cultured horses were grown in medium containing 2,4-D for 4 weeks in the dark at 28 °. C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Primary transformants were transferred from a tissue culture chamber to a greenhouse. After a quantitative PCR analysis to verify T-DNA insert copy number, only single copy transgenic plants exhibiting tolerance to the deletion agent were maintained for TL seed harvest. Seeds were then collected three to five months after transplantation. The method yields single locus transformants at a rate of up to 50% (Aldemita andHodges 1996, Chan et al 1993, Hiei et al. 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural Alpa (University of Minnesota) or Al88 hybrids as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are harvested from the maize plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm.

Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and transgenic plants are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium, containing the selection agent (eg imidazolinone but various selection markers may be used).

Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. The green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat transformation Wheat transformation is performed using the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybeans are processed according to a modification of the method described in Texas A&M Patent 5,164,310. Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. The epicotyl and the remnant cotyledon are further cultivated to develop axillary nodes. These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / canola transformation

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188). The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used. Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobacterium (containing the expression vector) bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobacterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K2 SO4, and 100 µm acetoseringone. Explantants are washed in half-strength Murashige-Skoog medium (Murashige and Skogog, 1962) and placed in the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobacterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 22: Evaluation methods of plants transformed with MADS15 inverted repeat under control of GOS2 dearroz promoter

Approximately 15 to 20 independent rice TO transformants were generated. Primary transformants were transferred from a tissue culture chamber to a greenhouse for TI seedlings. Eight inverted repeat construct events in which the progeny T1 secreted 3: 1 for presence / absence of the transgene were retained. For each of these events, approximately 10 seedlings containing the transgene (hetero and homozygous) and approximately 10 seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression. The selected T1 plants were transferred to a greenhouse. Each plant received an exclusive barcode to unambiguously link the corresponding phenotyping data to the plant. The selected Tl plants were grown in soil in 10 cm diameter pots in the following environmental settings: photoperiod = 11.5 h, daytime intensity = 30,000 Iux or more, daytime temperature = 28 ° C or more, night temperature = 22 ° C, humidity relative = 60-70%. Transgenic plants and the corresponding nullizigotes were grown side by side in random positions. From the sowing stage to the maturity stage the plants were passed several times through a digital image cabinet. At each moment digital images (2048x1536 pixels, 16 million colors) were taken from each plant from at least 6 different angles.

The plant shoot (or leaf biomass) was determined by counting the total number of pixels in the digital images of plant partisans discriminated from the background. This value was averaged for at-the-moment photos of different angles and converted to a physical surface value expressed in mm squared by calibration. Experiments show that the plant shoot measured in this way correlates with the plant shoot biomass . Areamax is the air area at the time the plant reached its maximum leaf biomass.

The mature primary panicles were collected, bagged, bar-coded and then oven dried at 37 ° C for three days.

The panicles were then traced and all seeds collected. The shells were separated from the empty ones using an air blower. After separation, both seed lots were then counted using a commercially available counting machine. The empty shells were discarded. The full shells were weighed on an analytical scale and the cross-sectional area of the seeds was measured using digital imaging. This procedure resulted in the establishment of the following seed related parameters:

Flowers per panicle is a parameter estimating the average number of rapiers per panicle in a plant, derived from the total seed number divided by the number of first panicles. The highest panicle and all panicles that overlap with the highest panicle when vertically aligned were considered the first panicles and were manually counted. The number of full grains was determined by counting the number of full husks that remained after the separation step. Total seed yield (total seed weight) was measured by weighing all full bark collected from a plant. Total number of seeds per plant was measured by counting the number of bark collected from a plant and corresponding to the number of rapiers per plant. These parameters were derived in an automated manner from digital images using the computational image analysis application and were statistically analyzed. Individual seed parameters (including width, length, area, weight) were measured using a custom-made device consisting of two main components, a weighing and imaging device, coupled with a computational application for image analysis. A two-factor ANOVA (analysis of variance ) corrected for the unbalanced model was used as a statistical model for the general evaluation of plant phenotypic characteristics. An F-test was performed on all measured parameters of all plants of all events transformed with that gene. The F test was performed to check for a gene effect on all transformation events and to verify for a general gene effect, also called in this place "global gene effect". If the value of the F test shows that the data is significant, then it was concluded that there was a "gene" effect, meaning that it was not just the presence or position of the gene that was causing the effect. The threshold for significance for a true global gene effect was adjusted to a 5% probability level for the F test.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" refer to plants treated in the same manner as the transgenic plant, but in which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test was adjusted to a 10% probability level.

Results for some events may be above or below this threshold. This is based on the hypothesis that a gene may only have an effect at certain positions in the genome, and that the occurrence of this deposition-dependent effect is not uncommon. This type of gene effect is also called nestlugar as a "gene strain effect". The value of ρ was obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the null hypothesis (ie, that there is no transgene effect). ) is correct.

Example 23: Measurement of production-related parameters for inverted repeat construction transformants:

Upon seed analysis as described above, applicants found that plants transformed with the MADS15 gene clamp-shaped construct had a higher seed yield, expressed as number of full grains, total seed weight, total number of seeds, and flowers per panicle, compared to plants lacking MADS15 dotransgene, whereas plants transformed with the sense MADS15 gene construct showed opposite effects. The values of ρ show that the increases were significant.

The results obtained for plants in generation T1 are summarized in Table H, which represent the mean values for all lines tested:

Table H:

<table> table see original document page 270 </column> </row> <table>

Example Section D: PLT

Example 24: Identification of Sequences Related to Nucleic Acid Sequence Used in the Methods of the Invention

Sequences (full length cDNAs, ESTs, or genomic) related to the nucleic acid sequence used in the methods of the present invention were identified among those maintained in the Entrez Nucleotides database at the National Center for Biotechnology Information (NCBI) using database sequence search tools. , such as the Basic Local Alignment Tool (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997) Nucleic Acids Res.25: 3389-3402). The program is used to find regions of local similarity between sequences by comparing nucleic acid or polypeptide sequences with sequence database and calculating the statistical significance of identities. For example, the nucleic acid-encoded polypeptide of the present invention was used for the TBLASTN algorithm, with standard settings and the filter for bypassing low complex sequence sequences. The result of the analysis was viewed by paired comparison, drawn according to the probability score (E value), where the score reflects the likelihood that a particular alignment will occur in the case (the lower the E value, the more significant the hit). In addition to E values, comparisons were also scored by percent identity.

Percent Identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. In some instances, the default parameters may be adjusted to modify the stringent stringency. For example, the value E can be increased to show less stringent pairings. In this way, practically exact short pairings can be identified.

Table I below provides a list of nucleic acid sequences related to the nucleic acid sequence used in the methods of the present invention.

Table I: Nucleic acid sequences related to nucleic acid sequence used in the methods of the present invention

<table> table see original document page 271 </column> </row> <table> <table> table see original document page 272 </column> </row> <table>

In some instances, related sequences have been experimentally grouped and publicly described by research institutions, such as The Institute for Genomic Research (TIGR). The Eukaryotic Gene Orthologs (EGO) database can be used to identify such related sequences either by keyword searching or by using the BLAST algorithm with the nucleic acid or polypeptide sequence of interest.

Example 25: Cloning of Nucleic Acid Sequences in the Methods of the Invention

The nucleic acid sequences used in the invention methods were PCR amplified using a custom Arabidopsis thaliana cDNA library starting from RNA extracted from different tissues (in pCMV Sport 6.0; Invitrogen, Paisley, UK) .PCR was performed using Hifi Taq DNA polymerase under standard conditions using 200 ng template in a 50 μΐ PCR mix. Primers used to amplify SEQ ID NO: 175 (PLT1) were prm08180 (SEQ IDNO: 195; sense, bold start codon, AttBl site in italics: 5'GGGGACAAGTTTGTA

CAAAAAAGCAGGCTTAAACAATGATCAATCCACACGGTG 3 ') eprm08181 (SEQ ID NO: 196; reverse, complementary, AttB2 site in italics: 5' GGGGA CCA CTTTGTA The primers used to amplify SEQ IDNO: 177 (PLT2) were prm08182 (SEQ ID NO: 197; direction, start codon in bold, AttBl site in italics: 5'GGGGA CAA GTTTGTA CAA AAAA GCA GGCTTAAA CA ATG AATTCTAAC AACTGGCTC 3 ') prm08183 (SEQ ID NO: 198; reverse, complementary, italicAttB2 site: 5'GGGGA CCA CTTTGTA CAA GAAA GC7GGG7TC ATCTTTTATTC ATTCC ACA 3 '),

Amplified PCR fragments were also purified using standard methods. The first step of the Gateway procedure, the BP reaction, was then performed, during which the serecombin PCR fragment in vivo with plasmid pDONR2 () 1 to produce, in accordance with Gateway terminology, an "input clone", pPLT1 for SEQ IDNO: 175 and pPLT2 for SEQ ID NO: 177. Plasmid pDONR2 () was purchased from Invitrogen as part of Gateway® technology.

Example 26: Expression Vector Construction

The input clones pPLT1 and pPLT2 were subsequently used in an LR reaction with destination vectors used for Oryza sativa transformation. A first vector contains as functional elements within the limits of T-DNA: a plant selectable marker, a traceable marker expression drawer; and a Gateway drawer intended for in vivo LR recombination with the nucleic acid sequence of interest already cloned into the input clone. A constructive rice promoter, a GOS2 promoter (SEQ ID NO: 210, alternatively SEQ ID NO: 194 is equally useful) was located before this Gateway drawer.

A second vector contains the same functional elements within the T-DNA boundaries: a selectable plant marker; a drawer traceable marker expression; and a Gateway drawer intended for in vivo LR combining with the nucleic acid sequence of interest already cloned into the input clone. A rice promoter for expression in multiple systems, an MT promoter (SEQ ID NO: 211) (PR00126) was located before this Gateway drawer.

After the LR5 recombination step the resulting expression vectors, pGOS2 :: PLT1, pGOS2; PLT2, pMT :: PLT1 and pMT :: PLT2 (Figure 16 shows the GOS2 promoter construct) were independently transformed into Agrobacterium strain LBA4044 and subsequently to Oryza sativa paraplants. Transformed rice plants were allowed to grow and were then examined for the parameters described below.

Example 27: Culture Transformation

Transformed Agrobacterium containing the expression vectors were used independently to transform Oryzasativa plants. Dried ripe seeds of the Japanese rice cultivar Nipponbare have been husked. Sterilization was performed by incubating for one minute in 70% ethanol followed by 30 minutes in 0.2% HgCl 2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus-inducing medium). After incubation in the dark for four weeks, scutellus-derived callosembriogenes were excised and even propagated. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic decal pieces were subcultured in fresh medium 3 days before co-cultivation (to stimulate cell division activity).

Agrobacterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobacterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD600) of about 1. The suspension was then transferred to a petri dish and the calluses immersed in the suspension for 15 minutes. Callus were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 25 ° C. Co-cultured horses were grown in medium containing 2,4-D for 4 weeks in the dark at 25 ° C. 28 ° C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Approximately 35 independent rice TO transformants were generated for a construction. The primary transformants were transferred from a tissue culture chamber to a greenhouse. After a quantitative PCR analysis to verify the number of copies of the T-DNA insert, only transgenic single copy plants exhibiting tolerance to the selection agent were maintained for TI seed harvest. Seeds were then collected three to five months after transplantation. The method produced single locus transformants at a rate of up to 50% (Aldemita and Hodges 1996, Chan et al. 1993, Hiei etal. 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural A188 (University of Minnesota) or hybrids with A188 as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are collected from the corn plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm. Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and plants GMOs are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium containing the selection agent (eg imidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. The green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat processing

Wheat processing is performed with the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybean is processed according to a modification of the method described in Texas A&M Patent 5,164,310.

Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. Epicotyl and remnant cotyledon are further grown to develop axillary nodes.

These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / canola transformation

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188).

The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used.

Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobacterium (containing the expression vector) by bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobacterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-3 medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

Alfalfa Transformation

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K 2 SO 4, and 100 µ m acetoseringone. Explantants were washed in half-strength Murashige-Skoog medium (Murashige andSkoog, 1962) and placed on the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobacterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 28: Phenotypic Evaluation Procedure

28.1 Evaluation Configuration

Assessment under normal cultivation conditions

Primary transformants were transferred from a tissue culture chamber to a greenhouse for cultivation and harvesting of IT. Six events, of which progeny T1 secreted 3: 1 for presence / absence of the transgene, were retained. For each of these events, approximately 10 Tl seedlings containing the transgene (hetero ehomozigotes) and approximately 10 Tl seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression. Transgenic plants and corresponding nullizigotes were grown side by side at random positions. . Greenhouse conditions were short days (12 hours of light), 28 ° C in the light and 22 ° C in the dark, and a relative humidity of 70%.

Evaluation under reduced availability conditions denitrogenRice plants were grown in potted soil under normal conditions except for nutrient solution. The pots were soaked from transplant to maturity with a specific nutrient solution containing reduced nitrogen (N) content, usually 7 to 8 times less. The rest of the crop (plant maturation, seed collection) was the same as for plants not cultivated under abiotic stress.

28.2 Statistical analysis: F test

A two-way ANOVA (analysis of variance) was used as a statistical model for the general evaluation of plant phenotypic characteristics. An F test was performed on all measured parameters of all plants of all events transformed with the gene of the present invention. The F test was performed to check for a gene effect on all transformation events and to check for a general gene effect, also known as the overall gene effect. The threshold for significance for a true overall gene effect has been adjusted to a 5% probability level for the F test. A significant Faponta test value for a gene effect means that it is not just the mere presence or position of the gene that is causing the differences. in phenotype.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" refer to plants treated in the same manner as the transgenic plant, but in which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test was adjusted to a 10% probability level.

Results for some events may be above or below this threshold. This is based on the hypothesis that a gene may only have an effect at certain positions in the genome, and that the occurrence of this deposition-dependent effect is not uncommon. This type of gene effect is also called nestlugar as a "gene strain effect". The value of ρ was obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the null hypothesis (ie, that there is no transgene effect). ) is correct.

28.3 Measured Parameters

Biomass related parameter measurement

From the sowing stage to the maturity stage the plants were passed several times through a digital imaging cabinet. At each moment digital images (2048x1536 pixels, 16 million decors) were taken from each plant from at least 6 different angles.

The plant aerial area (or leaf biomass, areamax) was determined by counting the total number of pixels in the digital images of plant partisans discriminated from the background. This value was averaged for at-the-same-time photo shoots and converted to a physical surface value expressed in mm squared by calibration.

Experiments show that the plant shoot measured in this way correlates with the plant shoot biomass. The shoot is the measured area at the moment the plant reached its maximum leaf biomass.

Early vigor is the aerial area of the plant (seedling) three weeks after germination. Increase in root biomass is expressed as an increase in total root biomass (measured as the maximum observed root biomass during the life of a plant); or as an increase in the index / aerial part (measured as the ratio of root mass to aerial departe mass in the active root and aerial part growth period).

Seed-related parameter measurements

The mature primary panicles were collected, counted, bagged, barcoded and then dried for three days in an oven at 37 ° C. The panicles were then traced and all seeds were collected and counted. The full shells were separated from the empty ones using an air-blowing device. The empty shells were discarded and the remaining fraction was counted again. The full shells were weighed on an analytical balance. The number of full grains was determined by counting the number of full husks that remained after the desparing step. Total seed yield was measured by weighing all bark collected from a plant. Total number of seeds per plant was measured by counting the number of bark collected from a plant. Millet seed weight (TKW) is extrapolated from the number of full grains counted and its total weight. The harvest index (HI) in the present invention is defined as the ratio of total seed production to shoot (mm2) multiplied by a factor 106. The total number of flowers per panicle as defined in the present invention is the ratio of the total number of seeds to the number of mature primary panicles. The grain fill rate as defined in the present invention is the ratio (expressed as a%) of the number of full grains to the total number of seeds (or rapiers).

Example 29: Results of Phenotypic Evaluation of Plantastransgenics

29.1 Results of phenotypic evaluation of plantastransgenic cultivated under normal cultivation conditions, expressing sequence of PLT1 or PLT2 nucleic acids under the control of a conductive promoter

The TKW measurement results of Tl seeds of transgenic PLT1 and PLT2 transgenic rice plants grown under normal growing conditions are shown in Table J, in absolute values (mean of events) and as a percentage compared to wild type plants. A substantial increase in TKW is observed for transgenic seeds containing any construct compared to wild type seeds.Table J: Results of TKW measurements of TL seeds from PLT1 and PLT2 transgenic plants grown under normal growing conditions.

<table> table see original document page 283 </column> </row> <table>

Individual seed parameters (including width, length, area, weight) were measured using a custom made device consisting of two main components, a weighing and imaging device, coupled with a computer application for image analysis.

Measurement results of seed area and seed length of Tl seeds of transgenic rice plantsPLT1 and PLT2 are shown in Table K in absolute values (mean of events) and as a percentage compared to wild type plants. A clear increase in both seed area and seed length is observed for transgenic seeds containing any construct compared to wild type seeds.

Table K: Results of seed area measurements and seed length of Tl seeds from PLT1 and PLT2 transgenic plants grown under normal cultivation conditions.

<table> table see original document page 283 </column> </row> <table>

Seed width was not significantly affected in Tl seeds of PLT and PLT2 transgenic plants (data not shown).

29.2 Results of phenotypic evaluation of plantastransgenic plants grown under reduced availability of denitrogen, expressing either PLT1 or PLT2 nucleic acid sequence under the control of a constitutive promoter.

The TKW measurement results of Tl seeds of transgenic PLT1 and PLT2 transgenic rice plants grown under reduced nitrogen availability are shown in Table L, absolute values (average of events) and as a percentage compared to wild type plants. A substantial increase in TKW is observed for transgenic asserts containing any construct compared to wild type seeds.

Table L: Results of seed TKW measurements

Tl of transgenic PLT1 and PLT2 plants grown under reduced nitrogen availability.

<table> table see original document page 284 </column> </row> <table>

29.3 Results of phenotypic evaluation of plant-transgenic plants grown under normal cultivation conditions, expressing PLT2 nucleic acid frequency under the control of a specific promoter-system.

The TKW measurement results of transgenic PLT2 cultivated rice seedlings Tl seeds under normal growing conditions, expressing PLT2 nucleic acid sequence under the control of a specific meristem promoter, are shown in Table M, in absolute values (event average) and as a percentage compared to wild plant type. A substantial increase in TKW is observed for transgenic assertions containing the construct compared to wild type seeds. Table ab: TKW Measurement Results of TL Seeds of Transgenic PLT2 Plants Grown Under Normal Growing Conditions, Expressing PLT2 Nucleic Acid Sequence Under Control of a specific meristem engine.

<table> table see original document page 285 </column> </row> <table>

Example Section E: bHLH

Example 30: Identification of SEQID NO: 213 bHLH Paralogues in Rice

SEQID 2 was used to search for parallels in the rice genome using a BLASTP algorithm used to perform query dissimilarity of a query protein sequence with a given database protein sequence. The sequence of database proteins sought corresponded to the rice proteome of the MIPS Institute, Oryza sativa MIPS (MOsDB) 5 database comprising 59,712 sequences; 27,051,637 total letters. Results were ranked according to highest similarity if determined from the highest score and lowest e. Hits identifying bHLH protein sequence dialogues with SEQID NO: 2 had a score of at least 50 and a lower value than e-05. Paired alignments between the query string and the newly identified dialogs are shown below.

Query: SEQID 2Smaller sum

High probability

Sequences producing High Score Segment Pairs: Score P (N) N

9629.m00093 | protein DNA-Helix-Helix DNA Binding Domain, 1132 1.3e-l 15 1

9629.m00090 | protein DNA-Helix-Helix DNA Binding Domain, 305 5.7e-28 1

9630.mO 1262 | protein Helix-Helix-DNA DNA Binding Domain, 189 1.1 and-15 1

9629.m07159 | protein Helix-Helix-DNA DNA Binding Domain, 112 3.4e-05 1

> 9629.m00093protein Putative helix-helix DNA binding domain

Length = 225Score = 1132 (403.5 bits), Expected = 1.3e-115, P = 1.3e-115Identities = 224/224 (100%), Positive = 224/224 (100%)

Consultation: 1 MKSRKNSTTSIKAAGSCHTSSSGGGGGGGNCYSSSSSKMERKDVEKNRRLHMKGLCLKLS 60

MKSRKNSTTSTXAAGSCHTSSSGGGGGGGNCYSSSSSKMERKDVEKNRRLHMKGLCLKLSSubject: 1 MKSRKNSTTSTKAAGSCHTSSSGGGGGGGNCYSSSSSKMERKDVEKNRRLHHQSKKHRQHRQHRQHRQHRQHHQKHLRQHHRQHRQHRQHRQHLQDQRQHRQHLQQHRQHHRQJHRQ

SLIP AAAPRRHHHHYSTS S S SPP S STKE A VTQLDHLEQ AAA YIKQLKGRIDELKKRKQQ Subject: 61 SLlPAAAPRRHHHYSTSSSSSPPSSTKEAVTQLDHLEQAAAYIKQLKGRIDELKKTTSVEGGEREVEEREVSGVEEREVGRTVEVGTLRVQG

AAALTTSTSNGGGGGMPWEVRCQDGTLDVVWSEAIREERERAVRLHEVIGVLEEEGAESujeito: 121 AAALTTSTSNGGGGGMPWEVRCQDGTLDVVWSEAIREERERAVRLHEVIGVLEEEGAE 180Consulta: 181 224 WNASFS WGDKIFYTLHSQALCSRIGLDASRVSHRLRNLLLQY

WNASFSWGDKIFYTLHSQALCSRIGLDASRVSHRLRNLLLQYSupport: 181 WNASFSWGDKIFYTLHSQALCSRIGLDASRVSHRLRNLLLQY 224> 9629.m000901protein Helix-loop-helix DNA binding domain

Length = 363 Score = 305 (112.4 bits), Expected = 5.7e-28, P = 5.7e-28Identities = 87/230 (37%), Positive = 126/230 (54%)

Consultation: 19 TSSSGGGGGGGNCYSSSSSKMERKDVEKNRRLHMKGLCLKLSSLIPAAAPRRHHHHYSTS 78TSSSG G +++++ ERK ++ E + RR MKGLC + KL + SLIP + HS

Subject: 18 TSSSGSGASS — TAAAAAAERKEMERRRRQDMKGLCVKLASLIP ----- KEHCSMSKM 68

Consultation: 79 SSSSPPSSTKEAVTQLDHLEQAAAYIKQLKGRIDELKKRKQQ ---------------- AA 122

++ S TQL L-H-AAAYIK + LK R + DEL ++ AA

Subject: 69 QAASR -------- TQLGSLDEAAAYIKKLKERVDELHHKRSMMSITSSRCRSGGGGGPAA 120

Consultation: 123 ALTTSTSNGGGGGMPWEVRCQDGTLDWWSEAIRE ------------ ERERAVRLHEV 170

STS GGGG ++ + VV V V R -h + H + VSujeito: 121 AAGQSTSGGGGGEEEEEDMTRTT AAAA VSL VRQHVQEGSLISLDVVLICSAARP VKFHDV 180Consulta: 171 IGVLEEEGAEVVNASFSVVGDKIFYTLHSQALCSRIGLDASRVSHRLRNL VLEEEGA 220I-HH-A FS + + ++ + YT + S-H-SRIG The ASR + S RLR LSujeito : 181 ITVLEEEGADIISANFSLAAHNFYYTIYSRAFSSRJGIEASRISERLRAL 230> 9630.m01262lprotein Putative Helix-Helix DNA Binding Domain

Length = 211 Score = 189 (71.6 bits), Expected = l.le-15, P = 1.1 and -15Identities = 66/214 (30%), Positive = 102/214 (47%)

Consultation: 21 SSGGGGGGGNCYSSSSSKMERKDVEKNRRLHMKGLCLKLSSLIPAAAPRRHH-HHYSTSS 79S + GGGGGGG K + RK E + RR M L L SL + + A P + + S S +

Subject: 7 SAGGGGGGG -------- KPDRKTTERIRREQMNKLYSHLDSLVRSAPPTVNSEPSHSNSN 58

Consultation: 80 SSSPPSSTK ------ EAVTQLDHLEQAAAYIKQLKGRIDELKKRKQQ — AAALTTSTSN 130

S + A T + D L YY + Q + R + D L +++ K ++ + S + S + Subject: 59 SKYHQRKLRILGGAAAATTRPDRLGVAAEYIRQTQERVDMLREKKRELTGGGGGGSSSSS 118Query: 131 GGGGGM VWE VRL VEH VRL VEHV

GG PVEV + L ++ + A + H + + E + G + VNAFSSubject: 119 GAGAATAAAPEVEVQHLGSGLHAILFTGAPPTD — GASFHRAVRAVEDAGGQVQNAHFS 175Reference: 188 WGDKIFYTLHSQALCSRIGLDASRVSHRL

VGK YT + H + G ++ RV RL + + Subject: 176 VAGAKAVYTIHAMIGDGYGGIE — RVVQRLKEAI 207

Example 31: Identification of bHLH orthologs of SEQIDNO: 213 in Arabidopsis thaliana

SEQID 2 was used to search for orthologs in the Arabidopsis genome using a BLASTP algorithm used to perform a dissimilarity search of a query protein sequence with a given database protein sequence. The database protein sequence sought corresponded to the IMPS Arabidopsis proteome, the MIPS Arabidopsis thaliana (MAtDB) database comprising 26,735 sequences; 11,317,104 total letters. Results were ranked according to greater similarity if determined from the highest score and lowest e. Correctly identifying orthologous bHLH protein mismatches with SEQID2 had a score of at least 50 and a value less than e-05. Paired alignments between query string and the newly identified orthologue are shown below.

Consultation: SEQID 2

Database: / home / data / blast / orgs_sets / arabi

26,735 Sequences; 11,317,104 total letters.

Lowest sum

High probability

Score P (N) N180 4.5e-15 1149 8.7e-12 1118 2.0e-06 1105 1.5e-05 1110 1. 5e-05 1

Sequences Producing High Score Segment Pairs: Atlgl0585 unknown proteinAt4g20970 hypothetical proteinAt4g25410 putative protein

At5g51780 putative bHLH transcription factor (bHLH036) At5g51790 putative protein> Atlgl0585 unknown protein

Length = 122 Score = 180 (68.4 bits), Expected = 4.5e-15, P = 4.5e-15Identities = 38/119 (31%), Positive = 72/119 (60%)

Consultation: 106 QLKGRIDELKKRKQQAAALTTSTSNGGGGGMPWEVRCQDGTLDWWSEAIREERERAV 165QLK ++ LK ++ K + G + P + + R + D T +++ +++++ RV

Subject: 3 QLKENVNYLKEKKRTLLQGELGNLYEGSFLLPKLSIRSRDSTIEMNLIMD-LNMKR --- V 58

Consultation: 166 RLHEVIGVLEEEGAEWNASFSWGDKIFYTLHSQALCSRIGLDASRVSHRLRNLLLQY 224

LHE ++ + EEEGA + V ++ A + + D + YT + + QA + SRIG + D SR + R + R ++ Y Subject: 59 MLHELVSIFEEEGAQVMSANLQNLNDRTTYTIIAQAIISRIGIDPSRIEERVRKIIYGY 117> At4g20970 hypothetical protein

Length = 167Score = 149 (57.5 bits), Expected = 8.7e-12, P = 8.7e-12Identities = 49/171 (28%), Positive = 86/171 (50%)

Consultation: 33 SSSSSKMERKDVEKNRRLHMKGLCLKLSSLIPAAAPRRHHHHYSTSSSSSPPSSTKEAVT 92+ S ++ RK VEKNRR + MK L + L SL + P HH ST + P EA

Subject: 8 TGQSRSVDRKTVEKNRRMQMKSLYSELISLLP -------- HHSSTEPLTLP-DQLDEAAN 58

Inquiry: 93 QLDHLEQAAAYIKQLKGRI --- DELKKRKQQ-AAALTTSTSNGGGGGMPWEVRCQDGTL 148

+ L + ++ K + L + K ++++++ S + P + E ++ + G ++ Subject: 59 YIKKLQVNVEKKRERKRNLVATTTLEKLNSVGSSSVSSSVDVSVPRKLPKIEIQ-ETGSI 117Query: 14 9 DVVVVSEAIREERERASEEVHVEHVVVVVHV

+ + ++ E E + I VL EE GAE + + A + S + V D + F + TLHS Subject: 118 FHIFLVTSL ---- EHKFMFCEIIRVLTEELGAEITHAGYSIVDDAVFHTLH 164

Example 32: Identification of a Medicagotruncatula bHLH

The issue to be discussed was whether the query sequence (Medicago truncatula SEQID NO: 227) was a bHLH polypeptide according to the definition applied here. SEQ ID 227 was compared to the Arabidopsis proteome database of MIPS institute.

Comparison was performed using the BLASTP algorithm.

2.0MP-WashU, which performs similarity searches of a query protein sequence with a given database protein sequence. (Reference: Gish, W. (1996-2002)). The parameters used in the comparison were value 10 Removal Score (S2): 56

The database protein sequence sought matched the MIPS Institute Arabidopsis proteome comprising 26,735 sequences (Database: / home / data / blast / orgs_sets / arabi 26,735 sequences; 11,317,104 total letters). Results were ranked according to highest similarity if determined from the highest score and lowest e. The first Hit identified (underlined in alignment) corresponded to SEQ ID NO: 225 (At4g20970) indicating that the sequence is a bHLH polypeptide according to the definition applied herein.

Paired alignments between the Medicago query sequence and the first hit corresponding to the newly identified bHLH are shown below.

BLASTP 2.OMP-WashU [09-Sep-2002] [decunix4.0-ev56-I32LPF64 2002-09-09ΊΊ7: 45: 09]

Query = SEQ ID NO: 227 (140 letters)

Database: /home/data/blast/orgs_sets/arabi26.735 Sequences; 11,317,104 total letters.

Seeking____10____20____ 30.. . .40 ____ 50____60____7 0____ 80 ---- 90 ---- 100% made

Lowest sumHigh probability

Sequences Producing High Score Segment Pairs: Score P (N) N

At4g20970 hypothetical protein_248 2.8e-22 1

At5g51780 Putative bHLH Transcription Factor (bHLH036) 103 6.5e-07 1

Atlgl0585 unknown protein 100 1.4e-06 1

At4g25400 Putative bHLH Transcription Factor (bHLH118) 99 7.3e-06 1

> At4g20970 hypothetical protein

Length = 167Score = 248 (92.4 bits), Expected = 2.8e-22, P = 2.8e-22Identities = 52/123 (42%), Positive = 82/123 (66%)

Query: 7 EAISVPDQLKEATNYIKKLQINLEKMKEKKNFLLG --- IQRPN ------ VNLNRNQKMGL 57

E +++ PDQL EA NYIKKLQ + N + EK + E + K L + +++ N V + + + + Subject: 45 EPLTLPDQLDEAANYIKKLQVNVEKKRERKRNLVATTTLEKLNSVGSSSVSSSVDVSVPR 104QueryLVVHVLVHVLVHVLVHVLVHVLQVLVVLVHVLVLRQVLQVLVLVHVLQVLVHVLQVLRQVLQVLVLQVLVLQVLVQVLVQVNLQE EQ + E

K PKI + IQ + G + + L + T LE + F + F E RVL EE G +1 + A Y + + D + VFH ++ HS Subject: 105 KLPKIEIQETGSIFHIFLVTSLEHKFMFCEIIRVLTEELGAEITHAGYSIVDDAVFHTLH 164Query: 117 CQV 119C +

Subject: 165 CKV 167

Example 33: Identification of Sequences Related to Nucleic Acid Sequence Used in the Methods of the Invention

BHLH sequences, either nucleotide (full-length cDNA, ESTs or genomic) or protein (full or partial-length polypeptides), were used to identify other bHLH nucleotide or protein sequences among those maintained in the Entrez Nucleotides database at the National Center for BiotechnologyInformation ( NCBI) using database sequence search tools such as the Basic Local Alignment Tool (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997 ) Nucleic Acids Res. 25: 3389-3402). The program was used to find regions of local similarity between sequences by comparing nucleic acid or polypeptide sequences with sequence database and calculating the statistical significance of identities. For example, the opolipeptide encoded by SEQ ID NO: 213 was used as a query for nr sequence database strings (non-redundant: (Databases: All GenBank + EMBL + DDBJ + PDB strings) (but not EST, STS strings , GSS, environmental samples, or HTGS phasing sequences 0, 1, or 2) 3,819,973 sequences; 16,928,533,343 total letters) using the TBLASTN algorithm, with default settings and the filter to ignore low complexity sequences. The result of the analysis (shown below) was viewed by paired comparison, and ranked according to the deprobability score (E value), where the score reflects the likelihood that a particular alignment will occur at random (the lower the E value, the more significant the hit). comparisons were also counted by percent identity. percent identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid sequences (or polypeptides) compared over a particular length.

Database hits identifying BHLH proteins yielded a score of at least 50 and a value equal to or less than e-05.

Punctuation

Value E

Sequences producing significant alignments: (Bits)

giI55765745Iref | NM_183508.2 I Oryza sativa (japonica cultivar-gro 440 le-121

giI42821746IdbjIAK109432.2 I Oryza sativa (japonica cultivar-g 440 le-121

gi I 55769744 IrefIXM_549862.11 Oryza sativa (japonica cultivar-gro 247 le-105

giI58530787IdbjIAP008207.11 Oryza sativa (japonica cultivar-g 263 2e-68

giI17385651IdbjIAP002845.3 I Oryza sativa (japonica cultivar-g 263 2e-68

giI32980356Idbj | AK070332.1 | Oryza sativa (japonica cultivar-g 251 9e-65

gi I 34894135Iref | NM_183504.1 I Oryza sativa (japonica cultivar-gro 137 3e-30

gi115293050 IgbIAY050959.11 Arabidopsis thaliana unknown prote 80.1 6e-16

giI79340923Iref | NM_100934.2 I Arabidopsis thaliana transcripti 87.8 2e-15

giI58531195IdbjIAP008214.1 I Oryza sativa (japonica cultivar-g 51.6 4e-13

giI42408246IdbjIAP004557.3 I Oryza sativa (japonica cultivar-g 51.6 4e-13

gi I 42408168IdbjIAP004376.3 I Oryza sativa (japonica cultivar-g 51.6 4e-13

giI22328837 | ref | NM_118215.2 | Arabidopsis thaliana DNAbinding 78.2 le-12

gi I 7268888IemblAL161554.2IATCHRIV54 Arabidopsis thaliana DNA chr 77.8 2e-12

gi I 5262774IembIAL080282.ilATT13K14 Arabidopsis thaliana DNA c 77.8 2e-12

giI50906596Iref | XM_464787.11 Oryza sativa (japonica cultivar-gro 77.0 3e-12

Example 34: Determination of global similarity and identity between bHLH transcription factors

Overall percentage similarity and identity between bHLH polypeptides was determined using the MatGAT computational application (BMC Bioinformatics. 2003 Jul 10; 4:29. MatGAT: an application that generates similarity / identity matrices using protein or DNA sequences.

Campanella JJ, Bitincka L, Smalley J.). MatGAT Computational Application Generates similarity / identity matrices for DNA or protein sequences without needing data alignment. The program performs a series of paired alignments, calculates similarity and identity, and then places the results in a distance matrix.

Parameters used in the comparison were:

Score Matrix: Blosum62

First Gap: 12

Extension Gap: 2

Results are shown in Figure 19a.

Example 35: Determination of Global Similarity and Identity between bHLH Domains in bHLH Polypeptides

BHLH domains were mapped using the SMART computational application (Schultz et al. (1998) Proc. Natl. Acad. Sci. USA 95,5857-5864; Letunic et al. (2006) Nucleic Aeids Res 34, D257-D260). SMART is available through EMBL Institute (European Molecular Biology Laboratory (EMBL)).

The bHLH domain sequences used are given below.Hv_BHLH

KESEKERRKRMKALCEKLASLIPREHCCSTTDTMTQLGSLDVGASYIKKLKERVDE0S_NP_9 0839 3_BHLH

KEME RRRRQ DMKGLCVKLASLIPKEHCSMSKMQAAS RTQLGS LDEAAAYIKKLKERVDEOS_NP_908397.1_BHLH

KDVEKNRRLHMKGLCLKLSSLIPAAAPRRHHHHYSTSSSSSPPSSTKEAVTQLDHLEQAAAYIKQLKGRIDE

Os_XP_4 64787.1_BHLH

KTTERIRREQMNKLYSHLDSLVRSAPPTVNSIPSHSNSNSKYHQRKLRILGGAAAATTRPDRLGVAAEYIRQTQERVDM

ATlG10585_bHLHnlrekdrrmrmkhlfsilsshvsptrklpvphlidqatsymiqlkenvnyAt 4 g2 0 9 7 0_bHLH

KTVEKNRRMQMKSLYSELISLLPHHSSTEPLTLPDQLDEAANYIKKLQVNVEKBhLH overall similarity and identity percentages of bHLH polypeptides were determined using the computational application and parameters described in Example 34.

Results are shown in Figure 19b.

Example 36: Cloning of Nucleic Acid Sequence in Methods of the Invention

The nucleic acid sequence used in the invention methods was amplified by PCR using a custom Oryza sativa seedling decDNA library (in pCMV Sport6.0; Invitrogen, Paisley, UK) as a template. PCR was performed using Hifi Taq DNA Polymerase under standard conditions using 200 ng of template in a 50 μΐ PCR mix. The primers used were prm06808 (SEQ ID NO: 231; sense, bold start codon, italic AttBl site: 5'-ggggacaagtttgtacaaaaaagcaggcttaaacaatgaagagccgagaacagc 3 ') andprm06809 (SEQ ID NO: 232; reverse 2 Att; 'ggggaccactttgtacaagaaagctgggtgcagagtgaaagagtggtgtg 3'), which includes AttB sites for Gateway recombination. The amplified PCR fragment was also purified using standard methods. The first step of the Gateway procedure, the BP5 reaction, was then performed during which PCR fragmentation recombines in vivo with plasmid pDONR201 to produce, according to Gateway terminology, an "input clone", pbHLH. Plasmid pDONR201 was purchased from Invitrogen, as part of Gateway® technology.

Example 37: Expression Vector Construction

The input clone p076 was subsequently used in an LR reaction with pGOS2, a destination vector used for Oryza sativa transformation. This vector contains as functional elements within the T-DNA limits: a selectable plant marker; a traceable marker expression drawer; and a Gateway drawer intended for in vivo LR combining with the nucleic acid sequence of interest already cloned into the input clone. A rice GOS2 promoter (SEQ LD NO: 233, alternatively, SEQ ID NO: 230 is equally useful) for constructive expression (internal reference PROO129) was located before this Gateway drawer.

After the LR recombination step, the resulting expression vector pGOS2 :: bHLH (Figure 20) was transformed into Agrobacterium strain LBA4044 and subsequently to Oryza sativa plants.

Example 38: Plant Transformation

Rice processing

Agrobacterium containing the expression vector was used to transform Oryza sativa plants. Dried ripe seeds of the Japanese Nipponbare rice cultivar were peeled. Sterilization was performed by incubating for one minute in 70% ethanol, followed by 30 minutes in 0.2% HgCl2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus induction medium). After incubation in the dark for four weeks, scutellus-derived embryogenic calli were excised and propagated in the same medium. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic callus pieces were subcultured in fresh medium 3 days prior to co-cultivation (to stimulate cell division activity).

Agrobacterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobaeterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD60) of about 1. The suspension was then transferred to a Petri dish and the calluses immersed in the suspension for 15 minutes. Callus were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 25 ° C. Co-cultured horses were grown in medium containing 2,4-D for 4 weeks in the dark at 25 ° C. 28 ° C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Approximately 30 independent rice TO transformants were generated for a construction. The primary transformants were transferred from a tissue culture chamber to a greenhouse. After a quantitative PCR analysis to verify the number of copies of the T-DNA insert, only transgenic single copy plants that exhibit selection agent tolerance were maintained for TL seed harvest. Seeds were then collected three to five months after transplantation. The method produced single locus transformants at a rate of up to 50% (Aldemita and Hodges 1996, Chan et al. 1993, Hiei etal. 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural Alpa 88 (University of Minnesota) or Al 88 hybrids as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are collected from the corn plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm. Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and plants GMOs are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium containing the selection agent (eg imidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2- 3 weeks, or until aerial parts develop. The green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat processing

Wheat processing is performed with the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybean is processed according to a modification of the method described in Texas A&M Patent 5,164,310.

Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. Epicotyl and remnant cotyledon are further grown to develop axillary nodes.

These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / Yola processing

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188).

The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used.

Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobaeterium (containing the expression vector) bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobacterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

Alfalfa Transformation

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K2SO4, and 100 μτη acetoseringone. Explantants are washed in half-strength Murashige-Skoog medium (Murashige and Skogog, 1962) and placed in the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobacterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 39: Evaluation Procedure39.1 Evaluation Configuration

Approximately 30 independent rice TO transformants were generated. Primary transformants were transferred from a tissue culture chamber to a greenhouse for TI seedlings. Seven events, of which progeny T1 secreted 3: 1 for presence / absence of transgene, were retained. For each of these events, approximately 10 Tl seedlings containing the transgene (hetero and homozygotes) and approximately 10 Tl seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression. Transgenic plants and corresponding nullizigotes were grown side by side at random positions. Greenhouse conditions were short days (12 hours of light), 28 ° C in the light and 22 ° C in the dark, and a relative humidity of 70%.

Four T1 events were additionally evaluated in generation T2 following the same evaluation procedure as for T1 generation but with more individuals per event. From the sowing stage to the maturity stage the plants were passed several times through a digital image cabinet. At each moment digital images (2048x1536 pixels, 16 million colors) were taken from each plant from at least 6 different angles.

39.2 Statistical analysis: t-test and F-test

A two-way ANOVA (analysis of variance) was used as a statistical model for the general evaluation of plant phenotypic characteristics. An F test was performed on all measured parameters of all plants of all events transformed with the gene of the present invention. The F test was performed to check for a gene effect on all transformation events and to check for a general gene effect, also known as the overall gene effect. The threshold for significance for a true overall gene effect is set at a 5% probability level for the F test. A significant F test value points to a gene effect, meaning that it is not just the mere presence or apposition of the gene that is causing the differences. in phenotype.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" refer to plants treated in the same manner as the transgenic plant, but in which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test has been adjusted to a 10% probability level. Results for some events may be above or below this threshold. This is based on the hypothesis that a gene can only have an effect on certain positions in the genome, and that the occurrence of this deposition dependent effect is not uncommon. This type of gene effect is also called nestlugar as a "gene strain effect". The value of ρ was obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the null hypothesis (ie, that there is no transgene effect). ) is correct.

Example 40: Evaluation Results

Early vigor was determined by counting the total number of plant shootings broken down from the bottom. This value was averaged for photos taken at the same time from different angles and converted to a physical surface value expressed in mm squared by calibration. The results described below are for plants three weeks after germination.

Early vigor (if determined by aerial area) was seen in seven IT generation events, with an overall increase in aerial area for transgenic seedlings of 22% compared to control plants. Four of these Tl events were further evaluated in the T2 generation, and all four of these events generated an increase in aerial area for transgenic seedlings compared to control plants, with an overall increase in aerial area for transgenic seedlings of 13% compared with control plants. The results were also shown to be statistically significant with the ρ value of the F test being 0.0007 (T2 generation) indicating that the observed effect is probably transgene-dependent rather than the gene position or a strain effect.

Example Section F: SPL15

Example 41: Identification of Sequences Related to Nucleic Acid Sequence Used in the Methods of the Invention

Sequences (full length cDNAs, ESTs, or genomic) related to the nucleic acid sequence used in the methods of the present invention have been identified among those maintained in the Entrez Nucleotides database at the National Center for Biotechnology Information using database sequence search tools, such as the tool. Local Alignment Basics (BLAST) (Altschul et al. (1990) J. Mol. Biol. 215: 403-410; and Altschul et al. (1997) Nucleic Acids Res.25: 3389-3402). The program is used to find regions of local similarity between sequences by comparing nucleic acid or polypeptide sequences with sequence database and calculating the statistical significance of identities. The nucleic acid encoded polypeptide of the present invention was used for the TBLASTN algorithm, with default settings and the filter to bypass low-complexity sequence counterweight. The result of the analysis was viewed by paired comparison, drawn according to the probability score (E value), where the score reflects the likelihood that a particular alignment will occur in the case (the lower the E value, the more significant the hit). In addition to E values, comparisons were also scored by percent identity.

Percent Identity refers to the number of identical nucleotides (or amino acids) between the two nucleic acid (or polypeptide) sequences compared over a particular length. In some instances, the default parameters may be adjusted to modify the stringent stringency.

Table N below provides a list of nucleic acid sequences related to the nucleic acid sequence used in the methods of the present invention.

Table N: Nucleic acid sequence related nucleic acid sequences (SEQ ID NO: 234) used in the methods of the present invention, and the corresponding deduced polypeptides

<table> table see original document page 303 </column> </row> <table> <table> table see original document page 304 </column> </row> <table> <table> table see original document page 305 < / column> </row> <table>

Example 42: Determination of global similarity and identity between SPL1 5 transcription factors and their SPL DBD.

Overall percentages of similarity and identity between SPL15 transcription factors were determined using one of the methods available in the art, the Matrix Global Matrix Alignment Tool (BMC Bioinformatics. 2003 4:29. MatGAT: an application that generates similarity / identity matrices using protein orDNA sequences Campanella JJ, Bitincka L, Smalley J, computer application hosted by Ledion Bitincka). Computational application MatGAT generates similarity / identity matrices for DNA or protein sequences without pre-alignment of data. The program performs a series of paired alignments using the Myers and Miller alignment algorithm (with a gap opening penalty of 12, a gap extension penalty of 2), calculating similarity and identity using eg Blosum 62 (for polypeptides), and then put the results in a distance matrix. Sequence similarity is shown on the bottom half of the dividing line and sequence identity is shown on the top half of the diagonal dividing line. The sequence of SEQ ID NO: 235 is indicated as number 5 in the matrix.

Parameters used in the comparison were:

Score Matrix: Blosum62

First Gap: 12

Extension Gap: 2

Results of computer application analysis are shown in Table O for similarity and overall identity over full length of the transcription factor polypeptides SPL1. Percent identity is given above diagonal and percentage similarity is given below diagonal. Percent identity between SPL15 transcription factor paralogues and orthologs ranges from 30 to 70%, reflecting the relatively low conservation of sequence identity between them outside the SPL DBD.

Table O: MatGAT results for similarity and overall identity over the full length of SPL15 transcription factor polypeptides.

<table> table see original document page 306 </column> </row> <table>

Results of computer application analysis are shown in Table P for SPLDBD similarity and overall identity of the SPL15 transcription factor polypeptides. Percentage identity is given above the diagonal and percentage similarity is given below the diagonal.

Percentage identity between SPL DBD of SPL15 transcription factor paralogues and orthologs ranges from 70% to 100%. Table Mat: MatGAT results for similarity and overall identity over SPL DBD of transcription factor polypeptidesSPL15.

<table> table see original document page 307 </column> </row> <table>

Example 43: Cloning of Nucleic Acid Sequence in Methods of the Invention

DNA Manipulation: Unless otherwise determined, recombinant DNA techniques are performed according to standard protocols described in (Sambrook (2001) Molecular Cloning: Alaboratory Manual, 3rd Edition Cold Spring Harbor Laboratory Press, CSH, New York) or in Volumes 1 and 2 by Ausubel et al. (1994), Current Protocolsin Molecular Biology, Current Protocols. Standard materials and methods for plant molecular work are described in Plant Molecular BiologyLabhase (1993) by R.D.D. Croy, published by BIOS Scientific PublicationsLtd (UK) and Blackwell Scientific Publications (UK).

The nucleic acid sequence used in the invention methods was PCR amplified using a custom Arabidopsis thaliana mixed tissue decDNA library (in pCMV Sport 6.0; Invitrogen, Paisley, UK) as a template. PCR was performed using Hi Taq DNA polymerase under standard conditions, using 200 ng of model in a 50 μΐ PCR mix. The primers used were prm07277 (SEQ ID NO: 280; sense, bold start codon, lowercase AttBl site: 5'-ggggacaagtttgtacaaaaaagcaggcttaaacaATGGAGTTGTTAATGTGTTCG3 ') and prm07278, minus reversal: Attention ID2: 281; 'ggggaccactttgtacaagaaagctgggtTGATGAAGATCTTAAAAGGTGA 3'), which includes the AttB sites for Gateway recombination. The amplified PCR fragment was also purified using standard methods. The first stepped Gateway procedure, the BP5 reaction was then performed, during which PCR fragmentation recombines in vivo with plasmid pDONR201 to produce, according to Gateway terminology, an "input clone", pSPL15. Plasmid pDONR201 was purchased from Invitrogen, as part of Gateway® technology.

Example 44: Expression Vector Construction

The input clone ρ 13075 was subsequently used in an LR reaction with p06659, a destination vector used for Oryza sativa transformation. This vector contains as functional elements within the limits of T-DNA: a selectable plant marker; a drawer traceable marker expression; and a Gateway drawer intended for in vivo LR combining with the nucleic acid sequence of interest already cloned into the input clone. A rice HMGB promoter (SEQ ID NO: 294) for constitutive expression was located before this Gateway drawer. Alternatively, the HMGB promoter represented by SEQ ID NO: 46 is equally useful. A similar construct was made with the SPL15 decoding sequence under the control of the constitutive promoter GOS2 (SEQ IDNO: 295).

Following the LR recombination step, the resulting expression vector pHMGB :: SPL15 (Figure 26), or pGOS2 :: SPL15, was transformed into Agrobacterium strain LBA4044 and subsequently to Oryza sativa plants.

Example 45: Plant Transformation

Rice processing

Agrobacterium containing the expression vector was used to transform Oryza sativa plants. Dried ripe seeds of the Japanese Nipponbare rice cultivar were peeled. Sterilization was performed by incubating for one minute in 70% ethanol, followed by 30 minutes in 0.2% HgCl2, followed by a 15-minute wash 6 times with sterile distilled water. The sterile seeds were then germinated in a medium containing 2,4-D (callus induction medium). After incubation in the dark for four weeks, scutellus-derived embryogenic calli were excised and propagated in the same medium. After two weeks, the corns were multiplied or propagated by subculture in the same medium for another 2 weeks. Embryogenic callus pieces were subcultured in fresh medium 3 days prior to co-cultivation (to stimulate cell division activity).

Agrobaeterium strain LBA4404 containing the expression vector was used for co-cultivation. Agrobacterium was inoculated into AB medium with the appropriate antibiotics and grown for 3 days at 28 ° C. The bacteria were then collected and suspended in liquid co-cultivation medium for a density (OD60) of about 1. The suspension was then transferred to a Petri dish and the calluses immersed in the suspension for 15 minutes. Callused, they were then dried on a filter paper and transferred to a solidified co-culture medium and incubated for 3 days in the dark at 25 ° C.

Co-cultured calli were cultured in medium containing 2,4-D for 4 weeks in the dark at 28 ° C in the presence of a selection agent. During this period, rapidly growing hard-shelled islands developed. After transfer of this material to a medium of regeneration and incubation in light, the embryogenic potential was released and aerial parts developed in the next four to five weeks. Aerial parts were excised from the callose incubated for 2 to 3 weeks in auxin-containing medium from which they were transferred to the soil. Hardened aerial parts were cultured in high humidity and short days in a greenhouse.

Approximately 35 independent rice TO transformants were generated for a construction. The primary transformants were transferred from a tissue culture chamber to a greenhouse. After a quantitative PCR analysis to verify the number of copies of the T-DNA insert, only transgenic single copy plants exhibiting tolerance to the selection agent were maintained for TI seed harvest. Seeds were then collected three to five months after transplantation. The method produced single locus transformants at a rate of up to 50% (Aldemita and Hodges 1996, Chan et al. 1993, Hie et al 1994).

Corn Processing

Corn transformation (Zea mays) is performed with a modification of the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. Transformation is genotype dependent in maize and only specific genotypes are receptive to transformation and regeneration. Natural Alpa (University of Minnesota) or Al88 hybrids as a central are good sources of donor material for transformation, but other genotypes can also be used successfully. Ears are harvested from the maize plant approximately 11 days after pollination (DAP) when the length of the immature embryo is about 1 to 1.2 mm.

Immature embryos are co-cultured with Agrobacterium tumefacienscontaining the expression vector, and transgenic plants are recovered through organogenesis. Excised embryos are cultured in decal induction medium, then maize regeneration medium containing the selection agent (eg imidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2- 3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to maize rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Wheat processing

Wheat processing is performed with the method described by Ishida et al. (1996) Nature Biotech 14 (6): 745-50. The Bobwhite cultivar (available from CIMMYT, Mexico) is commonly used in transformation. Immature embryos are co-cultured with Agrobacteriumtumefaciens containing the expression vector, and transgenic plants are recovered through organogenesis. After incubation with Agobacterium, the embryos are cultured in vitro in decalo induction medium, then regeneration medium containing the selection agent (egimidazolinone but various selection markers may be used). Petri dishes are incubated in light at 25 ° C for 2-3 weeks, or until aerial parts develop. Green shoots are transferred from each embryo to rooting medium and incubated at 25 ° C for 2-3 weeks until roots develop. Rooted shoots are transplanted parasol in the greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and contain a single copy of the T-DNA insert.

Soybean Processing

Soybeans are processed according to a modification of the method described in Texas A&M Patent 5,164,310. Several commercial soybean varieties are receptive to processing by this method. The Jack cultivar (available from the Illinois Seed foundation) is commonly used for processing. Soybean seeds are sterilized for in vitro sowing. The hypocotyl, radicle and cotyledon are excised from young seven-day-old seedlings. Epicotyl and remnant cotyledon are further grown to develop axillary nodes.

These axillary nodes are excised and incubated with Agrobacterium tumefacienscontaining the expression vector. After co-cultivation treatment, the explants are washed and transferred to selection medium. Regenerated aerial parts are excised and placed in a prolonged aerial departe. Aerial parts no larger than 1 cm are placed in rooting medium until roots develop. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants which exhibit tolerance to the selection agent and which contain a single copy of the T-DNA insert.

Rapeseed / canola transformation

Cotiledonary and hypocotyl petioles of 5-6 day old seedlings are used as explants for tissue culture and transformed according to Babic et al. (1998, Plant Cell Rep 17: 183-188).

The commercial cultivar Westar (Agriculture Canada) is the standard variety used for processing, but other varieties can also be used.

Canola seeds are surface-sterilized for in vitro sowing. Cotyledon petiole seedlings with coupled cotyledon are excised from seedlings in vitro, and inoculated with Agrobacterium (containing the expression vector) bathing the cutting end of the petiole explant on bacterial suspension. The explants are then cultured for 2 days in MSBAP-3 medium containing 3 mg / l BAP, 3% sucrose, 0.7% Phytagar at 23 ° C, 16 hr light. After two days of co-cultivation with Agrobaeterium, petiole explants are transferred to MSBAP-3 medium containing 3 mg / l deBAP, cefotaxime, carbenicillin, or timentin (300 mg / l) for 7 days, and then cultured in MSBAP-medium. 3 with cefotaxime, carbenicillin, or timentin selection agent until shoot regeneration. When the aerial parts are 5-10 mm long, they are cut and transferred to the aerial part elongation layer (MSBAP-0.5, containing 0.5 mg / l deBAP). Aerial parts about 2 cm long are transferred to rooting medium (MSO) for root induction. Rooted shoots are transplanted to soil in the greenhouse. T1 seeds are produced from plants that exhibit selection agent tolerance and contain a single copy of the T-DNA insert.

Alfalfa Transformation

A regenerating alfalfa clone (Medicago sativa) is transformed using the method of (McKersie et al., 1999 Plant Physiol 119: 839-847). Alfalfa regeneration and transformation is genotype dependent and therefore a regenerating plant is required. Methods to obtain regenerating plants have been described. For example, these may be selected from the cultivar Rangelander (Agriculture Canada) or any other commercial alfalfa variety as described by Brown DCW and A Atanassov (1985. Plant Cell Tissue Organ Culture 4: 111-112). Alternatively, RA3 (University of Wisconsin) avariety has been selected for use in tissue culture (Walker et al., 1978 Am J Bot 65: 654-659). Petioles explants co-cultured with a night culture of Agrobacterium tumefaciensC58C1 pMP90 (McKersie et al., 1999 Plant Physiol 119: 839-847) or LBA4404 containing the expression vector. Explants are co-cultured for 3 d in the dark in SH induction medium containing 288 mg / l Pro, 53 mg / l thioproline, 4.35 g / l K2SO4, and 100 μηι of acetoseringone. Explantants are washed in half-strength Murashige-Skoog medium (Murashige and Skogog, 1962) and placed in the same SH induction medium without acetoseringonamas with a suitable selection agent and suitable antibiotic to inhibit Agrobacterium growth. After several weeks, somatic embryos are transferred to BOY2 development medium containing no growth regulator, no antibiotic, and 50 g / L desucrose. Somatic embryos are subsequently germinated in half-force Murashige-Skoog. Rooted seedlings were transplanted into pots and grown in a greenhouse. T1 seeds are produced from plants that exhibit tolerance to the selection agent and that contain a single copy of the T-DNA insert.

Example 46: Evaluation Procedure

46.1 Evaluation Configuration

Approximately 35 independent rice TO transformants were generated. Primary transformants were transferred from a tissue culture chamber to a greenhouse for TI seedlings. Seven events, of which progeny T1 secreted 3: 1 for presence / absence of transgene, were retained. For each of these events, approximately 10 Tl seedlings containing the transgene (hetero and homozygotes) and approximately 10 Tl seedlings lacking the transgene (nullizigotes) were selected by monitoring visual marker expression.

Transgenic plants and corresponding nullizigotes were grown side by side in random positions. Greenhouse conditions were short days (12 hours of light), 28 ° C in the light and 22 ° C in the dark, and a relative humidity of 70%.

Five T1 events were additionally evaluated in generation T2 following the same evaluation procedure as for generation T1 but with more individuals per event. From the sowing stage to the maturity stage the plants were passed several times through a digital image cabinet. At each moment digital images (2048x1536 pixels, 16 million colors) were taken from each plant from at least 6 different angles.

Early vigor is the aerial area of the plant (seedling) three weeks after germination. Increase in root biomass is expressed as an increase in total root biomass (measured as maximum root biomass observed during the life of a plant); or as an increase in root / shoot index (measured as the ratio between massaricular and shoot mass in the active root and shoot growth period).

46.2 Statistical analysis: F test

A two-way ANOVA (analysis of variance) was used as a statistical model for the general evaluation of plant phenotypic characteristics. An F test was performed on all measured parameters of all plants of all events transformed with the gene of the present invention. The F test was performed to check for a gene effect on all transformation events and to check for a general gene effect, also known as the overall gene effect. The threshold for significance for a true overall gene effect is set at a 5% probability level for the F test. A significant F test value points to a gene effect, meaning that it is not just the mere presence or apposition of the gene that is causing the differences. in phenotype.

To verify for an effect of genes within an event, i.e., for a specific strain effect, a t-test was performed within each event using transgenic and corresponding null plant data sets. "Null plants" or "null segregants" or "nullizigotes" refer to plants treated in the same manner as the transgenic plant, but in which the transgene has been secreted. Null plants can also be described as homozygous negative transformed plants. The significance threshold for the t-test has been adjusted to a 10% probability level. Results for some events may be above or below this threshold. This is based on the hypothesis that a gene can only have an effect on certain positions in the genome, and that the occurrence of this deposition dependent effect is not uncommon. This type of gene effect is also called nestlugar as a "gene strain effect". The value of ρ was obtained by comparing the value of t with the distribution of t or alternatively by comparing the value of F with the distribution of F. The value of ρ then generates the probability that the null hypothesis (ie, that there is no transgene effect). ) is correct.

Example 4 7: Evaluation Results

The plant shoot (or leaf biomass) was determined by counting the total number of pixels in the digital images of plant partisans discriminated from the background. This value was averaged for at-the-same-time photo shoots and converted to a physical surface value expressed in mm squared by calibration.

Experiments show that the plant shoot measured in this way correlates with the plant shoot biomass. The aerial area is the moment in which the plant reached its maximum leaf biomass.

The mature primary panicles were collected, counted, bagged, barcoded and then dried for three days in an oven at 37 ° C. The panicles were then traced and all seeds were collected and counted. The full shells were separated from the empty ones using an air-blowing device. The empty shells were discarded and the remaining fraction was counted again. The full shells were weighed on an analytical balance. The number of full grains was determined by counting the number of full husks that remained after the desparing step. Total seed yield was measured by weighing all bark collected from a plant. Total number of seeds per plant was measured by counting the number of bark collected from a plant. Millet seed weight (TKW) is extrapolated from the number of full grains counted and its total weight. The harvest index (HI) in the present invention is defined as the ratio of total seed production to shoot (mm2) multiplied by a factor 106. The total number of flowers per panicle as defined in the present invention is the ratio of the total number of seeds to the number of mature primary panicles. The grain fill rate as defined in the present invention is the ratio (expressed as a%) of the number of full grains to the total number of seeds (or rapiers).

As shown in Tables Q to W, aerial biomass, number of flowers per panicle, seed yield, total number of seeds, number of full grains, one thousand seed weight (TKW), and harvest index are increased in transgenic plants. with increased expression of a nucleic acid encoding a SPL15 transcription factor polypeptide compared to appropriate control plants.

Results of generations T1 and T2 are shown.

Table Q shows the number of transgenic events with an increase in aerial biomass, the percentage of this increase, as well as the statistical significance of this increase according to the F test.

Table Q: Number of transgenic events with an increase in aerial biomass, the percentage of increase, and P value of the F1 generation test T1 and T2 of transgenic rice with increased expression of a nucleic acid encoding a transcription factor polypeptide SPL15.

<table> table see original document page 317 </column> </row> <table>

Table R shows the number of transgenic events with an increase in the total number of flowers per panicle, the percentage of the increase, as well as the statistical relevance of this increase according to the test F. Table R: Number of transgenic events with an increase in flowers per panicle, the percentage increase, and P value of the F1 test on Tgeneration and T2 generation of transgenic rice with increased expression of a nucleic acid encoding a transcription factor polypeptide SPL15.

<table> table see original document page 318 </column> </row> <table>

Table S shows the number of transgenic events with an increase in total seed yield (total seed weight), the percentage of this increase, as well as the statistical relevance of this increase according to the F test.

Table S: Number of transgenic events with an increase in seed yield, the percentage of increase, and P value of the Fem generation T1 and T2 test of transgenic rice with increased expression of a nucleic acid encoding an SPL15 transcription factor polypeptide.

<table> table see original document page 318 </column> </row> <table>

Table T shows the number of transgenic events with an increase in the total number of seeds, the percentage of this increase, as well as the statistical relevance of this increase according to the F test.

Table T: Number of transgenic events with an increase in total number of seeds, the percentage of increase, and P value of this test in generation T1 and T2 of transgenic rice with increased expression of a nucleic acid encoding a SPL15 transcription factor polypeptide.

<table> table see original document page 318 </column> </row> <table>

Table U shows the number of transgenic events with an increase in the number of full grains, the percentage of this increase, as well as the statistical relevance of this increase according to the F test.

Table U: Number of transgenic events with an increase in number of full grains, the percentage of increase, and P value of test F in generation T1 and T2 of transgenic rice with increased expression of a nucleic acid encoding a transcription factor polypeptide SPL15.

Number of Grains Filled Number of Events Showing Increase% Difference Test P Value FGeneration Tl 6 of 7 14 0.0031Generation T2 4 of 5 19 0.0003

Table V shows the number of transgenic events with an increase in the Harvest index, the percentage of this increase, as well as the statistical significance of this increase according to the F test.

Table V Number of transgenic events with an increase in Harvest index, the percentage of increase, and P value of the F1 Tgeneration test T and T2 of transgenic rice with increased expression of a nucleic acid encoding a transcription factor polypeptide SPL1.5. iite

Table W shows the number of transgenic events with an increase in the weight of one thousand seeds (TKW), the percentage of this increase, as well as the statistical relevance of this increase according to the F test.

Table W: Number of transgenic events with a one thousand seed weight increase (TKW), the percentage of the increase, and P value of this test T generation T1 and T2 of increased expression transgenic rice of a nucleic acid encoding a transcription factor polypeptideSPL15 .

Weight of one thousand seeds Number of events showing an increase% Difference Test P value FG Generation Tl 4 out of 7 2 0.0041Generation T2 4 out of 5 1 0.0259Example 48: Results of phenotypic evaluation of plantastransgenics expressing SEQID NO: 234 under control of a conductive promoter

The results of the evaluation of transgenic rice plants expressing the nucleic acid sequence useful for carrying out the inventive methods under the control of the constitutive promoter GOS2 are presented in Table X. The percentage difference between the transgenic and the corresponding nullizigotoscopes is also shown.

Transgenic plants expressing the nucleic acid sequence useful for carrying out the methods of the invention have increased early vigor and increased TKW compared to control plants (in this case, the nullizygotes).

Table X: Results of the evaluation of T1 generation transgenic rice plants expressing the nucleic acid sequence useful for carrying out the methods of the invention under the control of a strong constitutive GOS2 promoter.

<table> table see original document page 320 </column> </row> <table> SEO LISTING

<110> CropDesign N.V.

<120> Plants having improved yield characteristics and a method for making them

<130> PF58328 <160> 301

<170> PatentIn version 3.3 <210> 1 <211> 630 <212> DNA

<213> Arabidopsis thaliana <400> 1

atggagaatg ggaaaagaga cagacaagac atggaagtga ataccacacc gaggaagcct 60

cgtgtactac tcgctgcaag tggaagcgtc gctgctatca aattcggcaa tctctgccat 120tgcttcaccg aatgggcaga agtcagagcc gtcgttacga aatcatctct acatttcctc 180gataaactct ctctcccaca agaagtgact ctgtatactg atgaagatga atggtctagc 240tggaacaaga tcggtgatcc tgtccttcac atcgagctta gacgttgggc tgatgtttta 300gtcattgctc ctttgtctgc taacacctta ggcaagattg ctggtgggct ttgtgataat 360cttctgactt gcattatacg agcttgggac tataccaaac cactgtttgt tgctccagct 420atgaatactt tgatgtggaa caatcctttc actgaaaggc atcttttgtc tcttgatgaa 480ctgggaatca cacttattcc tcctatcaag aagagacttg cctgtggaga ctacggtaat 540ggagctatgg ctgagccctc tcttatctat tccactgtca gactcttctg ggagtctcag 600gctcatcagc aaaccggtgg aactagttaa 630

<210> 2

<211> 209

<212> PRT

<213> Arabidopsis thaliana

<400> 2

Met Glu Asn Gly Lys Arg Asp Arg Gln Asp Met Glu Val Asn Thr Thr1 5 10 15

Pro Arg Lys Pro Arg Val Leu Leu Wing Wing Be Gly Ser Val Wing Wing

20 25 30

Ile Lys Phe Gly Asn Leu Cys His Cys Phe Thr Glu Trp Wing Glu Val

35 40 45

Arg Wing Val Val Thr Lys Be Ser Read His Phe Read Asp Lys Read Ser

50 55 60

Leu Pro Gln Glu Val Thr Read Leu Tyr Thr Asp Glu Asp Glu Trp Ser65 70 75 80

Trp Asn Lys Ile Gly Asp Pro Val Leu His Ile Glu Leu Arg Arg Trp

85 90 95

Asp Wing Val Leu Val Ile Pro Wing Read Ser Wing Asn Thr Leu Gly Lys

100 105 110

Ile Wing Gly Gly Leu Cys Asp Asn Leu Leu Thr Cys Ile Ile Arg Wing

115 120 125

Trp Asp Tyr Thr Lys Pro Leu Phe Val Pro Wing Met Asn Thr Leu

130 135 140

Met Trp Asn Asn Pro Phe Thr Glu Arg His Read Leu Read Le Asp Glu145 150 155 160

Leu Gly Ile Thr Leu Ile Pro Pro Ile Lys Lys Arg Leu Wing Cys Gly

165 170 175

Asp Tyr Gly Asn Gly Wing Met Wing Glu Pro To Be Read Ile Tyr To Be Thr

180 185 190

Val Arg Read Phe Trp Glu Be Gln Wing His Gln Gln Thr Gly Gly Thr195 200 205

To be

<210> 3 <211> 37 <212> DNA

<213> Artificial sequence <220> <223> primer: prm00957 <400> 3

aaaaagcagg ctcacaatgg agaatgggaa aagagac <210> 4 <211> 33 <212> DNA

<213> Artificial sequence <220>

<223> initiator: prm00958 <400> 4

agaaagctgg gttggtttta actagttcca ccg <210> 5 <211> 1243 <212> DNA

<213> Oryza sativa <400> 5

aaaaccaccg agggacctga tctgcaccgg ttttgatagtgttttccgat cgagggacga aaatcggatt cggtgtaaagttattccgga gcatgattgg gaagggagga cataaggcccgtccagatct ccagatcact cagcaggatc ggccgcgttcttcggcttcc cgcaaggcgg cggccggtgg ccgtgccgccgcacgccgcc gccgccgacc cggctctgcg tttgcaccgcatagatagct actactctct ccgtttcaca atgtaaatcaatattgatgt taatgaatat agacatatat atctatttagatgtaggaaa tgctagaatg acttacattg tgaattgtgatggatggatg caggatcatg aaagaattaa tgcaagatcgttactaattg cgctgcatat atgcatgaca gcctgcatgcccattaggaa gtaaccttgt cattacttat accagtactatcatgagcaa atctacaaaa ctggaaagca ataaggaatacattaatcac caaatatttc gccttctcca gcagaatatatgtacacact gacagtgtac gcataaacgc agcagccagcgcacactggc cttccatctc aggctagctt tctcagccacgcgcgcgcac aggcacaaat tacgtacaaa acgcatgaccgaatcgctcc cgcgcgcggc ggcggcgcgc acgtacgaatccacgacacg atcgcgcgcg acgccggcga caccggccatccgactataa attackgtagcgc <a>

<213> Artificial sequence <220>

<223> Subject retained 1 <220>

<221> VARIANT <222> (1) .. (1) <223> / replace = "Arg" <220>

<221> VARIANT

<222> (4). . (4)

<223> / replace = "lie"

<220>

<221> VARIANT

<222> (9) .. (9)

<223> / replace = "Thr"

<220>

<221> VARIANT

<222> (14) .. (14)

<223> / replace = "Be"

<220>

<221> VARIANT <222> (15) .. (15)

<223> / replace = "Met" / replace = "Val '

37

33

tgagggaccc gttgtgtctg 60ttaagggacc tcagatgaac 120atgtcgcatg tgtttggacg 180gcgtagcacc cgcggtttga 240gtagcttccg ccggaagcga 300cttgcacgcg atacatcggg 360ttctactatt ttccacattc 420attcattaac atcaatatga 480aatggacgaa gtacctacga 540tatctgccgc atgcaaaatc 600gggcgtgtaa gcgtgttcat 660catactatat agtattgatt 720cgggactgga aaagactcaa 780tatctctcca tcttgatcac 840ttaactgtcg tctcaccgtc 900ccatcgtaca tgtcaactcg 960aaatcaaaac caccggagaa 1020gcacgcacgc acgcccaacc 1080ccacccgcgc cctcacctcg 1140catctcacca ccaaaaaaaa 1200caa 1243 <220> <221> <222 > <223> <220> <221> <222>

VARIANT (18) .. (18) / replace

"Glu" / replace = "Wing" / replace = "Be"

VARIANT (21) .. (21) <223> / replace = "Going" <220>

VARIANT (22). . (22) / replace

<221><222><223><220><221><222><223><220><221><223><220><221><222><223> <220> <221 > <222>

VARIANT (23) .. (23) / replace

VARIANT

(25) .. (25) / replace

VARIANT

(26). . (26) / replace

"Arg" / replace = "Gly"

= "Ser" / replace = "lie"

= "Be"

VARIANT (29). . (29) <223> / replace <220>

VARIANT (31) .. (31) / replace

<221><222><223><220><221><222><223> <400>

VARIANT (34) .. (34) / replace6

= "Asp"

"Asp"

= "Lys"

= "Wing"

Lys Pro Arg Val Leu Leu Wing Wing Ser Gly Ser Val Wing Wing Ile Lys15 10 15

Phe Gly Asn Leu Cys His Cys Phe Thr Glu Trp Wing Glu Val Arg Wing20 25 30

Val Val

<210> 7 <211> 98 <212> PRT

<213> Artificial sequence <220>

<223> Subject retained 2 <220>

<221> VARIANT <222> (8) .. (8) <223> / replace <220>

<221> VARIANT <222> (12) .. (12) <223> / replace = "lie" <220>

<221> VARIANT

<222> (13) .. (13)

<223> / replace = "Met"

<220>

<221> VARIANT

"Lys" / replace = "Gln" <222> <223> <220> <221> <222> <220> <221> <222> <223> <220> <221> <222> <223 > <220> <221> <222> <223> <220> <221> <222> <223> <220> <222> <223>

(14) .. (14) / replace

VARIANT (24). . (24) / replace

VARIANT (30) .. (30) / replace

VARIANT

(38). . (38)

/replace

VARIANT

(39) .. (39) / replace

VARIANT (44). . (44) / replace

= "lie"

= "Wing"

= "Met"

= "Go"

"Go"

= "Phe"

VARIANT (45) .. (45) / replace / replace = "Lys"

<220><221><222><223><220><221><222><222><221><222><223><220><221><222> <223> <220 > <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <220>

= "Be" / replace = "Asn" / replace

"Asp"

VARIANT (48). . (48) / replace

VARIANT (50) .. (50) / replace

VARIANT (57) .. (57) / replace

VARIANT (60) .. (60) / replace

VARIANT

(65) .. (65)

/replace

VARIANT

(66) .. (66) / replace

VARIANT (68). . (68) / replace

= "Phe" / replace = "Met" / replace = "lie"

= "Wing"

= "Phe"

= "Be"

"Be" / replace = "Gln" / replace = "Wing"

"Lys"

"Phe" / replace = "He"

<221> UNSAFE <222> (69) .. (70)

<223> Xaa can be any naturally occurring amino acid <220>

<221> VARIANT <222> (71) .. (71)

<223> / replace = "lie" / replace = "Met" <220>

<221> VARIANT <222> (72) .. (72)

<223> / replace = "Asn" / replace = "Be" <220>

<221> VARIANT <222> (73) .. (73)

<223> / replace = "Read" / replace = "Gln" <220>

<221> VARIANT

<222> (74) .. (74)

<223> / replace = "Met"

<220>

<221> VARIANT

<222> (76) .. (76)

<223> / replace = "Go"

<220>

<221> VARIANT <222> (77) .. (77)

<223> / replace = "Be" / replace = "Wing" / replace = "lie" <220>

<221> VARIANT

<222> (79) .. (79)

<223> / replace = "Go"

<220>

<221> VARIANT <222> (82) .. (82)

<223> / replace = "Go" / replace = "Thr" <220>

<221> VARIANT <222> (83) .. (83)

<223> / replace = "Thr" / replace = "Ser" <220>

<221> VARIANT <222> (85) .. (85)

<223> / replace = "Thr" / replace = "Lys" <220>

<221> VARIANT <222> (91) .. (91) <223> / replace = "His" <220>

<221> VARIANT <222> (93) .. (93) <223> / replace = "Thr" <220>

<221> VARIANT <222> (97) .. (97) <223> / replace = "Be" <400> 7

Val Leu His Ile Glu Leu Arg1 5

Pro Leu Ser Asa Asn Thr Leu20

Asn Leu Leu Thr Cys Ile Ile35

Phe Val Wing Pro Wing Met Asn

50 55

Glu Arg His Leu Xaa Xaa Leu65 70

Pro Ile Lys Lys Arg Leu Ala85

Arg Trp Ala10

Gly Lys Ile25

Arg Wing Trp40

Thr Leu Met

Asp Glu Leu

Cys Gly Asp90

Asp Val Leu

Wing Gly Gly

Asp Tyr Thr45

Trp Asn Asn60

Gly Ile Thr75

Tyr Gly Asn

Val Ile Ala15

Read Cys Asp30

Lys Pro Leu

Pro Phe Thr

Read Ile Pro80

Gly Wing Met95Ala Glu

<210> 8 <211> 171 <212> PRT

<213> Arabidopsis thaliana <400> 8

Lys Pro Arg Val Leu Leu Wing Wing Ser Gly Ser Val Wing

1 5 10

Phe Gly Asn Leu Cys His Cys Phe Thr Glu Trp Wing Glu

20 25

Val Val Thr Lys Being Ser Leu His Phe Leu Asp Lys Leu

35 40 45

Gln Glu Val Thr Read Leu Tyr Thr Asp Glu Asp Glu Trp Ser

50 55 60

Lys Ile Gly Asp Pro Val Leu His Ile Glu Leu Arg Arg65 70 75

Val Leu Val Ile Pro Wing Leu Ser Wing Asn Thr Leu Gly

85 90

Gly Gly Read Cys Asp Asn Read Leu Thr Cys Ile Ile Arg

100 105

Tyr Thr Lys Pro Read Phe Val Pro Wing Ala-Met Asn Thr115 120 125

Asn Asn Pro Phe Thr Glu Arg His Read Leu Be Read Asp

130 135 140

Ile Thr Leu Pro Ile Pro Ile Lys Lys Arg Leu Wing Cys145 150 155

Gly Asn Gly Wing Met Wing Glu Pro Ser Leu Ile165 170

<210> 9 <211> 606 <212> DNA

<213> Arabidopsis thaliana <400> 9

atgaatatgg aagtggatac agtaacaagg aagcctcgtaagtgtggctt caattaagtt cagtaatctc tgccattgttaaagccgtcg cttcaaaatc atctctcaat ttcgttgatagtgactctct atacagatga agatgaatgg tctagctggacttcatatcg agctcagacg ctgggctgat gttatgatcaacattagcca agattgctgg tgggttatgt gataatctattgggattata gcaaaccgtt gtttgttgca ccggcgatgacctttcacag aacggcacct tgtcttgctt gatgaacttgatcaagaaga aactggcctg tggagactac ggtaatggcgatttattcca ctgttagact gttctgggag tcacaagctcagttga <210> 10 <211> 201 <212> PRT

<213> Arabidopsis thaliana <4 00> 10

Met Asn Met Glu Val Asp Thr Val Thr Arg Lys Pro Arg

1 5 10 Wing Wing Be Gly Ser Val Wing Wing Ile Lys Phe Ser Asn

20 25

Cys Phe Ser Glu Trp Wing Glu Val Lys Wing Val Wing Be

35 40 45

Leu Asn Phe Val Asp Lys Pro To Be Leu Pro Gln Asn Val

50 55 60

Thr Asp Glu Asp Glu Trp Be Ser Trp Asn Lys Ile Gly65 70 75

Read His Ile Glu Read Le Arg Arg Trp Wing Asp Val Met Ile

85 90

Leu Ser Wing Asn Thr Leu Wing Lys Ile Wing Gly Gly Leu

Wing Ile Lys1 5

Val Arg Ala30

Be Leu Pro

To be trp asn

Trp Wing Asp80 *

Lys Ile Ala95

Trp Asp110 Wing

Read Met Trp

Glu Leu Gly

Gly Asp Tyr160

tcttactagctctcagaatgaaccttctctacaagattggttgctccttttgacatgtatacactttgatgaatcaccctcaatggctgagtaaacaaag

tgcaagtggaggctgaagtcacctcagaattgatcccgttgtctgctaacagtaagagcagtggaacaataattcctcccgccttctctgagatggaacc

Ile Leu Leu1 5

Read Cys His30

Lys Ser Ser

Thr Leu Tyr

Asp Pro Val80

Ile Wing Pro95

Cys Asp Asn

60120180240300360420480540600606105

Ile Val Arg Wing Trp Asp Tyr120

Val Wing Pro Wing Met Asn Thr Read Le Met Trp Asn

100

Leu Leu Thr Cys115

130

135

Arg His Leu Val Leu Leu Asp Glu Leu Gly Ile145 150 155

Ile Lys Lys Lys Leu Wing Cys Gly Asp Tyr Gly

165 170

Glu Pro Ser Leu Ile Tyr Ser Thr Val Arg Leu

180 185

Wing Arg Lys Gln Arg Asp Gly Thr Ser195 200

<210> 11

<211> 1128

<212> DNA

<213> Oryza sativa

<4 00> 11

caaccaaagt ccaaaggcta ttctgaaccg aagtcctcccccgtcgcggc accctcctcc gccgccggat ctccaccggatcctcccccg gatcttcacc gccggtgaag tactgaggtgacaagaaacc ttgggattgg atttccctca tcctagcaaactctggaagc gtcgctgcta taaaatttga gagcctttgcagaagtcaga gccgtcgcca ccaaggcttc attacatttttagcaatatt attctttaca ctgatgatga tgaatggtcttgaagttttg cacattgaac tgcgaaaatg ggcagatatcagctaatacc ctagctaaga ttgctggtgg tttatgtgacgagagcatgg gactacagca aaccactctt tgttgccccagaacaacccg ttcaccagtc gtcatcttga gacaatcaacccctcccatt accaaaaggc tggcctgtgg tgattatggtttctgtgatc gattccaccg tcaggcttgc ttgcaagagattcacctgtg gttcctgccg gcagaaacct cccatctagctcaagattaa actctggacc tagttttcta tggtaaagagaatgttaagt gatgtctatg tcggccaaca tagcacctttctattagtga tatggtagga cgtggagatt ggagaaaggcgttgcttttc caaatttctt gtgacataat gctaaggtgctagcactatt tttttctgca aatgtttgca aagactcgtg <210> 12 <211> 220 <212> PRT

<213> Oryza sativa <4 00> 12

Met Thr Thr Be Glu Be Val Gln Glu Thr Leu1 5

110

Ser Lys Pro Read Phe125

Asn Pro Phe Thr Glu140

Thr Leu Ile Pro Pro160

Asn Gly Ala Met Ala175

Phe Trp Glu Ser Gln190

acaccaaaacgtacggcgccatgactacatcctcggggtcccgtagcttctattgatagaaacctggaagaatggtgattgaacctcttgagctatgaacactgctaggtaaatggtgcaacagccacttatgatgcggcatacttgatgatgatgatgat

ttcgaggccc

ggccaccccc

cagagtcagt

tccttgctgc

cggaatgggc

cgtctctgcc

agatagggga

cgccattatc

catgcatagt

ccttcatgtg ■

tatctttggt

tggctgagcc

attack

actattctgt

cacaaaatga

tgttgtacta

gcactttaat

gttcatgttg

Gly Leu

His Pro Ser Lys Pro Arg Val Leu Leu Wing Wing Ser Gly

10

20

25

Wing Ile Lys Phe Glu To Be Read Cys Arg To Be Phe

35

IO

Val Arg Wing Val Wing Thr Lys Wing Ser Leu His

50

55

Be Leu Pro Be Asn Ile Ile Read Tyr Thr Asp

65 70 75

Thr Trp Lys Lys Ile Gly Asp Glu Val Leu His

85 90

Trp Wing Asp Ile Met Val Ile Wing Pro Read Ser

100 105

Lys Ile Wing Gly Gly Leu Cys Asp Asn Leu Leu

115 120

Trp Wing Asp Tyr Ser Lys Pro Read Phe Val Wing

130 135

Phe Met Trp Asn Asn Pro Phe Thr Be Arg His

145 150 155

Leu Leu Gly Ile Ser Leu Val Pro Pro Ile Thr Lys Arg

Be glu

45Phe Ile60

Asp Asp

Ile glu

Asn Wing

Thr Cys125Pro Ala140

Read Glu

Asp Phe Pro15

Ser Val Ala30

Trp Wing Glu

Asp Arg Thr

Glu Trp Ser80

Read Arg Lys95

Thr Leu Ala110

Ile Val Arg

Met asn thr

Thr Ile Asn160

Read Ala Cys

60120180240300360420480540600660720780840900960102010801128165 170

Gly Asp Tyr Gly Asn Gly Wing Met Wing Glu Pro

180 185

Thr Val Arg Leu Wing Cys Lys Arg Gln Pro Leu

195 200

Pro Val Val Pro Wing Gly Arg Asn Leu Pro Ser

210 215

<210> 13 <211> 1164 <212> DNA <213> Triti <400> 13gcacgagcagtccagttgaggtggtgcactgccgctataagtggccaccactttacactgatcgagctgcgctaagatcgtacagcaaacacggagcggcaaaaggctggacttccgtgacctgtcagtataggaatttgtagtgtttgttcctccctgcatgctacaatatagcttcctgaagtgtccactgtcctaaa <210> 14 <211> 220 <212> PRT <213> Triti <400> 14Met Ala THR1

Gln Pro Ser

175

Ser Val Ile Asp Ser190

Asn Thr Asn Ser Ser205

Ser220

ccgcggagcccggttcaagcactcacaaccaatttgagagcgccatccttatgacgatgaggaaatgggcccggtgggttcgctctttgtatcttcacaccctgtggcggggctcgcgtgataaccggccccttgtcttaaaaaaa

gccagagacaagagctgatgtagtaggccccctttgccgtgcacttcgttatggtctaccagacgttatgatgcgacaactgccccagccaatcaaccaattacgggaaccaagacgcaaatctagctgaagtgtgcatcattggggggtgtgtgtgtgtgt

gacgctgccggctacatcagcgggtcctccagcttctctggatagatcattggaagaagagtgatcgctcctcttgacctatgaacacgtctgggcatagggcgcaatggccgcacgatgtgcagcgttggggggggggggtgt

ccgtcggccgagccagtacattgctgcttcagtgggcggactctaccaagtaggagatgacattgtcagcgcatcgtgagtaatgtggaaccttgatcccccgaaacctccgagcagttcggccacttgtgggggggggggcggggggggg

tcctccgcccagagagtttgaggaagtgtatgtccgagcttggcatcgttagtcttacacaaataccctgagcgtgggaccaacccattccccagttaccgcagatccatactcgcgggtttgtcaagcttttttccaacaacaatagggagtagtag

Allah

Val

To be

65

Thr

Trp

Lys

Allah

Leu145Gln

Gly

To be

Ile Lys35

Arg Wing50

Read pro

Trp lys

Asp wing

Ile Ala115Trp Asp130

Met trp

Read gly

Asp Tyr

Val Arg195

cum aestivum

Be Glu Pro5

Arg Pro Arg20

Phe Glu Ser

Val Wing Thr

Ser Gly Ile70

Lys Ile Gly85

Val Met Val100

Gly Gly Leu

Tyr ser lys

Asn Asn Pro150

Ile Wing Leu

165Gly Asn Gly180

Read Ala Cys

Val gln

Val leu

Read Cys

40Thr Pro55

Val leu

Asp Glu

Ile wing

Cys Asp120Pro Leu135

Phe thr

Ile pro

Met wing

Lys Thr200

Glu Ser Leu10

Read Wing Ala25

Arg Ser Phe

Ser Leu His

Tyr Thr Asp75

Val Leu His90

Pro Leu Ser105

Asn Leu Leu

Phe Val Wing

Glu Arg His155

Pro Val Thr170

Glu Thr185 Wing

Gln Pro His

Val Val

Be gly

Be glu

45Phe Val60

Asp Asp

Ile glu

Asn Wing

Thr Cys125Pro Ala140

Read his

Lys arg

Be gln

Asp Wing205

His Tyr Ser15

Ser Val Ala30

Trp Wing Asp

Asp Arg Ser

Glu Trp Ser80

Read Arg Lys95

Thr Leu Ala110

Ile Val Arg

Met asn thr

Thr Ile Asn160

Read Ala Cys

175Ile His Thr190

To be to be

601201802403003604204805406006607207808409009601020108011401164Leu Wing Gly Pro Val Ser Asn Asn Arg Pro Being Ser210 215 220

<210> 15 <211> 1165 <212> DNA <213> Zea mays <400> 15

gcacgaggtc agatcgacct ggactttggc ggcagctact tctagaacgc ggcggcacggcagcaagcca gcaaggagaa cgcggcggca cgggcgtgct gctacttctg cttcctctccgacgcttgct gttgagcggt tcaagcagag ctgatggcta catcagagcc agtacaagagagtttggtgg tgcactactc acaacctagt aggccccggg tcctccttgc tgcttcaggaagtgtagccg ctataaaatt tgagagcctt tgccgtagct tctctgagtg ggcggatgtccgagctgtgg ccaccacgcc atccttgcac ttcgttgata gatcatctct accaagtggcatcgttcttt acactgatga cgatgaatgg tctacctgga agaagatagg agatgaagtcttacacatcg agctgcggaa atgggcagac gttatggtga tcgctccatt gtcagcaaataccctggcta agatcgccgg tgggttatgc gacaacctct tgacctgcat cgtgagagcgtgggactaca gcaaaccgct ctttgttgcc ccagccatga acacgttaat gtggaacaacccattcacgg agcggcatct tcacacaatc aaccaactgg gcatagcctt gatccccccagttaccaaaa ggctggcctg tggcgattac gggaacggcg caatggccga aacctcgcagatccatactt ccgtgaggct cgcgtgcaag acgcaaccgc acgatgcgag cagttcactcgcgggtcctg tcagtaataa ccggccatct agctgatgca gcagttggcc acttgattgtcaagcttagg aatttgtttt atatgcagtg tgcatctgga gtgttgtaac agattttttttccaactagt gtttgtgtgt att gaaattg ggggggaaag gctgttgtca caggataacaataacttcct ccctgctcag taattatgag tctattcagt ttgtaattgt cggtgggagtacaattatgc tacaatattg tttgtttgtg tgtgtgtggg agttggggag tgctgctgccacagagatag cttcctccct gcccagccag taatgatagt gattattcag tttgtaaaaaaaaaaaaaaa aaaaaaaaaa aaaaa <210> 16 <211> 220 <212> PRT <213> Zea mays <4 00> 16

Met Wing Thr Be Glu Pro Val Gln Glu Be Leu Va: 15 10

Gln Pro Be Arg Pro Arg Val Leu Leu Wing Wing]

20 25

Wing Ile Lys Phe Glu To Be Read Cys Arg To Be Phe If:

35 40

Val Arg Wing Val Wing Thr Thr Pro Be Read His Phi50 55 60

Ser Leu Pro Ser Gly Ile Val Leu Tyr Thr Asp Asi65 70 75

Thr Trp Lys Lys Ile Gly Asp Glu Val Leu His Ile

85 90

Trp Wing Asp Val Met Val Ile Pro Wing Read Ser Alc

100 105

Lys Ile Wing Gly Gly Leu Cys Asp Asn Leu Leu Thi

115 120 125

Wing Trp Asp Tyr Ser Lys Pro Read Phe Val Wing Pro Pro Wing Met Asn Thr

130 135 140

Read Met Trp Asn Asn Pro Phe Thr Glu Arg His Read His Thr Ile Asn145 150 155 160

Gln Leu Gly Ile Wing Leu Ile Pro Pro Val Thr Lys Arg Leu Wing Cys

165 170 175

Gly Asp Tyr Gly Asn Gly Wing Met Wing Glu Thr Be Gln Ile His Thr

180 185 190

Ser Val Arg Read Ala Cys Lys Thr Gln Pro His Asp Al Ser Be Ser

195 200 205

Read Gly Wing Pro Val Ser Asn Asn Arg Pro Ser Ser 210 215 220

<210> 17 <211> 631 <212> DNA

Val His Tyr Ser 15 Gly Ser Val Wing 30 Glu Trp Wing Asp45 Val Asp Arg SerAsp Glu Trp Ser 80Glu Read Arg Lys 95 Asn Thr Read Wing 110 Cys Ile Val Arg

601201802403003604204805406006607207808409009601020108011401165 <213> Picea abies <220>

<221> misc_feature <222> (546) .. (546) <223> η is a, c, g, or t <400> 17

ggacgcgtgg gtactgtact gcatatagaa ctccggcaatgctccactat cagcaaacac gcttgctaag atagcaggtgacttgcataa tacgtgcatg ggactttaac aagcctctctactttcatgt ggaacaaccc atttactcaa cggcatttgggtatcactta tcccccctat aacaaagacg ttagcttgtgatgtcagaac cttcctcaat agatacaact cttaggtttttgaagtatct gatctccctt tctccaattc ttgttatgaaggactgtggt atagactcag attcgtagct gggctagacatggctatgtt aagaaattct gatgataaac aatctcgaacgttccnccat tctggacact tacaagaact gtgtggatacgtcaatctac attgcatatc atctaatttg g <210> 18 <211> 120 <212> PRT <213> Picea abies <4 00> 18

Gly Arg Val Gly Thr Val Leu His Ile Glu Leu15 10

Met Val Wing Ile Pro Wing Read To Be Wing Asn Thr

20 25

Gly Gly Read Cys Asp Asn Read Leu Thr Cys Ile

35 40

Phe Asn Lys Pro Read Phe Val Wing Pro Wing Met

50 55

Asn Asn Pro Phe Thr Gln Arg His Leu Asp Ser65 70 75

Val Ser Leu Ile Pro Pro Ile Thr Lys Thr

85 90

Gly Asn Gly Wing Met Be Glu Pro Be Ser Ile

100 105

Phe Ser Leu Asp Pro Ile Lys115 120

<210> 19 <211> 769 <212> DNA

<213> Brassica napus <4 00> 19

gagatctgta agggttttca aaaagagcca tggagaacggtggaagtgca gctctcccct agaaagcccc gcgtactcctccgccatcaa attcggcaac ctctgccact gcttcacagatcgtctcgaa atcgtctctc cacttcctcg acaagctctctctacaccga cgaagacgag tggtcgagct ggaacaagattcgagctcag acgctgggct gacgtcatgg tcatcgctccccaagatagc tggtgggatg tgtgataatc ttctgacttgatagcaaacc gcttttcgtt gcgccggcta tgaatactttcggagaggca tcttttgtcg cttgatgagc ttggaatcacagaggttggc ttgtggtgac tatggtaatg gcgcgatggcccactgttag actcttctgg gagtctcagg ctcatcagcaaccatgatgg ttttgcactt tgcaatggtt ggtcagtgtctgctcgtcct tgtatatgtt aagaacagct gtctgggttg <210> 20 <211> 209 <212 > PRT

<213> Brassica napus <400> 20

Glu Met Asn Gly Lys Arg Asp Met Glu Asp Met15 10

gggctgatgcgcctatgcgattgtagctccactctatctcgtgattatggcactcgatcctctaacatgcgcatgaacccttatgatttatcctgcttat

tatggtgattcaatctactgtgcaatgaatggagatgggaaaatggtgcattcaattaaattagtactatagctcgtgtgtgtaacatttaagaaaatgt

Arg Gln

Read Wing

Ile arg

45Asn Thr60

Ile SerAla CysAsp Thr

Trp Wing Asp15

Lys Ile Ala30

Wing Trp Asp

Phe Met Trp

Glu Met Gly80

Gly Asp Tyr95

Thr Leu Arg110

gaaaagagaccgcagcaaccgtgggcggaatctcccacagcggcgatccctttgtctgcttatcataagagatgtggaactcttattccttgagccgtctaagtggtggaatagattgttatgtctctt

agagaagacaggaagcgtcggtgagagccggaagtgactcgtgcttcacaaacactttaggcttgggattaatccttttaccgatcaagacttatctattactagttaatctgtctgaaa

60120180240300360420480540600631

60120180240300360420480540600660720769

Glu Val Gln Leu Ser15Pro Arg Lys Pro 20 Arg Val Leu Leu Wing 25 Wing Thr Gly Ser Val 30 Wing AlaIle Lys Phe 35 Gly Asn Leu Cys His 40 Cys Phe Thr Glu Trp 45 Wing Glu ValArg Wing 50 Val Val Ser Lys Ser 55 Ser Leu His Phe Leu 60 Asp Lys Leu SerLeu Pro Gln Glu Val Thr Leu Tyr Thr Asp Glu Asp Glu Trp Ser Ser 70 70 75 80Trp Asn Lys Ile Gly 85 Asp Pro Val Leu His 90 Ile Glu Leu Arg Arg 95 TrpAla Asp Val Met 100 Val Ile Ala Pro Leu 105 Ser Ala Asn Thr Leu 110 Ala LysIle Ala Gly 115 Gly Met Cys Asp 120 Leu Leu Thr Cys Ile 125 Ile Arg AlaTrp Asp 130 Tyr Ser Lys Pro Leu 135 Phe Val Ala Pro Ala 140 Met Asn Thr LeuMet Trp Asn Asn Pro Phe Thr Glu Arg His Read Leu Be Read Asp Glu145 150 155 160Leu Gly Ile Thr Leu 165 Ile Pro Pro Ile Lys 170 Lys Arg Leu Wing Cys 175 GlyAsp Tyr Gly Asn 180 Gly Wing Met Wing Glu 185 Pro Be Leu Ile Tyr 190 Ser ThrVal Arg Leu 195 Phe Trp Glu Ser Gln 200 Wing His Gln Gln Ser 205 Gly Gly Thr

To be

<210> 21

<211> <212>

<222>

636DNA

<213> Brassica oleracea var. alboglabra <220>

<221> misc_feature (622) .. (622)

c, g, or t

<223> η is a, <400> 21

gacgcgtggg cggacgcgtg gggggttttc aaaaagagcc atggagaacg

cagagaagac atggaagtgc agctctcccc tagaaagccc cgcgtactcc

cggaagcgtc gccgccatca aattcggcaa cctctgccac tgcttcacag

agtgagagcc gtcgtctcga aatcgtctct ccacttcctc gacaagctct

ggaagtgact ctctacaccg acgaagacga gtggtcgagc tggaacaaga

cgtgcttcac atcgagctca gacgctgggc tgacgtcatg gtcatcgctc

taacacttta gccaagatag ctggtgggat gtgtgataat cttctgactt

agcttgggat tatagcaaac cgcttttcgt tgcgccggct atgaatactt

caatcctttt acggagaggc atcttttgtc gcttgatgag cttggaatca

tccgatcaag aagaggttgg cttgtggtga ctatggtaat ggcgcgatgg

tcttatctat tccactgtta gnctcttctg ggagtc <210> 22

ggaaaagagatcgcagcaacagtgggcggactctcccacatcggcgatccctttgtctgcgtatcataagtgatgtggaactcttattccctgagccgtc

<211> <212>

198PRT

alboglabra

<221> <222>

<213> Brassica oleracea var. <220>

INSECURE (194) .. (194)

<223> Xaa can be any naturally occurring amino acid <400> 22

Met Glu Asn Gly Lys Arg Asp Arg Glu Asp Met Glu Val Gln Leu Ser15 10 15

Pro Arg Lys Pro Arg Val Leu Leu Wing Wing Thr Gly Ser Val Wing Wing

20 25 30

Ile Lys Phe Gly Asn Leu Cys His Cys Phe Thr Glu Trp Wing Glu Val

35 40 45

Arg Wing Val Val Ser Lys Ser Ser Leu His Phe Le Ser Asp Lys Ser Leu Ser50 55 60

60120180240300360420480540600636Leu Pro Gln Glu Val Thr Read Tyr Thr Asp Glu65 70 75

Trp Asn Lys Ile Gly Asp Pro Val Leu His Ile

85 90

Wing Asp Val Met Val Ile Pro Wing Read Sera Wing

100 105

Ile Wing Gly Gly Met Cys Asp Asn Leu Leu Thr

115

120

Trp Asp Tyr Ser Lys Pro Read Phe Val Wing Pro

130

135

Met Trp Asn Asn Pro Phe Thr Glu Arg His Leu145 150 155

Read Gly Ile Thr Read Ile Pro Pro Ile Lys Lys

165 170

Asp Tyr Gly Asn Gly Wing Met Wing Glu Pro Ser

180 185

Val Xaa Leu Phe Trp Glu195

<210> 23 <211> 958 <212> DNA

<213> Nicotiana tabacum <400> 23

ggagctgtgt tgccgcagaa ggagcaagaa ttgttggtgcatttaattcc tagcagattt gatttagatt agggtagataccggttcaga ttaacaatgc accgaggaga cctcgcattcgtggctgcta tcaagtttgc cagtctatgc cgttcctttagcggttgcta caaaagcttc tcttcatttc atagacaaagattctttata ctgatgagga tgaatggtca acttggacgacacatcgagc tccggaggtg ggctgatatt atgattattgcttgggaaga ttgctggtgg actatgtgat aacttgttaagactacgata aacccctttt cgtggcacca gcaatgaatattcacagaaa agcaccttat ggcaattgat gagcttgggatcaaagagac tagcctgtgg agattacggg aatggagcaattccaagctg taagactcta ttatgacgca caattacgattgatccacag gtcatgaaat ttcattcatc ggtttgtacattcaggacca ggagcccagc tgtgttattt ttctcctaaatttcttgtgt cctagagcta ctttcaatca aggttcatgtagaatatcac gtaactgctg ttactaatgg atgtgatcat < 210> 24 <211> 207 <212> PRT

<213> Nicotiana tabacum <400> 24

Met Glu Thr Be Glu Met Glu Pro Val Gln Ile1 5 10

Arg Pro Arg Ile Leu Leu Wing Wing Ser Gly Ser

Asp

Glu

Asn

Cys

Ala140Leu

Arg

Read

Glu Trp

Read arg

Thr Leu110Ile Ile125

Met asn

To be read

Read Wing

Ile Tyr190

Ser Ser80Arg Trp95

Wing Lys

Arg Wing

Thr leu

Asp Glu160Cys Gly175

To be thr

ttcaaggatatggagacttctgcttgcagcctgattgggccttcacttccagataggtgacccctttgtccctgcatcgtcattgatgtgtctctctcattggctgaacccaggtggcagagtgatagaattcttcaccatcatcatcat

attggtgttgagagatggaaaagtggaagttgaagttaaaggaagatgtcccgtgtgctaagcaaatacaacgagcatgggaataatccaaccaccagtagtctctgatccaacgtggcggttgttgaaaccctagtatccggaaaaat

20

25

Phe Ala Be Read Cys Arg Be Phe Thr Asp Trp

35

40

Val Wing Thr Lys Wing Being Read His Phe Ile Asp

50

55

Glu Asp Val Ile Read Tyr Thr Asp Glu Asp Glu

Asn Asn Wing

Val Wing Ala30

Glu Val45 Wing

Lys Ala Ser60

Trp ser thr

65

70

75

Lys Ile Gly Asp Arg Val Leu His Ile Glu Leu Arg Arg Trp

85

90

Ile Met Ile Ile Pro Wing Read Ser Wing Asn Thr

100

105

Gly Gly Read Cys Asp Asn Read Leu Thr Cys Ile

115

120

Tyr Asp Lys Pro Read Phe Val Wing Pro Wing Met

130

135

Asn Pro Phe Thr Glu Lys His Leu Met Wing

Read Gly Lys110

Val Arg Wing125

Asn Thr Leu140

Ile Asp Glu

Pro Arg15

Ile lys

Lys Wing

Read pro

Trp Thr80Ala Asp95

Ile AlaTrp AspMet TrpLeu Gly

60120180240300360420480540600660720780840900958145 150 155 160

Ile Ser Leu Ile Pro Pro Val Ser Lys Arg Leu Wing Cys Gly Asp Tyr

165 170 175

Gly Asn Gly Wing Met Wing Glu Pro To Be Read Ile Phe Gln Wing Val Arg

180 185 190

Read Tyr Tyr Asp Wing Gln Read Arg Be Gly Gly Ser Asn Val Wing195 200 205

<210> 25 <211> 1016 <212> DNA

<213> Solanum tuberosum <400> 25

gaattgatcg gcgccggact tcgatcaccg tcggcgtttc taagccgtcg ccggaactaa 60

gccggagaaa atctcaatca agtgagctaa gcactcggcg tttcaacttc tggttataaa 120tttattaaag cctttgctcc attaggttag gtagatacgg atcctatgac ttcagagatg 180gaaccagttc agattaatgg tgcacctagg agacctcgta ttctgctggc agcaagtgga 240agtgtggctg caattaagtt tgcaaatcta tgtggttgtt tttctgaatg ggcagaagtt 300aaagcagttg caacaaaacc ttctcttcat ttcatagaca aagcttcact tccggaagat 360gccattctat atactgatga ggaggaatgg tccacttgga agaaaattgg tgatagtgtg 420ctacacattg agctccgcag gtgggctgat attatggtta ttgccccttt gtcagcaaac 480acacttggga agattgcagg tggactatgt gataacttgt taacctgcat cgtacgagca 540tgggactaca ataaacccct ttttgtggca ccagccatga atacattgat gtggaataat 600ccattcacag aacgacacct tatggtaatt gatgagcttg gaatctctct cataccacca 660gtttctaaaa gactagcttg tggagattat ggaaacggcg ctatggctga accttctctc 720atctactcaa ctgtaagact cttctatgag tcacggtcac aatcaggtgg catcaacttg 780gcttgatcca cggatcatta aattttattc gtcggtttgt acaagtggta gaaattgttg 840aaattcagga cctggaacag tgttacttag catacaccag ttgtgtttat ttttctcctt 900aattagtgtc atgtttgtat the ccaagagctg ctttcaatc aagtttcatg ttcattgtag 960tctattgact ctgaatatta tgcaactcta tatagtaccc gctactaatt atgtat 1016

<210> 26 <211> 206 <212> PRT

<213> Solanum tuberosum <400> 26

Met Thr Be Glu Met Glu Pro Val Gln Ile Asn Gly Wing Pro Arg Arg1 5 10 15 Pro Arg Ile Leu 20 Leu Wing Wing Ser Gly 25 Ser Val Wing Ile 30 Lys PheAla Asn Leu 35 Cys Gly Cys Phe Ser 40 Glu Trp Wing Glu Val 45 Lys Wing ValAla Thr 50 Lys Pro Be Read His 55 Phe Ile Asp Lys Wing 60 Be Read Pro GluAsp Wing Ile Leu Tyr Thr Asp Glu Glu Glu Trp Be Thr Trp Lys65 70 75 80Ile Gly Asp Ser Val 85 Leu His Ile Glu Leu 90 Arg Arg Trp Wing Asp 95 IleMet Val Ile Wing 100 Pro Leu Ser Wing Asn 105 Thr Leu Gly Lys Ile 110 Wing GlyGly Leu Cys 115 Asp Leu Leu Thr 120 Cys Ile Val Arg Wing 125 Trp Asp TyrAsn Lys 130 Pro Leu Phe Val Wing 135 Pro Wing Met Asn Thr 140 Leu Met Trp AsnAsn Pro Phe Thr Glu Arg His Leu Met Val Ile Asp Glu Leu Gly Ile145 150 155 160Ser Leu Ile Pro Pro 165 Val Lys Arg Leu 170 Ala Cys Gly Asp Tyr 175 GlyAsn Gly Wing Met 180 Wing Glu Pro Ser Leu 185 Ile Tyr Ser Thr Val 190 Arg LeuPhe Tyr Glu 195 Ser Arg Ser Gln Ser 200 Gly Gly Ile Asn Leu 205 Wing

<210> 27 <211> 967 <212> DNA <213> Glycine max <400> 27

ggttgaacag aagagattta ggcagcccga accgacgatg ttgtttttaa ctgacagcag 60

cacaaagcga acactaacac taacgtgaaa cgcgttgcgt tgtcggaaaa tcaccctcac 120

ctgatcactg tggaagtctc taggtgatgg ccggttcaga acctgttagg gcagagggag 180

agactatggc tgtggatgct gccccaagga agccccggat tctacttgct gctagtggga 240

gtgttgctgc tgtcaaattt gcaaatcttt gtcactgttt ctctgaatgg gcagatgtaa 300

gagcagtttc cacaagtgca tctttgcatt tcattgatag agcagcaatg cccaaggatg 360

taattctata cacggatgac aatgaatggt ctagttggaa gaaattaggt gatagcgtgc 420

ttcacattga gcttcgcaaa tgggctgata tcatggtcat cgctccatta tcagcaaaca 480

cccttggcaa gattgctgga gggttgtgtg acaatctact gacatgcatc gttcgagcct 540

gggactacag caagccattc tttgttgcac cagccatgaa cactttgatg tggaacaatc 600

ctttcactga gcggcatttc atctccattg atgagcttgg catttctctc atcccgcctg 660

ttacaaagag gttagcttgt ggggattatg gcaatggtgc catggctgaa ccctctacca 720

tttactcaac tgtaaggctc ttctatgagt caaaggctca gcaaggtaga gctgtggtaa 780

ccttaaggtg aatgaagggg tagtcctacg cccccattca tggtagtcaa tagcactcta 840

tagcaggata acggagcgca gcagccctga gttactatgg aagtcgaaat cgctgagcga 900

ttttcaataa ccgctgtagc ggctgcaata gtggtctaaa tacagctttt cgggtgctac 960

agcgcac 967 <210> 28 <211> 214

<212> PRT

<213> Glycine max

<400> 28

Met Wing Gly Ser Glu Pro Val Arg Wing Glu Gly Glu Thr Met Wing Val1 5 10 15 Asp Wing Pro Arg Wing Lys Pro Arg Ile Leu Wing Wing Wing Being Gly Ser 20 25 30 Val Wing Wing Val Lys Phe Wing Asn Leu Cys His Cys Phe Ser Glu Trp 35 40 45 Wing Asp Val Arg Wing Wing Val Ser Thr Wing Be Seru His Phe Ile Asp 50 55 60 Arg Wing Wing Met Pro Lys Asp Val Ile Leu Tyr Thr Asp Asn Glu65 70 75 80Trp Being Ser Trp Lys Lys Leu Gly Asp Ser Val Leu His Ile Glu Leu 85 90 95 Arg Lys Trp Wing Asp Ile Met Val Ile Pro Wing Leu Being Wing Asn Thr 100 105 110 Leu Gly Lys Ile Wing Gly Gly Leu Cys Asp Leu Leu Thr Cys Ile 115 120 125 Val Arg Wing Trp Asp Tyr Ser Lys Pro Phe Phe Val Wing Pro Ala Met 130 135 140 Asn Thr Leu Met Trp Asn Pro El Phe Thr Glu Arg His Phe Ile Ser145 150 155 160Ile Asp Glu Leu Gly Ile Ser Leu Ile Pro Val Thr Lys Arg Read 165 165 175 Cys Wing Gly Asp Tyr Gly Asn Thr Ile 180 185 190 Tyr Ser Thr Val Arg Leu Phe Tyr Glu Ser Lys Wing Gln Gln Gly Arg 195 200 205 Wing Val oin Val Thr Leu Arg

<210> 29 <211> 1067 <212> DNA

<213> Vitis vinifera <400> 29

ttgggagaga tgccacggcc ctcattcgct ctcgatcccc gacctcggct tcactctctc 60

taattttccc gagcaattct ccgagttgag gctgatttga aagttaatga tgatgacata 120tgcagaacct ttgagtccag aagttgatgc gataccagtc aacattgctc ccagaagacc 180ccggattcta cttgctgcta gtgggagtgt agctgctatg aagtttggga atctcgtcca 240ttctttttgt gaatgggcag aagtaagagc agttgtcaca aaggcttctt tacacttcat 300tgatagagca gcactgccta aggatttata tctttacact gatgatgatg aatggtccag 360ttggacaaaa ttaggagaca gtgtgcttca cattgagctc cgcaggtggg 420 ctgatgtcat

ggtaatcgcc ccattatcag caaatacact tggcaagatt gccgggggac tgtgtgacaa 480

cctgctgaca tgcattgtgc gagcgtggga ctacagcaag ccaatgtttg ttgcgccagc 540

tatgaacacc ttcatgtgga ccaatccttt cacagaacgc catcttatga caattgatga 600

acttggaatt tctcttattc cacctgtcac taaaaggctg gcttgcggag attatgggac 660

tggtgcaatg gctgaacctt ttctcatcca ctcaaccgta agactcttct tggagacacg 720

ggctcaatca agtagcagta atgtgcagta attgggtatg gttaatctgt ctgttggaga 780

agtcaccaga gagttggctg aaatggcatg ttggaaatgc taaacgtagt tcacttggac 840

cgcattgatt ttgttatgac cgtccattaa acttactacg tcttgtagat tgttggtatc 900

ggtggatttg catatcctgt ttctcaatat ttctagtagc gccttttgaa tgttatgtgc 960

cttgtgtatt aatggtgagg gagaatgtat cagtctccta aatttctcaa tctctgtgta 1020

gacatgcttg atgatacatt cctataaata gactctgatt tctgggc 1067 <210> 30 <211> 214 <212> PRT

<213> Vitis vinifera <400> 30

Met Met Met Tyr Tyr Wing Glu Pro Leu Ser Pro Glu Val Asp Wing Ile1 5 10 15 Pro Val Asn Ile Wing Pro Arg Arg Pro Arg Ile Leu Wing Wing Ser 20 25 30 Gly Ser Val Wing Met Lys Phe Gly Asn Leu Val His Ser Phe Cys 35 40 45 Glu Trp Wing Glu Val Arg Wing Val Val Thr Thr Wing Lys Ser Read His Phe 50 55 60 Ile Asp Arg Wing Wing Leu Pro Lys Asp Leu Tyr Leu Tyr Thr Asp65 70 75 80Asp Glu Trp Ser Ser Trp Thr Lys Leu Gly Asp Be Val Leu His Ile 85 90 95 Glu Leu Arg Arg Trp Wing Asp Val Met Val Ile Wing Pro Leu Be Wing 100 105 110 Asn Thr Leu Gly Lys Ile Wing Gly Gly Leu Cys Asp Leu Thr 115 120 125 Cys Ile Val Arg Wing Trp Asp Tyr Be Lys Pro Met Phe Val Wing Pro 130 135 140 Wing Met Asn Thr Phe Met Trp Thr Asn Light Phe Thr Glu Arg His Leu145 150 155 160Met Thr Ile Asp Glu Read Gly Ile Be Leu Ile Pro Pro Val Thr Lys 165 170 175 Arg Read Leu Cys Wing Gly Asp Pro Phe 180 185 190 Leu Ile His Ser Thr Val Arg Leu Phe Leu Glu Thr Arg Wing Gln Ser 195 200 205 Ser Ser As Asn Val Gln 210 <210> 31 <211> 1081 <212> DNA <213> Hordeum vulgare <400 > 31

cggcacgagg ctccccaatt ccctccgccc gcggcgcgcc gctccgggca gcctcggtcg 60

ccgccgccgc cgccgctgcc tccacctacc gccggccgac gacgcccttc gaacctaccg 120

cctccgccgc catcttcaac cgcttatttg cctgcgctcc taccagatcg cctctgagtt 180

gagggctcca agtgaagcag atggctacat cagaaccggt acagcaaagc tgggagctgg 240

aatccagcag gcctcgggtc ctccttgctg cgtcagggag tgtagctgct ataaaattcg 300

agagcctctg ccgtatcttc tccgagtggg cggaagtccg agctgtggcg accaagtcag 360

cattgcactt tgttgacaga tcatctctgc caagcgacgt cgtcctttac actgatgatg 420

atgagtggtc tacctggaca aagataggag acgaggttct gcacatagag ctgcgaaagt 480

gggcagacat catggtgatc gcccccttat cagcaaacac tctggccaag atcgccggcg 540

ggttatgcga caacctcctg acgtgcatta tccgagcgtg ggactacaag aagccgatct 600

tcgccgcgcc agccatgaac accttcatgt ggaacaaccc attcacggcg cgccacatcg 660

agaccatgaa ccaactgggc atctccctgg tcccgcccac cacgaaacgg ctggcctgcg 720

gcgactacgg gaacggcgcg atggctgagc cctcgcagat ccacacgact gtgaggctcg 780cgtgcaagtc gcagacgttt ggcacgggca tttcgcccgc gcacccttcc agcggccacc 84 0ccgtctagcc gatgcggtga tggtcactat gttcaggtgg tctagtactc gaggtttttc 900tatgccgccc acatcatcag ggtttgagaa agtgaagggt atcaccaggt gttctgttca 960tcggtggtgt ctgtattaac cgaagtatcg tacctgtggg atatggatca gcttatttgt 1020tatgtggtag tagacgttag ggcgtagacg tagagattgg ggaattgctg 10801081 ttgctccagc

<210> 32 <211> 215 <212> PRT

<213> Hordeum vulgare <400> 32

Met Wing Thr Be Glu Pro Val Gln Gln Be Trp Glu Leu Glu15 10

Arg Pro Arg Val Leu Leu Wing Wing Ser Gly Ser Val Wing Wing

20 25 30

Phe Glu Be Leu Cys Arg Ile Phe Glu Be Trp Wing Glu Val

35 40 45

Val Wing Thr Lys Ser Wing Read His Phe Val Asp Arg Ser Ser

50 55 60

Ser Asp Val Val Leu Tyr Thr Asp Asp Asp Glu Trp Ser Thr65 70 75

Lys Ile Gly Asp Glu Val Leu His Ile Glu Leu Arg Lys Trp

85 90

Ile Met Val Ile Pro Wing Read Ser Wing Asn Thr Read Wing Wing Lys100 105 110

Gly Gly Read Cys Asp Asn Read Leu Thr Cys Ile Ile Arg Wing

115 120 125

Tyr Lys Lys Pro Ile Phe Wing Pro Wing Wing Met Asn Thr Phe

130 135 140

Asn Asn Pro Phe Thr Wing Arg His Ile Glu Thr Met Asn Gln145 150 155

Ile Ser Leu Pro Val Pro Thr Thr Lys Arg Leu Wing Cys Gly

165 170

Gly Asn Gly Wing Met Wing Glu Pro Be Gln Ile His Thr Thr180 185 190

Read Ala Cys Lys Be Gln Thr Phe Gly Thr Gly Ile Be Pro

195 200 205

Pro Ser Ser Gly His Pro Val210 215

<210> 33 <211> 1175 <212> DNA

<2l3> Gossypium hirsutum <400> 33

cccattgtca cctttagtgc tggtgagcat ttaagagagg gcagagcaac ggaggaaatc 60

cccttttttg tgcagtggaa gtaaaaatta cctaccatta gaaaaggaga aggattggta 120aagtattgca gcgctccgta aaggcatttt ggtatagcaa gtagcagcaa ttcatcaaaa 180ggcagcattt ctttttgcac caggggccct ttaaaaagct caactctcgc gcttctttgc 240gtttcaaatc tctctgcaag tcgcaaggac atctgaagca gaaagcccag attcctctct 300aatcttagat tccagatttg cataggttat ggcgtatcct gagcctcaag gtgcagatag 360ggagatggtt aaggtcaatc ctaccccaag aaaaccccgg gttttactcg ctgccagtgg 420aagtgtagct gccataaagt ttgggaatct ctgccattgt ttctctgaat gggcagaagt 480aaaagcagtt gccacgaaag cttctttgca tttcattgac agagcatcac ttcctaagga 540tctaaagctt tacactgatg aggaggaatg gtctagttgg gggaaaatag gtgacagtgt 600gcttcacatt gagcttcgtc gatgggctga tattatggtc attgccccat tgtcagcaaa 660cacacttggc aagattgctg gaggattatg tgacaatttg ttaacttgtg tcgtacgagc 720atgggactac agcaagccaa tgtttgttgc accagctatg aacactttca tgtggagcaa 780ccctttcaca gaaaagcatc tcatgacaat tgatgagctt ggtatttctc tcatcccccc 840tgtctccaaa agactagctt gtggggacta tggaaacggc gcaatggcag aaccttctct 900aatccactcg actgtaagat g tattcttgga tcacgacct caaccaagtg actgaagatc 960tatttattcg ccatgaatta taaATactat attagttgta tggtagccca

Ser Ser15

Ile lys

Arg Wing

Read pro

Trp Thr80Ala Asp95

Ile wing

Trp Asp

Met trp

Read Gly160Asp Tyr175

Val ArgAla Histcctgtaagc tgatgatact gatgagagtt ggcag <210> 34 <211> 208 <212> PRT

<213> Gossypium hirsutum <400> 34

1175

Met Tyr Pro Wing Glu Pro Gln Gly Wing Asp Arg Glu Met Val Lys Val1 5 10 15 Asn Pro Thr Pro 20 Arg Lys Pro Arg Val 25 Leu Leu Wing Wing Ser 30 Gly SerVal Wing 35 Ile Lys Phe Gly Asn 40 Leu Cys His Cys Phe 45 Ser Glu TrpAla Glu 50 Val Lys Wing Val Wing 55 Thr Lys Wing Ser Leu 60 His Phe Ile AspArg Wing Ser Leu Pro Lys Asp Leu Tyr Thr Asp Glu Glu65 70 75 80Trp Ser Ser Trp Gly 85 Lys Ile Gly Asp Ser 90 Val Leu His Ile Glu 95 LeuArg Arg Trp Wing 100 Asp Ile Met Val Ile 105 Wing Pro Leu Ser Wing 110 Asn ThrLeu Gly Lys 115 Ile Wing Gly Gly Leu 120 Cys Asp Asn Leu Read 125 Thr Cys ValVal Arg 130 Wing Trp Asp Tyr Ser 135 Lys Pro Met Phe Val 140 Ala Pro Wing MetAsn Thr Phe Met Trp Ser Asn Pro Phe Thr Glu Lys His Leu Met Thr145 150 155 160Ile Asp Glu Leu Gly 165 Ile Ser Leu Ile Pro 170 Pro Val Ser Lys Arg 175 LeuAla Cys Gly Asp 180 Tyr Gly Asn Gly Wing 185 Met Wing Glu Pro Ser 190 Leu IleHis Ser Thr 195 Val Arg Leu Phe Leu 200 Glu Ser Arg Pro Gln 205 Pro Ser Asp

<210> 35 <211> 1119 <212> DNA

<213> Sorghum bicolor <400> 35

gcacgagggc tctctccgct cacctccact tccccgcccc

acacgggcgc gcacccgtcc gcctccgact gacccggagcgggagggaga gcccgaaccg cggcggggcg ggagggctccgggccagccg ccccatgggc cggtcgcctg agcgcgctac

ggaaagctga gcgttagcgg agctgatggc tacatcagaggatggactac tcacaaccta gtaagcctcg ggtactccttcgctataaaa tttgagaacc tttgtcgtag tttctctgag

ggccaccgca tcatctttgc actttattga tagatcatct

ttacactgat gatgatgagt ggtctacctg gaagaagata

tgagctgcgt aaatgggcag atattatggt gattgctccg

taagatcgcc ggtgggttat gtgacaacct cttaacatgc

cagcaaaccg ctctttgttg ccccagctat gaacacatta

agagcgtcat cttcaaacga tcaaccaact gggcataatc

aaggttggct tgtggcgatc acgggaatgg tgcaatggct

ttctgtgagg cttgcatgga agacgcaacc acatgatgca

tgtcagtaat aaccgcccat ctagctggtg tttgacctct

tgttttatct gcagtgtgca tcttgatgtt agaaatgttg

tgttttagtg tgtattgtga tgagaatgtg gggaaaggaa

ataaattact gccagctcag tatcgatagt gaggatgag <210> 36 <211> 225 <212> PRT

<213> Sorghum bicolor <400> 36

Met Wing Thr Be Glu Pro Val Gln Glu Asn Leu15 10

cgccccggtccgactcgaccttttcgtttggtatcagctcccagtacaaggctgcttcagtgggcggatgcttccaagtgggagatgaagttatcagcaaatcgtgagagatgtggaacattgatcccccagagagtagtagtagctag

tccgtccttgttgttcaggaccccgccggaatcgaccggcagaatttggcgaagtgtagctccgagccgtacattgttctttttacacatataccctggccgtgggactaacccattcaccagttaccaaagatctatactgtgcgtgtgtcgtg

60120180240300360420480540600660720780840900960102010801119

Met Asp Tyr Ser Wing

15Gln Pro Be Lys Pro Arg Val Leu Read Wing Wing Be Gly Be Val Wing

20

25

30

Wing Ile Lys Phe Glu Asn Leu Cys Arg Be Phe Ser Glu Trp Wing Asp

35

40

45

Val Arg Wing Val Wing Thr Wing Be Ser Read His Phe Ile Asp Arg Ser

50

55

60

Ser Leu Pro Ser Asp Ile Val Leu Tyr Thr Asp Asp Asp Glu Trp Ser

65

70

75

80

Thr Trp Lys Lys Ile 85 Gly Asp Glu Val Leu 90 His Ile Glu Leu Arg 95 LysTrp Wing Asp Ile 100 Met Val Ile Wing Pro 105 Leu Ser Wing Asn Thr 110 Leu AlaLys Ile Wing 115 Gly Gly Leu Cys Asp 120 Asn Leu Leu Thr Cys 125 Ile Val ArgAla Trp 130 Asp Tyr Ser Lys Pro 135 Leu Phe Val Ala Pro 140 Wing Met Asn ThrLeu Met Trp Asn Pro Phe Thr Glu Arg His Leu Gln Thr Ile Asn145 150 155 160Gln Leu Gly Ile Ile Pro Leu Ile Pro Pro Val 170 Thr Lys Arg Leu Wing 175 CysGly Asp His Gly 180 Asn Gly Wing Al Met 185 Wing Glu Thr Ser Gln Ile 190 Tyr ThrSer Val Arg 195 Leu Wing Trp Lys Thr 200 Gln Pro His Asp Wing 205 Ser SerLeu Val 210 Val Pro Val Ser Asn 215 Asn Arg Pro Ser Ser 220 Trp Cys Leu Thr

Ser225

<210> 37 <211> 975 <212> DNA

<213> Lycopersicon esculentum <400> 37

ctacaattga tcggcgccgg actttgatca ccgtcggcgt ttctaagccg ttgccggact 60

atgccggagc aaatctcaat caagtgagct aaccactctg tgtttcaact tctggttata 120aattttttaa agcctttgct ccgttaggtt aggtagatac ggatcctatg acttcagaga 180tggaaccagt tcagattaat ggtgcaccta ggagacctcg tattctgctg gcagcaagtg 240gaagtgtggc ttcaattaag tttgctaatc tatgtcgttg tttttctgaa tgggcagaag 300ttaaagcagt tgcaacgaaa ccttctcttc atttcataga caaagcttcg cttccggaag 360atgtcattct ttatactgat gaggaggaat ggtccacttg gtgcagattg gtgatagtgt 420gctacacatt gagctccgca gatgggctga tattatggtt attgcccctt tgtcagcaaa 480cacacttggg aagattgcag gtggactatg tgataacttg ttaacctgca tcgtacgagc 540atgggactac aataaacccc tttttgtggc accagccatg aatacattga tgtggaataa 600tccattcaca gaacgacacc ttatggtaat tgatgagctt ggaatctctc tcataccccc 660agtttcaaaa agactagcat gtggagatta tggaaatggc gctatggctg aaccttctct 720catttactca actgtaagac tcttttatga gtcacggtca caatcaggtg gcatcaactt 780ggcttgattc gcagatcatt aaattttatt catcggtttg tacaagtggt aaaaattgtt 840gaaattcagg acctggaaca gtgttactta gcatacacca gttgggttta tttttctcct 900aaattcttca cctctttaac t ttgtgtcatg ttgtatcca agcggtaact ttcaatcaag 960tttcatgttc attgt 975

<210> 38 <211> 206 <212> PRT

<213> Lycopersicon esculentum <220>

<221> UNSAFE <222> (74) .. (74)

<223> Xaa can be any naturally occurring amino acid <400> 38

Met Thr Be Glu Met Glu Pro Val Gln Ile Asn Gly Wing Pro Arg Arg1 5 10 15

Pro Arg Ile Leu Read Wing Ala Wing Gly Ser Val Wing Wing Ile Lys Phe20 25 30Ala Asn Leu Cys35

Thr Lys Pro50 Wing

Asp Val Ile Leu65

Ile Gly Asp Ser

Met Val Ile Ala100

Gly Leu Cys Asp115

Asn Lys Pro Leu130

Asn Pro Phe Thr145

Ser Leu Ile Pro

Asn Gly Ala Met180

Phe Tyr Glu Ser195

<210> 39 <211> 1395 <212> DNA <213> Arabidopsis thaliana <400> 39

atggcgaaaa cagctcttac tcctcccgcg tctggttccg aagttccaag atccggtaca 60

cctggagatg cgtctggcaa caaacctcaa acggatgcca ccggcgtctc agctactgat 120actgcttctc agaaacgcgg tcgtggtcga ccgccaaagg ctaaatctga ctcttcccaa 180atcggtgccg tttctgcgaa ggcgagtact aaaccaagtg gtcgtccgaa aagaaacgta 240gctcaggctg ttccttctac gtctgtggcg gcggctgtga agaaacgtgg tagggcaaag 300aggtcgactg taacggcggc tgtggttact actgctactg gagagggttc tagaaaacga 360gggaggccaa agaaagatga cgtggcggct gcaactgttc cagcagaaac tgtggtggct 420ccagctaaga gacgtggaag gaaacctact gtcgaagtag ctgcacagcc tgtgcgcagg 480actaggaagg tattcgggtt ttctatgcat gaacaaaaga gcacttcagt ggcaccggta 540gctgcaaacg tcggagatct caagaaaaga accgcactct tacaaaagaa agtgaaggaa 600gctgcagcta agttgaaaca agcagtaaca gcaattgacg aggtccagaa gttagcggat 660ggaatattga ccagcgatga cgtcgacttc tctgttttgt tttcaaactt aaagaccctc 720gcggcatttg atttcgattt ccgattaggg ttcctcgcag atcctccttc ctcggctata 780gagaagaaga tgaatatgga agtggataca gtaacaagga agcctcgtat cttactagct 840gcaagtggaa gtgtggcttc aattaagttc agtaatctct gccattgttt ctcagaatgg 900gctgaagtca aagccgtcgc t ttcaaaatca ctctcaatt tcgttgataa accttctcta 960cctcagaatg tgactctcta tacagatgaa gatgaatggt ctagctggaa caagattggt 1020gatcccgttc ttcatatcga gctcagacgc tgggctgatg ttatgatcat tgctcctttg 1080tctgctaaca cattagccaa gattgctggt gggttatgtg ataatctatt gacatgtata 1140gtaagagcat gggattatag caaaccgttg tttgttgcac cggcgatgaa cactttgatg 1200tggaacaatc ctttcacaga acggcacctt gtcttgcttg atgaacttgg aatcacccta 1260attcctccca tcaagaagaa actggcctgt ggagactacg gtaatggcgc aatggctgag 1320ccttctctga tttattccac tgttagactg ttctgggagt cacaagctcg taaacaaaga 1380gatggaacca 1395 gttga

<210> 40 <211> 464 <212> PRT

<213> Arabidopsis thaliana <400> 40

Met Wing Lys Thr Wing Leu Thr Pro Pro Wing Be Gly Be Glu Val Pro15 10 15

Arg Be Gly Thr Pro Gly Asp Wing Be Gly Asn Lys Pro Gln Thr Asp

20 25 30

Wing Thr Gly Val Be Wing Thr Asp Thr Wing Be Gln Lys Arg Gly Arg

35 40 45

Gly Arg Pro Pro Lys Wing Lys Be Asp Be Gln Ile Gly Val Wing

Arg cys

To be read

Tyr thr

70Val Leu85

Pro leu

Asn leu

Phe val

Glu Arg150Pro Val165

Glu Wing

Phe

His

55

Asp

His

To be

Read

Ala135His

SerProArg Ser Gln

Be Glu40

Phe Ile

Glu Glu

Ile glu

Asn105Thr Cys120 Wing

To the wing

Read met

Lys arg

Ser Leu185Ser Gly200

Trp Wing GluAspXaa

Read

90

Thr

Ile

Met

Val

Leu170Ile

Gly

Lys Ala60

Met val75

Arg Arg

Read gly

Val arg

Asn Thr140Ile Asp155

CysTyr SerIle Asn Wing

Val Lys Ala45

Be Leu Pro

His Leu Val

Trp Wing Asp95

Lys Ile Ala110Ala Trp Asp125

Read Met Trp

Glu Leu Gly

Gly Asp Tyr175

Thr val arg

190Leu Ala205

Val

Glu

Gln80Ile

Gly

Tyr

Asn

Ile160Gly

Leu50 55 60 Be Wing Lys Wing Be Thr Lys Pro Be Gly Arg Pro Lys Arg Asn Val65 70 75 80Ala Gln Wing Val Pro Be Thr Be Val Wing Wing Val Lys Lys Arg 85 90 95 Gly Arg Wing Lys Arg Be Thr Val Thr Wing Val Wing Val Thr Thr Wing 100 105 110 Thr Gly Glu Gly Be Arg Lys Arg Gly Arg Pro Lys Lys Asp Asp Val 115 120 125 Wing Wing Wing Thr Thr Pro Wing Glu Thr Val Val Wing Pro Lys Arg Wing 130 135 140 Arg Gly Arg Lys Pro Thr Val Glu Val Wing Wing Gln Pro Val Arg Arg145 150 155 160Thr Arg Lys Val Phe Gly Phe Be Met His Glu Glys Lys Be Thr Be 165 170 175 Val Wing Pro Val Wing Asn Val Gly Asp Leu Lys Lys Arg Thr Wing 180 185 190 Leu Leu Gln Lys Lys Val Lys Glu Wing Wing Lys Wing Lys Leu Lys Gln Wing 195 200 205 Val Thr Wing Ile Asp Glu Lys Leu Wing Asp Gly Ile Leu Thr 210 215 220 Phe Ser Asn Leu Lys Thr Leu225 230 235 240Ala Wing Phe Asp Ph and Asp Phe Arg Leu Gly Phe Leu Wing Asp Pro Pro 245 250 255 Ser Be Ala Ile Glu Lys Met Asn Met Glu Val Asp Thr Val Thr 260 265 270 Arg Lys Pro Arg Ile Leu Wing Ala Ser Gly Ser Val Ala Ser Ile 275 280 285 Lys Phe Be Asn Leu Cys His Cys Phe Be Glu Trp Wing Glu Val Lys 290 295 300 Wing Val Wing Be Lys Be Being Read Asn Phe Val Asp Lys Pro Be Leu305 310 315 320Pro Asp Glu Trp Be Ser Trp 325 330 335 Asn Lys Ile Gly Asp Pro Val Leu His Ile Glu Leu Arg Arg Trp Wing 340 345 350 Asp Val Met Ile Ile Wing Pro Read Ser Wing Asn Thr Leu Wing Lys Ile 355 360 365 Wing Gly Gly Leu Cys Asp Asn Leu Leu Thr Cys Ile Val Arg Wing Trp 370 375 380 Asp Tyr Ser Lys Pro Leu Phe Val Wing Ala Met Asn Thr Leu Met385 390 395 400Trp Asn Asn Pro Phe Thr Glu Leu His Leu Val Leu Asp Glu Leu 405 410 415 Gly Ile Thr Ile Ile Pro Pro Ile Lys Lys Lys Leu Cys Wing Gly Asp 420 425 430 Tyr Gly Asn Gly Wing Met Wing Glu Pro Be Leu Ile Tyr Be Thr Val 435 440 445 Arg Leu Phe Trp Glu Be Gln Wing Arg Lys Gln Arg Asp Gly Thr Be 450 455 460

<210> 41 <211> 1182 <212> DNA <213> Pinus sp. <400> 41

atattgaaac tcatttttaa acacttattc tgcactataa gaagcagcag gcccccagaa 60

atatgagctt catacacaca acgggggagt aatcagcacc atcaaagggg ggatttgcgg 120caattgtgag atcaagtgga ggaagctgga atagatctaa atctaggccc ttgaggagaa 180tgctctccta atggtggttt taaaaatcaa tactctagtg caaaacatgg atgcaacaaa 240ttctcaacct gatgaacctg gaagggatgc taattcatct cagaaaccac gaatcattct 300agcagccagc gggagtgtgg cagcaataaa atttggaatt cttgcccatt gtctatgtca 360atgggcagaa gtcaaagcag tggtcacaaa atctgctttg catttcattg acaagatgtc 420tcttccggct aatgttaagc tctacactga tgaaaatgaaaggtgatact gtactgcata tagaactccg gcaatgggctactctcagca aacacgcttg ctaagatagc aggtggcctacatagtacgt gcatgggact ttaacaagcc tctctttgtacatgtggaac aatccattta ctcaacggca tttggactcaacttatcccc ccaataacaa agaagttagc ttgtggtgattgaaccttct acgatagata caactcttag gttttcactttgtctgatgt cccgttcaac aattcttgtt atgaatctaaatggtataga cccagatagt gtttcttggt agctgggttatgtgtggtga agttaagttc tgatgataaa caatcatgaattgctcattt attctggaca cttacaagaa ctgtgtggatgtgtcaaact actatgcaaa tcatctaatt tggaacttgactttattttc ttccctgtga taattattaa tctctatgat <210> 42 <211> 215 <212> PRT < 213> Pinus sp. <400> 42

Met Val Val Leu Lys Ile Asn Thr Leu Val Gln1 5 10

Asn Be Gln Pro Asp Glu Pro Gly Arg Asp Wing

20 25

Pro Arg Ile Ile Read Wing Wing Ser Gly Ser Val

35 40

Gly Ile Leu Wing His Cys Leu Cys Gln Trp Wing

50 55

Val Thr Lys Ser Ala Read His Phe Ile Asp Lys65 70 75

Asn Val Lys Leu Tyr Thr Asp Glu Asn Glu Trp

tggtctagctgatgctatggtgcgacaatcgctcctgcaaatctcagagctatggaaatggatccttcaacatgcttagtgacagcatgacttgtgatttactcctgctttgatgtaaga

tg

ggagcaaaattgattgctcctacttacttgtgaatacttttcggactatcgtgcaatggcttgtatgaagacaatggactactcagcttgtatgtaacatataagagaatggagtatgca

85

90

Ile Gly Asp Thr Val Leu His Ile Glu Leu Arg

100 105

Met Val Ile Pro Wing Read To Be Wing Asn Thr Leu

115 120

Gly Leu Cys Asp Asn Leu Leu Thr Cys Ile Val

130 135

Asn Lys Pro Read Phe Val Pro Wing Met Wing Asn145 150 155

Asn Pro Phe Thr Gln Arg His Read Asp Ser Ile

165 170

Be Leu Ile Pro Pro Ile Thr Lys Lys Leu Wing

180 185

Asn Gly Wing Met Wing Glu Pro Be Thr Ile Asp

195 200

Ser Asu Pro Asp Ile Val210 215

<210> 43 <211> 804 <212> DNA <213> Oryza sativa <400> 43

atggggcggg ggaaggtgca gctgaagcgg atagagaacattctccaaga ggaggaatgg attgctgaag aaggcgcacggccgaggtcg ccgccatcgt cttctccccc aagggcaagctccaggatgg acaaaatcct tgaacgttat gagcgctattatttcagctg aatccgagag tgagggaaat tggtgccatgaagattgaga ccatacaaaa atgtcacaaa cacctcatggaatctcaaag aactccaaca gctagagcag cagctggagatcaagaaaga gccaccttat gcttgagtcc atttccgagcctgcaggagg agaacaaggc tctgcagaag gaactggtggggccagcagc aagtagggca gtgggaccaa acccaggtccccccaagccc agacaagctc ctcctcctcc tccatgctgaccaccacaaa atatctgcta cccgccggtg atgatgggcg

Asn met

Asn ser

Wing wing

45Glu Val60

Met ser

To be to be

Gln Trp

Wing Lys125Arg Wing140

Thr phe

Be glu

Cys gly

Thr Thr205

Asp

To be

30

Ile

Lys

Read

Trp

Ala110Ile

Trp

Met

Read

Asp190Leu

Thr15 wing

Gln lys

Lys phe

Val Wing

Pro Ala80Ser Lys95

Asp Wing

Gly wing

Asp Phe

Trp Asn160Gly Leu175

Tyr GlyArg Phe

agatcaacagagatctccgttctacgagtacatatgctgaaatacaggaagagaggatctgttcattgaatgcagaaaaaagaggcagaaaggcccaggcgggatcagcaagagaaatga

gcaggtgacgcctctgcgaccgccactgacaaaggctcttacttaaggcaagaatccctggcacataataggagaggtcagaatgtgaggccaagcccaaggcacttctttgcggcggcg

4805406006607207808409009601020108011401182

60120180240300360420480540600660720gcggccggcgg tggcggcgca gggccaggtg caactccgca tcggaggtct tccgccatgg 780atgctgagcc acctcaatgc ttaa 804

<210> 44 <211> 267

<212> PRT <213> Oryza sativa <400> 44 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Asn Gly Leu Read Lys Lys Wing 20 25 30 His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Wing Ala Ile Val Phe 35 40 45 Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Ser Arg Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Glu Lys Wing Wing Leu65 70 75 80Ile Be Glu Wing Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Leu Glu Be Ser Leu Lys His Ile Ile Be Arg Lys Ser 130 135 140 His Leu Met Leu Glu Be Ile Ser Glu Leu Gln Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val Glu Arg Gln 165 170 175 Lys Asn Val Arg Gly Gln Gln Gln Val Gly Gln Trp Asp Gln Thr Gln 180 185 190 Val Gln Wing Gln Wing Gln Wing Gln Pro Gln Wing Gln Thr Be Ser Ser 195 200 205 Ser Ser Ser Met Leu Arg Asp Gln Gln Wing Leu Leu Pro Pro Gln Asn 210 215 220 Ile Cys Tyr Pro Val Val Met Met Gly Glu Arg Asn Asp Wing Wing Ala225 230 235 240Ala Wing Wing Val Wing Wing Gln Gly Gln Valu Leu Arg Ile Gly Gly 245 250 255 Trp Met Leu Be His Leu Asn Ala 260 265 <210> 45 <211> 52

<212> DNA

<213> Artificial sequence <220>

<223> primer <400> 45

ggggacaagt ttgtacaaaa aagcaggctt aaacaatggg gcgggggaag gt 52

<210> 46 <211> 47 <212> DNA

<213> Artificial sequence <220>

<223> primer <400> 46

ggggaccact ttgtacaaga aagctgggtt tggccgacga cgacgac 47

<210> 47

<211> 2193

<212> DNA

<213> Oryza sativa

<400> 47

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgccatccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctacatttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatatatctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactcaaaaaatctt tctagctgaa ctcaatgggt aaagagagag attttttttaatgaagatat tctgaacgta ttggcaaaga tttaaacata taattatatattgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatccttagtaatta aagacaattg acttattttt attatttatc ttttttcgatgtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctccttcaactagca acacatctct aatatcactc gcctatttaa tacatttaggtgaattcaag cactccacca tcaccagacc acttttaata atatctaaaaaattttacag aatagcatga aaagtatgaa acgaactatt taggtttttcaaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcacacagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacggatccgcaacaa ccttttaaca gcaggctttg cggccaggag agaggaggagaaccaagcat cctcctcctc ccatctataa attcctcccc ccttttccccggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagccgaccgcctt ctt cgatcca tatcttccgg tcgagttctt ggtcgatctccacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattgtgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgattgatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttgaggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtacgattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgatttaataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcgactatcctttg tttattccct attgaacaaa aataatccaa ctttgaagacatgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggcacagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaacctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatctctttctggtt cagttcaatg aattgattgc tacaaataat gcttttataggctgtagttc agttaatagg taatacccct atagtttagt caggagaagatttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagtattatttttt ttattagctc tcaccccttc attattctga gctgaaagtctgtcctcaat tttgttttca aattcacatc gattatctat gcattatcctcctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaagagctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttttggtgtagct tgccactttc accagcaaag ttc <210> 48 <211> 42 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (2) .. (2) <223> / replace = "Going" <220>

<221> VARIANT

<222> (6) .. (6)

<223> / replace = "Asn"

<220>

<221> VARIANT

<222> (ΙΟ) -. (ΙΟ)

<223> / replace = "He"

<220>

<221> VARIANT <222> (12) .. (12) <223> / replace = "Tyr" <220>

<221> VARIANT <222> (16) .. (16)

<223> / replace = "lie" / replace = "read" <220>

aagaaaaact 120ctagtttcgt 180cgttcacatc 240ttcttgaata 300aaaaaataga 360attttatagt 420caatttttat 480tagatgcaag 540caatacacgt 600tagcaatatc 660tacaaaaaat 720acatacaaaa 780acaggcaaca 840ggacagcaag 900aggcaaagaa 960tctctatata 1020agaagccgag 1080ttccctcctc 1140ttctaggttg 1200cctgttcttg 1260ttagtagtat 1320ggaatcttgc 1380tgcttggtgt 1440tttgacgaag 1500ggtcccgttg 1560ttgtttagat 1620caggggattc 1680taaaaagtca 1740cgttatccta 1800acttatccga 1860attcatttgg 1920tggcatgaac 1980cttgtatcta 2040aaatttatga 2100gtgcagttct 2160 2193 <221> <222> <223> <220> < 221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <223> <220> <221> <222><223><220><221><222><223><220><221><222><220><221><222><223><220> <221> <222 > <223> <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <223> <4 00>

VARIANT

(17) .. (17) / replace

VARIANT

(18) .. (18)

/replace

VARIANT

(19) ·· (19) / replace

VARIANT

(20) .. (20) / replace

VARIANT (23) .. (23) / replace

VARIANT

(30) .. (30) / replace

VARIANT

(31) .. (31) / replace

VARIANT

(32) .. (32) / replace

VARIANT

(33) .. (33) / replace

VARIANT (35) .. (35) / replace

VARIANT

(37) .. (37)

/replace

VARIANT

(38). . (38)

/replace

VARIANT (41) .. (41) / replace48

= "Gly"

"Wing" / replace = "Go"

= "Go"

= "Go"

= "Pro" / replace = "Wing" / replace = "Asn"

= "Phe"

"To be"

"To be"

= "Asn" / replace = "Glu"

= "Cys" / replace = "Arg" / replace = "Ser"

= "Glu"

"lie" / replace = "Arg" / replace = "Lys"

= "Glu"

Leu1

Allah

Read Lys Lys

Read Ile Ile20

Asp Ser Lys Met35

<210> 49 <211> 19 <212> PRT <213> String <220>

Wing His Glu Ile Ser Val Leu Cys Asp Wing Glu Val5 10 15

Phe Ser Thr Lys Gly Lys Read Tyr Glu Tyr Wing Thr

25 30

Asp Asn Ile Leu Glu Arg40

artificial <223> Artificial sequence <220>

<221> VARIANT

<222> (4) .. (4)

<223> / replace = "Be"

<220>

<221> VARIANT

<222> (5) .. (5)

<223> / replace = "Arg"

<220>

<221> VARIANT

<222> (6) .. (6)

<223> / replace = "lie"

<220>

<221> VARIANT <222> (8) .. (8)

<223> / replace = "Thr" / replace = "Ser" <220>

<221> VARIANT

<222> (9) .. (9)

<223> / replace = "lie"

<220>

<221> VARIANT <222> (10) .. (10) <223> / replace = "Asn" <220>

<221> VARIANT <222> (11) .. (11)

<223> / replace = "Arg" / replace = "Asn" <220>

<221> VARIANT <222> (12) .. (12)

<223> / replace = "Cys" / replace = "Arg" <220>

<221> VARIANT

<222> (13) .. (13)

<223> / replace = "His"

<220>

<221> VARIANT <222> (14). . (14) <223> / replace = "Lys" <400> 49

Lys -Leu Lys Wing Lys Val Glu Wing Leu Gln Lys Ser Gln Arg His Leu15 10 15

Met Gly Glu

<210> 50 <211> 11 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (2) .. (2)

<223> / replace = "Gln" / replace = "Go" / replace = "Wing" <220>

<221> VARIANT

<222> (6) .. (6)

<223> / replace = "Phe"

<220>

<221> VARIANT <222> (7) .. (7) <223> / replace = "Phe" <220>

<221> VARIANT

<222> (8) .. (8)

<223> / replace = "Phe"

<220>

<221> VARIANT

<222> (9) .. (9)

<223> / replace = "Phe"

<220>

<221> VARIANT <222> (10) .. (10)

<223> / replace = "Cys" / replace = "Phe" <220>

<221> VARIANT <222> (11) .. (11) <223> / replace = "Met" <400> 50

Gln Pro Gln Thr Be Ser Ser Ser Ser Ser Phe15 10

<210> 51 <211> 7 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (1). . (1)

<223> / replace = "Wing" / replace = "Go" / replace = "Read" <220>

<221> VARIANT

<222> (2) .. (2)

<223> / replace = "Pro"

<220>

<221> UNSAFE <222> (3) .. (3)

<223> Xaa can be any naturally occurring amino acid, preferably

Leu, Pro or His

<220>

<221> UNSAFE <222> (3) .. (3)

<223> Xaa can be any naturally occurring amino acid, preferably

Leu, Pro or His

<220>

<221> UNSAFE <222> (6) .. (6)

<223> Xaa can be any naturally occurring amino acid, preferably

Leu, Pro or His

<220>

<221> VARIANT <222> (7) .. (7) <223> / replace = "His" <400> 51

Gly Leu Xaa Trp Met Xaa Ser

1 5

<210> 52

<211> 735

<212> DNA

<213> Hordeum vulgare <400> 52

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg ccaggtcacc 60

ttctccaagc gccgctcggg gctgctcaag aaggcgcacg agatctccgt gctctgcgac 120gccgaggtcg gcctcatcat cttctccacc aagggaaagc tctacgagtt ctccaccgag 180tcatgtatgg acaaaattct tgaacggtat gagcgctact cttatgcaga aaaggttctc 240gtttcaagtg aatctgaaat tcagggaaac tggtgtcacg aatataggaa actgaaggcg 300aaggttgaga caatacagaa atgtcaaaag catctcatgg gagaggatct tgaatctttg 360aatctcaagg agttgcagca actggagcag cagctggaaa gctcactgaa acatatcaga 420gccaggaaga accaacttat gcacgaatcc atttctgagc ttcagaagaa ggagaggtca 480ctgcaggagg agaataaagt tctccagaag gaacttgtgg agaagcagaa ggcccaggcg 540gcgcagcaag atcaaactca catgatgagggggggggggggggggggggggggc

<210> 53 <211> 244 <212> PRT

<213> Hordeum vulgare <400> 53

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Be Thr Glys Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Val Be Glu Be Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Be Read Lys His Ile Arg Wing Arg Lys Asn

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln

165 170 175

Lys Wing Gln Wing Wing Gln Gln Asp Gln Thr

180 185 190

Be Ser Be Ser Phe Met Met Asp Pro Wing Pro Val Wing Asp Thr

195 200 205

Ser Asn His Pro Wing Wing Wing Gly Glu Arg Wing Glu Asp Val Wing Val

210 215 220

Gln Pro Gln Val Pro Leu Arg Thr Wing Leu Pro Leu Trp Met Val Ser225 230 235 240

His Ile Asn Gly

<210> 54 <211> 1196 <212> DNA

<213> Hordeum vulgare <400> 54

ctctcccctc ccacttcacc caaccacctg acagccatgg ctccgccacc tcgcctccgc 60

ccgcgcctct gagagtagcc gtcgcggtcg ctcgctcgct cgctcgctgc tgccggtgtt 120

ggcccggtcc tcgagcggag atggggcgca ggaaggtgca gctgaagcgg atcgagaaca 180

agatcaaccg ccaggtcacc ttctccaagc gccgctcggg gctgctcaag aaggcgcacg 240

agatctccgt gctctacgac gccgaggtcg gcctcatcat cttctccacc aagggaaagc 300

tctacgagtt ctccaccgag tcatgtatgg acaaaattct tgaacggtat gagcgctact 360

cttatgcaga aaaggttctc gtttcaagtg aatctgaaat tcagggaaac tggtgtcacg 420

aatataggaa actgaaggcg aaggttgaga caatacagaa atgtcaaaag catctcatgg 480

gagaggatct tgaatctttg aatctcaagg agttgcagca actggagcag cagctggaaa 540

gctcactgaa acatatcaga gccaggaaga accaacttat gcacgaatcc atttctgagc 600ttcagaagaa ggagaggtca ctgcaggagg agaataaagt tctccagaagagaagcagaa ggcccaggcg gcgcagcaag atcaaactca gcctcaaacccttcttcctt catgatgagg gatgctcccc ctgtcgcaga taccagcaatcggcaggcga gagggcagag gatgtggcag tgcagcctca ggtcccactcttccactgtg gatggtgagc cacatcaacg gctgaagggc ttccagcccactattcagta cgagtaacaa gttgcagcgg ccagcctggt gtatcatgcgtgctaacccc atggagggga gaggaaaaga aatcagagta aagcagcaaggtgtatattt cacttcgtcc acctcagttt cctttccacc tgggctgagaagtaatctac catgtaattt atatgtagca tgagtgacga attttcaacttccgttgctc ctgggtgttg tttctgtgaa ttaacctatc gaatatgagc <210> 55 <211> 244 <212> PRT

<213> Hordeum vulgare <400> 55

Met Gly Arg Arg Lys Val Gln Read Lys Arg Ile Glu Asn1 5 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Read

20 25

His Glu Ile Ser Val Leu Tyr Asp Wing Glu Val Gly Leu

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Be Thr Glu Ser

50 55 60

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu65 70 75

Val Being Glu Being Glu Ile Gln Gly Asn Trp Cys His

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln

100 105

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu115 120 125

Glu Gln Gln Read Glu Be Be Read Lys His Ile Arg Wing

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys145 150 155

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val

165 170

Lys Wing Gln Wing Wing Gln Gln Asp Gln Thr Gln Pro Gln

180 185

Ser Ser Ser Ser Ser Phe Met Met Asp Wing Pro Pro Val195 200 205

Ser Asn His Pro Wing Wing Wing Gly Glu Arg Wing Wing Glu Asp

210 215 220

Gln Pro Gln Val Pro Read Arg Thr Wing Read Pro Leu Trp225 230 235

His Ile Asn Gly

gaacttgtggagctcttcttcacccagcggcggacggcgctgtaagcgtagttgcgaacactgcaggaattggctgtacgttcgatgatagttgtg

Lys Ile Asn15

Lys Lys Ala30

Ile Ile Phe

Cys Met Asp

Lys Val Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Asn

Glu Arg Ser160

Glu Lys Gln175

Thr Ser Ser190

Asp Thr Wing

Val Wing Val

Met Val Ser240

6607207808409009601020108011401196

<210> 56 <211> 1161 <212> DNA <213> Trit <400> 56ggcacgagctcggcacgagccccggcaggtccgtgctctgagttctccaccagaaaaggtggaaactgaaatcttgaatctgaaacatatagaggcagagagaga

icum aestivum

ctctctctctggagatgggggaccttctcccgacgccgagcgagtcatgttctcgtttcaggcgaaggtttttgaatctccagatccagggtcactgcagggcggcgcaa

ctctctctctcgcgggaaggaagcgccgctgtcggcctcaatggacaaaaagtgaatctggagacaatacaaggagttgcaagaaccaacgaggagaatacaagatcaga

ctctctctcttgcagctgaacggggctgcttcatcttctcttcttgaacgaaattcagggagaaatgtcaagcaactggattatgcacgaaagttctccactcagcctca

ctctctcctcgcggatcgagcaagaaggcgcaccaagggagtatgagcgcaaactggtgtaaagcatctggcagcagctgatccatttctgaaggaactcaacaagctct

gtgccgaattaacaagatcacacgagatctaagctctacgtactcttatgcacgaatataatgggagagggaaagctcacgagcttcagagtcgagaagctcttcttctt

60120180240300360420480540600660ccttcatgat gagggatgct ccccctgccg cagctaccag cattcatcca gcggcggcag 720gcgagagggc aggggatgcg gcagtgcagc cgcaggcccc accccggacg gggcttccac 780tgtggatggt gagccacatc aacggctgaa gggcttccag cccatataag cgtactattc 840agtagagagt aacaagttgc accggccagt ctggtgtatg ttgcggttgc tagcacgcct 900gaccccttgg aggggaaagg aaaagaaatc agagtaaagt agcaagctgc agcgatgtgt 960atatttcact ttgtccaccc cagtttccct cccagctggg ctcaatttac catgtaatct 1020atatgtagct tgagtgatga attttcaagt ttccatgata cccgtctcta gtgggatgtt 1080gtttatgtga attaacctat caaatatgag cattgtgtat attgtgattc ttgaaaataa 1140ataaatcagg atctttgtct t 1161

<210> 57 <211> 244 <212> PRT

<213> Triticum aestivum <400> 57

Met Gly Arg Gly Lys Val Gln Read Le Lys Arg Ile Glu Asn Lys Ile Asn15 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Be Thr Glys Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Val Be Glu Be Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be Arg Lys Asn

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln

165 170 175

Lys Wing Gln Wing Wing Gln Gln Asp Gln Thr

180 185 190

Be Be Be Be Phe Met Met Asp Pro Wing Pro Wing Pro Wing

195 200 205

Ser Ile His Pro Wing Gly Wing Gly Glu Arg Wing Gly Asp Wing Val Wing

210 215 220

Gln Pro Gln Pro Pro Arg Thr Gly Leu Pro Leu Trp Met Val Ser225 230 235 240

His Ile Asn Gly

<210> 58 <211> 1210 <212> DNA

<213> Triticum aestivum <400> 58

tccctctcct ccctctcttc cgcctcaccc aaccacctga cagccatggc tccgcccccc 60

cgcccccgcc tgcgcctgtc ggagtagccg tcgcggtctg ccggtgttgg aggcttgggg 120

tgtagggttg gccccgttct ccagcggaga tggggcgcgg gaaggtgcag ctgaagcgga 180

tcgagaacaa gatcaaccgg caggtgacct tctccaagcg ccgctcgggg ctgctcaaga 240

aggcgcacga gatctccgtg ctctgcgacg ccgaggtcgg cctcatcatc ttctccacca 300

agggaaagct ctacgagttc tccaccgagt catgtatgga caaaattctt gaacggtatg 360

agcgctactc ttatgcagaa aaggttctcg tttcaagtga atctgaaatt cagggaaact 420

ggtgtcacga atataggaaa ctgaaggcga aggttgagac aatacagaaa tgtcaaaagc 480

atctgatggg agaggatctt gaatctttga atctcaagga gttgcagcaa ctggagcagc 540

agctggaaag ctcactgaaa catatcagat ccaggaagaa ccaacttatg cacgaatcca 600

tttctgagct tcagaagaag gagaggtcac tgcaggagga gaataaagtt ctccagaagg 660

aactcgtcga gaagcagaag gcccaggcgg cgcaacaaga tcagactcag cctcaaacaa 720gctcttcttc ttcttccttc atgatgaggg atgctccccc tgccgcaact accagcattc 780

atccagcggc atcaggagag agggcagagg atgcggcagt gcagccgcag gccccacccc 840

ggacggggct tccactgtgg atggttagcc acatcaacgg ctgaagggct tccagcccat 900

ataagcgtac tattcagtag agagtaacaa gttgcaccgg ccagcctggt gtatgttgcg 960

gttgctagca tgcctgaccc cttggagggg aaaggaaaag aaatcagagt aaagtagcaa 1020

gctgcagtga tgtgtatatt tcactttgtc cacctcagtt tccctcccag ctgggctcaa 1080

tttaccatgt aatctatatg tagcttgagt gatgaatttt caagtttcca tgatacccgt 1140

ctcgagcggg tgttgtttat gtgaattaac ctatcaaata tgagcattgt gtaaaaaaaa 1200

aaaaaaaaaa 1210 <210> 59 <211> 244

<212> PRT <213> Triticum, aestivum <400> 59 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys In Asn1 5 10 15 Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Lys Lys Wing 20 25 30 His Glu Ile Being Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe 35 40 45 Being Thr Lys Gly Lys Leu Tyr Glu Phe Being Thr Glu Being Cys Met Asp 50 55 60 Lys Ile Leu Glu. Arg Tyr Glu Arg Tyr Be Tyr Ala Glu Lys Val Leu65 70 75 80Val Be Glu Be Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Leu Glu Be Ser Leu Lys His Ile Arg Ser Arg Lys Asn 130 135 140 Gln Leu Met His Glu Ser Ile Ser Glu Leu Gln Lys Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln 165 170 175 Lys Wing Gln Wing Wing Gln Asp Gln Thr Gln Pro Gln Thr Ser Ser Ser 180 Phe Met Met Arg Asp Wing Pro Pro Wing Wing Thr Thr 195 200 205 Ser Ile His Pro Wing Wing Being Gly Glu Arg Wing Glu Wing Asp Wing Wing Val 210 215 220 Gln Pro Wing Gln Pro Wing Thr Gly Leu Pro Leu Trp Met Val Ser225 230 235 240His Ile Asn Gly

<210> 60 <211> 735 <212> DNA

<213> Triticum monococcum <400> 60

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaggtgacc 60

ttctccaagc gccgctcggg gcttctcaag aaggcgcacg agatctccgt gctctgcgac 120

gccgaggtcg gcctcatcat cttctccacc aagggaaagc tctacgagtt ctccaccgag 180

tcatgtatgg acaaaattct tgaacggtat gagcgctatt cttatgcaga aaaggttctc 240

gtttcaagtg aatctgaaat tcagggaaac tggtgtcacg aatataggaa actgaaggcg 300

aaggttgaga caatacagaa atgtcaaaaa catctcatgg gagaggatct tgaatctttg 360

aatctcaagg agttgcagca actggagcag cagctggaaa gctcactgaa acatatcaga 420

tccaggaaga accaacttat gcacgaatcc atttctgagc tgcagaagaa ggagaggtca 480

ctgcaggagg agaataaagt tctccagaag gaactcgtgg agaagcagaa ggcccatgcg 540

gcgcagcaag atcaaactca gcctcaaacc agctcttctt cttcttcctt catgctgagg 600

gatgctcccc ctgccgcaaa taccagcatt catccagcgg cggcaggcga gagggcagag 660

gatgcggcag tgcagccgca ggccccaccc cggacggggc ttccaccgtg gatggtgagc 720cacatcaacg ggtga <210> 61 <211> 244 <212> PRT

<213> Triticum monococcum <400> 61

Met Gly Arg Gly Lys Val Gln Leu1 5

Arg Gln Val Thr Phe Ser Lys Arg20

His Glu Ile Ser Val Leu Cys Asp

35 40

Being Thr Lys Gly Lys Leu Tyr Glu

50 55

Lys Ile Read Glu Arg Tyr Glu Arg65 70

Val Ser Ser Glu Ser Glu Ile Gln85

Lys Leu Lys Wing Lys Val Glu Thr100

Met Gly Glu Asp Leu Glu Ser Leu115 120

Glu Gln Gln Leu Glu Be Ser Leu

130 135

Gln Read Met His Glu Ser Ile Ser145 150

Leu Gln Glu Glu Asn Lys Val Leu165

Lys Wing His Wing Wing Gln Gln Asp180

Ser Ser Ser Ser Phe Met Leu Arg195 200

Ser Ile His Pro Wing Wing Wing Gly

210 215

Gln Pro Gln Pro Pro Arg Thr225 230

His Ile Asn Gly

735

Lys Arg Ile Glu Asn Lys Ile Asn 10 15 Arg Ser Gly Leu Read Le Lys Lys Ala25 30 Wing Glu Val Gly Leu Δ R Ile Ile PhePhe Be Thr Glu 4 D Be Cys Met Asp 60 Tyr Be Tyr Wing Glu Lys Val Leu 75 80Gly Asn Trp Cys His Glu Tyr Arg 90 95 Ile Gln Lys Cys Gln Lys His Leu105 110 Asn Leu Lys Glu Leu Gln Leu 125 Lys His Ile Arg Be Arg Lys Asn 140 Glu Leu Gln Lys Lys Glu Arg Ser 155 160Gln Lys Glu Leu Val Glu Lys Gln 170 175 Gln Thr Gln Pro Gln Thr Ser185 190 Asp Pro Wing Pro Wing Asn Thr 205 Wing Glu Arg Wing Glu Asp Wing Wing Val 220 Gly Leu Pro Pro Trp Met Val Ser 235 240

<210> 62 <211> 735 <212> DNA <213> Triti <400> 62atggggcgggttctccaagcgccgaggtcgtcatgtatgggtttcaagtgaaggttgagaaatctcaaggtccaggaagactgcaggagggcgcagcaaggatgctccccgatgcggcagcacatcaacg <210> 63 <211> 244 <212> PRT <213> Triti <400> 63Met Gly Arg1

cum aestivum

ggaaggtgcagccgctcggggcctcatcatacaaaattctaatctgaaatcaatacagaaagttgcagcaaccaacttatagaataaagtatcaaactcactgccgcaaatgcagccgcaggtga

gctgaagcgggcttctcaagcttctccacctgaacggtattcagggaaacatgtcaaaagactggagcaggcacgaatcctctccagaaggcctcaaacctaccagcattggccccaccc

atcgagaacaaaggcgcacgaagggaaagcgagcgctatttggtgtcacgcatctcatggcagctggaaaatttctgagcgaactcgtggagctcttcatcatccagcggcggacggggg

agatcaaccgagatctccgttctacgagttcttatgcagaaatataggaagagaggatctgcctcactgaattcagaagaaagaagcagaacttcttccttcaacaggcgattccaccgtg

gcaggtgaccgctctgcgacctccaccgagaaaggttctcactgaaggcgtgaatctttgacatatagaggagaggtcaggcccatgcgcatgctgagggagggcagaggatggtgagc

cum aestivum

Gly Lys Val Gln Read Lys Arg Ile5 10

60120180240300360420480540600660720735

Glu Asn Lys Ile Asn15His Ile Asn Gly <210> 64 <211> 738 <212> DNA

<213> Lolium perenne <400> 64

atggggcgcg gcaaggtgca gctcaagcgg atcgagaacattctccaagc gccgctcggg gctgctcaag aaggcgcacggccgaggtcg ggctcatcat cttctccacc aagggaaagctcatgtatgg acaaaattct tgagcggtat gagcgctactatttcaaccg aatctgaaat tcagggaaac tggtgtcatgaaggttgaga caatacagag atgtcaaaag catctaatggaatctcaagg agttgcagca actagagcag cagctggaaagccagaaaga accagcttat gcacgaatcc atatctgagcctgcaggagg agaataaaat tctccagaag gaactcatagcagcaagcgc agtgggagca aactcagccc caaaccagctatgggggaag ctaccccagc aacaaattgc agtaatccccgcagaggatg cgacggggca gccttcagct cgcacggtgccacatcaaca atggctga <210> 65 <211> 245 <212> PRT

<213> Lolium perenne <400> 65

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

Ser Gly Leu Leu Lys Lys Wing 30 Glu Val Gly Leu δ ^ Ile Ile PheSer Thr Glu * ± 3 Ser Cys Met Asp 60 Ser Tyr Wing Glu Lys Val Leu 75 80Asn Trp Cys His Glu Tyr Arg90 95 Gln Lys Cys Gln Lys His Leu 110 Leu Lys Glu Leu Gln Gln Leu 125 His Ile Arg Be Arg Lys Asn 140 Leu Gln Lys Lys Glu Arg Be 155 160Lys Glu Leu Val Glu Lys Gln170 175 Thr Gln Pro Gln Thr Be Ser 190 Wing Pro Pro Wing Asn Thr 205 Arg Glu Wing Asp Wing Val Wing 220 Leu Pro Pro Trp Met Val Ser 235 240

agatcaaccgagatctccgttctacgagttcctatgcagaaatataggaagagaggatctgttcactgaattcaaaagaaagaagcagaacttcctcctccagcagcggcttccaccatg

ccaggtcaccgctctgcgaccgcaaccgacgaaagtgctcactgaaggcgtgaatcattgacatattagaggagaggtcaggcccacacgctcctttatgcagcgacagagatggtgagt

20

25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35

40

Being Thr Lys Gly Lys Read Tyr Glu Phe Wing Thr

50

55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr

65

70

75

Ile Be Thr Glu Ile Be Glu Ile Gln Gly Asn Trp

Glu Asn Lys Ile Asn15

Leu Leu Lys Lys Ala30

Gly Leu Ile Ile Phe45

Asp Ser Cys Met Asp60

Glu Wing Lys Val Leu80

Cys His Glu Tyr Arg

6012018024030036042048054060066072073885 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Arg Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Be Read Lys His Ile Arg Wing Arg Lys Asn

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Ile Glu Lys Gln

165 170 175

Lys Wing His Thr Gln Gln Wing Gln Trp Glu Gln Thr Gln Pro Gln Thr

180 185 190

Be Ser Ser Ser Ser Ser Phe Met Met Gly Glu

195 200 205

Asn Cys Be Asn Pro Pro Wing Wing Wing Be Asp Arg Wing Glu Asp Wing

210 215 220

Thr Gly Gln Pro Ser Wing Arg Thr Val Leu Pro Pro Trp Met Val Ser225 230 235 240

His Ile Asn Asn Gly245

<210> 66 <211> 738 <212> DNA

<213> Lolium temulentum <400> 66

atggggcgcg gcaaggtgca gctcaagcgg atcgagaaca agatcaaccg ccaggtcacc 60

ttctccaagc gccgctcagg cctgctcaag aaggcgcacg agatctccgt gctctgcgac 120gcagaggtcg ggctcatcat cttctccacc aagggaaagc tctacgagtt cgccaccgac 180tcatgtatgg acaaaattct tgagcggtat gagcgctact cctatgcaga gaaagtgctc 240atttcaactg aatctgaaat tcagggaaac tggtgtcatg aatataggaa actgaaggcg 300aaggttgaga caatacagag atgtcaaaag catctaatgg gagaggatct tgaatcattg 360aatctcaagg agttgcagca actagagcag cagctggaaa gttcactgaa acatattaga 420tccagaaaga gccagcttat gcacgaatcc atatctgagc ttcaaaagaa ggagaggtca 480ctgcaagagg agaataaaat tctccagaag gaactcatag agaagcagaa ggcccacacg 540cagcaagcgc agttggagca aactcagccc caaaccagct cttcctcctc ctcctttatg 600atgggggaag ctaccccagc aacaaatcgc agtaatcccc cagcagcggc cagcgacaga 660gcagaggatg cgacggggca gcctccagct cgcacggtgc ttccaccatg gatggtgagt 720cacctcaaca atggctga 738

<210> 67 <211> 245 <212> PRT

<213> Lolium temulentum <400> 67

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Wing Thr Asp Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Ile Be Thr Glu Be Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Arg Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be Arg Lys Be

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Ile Glu Lys Gln

165 170 175

Lys Wing His Thr Gln Gln Wing Gln Read Glu Gln Thr Gln Pro Gln Thr

180 185 190

Be Ser Ser Ser Ser Ser Phe Met Met Gly Glu

195 · 200 205

Asn Arg Be Asn Pro Pro Wing Wing Wing Be Asp Arg Wing Glu Asp Wing

210 215 220

Thr Gly Gln Pro Pro Arg Wing Thr Val Leu Pro Pro Trp Met Val Ser225 230 235 240

His Leu Asn Asn Gly245

<210> 68 <211> 738 <212> DNA <213> Zea mays <400> 68

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg ccaggtgacc 60

ttctccaagc gccgctcggg gctgctcaag aaggcgcacg agatctccgt gctctgcgac 120gccgaggtcg cgctcatcat cttctccacc aaagggaagc tctacgagta ttccaccgat 180tcatgtatgg acaaaattct tgaccggtac gagcgctact cctatgcaga aaaggttctt 240atttcagcag aatctgaaac tcagggcaat tggtgccacg agtatagaaa actaaaggcg 300aaggtcgaga caatacaaaa atgtcaaaag cacctcatgg gagaggatct tgaaacgttg 360aatctcaaag agcttcagca actagagcag cagctggaga gttcactgaa acatatcaga 420accaggaaga accaacttat gctcgagtca atttcggagc tccaacggaa ggagaagtcg 480ctgcaggagg agaacaaggt tctgcagaag gagctcgcgg agaagcagaa agcccagcgg 540aagcaagtgc aatggggcca aacccaacag cagaccagtt cgtcttcctc gtgcttcgtg 600ataagggaag ctgccccaac aacaaatatc agcatttttc ctgtggcagc aggcgggagg 660ttggtggaag gtgcagcagc gcagccacag gctcgcgttg gactaccacc atggatgctt 720agccacctga gcagctga 738

<210> 69 <211> 245 <212> PRT <213> Zea mays <400> 69

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Lys Lys Wing 20 25 30 His Glu Ile Be Val Leu Cys Asp Wing Glu Val Wing Leu Ile Ile Phe 35 40 45 Be Thr Lys Gly Lys Read Tyr Glu Tyr Be Thr Asp Be Cys Met Asp 50 55 60 Lys Ile Read Asp Arg Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Val Leu65 70 75 80Ile Be Wing Glu Be Glu Thr Gln Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Thr Leu Asn Leu Lys Glu Leu Gln Leu 115 120 125 Glu Gln Gln Glu Leu Glu Being Ser Liu His Ile Arg Thr Arg Lys Asn 130 135 140 Gln Leu Met Leu Glu Being Ile Being Glu Leu Gln Arg Lys Glu Lys Ser145 150 155 160Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Glu Wing Lys Gln 165 170 175 Lys Wing Gln Arg Lys Gln Val Gln Gln Thr 180 185 190 Ser Ser Ser Ser Ser Ser Cys Phe Val Ile Arg Glu Wing Pro Pro Thr Thr 195 200 205 Asn Ile Being Ile Phe Pro Val Wing Gly Gly Arg Leu Val Glu Gly210 215

Wing Wing Wing Gln Pro Wing Gln Arg Wing Val Gly Leu225 230 235

Be His Leu Ser Ser245

<210> 70 <211> 738 <212> DNA <213> Zea mays <400> 70

atggggcgcg ggaaggtgca gctgaagcgg atcgagaacattctccaagc gccgctcggg gctactcaag aaggcgcacggccgaggtcg cgctcatcat cttctccacc aagggcaagctcatgtatgg acaaaattct tgaacggtat gagcgctactatttccgcag aatatgaaac tcagggcaat tggtgccatgaaggtcgaga caatacagaa atgtcaaaag cacctcatggaatctcaaag agcttcagca actagagcag cagctggagaacaaggaaga gccagcttat ggtcgagtca atttcagcgcctgcaggagg agaacaaggt tctgcagaag gagctcgcggcagcaagtgc aacgggacca aactcaacag cagaccagttttaagggaag ctgccccaac aacaaatgtc agcatcttccgtggtggaag gggcagcagc gcagccgcag gctcgcgttgagccatctga gctgctga <210> 71 <211> 245 <212> PRT <213> Zea mays <400> 71

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Ser Thr Lys Gly Lys. Read Tyr Glu Tyr Ser Thr

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Ile Ser Wing Glu Tyr Glu Thr Gln Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys

100 105

Met Gly Glu Asp Leu Glu Thr Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130 135

Gln Leu Met Val Glu Ser Ile Ser Wing Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Val

165 170

Lys Asp Gln Arg Gln Gln Val Gln Arg Asp Gln

180 185

Be Ser Be Ser Thr Be Phe Met Leu Arg Glu

195 200

Asn Val Ser Ile Phe Pro Val Wing Gly Gly Wing

210 215

Wing Wing Wing Gln Pro Wing Gln Arg Wing Val Gly Leu225 230 235

Be His Leu Ser Cys245

<210> 72 <211> 762 <212> DNA

220

Pro Pro Trp Met Leu240

agatcaaccgagatctccgttctacgagtacctatgcagaaatatagaaagagaggatctgttcactgaatccaacggaaagaagcagaacgtcttccacctgtggcagcgactgccacc

ccaggtgacagctctgcgacctctaccgataaaggttctcactaaaggcgtgaaactttgacatatcagaggagagtcaagaccagcgggcccccatcatagaggcgggaggatggatgctt

Glu Asn

Leu Leu

Wing Leu

45Asp Ser60

Glu Wing

Cys his

Cys Gln

Glu Leu125Arg Thr140

Arg lys

Read Wing

Thr gln

Wing Wing205Arg Val220

Pro pro

Lys ile

15Lys Lys30

Ile Ile

Cys met

Lys val

Glu Tyr

95Lys His110

Gln Gln

Arg lys

Glu Lys

Glu Lys175Gln Gln190

Pro ThrVal GluTrp Met

Asn

Allah

Phe

Asp

Leu80Arg

Read

Read

To be

Ser160Gln

Thr

Thr

Gly

Leu240

60120180240300360420480540600660720738 <213> Oryza sativa <400> 72

atggggcggg gcaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaggtgaccttctccaagc gcaggtcggg gctgctcaag aaggcgaatg agatctccgt gctctgcgacgccgaggtcg cgctcatcat cttctccacc aagggcaagc tctacgagta cgccaccgactcatgtatgg acaaaatcct tgaacgttat gagcgctact cctatgcaga aaaggtccttatttcagctg aatctgacac tcagggcaac tggtgccacg aatataggaa actgaaggctaaggttgaga caatacagaa atgtcaaaag cacctcatgg gagaggatct tgaatctttgaatctcaaag agctgcagca gctggagcag cagctggaaa attcgttgaa acatatcagatccagaaaga gccaactaat gctcgagtcc attaacgagc ttcaacggaa ggaaaagtcactgcaggagg agaataaggt cctacagaaa gaaaaccctt gctccttcct acagctggtggagaagcaga aagtccagaa gcaacaagtg caatgggacc agacacaacc tcaaacaagttcctcatcat cctccttcat gatgagggaa gcccttccaa caactaatat cagtaactaccctgcagcag ctggcgaaag gatagaggat gtagcagcag ggcagccaca gcatgttcgcattgggctgc caccatggac <t2> 25 <2>

<213> Oryza sativa <400> 73

Gly Arg Gly Lys Val Gln Read Lys Arg Ile Glu Asn5 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Read

Met1

20

25

Asn Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35

40

Being Thr Lys Gly Lys Read Tyr Glu Tyr Wing Thr

50

55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr

65

70

75

Ile Be Wing Glu Be Asp Thr Gln Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys

100 105

Met Gly Glu Asp Leu Glu Ser Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Asn Be Read Lys His Ile

130 135

Gln Leu Met Leu Glu Ser Ile Asn Glu Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Val

165 170

Leu Gln Leu Val Glu Lys Gln Lys Val Gln Lys

180 185

Asp Gln Thr Gln Pro Gln Thr Ser Ser Ser Ser

195 200

Arg Glu Wing Read Pro Thr Thr Asn Ile Ser Asn

210 215

Gly Glu Arg Ile Glu Asp Val Wing Wing Gly Gln225 230 235

Ile Gly Leu Pro Pro Trp Met Leu Be His Ile245 250

<210> 74

<211> 741

<212> DNA

<213> Oryza sativa

<400> 74

atggggcggg gcaaggtgca gctgaagcgg atcgagaacattctccaagc gcaggtcaaa actgctcaag aaggcgaatggccgaggtcg cgctcatcat cttctccacc aagggcaagctcatgtatgg acaaaatcct tgaacgttat gagcgctactatttcagctg aatctgacac tcagggcaac tggtgccacgaaggttgaga caatacagaa atgtcaaaag cacctcatgg

Leu45 Wing

Asp Ser60

Glu Wing

Cys his

Cys Gln

Glu Leu125Arg Ser140

Arg lys

Asn pro

Gln Gln

Ser Ser205Tyr Pro220

Pro GlnAsn Gly

Lys

Lys

30

Ile

Cys

Lys

Glu

Lys110Gln

Arg

Glu

Cys

Val190Phe

Allah

His

Ile Asn15

Lys Wing

Ile Phe

Met asp

Val Leu80Tyr Arg95

His leu

Gln Leu

Lys Ser

Lys Ser160Ser Phe175

Gln Trp

Met met

Wing wing

Val Arg240

agatcaaccgagatctccgttctacgagtacctatgcagaaatataggaagagaggatct

gcaggtgaccgctctgcgaccgccaccgacaaaggtccttactgaaggcttgaatctttg

60120180240300360420480540600660720762

60120180240300360aatctcaaag agctgcagca gctggagcag cagctggaaatccagaaaga gccaactaat gctcgagtcc attaacgagcctgcaggagg agaataaggt cctacagaaa gaactggtggcaacaagtgc aatgggacca gacacaacct caaacaagttatgagggaag cccttccaac aactaatatc agtaactaccatagaggatg tagcagcagg gcagccacag catgaacgcactgagccaca the tcaacggcta <210> 75 <211> 246 <212> PRT

attcgttgaattcaacggaaagaagcagaacctcatcatcctgcagcagcttgggctgcc

acatatcagaggaaaagtcaagtccagaagctccttcatgtggcgaaaggaccatggatg

420480540600660720741

<400> 75 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Be Lys Arg Arg Be Lys Leu Read Lys Lys Ala 20 25 30 Asn Glu Ile Be Val Leu Cys Asp Glu Val Wing Wing Leu Ile Ile Phe 35 40 45 Ser Thr Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Ser Cys Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80Ile Ser Glu Wing Be Asp Thr Gln Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Val Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Be Leu Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Leu Glu Asn Ser Leu Lys His Ile Arg Ser Arg Lys Ser 130 135 140 Gln Leu Met Leu Glu Ser Ile Asn Glu Leu Gln Arg Lys Glu Lys Ser145 150 155 160 160Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Read Val Glu Lys Gln 165 170 175 Lys Val Gln Lys Gln Gln Val Gln Trp Asp Gln Thr Gln Pro Gln Thr 180 185 190 Ser Ser Ser Ser Ser Ser Ser P P Met Met Glu Wing Leu Pro Thr Thr 195 200 205 Asn Ile Be Asn Tyr Pro Wing Wing Gly Glu Arg Ile Glu Asp Val 210 215 220 Wing Gly Wing Gln Pro Gln His Glu Arg Ile Gly Leu Pro Pro Trp Met

225

235

230

Read His Ile Asn Gly245

<210> 76 <211> 735 <212> DNA

<213> Dendrocalamus latiflorus <400> 76

atggggcgcg ggaaggtgca gctgaagcgg atcgagaacattctccaagc gccggtcggg gctgctcaag aaggcgcatggccgaggtcg gccttatcat cttctccacc aagggcaagctcatgtatgg acaaaattct tgaacggtac gagcgttactatttcagccg aatctgaaac tcagggcaac tggtgtcacgaaggttgaga cgatacagaa atgtcaaaag cacctcatggaatctcaagg agctgcagca actcgagcag cagctggaaatccagaaaga gccagcttat gctcgagtcc atttccgagcctgcaggagg agaacaaggt tctgcagaag gaactcgtggcggttagtgc aatgggacca aactcagccg caaactagttatgagggaag ctctcccaac aacaaatatc agtatttacggcagaggacg cagcagggca gcctcagatt cacattgggccacatcaacg gctaa <210> 77

240

agatcaaccgagatctccgttctacgagtacctatgcagaaatataggaagagaggatccgttcagtgaattcaaaagaaagaagcagcacctcttcctcctgcggcagctgccgccatg

gcaagtgactcctctgcgaccgccaccgacaaaggttcttactgaaggcgtgaatctttgacatatcagaggagaagtcaggtccataaagtccttcatgcggcgagagggatggtgagc

60120180240300360420480540600660720735 <211> 244 <212> PRT

<213> Dendrocalamus <400> 77

Iatiflorus

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Leu Lys Lys Wing 20 25 30 His Glu Ile Be Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe 35 40 45 Ser Thr Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Ser Cys Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Glu Lys Val Leu65 70 75 80Ile Ser Ala Glu Ser Glu Thr Gln Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu 100 105 110 Met Gly Glu Asp Pro Glu Being Leu Asn Leu Lys Glu Leu Gln Leu 115 120 125 Glu Gln Gln Glu Leu Glu Ser Be Val Lys His Ile Arg Be Arg Lys Ser 130 135 140 Gln Leu Met Leu Glu Be Ile Be Glu Leu Gln Lys Glu Lys Ser145 150 155 160Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln 165 170 175 Gln Val His Lys Arg Read Val Val Gln Trp Asp Gln Thr Gln Pro Gln Thr 180 185 190 Be Be Be Be Be Be Phe Met Met Arg Glu Wing Leu Pro Thr Thr 195 200 205 Asn Ile Be Ile Tyr Wing Wing Gly Wing Wing Glu Arg Wing Glu Wing Asp Wing 210 215 220 Wing Gly Gln Pro Gln Ile His Ile Gly Read Pro Trp Met Val Ser225 230 235 240His Ile Asn Gly

<210> 78 <211> 735 <212> DNA

<213> Dendrocalamus Iatiflorus <400> 78

atggggcgcg ggaaggtgca gctgaagcgg atcgagaacattctccaagc gccggtcggg gctgctcaag aaggcgcatggccgaggtcg gccttatcat cttctccacc aagggcaagctcatgtatgg acaaaattct tgaacggtac gagcgttactatttcagccg aatctgaaac tcagggcaac tggtgtcacgaaggttgaga cgatacagaa atgtcaaaag cacctcatggaatctcaagg agctgcagca actcgagcag cagctggaaatccagaaaga gccagcttat gctcgagtcc atttccgagcctgcaggagg agaacaaggt tctgcagaag gaactcgtggcggttagtgc aatgggacca aactcagccg caaactagttatgagggaag ctctcccaac aacaaatatc agtatttacggcagaggacg cagcagggca gcctcagatt cacattgggccacatcaacg gctaa <210> 79 <211> 244 <212> PRT

<213> Dendrocalamus Iatiflorus <400> 79

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly20 25

agatcaaccgagatctccgttctacgagtacctatgcagaaatataggaagagaggatctgttcagtgaattcaaaagaaagaagcagca

CCtCttCCtC

ctgcggcagctgccgccatg

gcaagtgactcctctgcgaccgccaccgacaaaggttcttactgaaggcgtgaatctttgacatatcagaggagaagtcaggtccataaagtccttcatgcggcgagagggatggtgagc

Glu Asn Lys Ile Asn15

Leu Leu Lys Lys Ala30

60120180240300360420480540600660720735His Glu Ile Ser35

Ser Thr Lys Gly50

Lys Ile Leu Glu65

Ile Ser Ala Glu

Lys Leu Lys Ala100

Met Gly Glu Asp115

Glu Gln Gln Leu130

Gln Leu Met Leu145

Read Gln Glu Glu

Gln Val His Lys180

Be Ser Ser Ser195

Asn Ile Ser Ile210

Gly Gln Pro225 Wing

His Ile Asn Gly

80

Val Leu CysLysArg

40

To be

85

Lys

Read

Glu

Glu

Asn165Arg

To be

Tyr

Gln

Read tyr

55Tyr Glu70

Glu thr

Val glu

Glu Ser

Ser Ser135Ser Ile150

Lys val

Read val

To be Phe

Wing Ala215Ile His230

120

200

Glu Wing Val Gly Leu 45 Ile Ile PheTyr Wing Thr Asp 60 Ser Cys Met AspTyr Ser Tyr 75 Wing Glu Lys Val Leu 80Gly Asn 90 Trp Cys His Glu Tyr 95 ArgIle Gln Lys Cys Gln Lys His Leu105 110 Asn Leu Lys Glu Leu 125 Gln Gln LeuLys His Ile Arg 140 Ser Arg Lys SerGlu Leu Gln 155 Lys Lys Glu Lys Ser 160Gln Lys 170 Glu Leu Val Glu Lys 175 GlnTrp Asp Gln Thr Gln Pro Gln Thr185 190 Met Arg Glu Wing Leu 205 Pro Thr ThrAla Gly Glu Arg 220 Wing Glu Asp AlaGly Leu Pro 235 Pro Trp Met Val Ser 240

<210> <211> 813 <212> DNA <213> Zea mays <400> 80

atggggcgcg gcaaggtgca gctgaagcgg atagagaacattctccaagc gccggaacgg gctgctgaag aaggcgcacggccgaggtcg ccgtcatcgt cttctccccc aagggcaagctcccgcatgg acaaaattct agaacgttat gagcgatattatttcagctg aatctgaaag tgagggaaat tggtgccacgaaaattgaga ccatacaaag atgccacaag cacctgatggaatccaaaag agctccaaca actagagcag cagctggagatcaagaaaga gccaccttat ggccgagtca atttctgagcctgcaggagg agaacaagat tctacagaag gaactttcagagccggcagc agcagcagca gcaggtgcag tgggaccagcacaagctcat cgtcttcttc cttcatgatg aggcaggatccaaaacatct gcttcccgcc gttgagcatc ggagagagaggcgcagcagc agctgcctcc tccggggcag gcgcaaccacccgccatgga tgctgagcca cctcaatgca taa <210> 81 <211> 270 <212> PRT <213> Zea mays <400> 81

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile1 5 10

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly

agataaaccgagatctccgttctacgagtacctatgctgaaatacaggaagaggatctgctcactgaatacagaagaagaggcagaaagacacaggtagcagggactgcgaagaggtagctccgcat

gcaggtgacccctctgcgaccgcctccgacaaaggctcttactgaaggccagagtctttggcacatcagaggagaggtcaggcggtcgctccaggtccaggccacctccaggctgcggcgcgcaggtctg

20

25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35

40

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Ser

50

55

Lys

65

Ile

Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr

70 75

Be Glu Wing Be Glu Be Glu Gly Asn Trp

Glu Asn Lys Ile Asn1 5

Leu Leu Lys Lys Ala30

Val Ile Val Phe45 Ward

Asp Be Arg Met Asp60

Glu Lys Wing Wing Leu80

Cys His Glu Tyr Arg

6012018024030036042048054060066072078081385 90 95

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Arg Cys His Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Pro Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be Arg Lys Be

130 135 140

His Leu Met Wing Glu Ser Ile Be Glu Leu Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Be Glu Arg Gln

165 170 175

Lys Wing Val Wing Be Arg Gln Gln Gln Gln Gln Gln Val Gln Trp Asp

180 185 190

Gln Gln Thr Gln Val Gln Val Gln Thr Be Ser Ser Ser Ser Ser Phe

195 200 205

Met Met Arg Gln Asp Gln Gln Gly Read Pro Pro Gln Asn Ile Cys

210 215 220

Phe Pro Pro Read It Ile Gly Glu Arg Gly Glu Glu Val Wing Wing Wing225 230 235 240

Gln Wing Gln Gln Leu Pro Pro Gly Gln Gln Wing Pro Gln Leu Arg

245 250 255

Ile Ala Gly Leu Pro Pro Trp Met Leu Be His Leu Asn Ala260 265 270

<210> 82 <211> 689 <212> DNA

<213> Sorghum bicolor <400> 82

gcaaggtgca gctcaagcgg atagagaaca agataaaccg gcaggtgacc ttctccaagc 60

gccgcaacgg gctgctcaag aaggcgcacg agatctccgt cctctgcgac gccgaggtcg 120ccgtcatcgt cttctccccc aagggcaagc tctatgagta cgccaccgac tcccgcatgg 180acaaaattct cgaacgttat gagcgatatt cctatgctga aaaggctctt atttcagctg 240aatctgaaag tgagggaaac tggtgccacg aatacaggaa actgaaggcc aaaattgaga 300ccattcaaaa atgccacaag cacctgatgg gagaggatct agagtctttg aatcccaaag 360agctccaaca actagagcag cagctggaga gctcactgaa gcacatcaga tcaagaaaga 420gccaccttat ggctgagtct atttctgaac tacagaagaa ggagaggtca ctgcaggagg 480agaacaaggc tctacagaag gaacttgcgg agaggcagaa ggcggccgcg agcaggcagc 540agcagcaagg tgcagtggga ccagcagaca cagacccagg cccagacaag ctcatcatcg 600tcctccttca tgatgaggca ggatcagcag ggtctgccgc ctccacaaaa catatgcttc 660ccgccgctga taatcggaga gagaggtga 689

<210> 83 <211> 228 <212> PRT

<213> Sorghum bicolor <400> 83

Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn Arg Gln Val Thr15 10 15

Phe Ser Lys Arg Asn Gly Leu Leu Read Lys Lys Wing His Glu Ile Ser

20 25 30

Val Leu Cys Asp Glu Wing Val Wing Val Ile Val Phe Ser Pro Lys Gly

35 40 45

Lys Leu Tyr Glu Tyr Wing As Thr Be Arg Met Asp Lys Ile Leu Glu

50 55 60

Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Wing Leu Ile Ser Wing Glu65 70 75 80

Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg Lys

85 90 95

Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu Met Gly Glu Asp

100 105 110

Leu Glu Be Leu Asn Pro Lys Glu Leu Gln Gln Leu Glu Gln Gln Leu

115 120 125

Glu Ser Be Leu Lys His Ile Arg Be Arg Lys Be His Leu Met Ala130 135 140Glu Be Ile Be Glu Read Le Gln Lys Lys Glu Arg145 150 155

Asn Lys Wing Leu Gln Lys Glu Leu Wing Glu Arg

165 170

Be Arg Gln Gln Gln Gln Gly Wing Val Gly Pro

180 185

Gly Pro Asp Lys Leu Ile Ile Val Leu Leu His

195 200

Wing Gly Be Wing Wing Be Thr Lys His Met Leu

210 215

Arg Arg Glu Arg225

<210> 84 <211> 822 <212> DNA <213> Zea mays <400> 84

atggggcgcg gcaaggtaca gctgaagcgg atagagaacattctccaagc gccggaacgg cctgctcaag aaggcgcacggccgaggtcg ccgtcatcgt cttctccccc aagggcaagctcccgcatgg acaaaattct tgaacgctat gagcgatattatttcagctg aatctgaaag tgagggaaat tggtgccacgaaaattgaga ccatacaaaa atgccacaag cacctgatggaatcccaaag agctccagca actagagcag cagctggatatcaaggaaga gccaccttat ggccgagtct atttctgagcctgcaggagg agaacaaggc tctgcagaag gaacttgcggagccggcagc agcagcaaca gcagcaggtg cagtgggacccagacaagct catcatcgtc ctccttcatg atgaggcaggccacacaaca tctgcttccc gccgttgaca atgggagatagcggcggcgg cgcagcagca gcagccactg ccggggcagggcaggtctgc caccatggat gctgagccac ctcaatgcat <210> 85 <211> 273 <212> PRT <213> Zea mays <400> 85

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Thr

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys

100 105

Met Gly Glu Asp Read Glu Be Read Asn Pro Lys

115 120

Glu Gln Gln Read Asp Ser Be Read Lys His Ile

130 135

His Leu Met Wing Glu Ser Ile Ser Glu Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu

165 170

Lys Wing Val Wing Be Arg Gln Gln Gln Gln Gln

180 185

Asp Gln Gln Thr His Wing Gln Wing Gln Thr Be

195 200

Phe Met Met Arg Gln Asp Gln Gln Gly Leu Pro

Serve Leu Gln Glu Glu160

Gln Lys Wing Wing Wing175

Wing Asp Thr Asp Pro190

Asp Glu Wing Gly Ser205

Pro Wing Asp Wing Asn220

agataaaccgagatctccgttctacgagtacctatgctgaaatacaggaagaggatctgctcactgaatacagaagaaagaggcagaaagcagacacaatcagcaggggggtgaagggacaccgcaaa

gcaggtgacccctctgcgatcgccaccgacaaaggctcttactgaaggccagagtctttggcacatcagagagaggtcaggccgtcgcgtgcccaggccactgccgcctgctggctgcggctccgcatc

Glu Asn

Leu Leu

Val45 wing

Asp Ser60

Glu Wing

Cys his

Cys his

Glu Leu125Arg Ser140

Lys Lys

Read Wing

Gln Gln

Ser Ser205Pro Pro

Lys ile

15Lys Lys30

Ile val

Arg Met

Lys Wing

Glu Tyr

95Lys His110

Gln Gln

Arg lys

Glu Arg

Glu Arg175Val Gln190

To be to be

Asn

Allah

Phe

Asp

Leu80Arg

Read

Read

To be

Ser160Gln

60120180240300360420480540600660720780822

TrpSerHis Asn Ile210 215

Cys Phe Pro Pro Read Thr Met Gly Asp Arg Gly225 230 235

Wing Wing Wing Wing Gln Gln Gln Gln Pro Leu Pro

245 250

Gln Leu Arg Ile Wing Gly Leu Pro Pro Trp Met260 265

Allah

220

Glu Glu Leu Wing Ala240

Gly Gln Wing Gln Pro255

Leu Being His Leu Asn270

<210> 86 <211> 786 <212> DNA

<213> Lolium temulentum <400> 86

atgggtcgcg gcaaggtgca gctgaagcgg atagagaaca agataaaccgttctccaagc gccgcaacgg gctactcaag aaggcgcacg agatctccgtgccgaggtcg ccgtcgtcgt cttctccccg aaagggaagc tctatgagtatccagcatgg acaaaattct tgaacgttat gaacgctact cttatgctgaatttcagctg aatctgaaag tgagggaaat tggtgccatg aatacaggaaaagattgaga ctatacaaaa atgtcacaag cacctcatgg gggaggatctaacctgaaag agctccaaca actagagcag cagctggaga gttcattgaatcgagaaaga gccaccttat gatggagtcc atttctgagc tacagaagaactccaggagg agaacaaggc tctacagaag gaactggtgg agaggcagaacagcagcagc aagagcagtg ggaccgtcag acccaaacac aacaagcccacaggcccaga cgagctcatc atcttcctcc ttcatgatga gggatcagcacaacaaaaca tctgttaccc gctggtgaca atgggtggag aggctgtggcgggcagcagg ggcagcttcg catcggaggc ctgccaccat ggatgctgaggcttga <210> 87 <211> 261 <212> PRT

<213> Lolium temulentum <400> 87

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile Glu Asn15 10

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Wing Wing

35 40 45

Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Ser

50 55 60

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu65 70 75

Ile Ser Ala Glu Ser Glu Ser Glu Gly Asn Trp Cys His

85 90

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys Cys His

100 105

Met Gly Glu Asp Leu Glu Cys Leu Asn Leu Lys Glu Leu115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be

130 135 140

His Leu Met Met Glu Be Ile Be Glu Read Gln Lys Lys145 150 155

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val

165 170

Lys Wing Wing Wing Gln Gln Gln Gln Gln Glu Gln Trp Asp Arg

180 185

Thr Gln Gln Wing Gln Asn Gln Pro Gln Wing Gln Thr Ser195 200 205

Be Ser Phe Met Met Asp Gln Gln Wing His Wing Gln

210 215 220

Cys Tyr Pro Read Val Val Met Gly Gly Glu Wing Val Wing Wing225 230 235

tcaggtgacacctctgcgaccgccactgacaaaggctttggctgaaggcgggagtgtctagcacatagaggagcggtcaggcggccaggaaaccaacctggcccatgctcgcggcgccaccacctcaac

Lys Ile Asn15

Lys Lys Ala30

Val Val Phe

Be Met Asp

Lys Wing Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Ser

Glu Arg Ser160

Glu Arg Gln175

Gln Thr Gln190

To be to be

Gln Asn Ile

Wing Wing Pro240

60120180240300360420480540600660720780786Gly Gln Gln Gly Gln Read Arg Ile Gly Gly Read

245 250

Being His Leu Asn Ala260

<210> 88 <211> 786 <212> DNA

<213> Lolium perenne <400> 88

atgggtcgcg gcaaggtgca gctgaagcgg atagagaacattctccaagc gccgcaacgg gctactcaag aaggcgcacggccgaggtcg ccgtcgtcgt cttctccccg aaagggaagctccagcatgg acaaaattct tgaacgttat gaacgctactatttcagctg aatctgaaag tgagggaaat tggtgccatgaagattgaga ctatacaaaa atgtcacaag cacctcatggaacctgaaag agctccaaca actagagcag cagctggagatcgagaaaga gccaccttat gatggagtcc atttctgagcctccaggagg agaacaaggc tctacagaag gaactggtggcagcagcagc aagagcagtg ggaccgtcag acccaaacaccaggcccaga cgagctcatc atcttcctcc ttcatgatgacaacaaaaca tctgttaccc gccggtgaca atgggtggaggggcagcagg ggcagcttcg catcggaggc ctgccaccatgcttga <210> 89 <211> 261 <212 > PRT

<213> Lolium perenne <400> 89

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Thr

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys

100 105

Met Gly Glu Asp Leu Glu Cys Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130 135

His Leu Met Met Glu Ser Ile Ser Glu Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu

165 170

Lys Wing Arg Wing Gln Gln Gln Gln Gln Glu Gln Trp

180 185

Thr Gln Gln Wing Gln Asn Gln Pro Gln Wing Gln

195 200

Ser Be Phe Met Met Asp Gln Gln Wing His

210 215

Cys Tyr Pro Val Thr Met Gly Gly Glu Ala225 230 235

Gly Gn Gln Gly Gnu Leu Arg Ile Gly Gly Gnu

245 250

Being His Leu Asn Ala260

<210> 90

Pro Pro Trp Met Leu255

agataaaccg tcaggtgacc 60agatctccgt cctctgcgac 120tctatgagta cgccactgac 180cttatgctga aaaggctttg 240aatacaggaa actgaaggcg 300gggaggatct ggagtgtcta 360gttcattgaa gcacatcaga 420tacagaagaa ggagcggtca 480agaggcagaa ggcggccagg 540aacaagccca aaaccaacct 600gggatcagca ggcccatgct 660aggctgtggc cgcggcgcca 720ggatgctgag ccacctcaac 780 786

Glu Asn Lys

Leu Leu Lys30

Val Val Wing45

Asp Ser Ser60

Glu Lys Wing

Cys His Glu

Cys His Lys110

Glu Leu Gln125

Arg Ser Arg140

Lys Lys Glu

Read Val Glu

Asp Arg Gln190

Thr ser ser

205Ala Gln Gln220

Val Wing AlaPro Pro Trp

Ile Asn15

Lys Wing

Val phe

Met asp

Leu80 Wing

Tyr Arg95

His leu

Gln Leu

Lys Ser

Arg Ser160Arg Gln175

Thr gln

To be to be

Asn Ile

Pro240Met Wing Leu255 <211> 831 <212> DNA

<213> Hordeum vulgare <400> 90

atgggtcgcg gtaaggtgca gctgaagcgg atagagaacattctccaagc gccgcaacgg gctcctgaag aaggcgcacggcagaggtcg ccgtcatcgt cttctccccc aaaggcaagctccagcatgg acaaaattct tgaacgttat gagcgctactatttcagctg aatctgaaag tgaggggaat tggtgtcatgaagattgaga ccatacagaa gtgtcacaag cacctcatggaacctcaaag aactccaaca actggagcag cagctggagatcgagaaaga gccatcttat gatggagtcc atttctgagcctgcaggagg agaacaaggc cctacagaag gaactggtggaggcagcagc agctgcagca gcagcaacaa caacaacaaacagacccaaa cccataccca tactcaaaac cagccccaagtcctctttca tgatgaggga tcagcaggcc catgcccctcccaccggtga cgatgggtgg ggaggcgacg gcggcggcggcagcttcgca tatgcctacc gccatggatg ctgagccacc <210> 91 <211> 276 <212> PRT

<213> Hordeum vulgare <400> 91

agataaatcgagatctccgttctatgagtacttatgctgaaatacaggaagagaggatctgttcattgaatacagaagaaagaggcagaatgcaatgggacccagactagaacagaacatcggcgccggatcaacgcttg

gcaggtgacccctctgtgaccgccaccgacaaaggctcttacttaaggcgggattctctggcacatagagagaggtcaggcggccagcgcaccaagccctcatcatctttgtagctacgcagcaggcta

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu Lys Lys Wing 20 25 30 His Glu Ile Be Val Leu Cys Asp Wing Glu Val Wing Val Ile Val Phe 35 40 45 Be Pro Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Be Ser Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Wing Leu65 70 75 80Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu 100 105 110 Met Gly Glu Asp Leu Asp Ser Leu Asn Leu Lys Glu Leu Gln Leu 115 120 125 Glu Gln Gln Glu Leu Glu Be Being Read Lys His Ile Arg Be Arg Lys Be 130 135 140 His Met Le Met Met Glu Be Ile Be Glu Leu Gln Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val Glu Arg Gln 165 170 175 Lys Wing Wing Be Arg Gln Gln Gln Leu Gln Gln Gln Gln Gln Gln Gln 180 185 190 Gln Met Gln Trp Glu His Gln Wing Gln Thr Gln Thr His Thr His Thr 195 200 205 Gln Asn Gln Pro Gln Wing Gln Thr Being Ser Ser Ser Ser Ser 210 210 220 Met Arg Asp Gln Gln His Wing Pro Wing Gln Gln Asn Ile Cys Ser Tyr225 230 235 240Pro Pro Val Thr Met Gly Gly Glu Wing Thr Wing Wing Wing Pro Wing 245 250 255 Glu Gln Gln Wing Gln Leu Arg Ile Cys Leu Pro Trp Met Leu Ser 260 265 270 His Read Asn Wing

60120180240300360420480540600660720780831

<210><211> <212>

275

92

804

DNA

<213> Oryza sativa <400> 92

atggggcggg ggaaggtgca gctgaagcgg atagagaactttctccaaga ggaggaatgg attgctgaag aaggcgcacggccgaggtcg ccgccatcgt cttctccccc aagggcaagctccaggatgg acaaaatcct tgaacgttat gagcgctattatttcagctg aatctgagag tgagggaaat tggtgccatgaagattgaga ccatacaaaa atgtcacaaa cacctcatggaatctcaaag aactccaaca gctagagcag cagctggagatcaagaaaga gccaccttat gcttgagtcc atttccgagcctgcaggagg agaacaaggc tctgcagaag gaactggtggggccagcagc aagtagggca gtgggaccaa acccaggtccccccaagccc agacaagctt cttcttcttc ttcatgctgatcaccacaaa atatctgcta cccgccggtg atgatgggcccggcggcggt ggcggcccaa ggccaggtgc aacttccgcaatgctgagca ccttcaaggc ttaa <210> 93 <211> 267 <212> PRT <213> Oryza sativa <400> 93

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Thr Asp

cgatgaaccgagatctccgttctacgagtacatatgctgaaatacaggaagagaggatctgttcattgaatgcagaaaaaagaggcagaaaggcccaggcgggatcagcaagagaaatgattggaggctt

gcaggtgacgcctctgcgaccgccactgacaaaggctcttgcttaaggcaagaatccctggcacataagaggagaggtcagaatgtgaggccaagcccaaggcacttctttgcggcggcgtccgccatgg

Glu Asn Ser

Leu Leu Lys30

Wing Wing Ile45

To be Arg

Met Asn15

Lys AlaVal PheMet Asp

50

55

60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Wing Leu65 70 75 80Ile Be Wing Glu Be 85 Glu Be Glu Gly Asn 90 Trp Cys His Glu Tyr 95 ArgLys Leu Lys Wing 100 Lys Ile Glu Thr Ile 105 Gln Lys Cys His Lys 110 His LeuMet Gly Glu Asp Leu Glu Ser Leu Asn Leu Lys Glu Leu Gln Gln Leu

115

120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130

135

His Leu Met Leu Glu Ser Ile Ser Glu Leu Gln

Arg140Lys

125

Be Arg Lys Be

145

150

155

165

170

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu

TrpGln

Lys Asn Val Gly Gln Gln Gln Gln Val Gly Gln

180 185

Val Gln Wing Gln Wing Gln Wing Gln Pro Gln Wing

195 200

Phe Phe Phe Met Leu Arg Asp Gln Gln Wing Leu

210 215

Ile Cys Tyr Pro Pro Val Met Met Gly Gln Arg225 230 235

Arg Arg Arg Trp Arg Pro Lys Wing Arg Cys Asn

245 250

Phe Pro Pro Trp Met Read It Thr Thr Phe Lys Ala260 265

<210> 94

<211> 804

<212> DNA

<213> Oryza sativa

<400> 94

atggggcggg ggaaggtgca gctgaagcgg atagagaacattctccaaga ggaggaatgg attgctgaag aaggcgcacggccgaggtcg ccgccatcgt cttctccccc aagggcaagctccaggatgg acaaggatt

Leu220Asn

Phe

Lys glu

Val glu

Asp Gln190Thr Ser205

Be ProAsp AlaArg Ile

Arg Ser160Arg Gln175

Thr gln

Phe Phe

Gln asn

Wing Ala240Gly Gly255

agatcaacag gcaggtgacgagatctccgt cctctgcgactctacgagta cgccactgaccatatgctga aaaggctctt

60120180240300360420480540600660720780804

60120180240atttcagctg aatccgagag tgagggaaat tggtgccatg aatacaggaa acttaaggca

aagattgaga ccatacaaaa atgtcacaaa cacctcatgg gagaggatca tgaatccctg

aatctcaaag aactccaaca gctagagcag cagctggaga gttcattgaa gcacataata

tcaagaaaga gccaccttat gcttgagtcc atttccgagc tgcagaaaaa ggagaggtca

ctgcaggagg agaacaaggc tctgcagaag gaactggtgg agaggcagaa gaatgtgagg

ggccagcagc aagtagggca gtgggaccaa acccaggtcc aggcccaggc ccaagcccaa

ccccaagccc agacaagctc ctcctcctcc tccatgctga gggatcagca ggcacttctt

ccaccacaaa atatctgcta cccgccggtg atgatgggcg agagaaatga tgcggcggcg

gcggcggcgg tggcggcgca gggccaggtg caactccgca tcggaggtct tccgccatgg

atgctgagcc acctcaatgc ttaa <210> 95 <211> 267 <212> PRT

<213> Oryza sativa <400> 95

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr 20 Phe Ser Lys Arg Arg 25 Asn Gly Leu Leu Lys 30 Lys AlaHis Glu Ile 35 Ser Val Leu Cys Asp 40 Ala Glu Val Wing 45 Wing Ile Val PheSer Pro 50 Lys Gly Lys Leu Tyr 55 Glu Tyr Ala Thr Asp 60 Ser Arg Met AspLys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Leu65 70 75 80Ile Ser Ala Glu Ser 85 Glu Ser Glu Gly Asn 90 Trp Cys His Glu Tyr 95 ArgLys Leu Lys Wing 100 Lys Ile Glu Thr Ile 105 Gln Lys Cys His Lys 110 His LeuMet Gly Glu Asp His Glu Ser Leu Asn Leu Lys Glu Leu Gln Leu

115

120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130

135

His Leu Met Leu Glu Ser Ile Ser Glu Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu

165 170

Lys Asn Val Gly Gln Gln Gln Gln Val Gly Gln

180 185

Val Gln Wing Gln Wing Gln Wing Gln Pro Gln Wing

195 200

Being Being Being Met Leu Arg Asp Gln Gln Wing Leu

210 215

Ile Cys Tyr Pro Val Met Met Gly Glu Arg225 230 235

Wing Wing Wing Val Wing Wing Wing Gln Gly Gln Val Gln

245 250

Leu Pro Pro Trp Met Leu Be His Leu Asn Ala260 265

<210> 96 <211> 668 <212> DNA

<213> Virgin Tradescantia <400> 96

gggctggtga agaaggctca tgagatctcd atkytrtgtgattttctcta ctaaaggcaa actatacgag tatgccactgcttgaacgct atgaacgtta ctcatatgct gagaaggcttttacagggaa attggtgcca agagtatgtt aaacttaaggaaaagccaaa ggcatcttat gggagagcaa ctagaagcgtcaactagagc atcaacttga aggttctttg aggcttgtcaatgttggact ccatttccga acttcagagg aaggaaaagtaacctagaga aggagatttt ggagaagcag aaagaaaaggtgggaacagc agaatcagcc actacaaagc actaattcgc

125

Ile Ser Arg Lys Ser140

Lys Lys

Read val

Trp Asp

Gln Thr205Leu Pro220

Asn AspLeu Arg

Glu Arg Ser160

Glu Arg Gln175

Gln Thr Gln190

To be to be

Pro Gln Asn

Wing Wing Wing240

Ile Gly Gly255

atgcygaggtattccaaaattaacttcatcctaaggttgatggatctcaaggtcaagaaactctggaagactctggcacactccaaggcc

tgcgcttattggaaaatatcagatcctgaaggccttacatagaattgcaggactcaaatggcaaaacaagccaagctcaccttcgtgatt

300360420480540600660720780804

60120180240300360420480540gcagaaactc atccaacact aaacattgga aatttccaag gtagaacaaa taccgtccat 600gcagaagaaa gtctgcagcg tcagatgagg atcagcagca gcctactgcc mycmtggatg 660mttc

<210> 97 <211> 222 <212> PRT

<213> Virgin Tradescantia <220>

<221> VARIANT <222> (11) .. (11)

<223> Xaa can be any naturally occurring amino acid <220>

<221> VARIANT <222> (218) .. (218)

<223> Xaa can be any naturally occurring amino acid <220>

<221> VARIANT <222> (221) .. (221)

<223> Xaa can be any naturally occurring amino acid <400> 97

Gly Leu Val Lys Lys Wing His Glu Ile Ser Xaa Leu Cys Asp Wing Glu15 10 15

Val Wing Leu Ile Ile Phe Ser Thr Lys Gly Lys Leu Tyr Glu Tyr Wing

20 25 30

Thr Asp Be Lys Met Glu Asn Ile Read Glu Arg Tyr Glu Arg Tyr Ser

35 40 45

Tyr Wing Glu Lys Wing Read Thr Be Being Asp Pro Glu Read Gln Gly Asn

50 55 60

Trp Cys Gln Glu Tyr Val Lys Leu Lys Wing Lys Val Glu Wing Leu His65 70 75 80

Lys Ser Gln Arg His Leu Met Gly Glu Gln Leu Glu Wing Leu Asp Leu

85 90 95

Lys Glu Leu Gln Gln Leu Glu His Gln Leu Glu Gly Ser Leu Arg Leu

100 105 110

Val Arg Be Arg Lys Thr Gln Met Met Asu Asp Be Ile Be Glu Leu

115 120 125

Gln Arg Lys Glu Lys Being Leu Glu Glu Gln Asn Lys Asn Leu Glu Lys

130 135 140

Glu Ile Leu Glu Lys Gln Lys Glu Lys Wing Leu Wing His Gln Wing His145 150 155 160

Trp Glu Gln Gln Asn Gln Pro Read Gln Be Thr Asn Be Pro Pro Arg

165 170 175

Pro Phe Val Ile Wing Glu Thr His Pro Thr Read Asn Ile Gly Asn Phe

180 185 190

Gln Gly Arg Thr Asn Thr Val His Wing Glu Glu Being Read Gln Arg Gln

195 200 205

Met Arg Ile Ser Ser Ser Leu Pro Xaa Trp Met Xaa His

210 215 220

<210> 98 <211> 723 <212> DNA

<213> Virgin Tradescantia <400> 98

gtgcagctga aacggatgga gaacaagatt aacaggcagg tgacgttttc taaacgtcga 60

ggagggctgc tgaagaaagc tcatgagatc tctattctat gtgatgctga gattgctctt 120attattttct ctactaaagg gaagctctat gagtatgcca ccaattccaa aatggacaat 180attcttgaac gctatgagcg ttactcatat gctgaaaagg ctctaacttc atcagatcct 240gatatacagg gaaattggtg ccaagagtat gctaaactta agtctaaggt tgaggcttta 300tgtaaaagcc aaaggcatct tatgggagag cagcttgaaa cattgaatct caaagaattg 360cagcaactag agcaacagct cgaaggttct ctaaagcatg tcaggtcaag aaagactcaa 420gttatgctgg actctatttc tgaacttcag aggaaggaaa agtcactaga ggagcaaaac 480aagaacctag agaaggagat tttggagaag cagaaaatca aggctcttgc acagcaggct 540cactgggaac accagaatca accagcacca aggggttcac ctcctaggcc atttgtgatt 600gcagagttctc atccgacact aaatattgga catttccaag gcaggacaaa tgcagtcgaa 660gcagaagaaa atcagcagcc tcakatgaga atttgcagta gcctccggcccccccc23gcccccccc

<210> 99 <211> 241 <212> PRT

<213> Virgin Tradescantia <220>

<221> UNSAFE <222> (228) .. (228)

<223> Xaa can be any naturally occurring amino acid <400> 99

Val Gln Leu Lys Arg Met Glu Asn Lys Ile Asn Arg Gln Val Thr Phe1 5 10 15 Ser Lys Arg Arg Gly Gly Leu Leu Lys Lys Wing His Glu Ile Ser Ile 20 25 30 Leu Cys Asp Wing Glu Ile Wing Leu Ile Ile Phe Ser Thr Lys Gly Lys 35 40 45 Leu Tyr Glu Tyr Wing Thr Asn Ser. Lys Met Asp Asn Ile Leu Glu Arg 50 55 60 Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Wing Leu Thr Be Being Asp Pro65 70 75 80Asp Ile Gln Gly Asn Trp Cys Gln Glu Tyr Wing Lys Leu Lys Ser Lys 85 90 95 Val Glu Ala Leu Cys Lys Be Gln Arg His Leu Met Gly Glu Gln Leu 100 105 110 Glu Thr Leu Asn Leu Lys Glu Leu Gln Leu Glu Leu Gln Leu Glu 115 120 125 Gly Ser Leu Lys His Val Arg Be Lys Thr Gln Val Met Leu Asp 130 135 140 Be Ile Be Glu Leu Gln Arg Lys Glu Lys Be Leu Glu Glu Gln Asn145 150 155 160Lys Asn Leu Glu Lys Glu Ile Leu Glu Lys Gln Lys Ile Lys Wing Leu 165 170 175 Wing Gln Gln Wing His Trp Glu His Gln Asn Gln Pro Wing Pro Arg Gly

180 185 190

Be Pro Pro Arg Pro Phe Val Ile Glu Wing Be His Pro Thr Leu Asn

195 200 205

Ile Gly His Phe Gn Gly Arg Thr Asn Wing Val Glu Wing Glu Glu Asn

210 215 220

Gln Gln Pro Xaa Met Arg Ile Cys Ser Be Read Leu Pro Pro Trp Met225 230 235 240

Read

<210> 100 <211> 599 <212> DNA

<213> Virgin Tradescantia <4 00> 100

gggctkgtga agaaagctca tgagatctcg gtactttgtg atgctgagct tgctcttatt 60

atcttctctc ccaaaggcaa gctctatgag tatgccaccg attccaaaat ggaaattatt 120cttgaacgct atgaacgtta cacctacgct gaaaaagctt taattgcatc agatcctgat 180gtacagggaa actggtgtca tgagtacatt aagcttaaag ctaaatttga ggccttgaat 240aaaagccaga ggcatcttat gggagaacaa ctagatacgt tgaaccaaaa ggaattgctg 300caactagaga ctaagcttga aggttctctg aaaaacgtca ggtcaagaaa gactcaactt 360atgttggatt ccatttctga gcttcaagaa aagggaaagt cactccagga gcaaaacacc 420tgcctagaaa aggagatttt gggaaaacag aaagacaagg ctcccaaaca gcatgttcag 480tgggaaaaac agaatcaacc accacctacc tcttctgcgc caatgccatt cctcattggt 540gatattcacc caacccctaa tatcagaaat ttccaaggca gaacagtagc tgatgcaga 599

<210> 101 <211> 199 <212> PRT

<213> Virgin Tradescantia <400> 101

Gly Leu Val Lys Lys Wing His Glu Ile Be Val Leu Cys Asp Wing Glu1 5 10 15 Leu Wing Leu Ile Ile Phe Be Pro Lys Gly Lys Leu Tyr Glu Tyr Wing 20 25 30 Thr Asp Ser Lys Met Glu Ile Leu Glu Arg Tyr Glu Arg Tyr Thr 35 40 45 Tyr Wing Glu Lys Wing Leu Ile Wing Ser Asp Pro Asp Val Gln Gly Asn 50 55 60 Trp Cys His Glu Tyr Ile Lys Wing Leu Lys Phe Glu Wing Leu Asn65 70 75 80Lys Ser Gln Arg His Leu Met Gly Glu Gln Leu Asp Thr Leu Asn Gln 85 90 95 Lys Glu Leu Leu Gln Leu Glu Leu Glu Thr Lys Leu Glu Gly Leu Lys Asn 100 105 110 Val Arg Be Arg Lys Thr Gln Leu Met Leu Asp Ser Ile Ser Glu Leu 115 120 125 Gln Glu Lys Gly Lys Ser Read Gln Glu Gln Asn Thr Cys Wing Pro Met Pro 165 170 175 Phe Leu Ile Gly Asp Ile His Pro Thr Asn Ile Arg Asn Phe Gln 180 185 190 Gly Arg Thr Val Wing Asp Wing 195 <210> 102 <211> 753

<212> DNA

<213> Elaeis guineensis <4 00> 102

atggggagag ggagggtgca gctgaggcgg atcgagaaca agataaaccg gcaggtgacg 60

ttctcgaagc gccggtcggg gctcctgaag aaagcccacg agatctccgt cctctgcgac 120

gccgaggtcg ccctcatcat cttctcgacc aagggcaagc tctacgagta cgccaccgac 180

tcctgcatgg aaaggattct tgaacgctat gaacgttaca cctatgcaga aaaagcacta 240

atttcatctg gacccgaatt gcagggtaac tggtgccatg aatttggcaa actcaaagct 300

aaggttgagg ctttacaaaa aagccaaagg catctcatgg gtgagcaact tgagcccttg 360

aatctcaaag aactccagca actagagcaa cagcttgaaa gttctttaaa gcatataaga 420

accagaaagt gccaactcat gtttgaatcc atctctgagc ttcaaaaaaa ggaaaagtca 480

ctgcaggagc agaacaagat gctggagaag gagctcatgg agaagcagaa ggtgaaggca 54 0

ctaaaccagc aggcaccttg ggagcagcaa ggcccgccgc agacaagctc atcatcccca 600

acctccttcc tgatcggaga ctctctcccc accctgaata ttgggacata ccaatgtagc 660

ggaaatgaac atggggagga agcagcacaa ccccaggttc gtataggaaa cagcctgtta 720

ccaccttgga tgcttagcca cttgaacggg tag 753 <210> 103

<211> 250 <212> PRT <213> Elaeis guineensis <4 00> 103 Met Gly Arg Gly Arg Val Gln Leu Arg Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Gly Leu Leu Lys Lys Wing 20 25 30 His Glu Ile Be Val Leu Cys Asp Wing Glu Val Wing Leu Ile Ile Phe35 40 45 Ser Thr Tyr Thr Tyr Wing Glu Lys Wing Leu65 70 75 80Ile Being Ser Gly Pro Glu Leu Gln Gly Asn Trp Cys His Glu Phe Gly 85 90 95 Lys Leu Lys Ala Lys Val Glu Ala Leu Gln Lys Ser Gln Arg His Leu100 105 110

Met Gly Glu Gln Leu Glu Pro Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Be Read Lys His Ile Arg Thr Arg Lys Cys

130 135 140

Gln Read Met Phe Glu Be Ile Be Glu Read Gln Lys Lys Glu Lys Ser145 150 155 160

Leu Gln Glu Gln Asn Lys Met Leu Glu Lys Glu Leu Met Glu Lys Gln

165 170 175

Lys Val Lys Wing Read Asn Gln Gln Pro Wing Trp Glu Gln Gln Gly Pro

180 185 190

Pro Gln Thr Be Be Ser Be Pro Thr Be Phe Read Ile Gly Asp Ser

195 200 205

Read Pro Thr Read Asn Ile Gly Thr Tyr Gln Cys Ser Gly Asn Glu His

210 215 220

Gly Glu Glu Wing Gln Pro Wing Gln Val Arg Ile Gly Asn Ser Leu Leu225 230 235 240

Pro Pro Trp Met Leu Be His Leu Asn Gly245 250

<210> 104 <211> 699 <212> DNA <213> Allium sp. <400> 104

gtgcaattga agaggatgga aaacaagatt aatagacaag tgaccttctc aaaaagaaga 60

aatggtttgt tgaagaaagc tcatgagatt tcwgtgcttt gtgatgcaga agttgcactt 120attgttttct ctgctaaagg aaaactctat gaatattcaa ctgattcaag tatggaaaaa 180attctggaga ggtatgaacg ttattgcttt gcggagaaat catcaacaat gagtgacatt 240gactcccagg aggattggag ccttgaatat cacaaactga aggctaaggt tgagagttta 300aacaacaggc aaaggcatct tatgggagag caacttgaat ctctgagtct tcgagaaatt 360ggacagcttg agcaacaact tgagaattct ctcaaaactg ttcggacgcg caagagccaa 420gaattgttaa gttctatttc agagcttcag gacaaggaga aaactttgcg agatgagaac 480aaagctttag aaaatgagct tatgaaaagg gccagggcaa aagctattct ggaacaacaa 540gcacgatgga agcatcataa tcataaacaa caggataatc ttcataatcc aaatatcaac 600attggaaatt accaaacaag gaacaatgag ggaggagttg agccagcaac ggatgttcaa 660gtacgtgttg ttagaaattt gttgccccac tggatgctt 699

<210> 105 <211> 233 <212> PRT <213> Allium sp. <4 00> 105

Val Gln Read Lys Arg Met Glu Asn Lys Ile Asn Arg Gln Val Thr Phe15 10 15

Ser Lys Arg Arg Asn Gly Leu Read Lys Lys Wing His Glu Ile Ser Val

20 25 30

Leu Cys Asp Wing Glu Val Wing Wing Leu Ile Val Phe Ser Wing Lys Gly Lys

35 40 45

Leu Tyr Glu Tyr To Be Thr Asp To Be Met Glu Lys Ile Leu Glu Arg

50 55 60

Tyr Glu Arg Tyr Cys Phe Ala Glu Lys Ser Be Thr Met Ser Asp Ile65 70 75 80

Asp Be Gln Glu Asp Trp Be Read Glu Tyr His Lys Read Lys Ala Lys

85 90 95

Val Glu Ser Leu Asn Asn Arg Gln Arg His Leu Met Gly Glu Gln Leu

100 105 110

Glu Be Leu Be Leu Arg Glu Ile Gly Gln Leu Glu Gln Gln Leu Glu

115 120 125

Asn Be Leu Lys Thr Val Arg Thr Arg Lys Be Gln Glu Leu Read Le Be

130 135 140

Ser Ile Ser Glu Leu Gln Asp Lys Glu Lys Thr Leu Arg Asp Glu Asn145 150 155 160

Lys Wing Leu Glu Asn Glu Leu Met Lys Arg Wing Arg Wing Lys Wing Ile165 170 175Leu Glu Gln Wing Gln Arg Trp Lys His His Asn

180 185

Asn Read His Asn Pro Asn Ile Asn Ile Gly Asn

195 200

Asn Glu Gly Gly Val Glu Pro Wing Thr Asp Val

210 215

Arg Asn Leu Leu Pro His Trp Met Leu225 230

<210> 106 <211> 744 <212> DNA

<213> Dendrobium grex Madame Thong-IN <4 00> 106

atgggtcgtg gcagggtgca gctgaagcga atcgagaatattctcgaagc ggagatctgg tttgcttaag aaggcgcacggctgaagttg ctctgatcgt tttttccaat aagggaaagctccagcatgg agaaaattct tgaacggtat gagcgttattttttccaatg aggccaaccc ccaggctgat tggcgccttgagggttgaaa gcttacagaa gagccaaagg caccttatggagcattaaag aactccaacg tctagagcaa cagcttgaaatccagaaaga cacagctcat actacattca atttccgagcttgctggagc aaaacaagac cttagagaag gagattatagttggtgcagc atgccccatg ggagaagcaa aaccagtccccctgtgattt cggattctgt cccaactccc accagcagaagaagaagaat cacctcagcc acagttaaga gtaagcaacactcagtcata tgaatggaca ATAA <210> 107 <211> 247 <212> PRT

<213> Dendrobium grex Madame Thong-IN <4 00> 107

Met Gly Arg Gly Arg Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Be Asn Lys Gly Lys Read Tyr Glu Tyr Be Thr

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Phe Ser Asn Glu Wing Asn Pro Gln Wing Asp Trp

85 90

Lys Leu Lys Wing Arg Val Glu Ser Leu Gln Lys

100 105

Met Gly Glu Gln Read Asp Ser Read Ile Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys Phe Ile

130 135

Gln Leu Ile Leu His Be Ile Ser Glu Leu Gln145 150 155

Leu Leu Glu Gln Asn Lys Thr Leu Leu Glu Lys Glu

165 170

Lys Wing Lys Wing Read Val Gln His Wing Pro Trp

180 185

Be Gln Tyr Ser Serve Leu Pro Pro Val Ile

195 200

Thr Pro Thr Be Arg Thr Phe Gln Wing Arg Wing

210 215

Pro Gln Pro Gln Leu Arg Val Ser Asn Thr Leu225 230 235

Leu Being His Met Asn Gly Gln245

His Lys Gln 190 Gln AspTyr Gln 205 Thr Arg

aaataaaccgagatctccgttttatgagtacatatgctgaaatataataagggagcaactgttccttgaatacaaaagatctaaagagaaaatatagctccgtttcaagcctctgctgcc

gcaggtgacggctctgtgacttccaccgataagagcattaactgaaggcatgactccttggtttatacgaggaaaaataagccaaagcttgcactcccgcagagccaatcccatggatg

Glu Asn Lys

Leu Leu Lys30

Wing Leu Ile45

Asp Ser Ser60

Glu Arg Wing

Arg Leu Glu

Ser Gln Arg110

Glu Leu Gln125

Arg Ser Arg140

Lys Met Glu

Ile Ile Wing

Glu Lys Gln190

Be Asp Be

205Asn Glu Glu220

Read pro

Ile Asn15

Lys Wing

Val phe

Met glu

Wing Leu80Tyr Asn95

His leu

Arg Leu

Lys thr

Lys Ile160Lys Glu175

Asn gln

Val pro

Glu Ser

Trp Met240

60120180240300360420480540600660720744 <210> 108

<211> 2194

<212> DNA

<213> Oryza sativa

<400> 108

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 àtatctaaaa 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> 109 <211> 804 <212> DNA <213> Oryza sativa <400> 109

atggggcggg ggaaggtgca gctgaagcgg atagagaaca agatcaacag gcaggtgacg 60

ttctccaaga ggaggaatgg attgctgaag aaggcgcacg agatctccgt cctctgcgac 120

gccgaggtcg ccgccatcgt cttctccccc aagggcaagc tctacgagta cgccactgac 180

tccaggatgg acaaaatcct tgaacgttat gagcgctatt catatgctga aaaggctctt 240

atttcagctg aatccgagag tgagggaaat tggtgccatg aatacaggaa acttaaggca 300

aagattgaga ccatacaaaa atgtcacaaa cacctcatgg gagaggatct agaatccctg 360

aatctcaaag aactccaaca gctagagcag cagctggaga gttcattgaa gcacataata 420

tcaagaaaga gccaccttat gcttgagtcc atttccgagc tgcagaaaaa ggagaggtca 480

ctgcaggagg agaacaaggc tctgcagaag gaactggtgg agaggcagaa gaatgtgagg 540

ggccagcagc aagtagggca gtgggaccaa acccaggtcc aggcccaggc ccaagcccaa 600

ccccaagccc agacaagctc ctcctcctcc tccatgctga gggatcagca ggcacttctt 660

ccaccacaaa atatctgcta cccgccggtg atgatgggcg agagaaatga tgcggcggcg 720

gcggcggcgg tggcggcgca gggccaggtg caactccgca tcggaggtct tccgccatgg 780

atgctgagcc acctcaatgc ttaa 804 <210> 110 <211> 267 <212> PRT

<213> Oryza sativa <400> 110

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Asn Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Wing Wing Ile Val Phe

35 40 45

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Thr Asp Be Arg Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Wing Leu65 70 75 80

Ile Be Glu Wing Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Ile Be Arg Lys Be

130 135 140

His Leu Met Leu Glu Ser Ile Be Glu Leu Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val Glu Arg Gln

165 170 175

Lys Asn Val Gly Gln Gln Gln Gln Val Gly Gln Trp Asp Gln Thr Gln

180 185 190

Val Gln Wing Gln Wing Gln Wing Gln Pro Gln Wing Gln Thr Be Ser Ser

195 200 205

Be Being Be Met Met Leu Arg Asp Gln Gln Wing Read Leu Pro Pro Gln Asn

210 215 220

Ile Cys Tyr Pro Pro Val Met Met Gly Glu Arg Asn Asp Asa Wing Ala225 230 235 240

Wing Wing Wing Wing Val Wing Wing Gln Gly Gln Val Gln Leu Arg Ile Gly Gly

245 250 255

Leu Pro Pro Trp Met Leu Be His Leu Asn Ala260 265

<210> 111 <211> 52 <212> DNA

<213> Artificial sequence <220>

<223> primer <4 00> 111 '

ggggacaagt ttgtacaaaa aagcaggctt aaacaatggg gcgggggaag gt 52

<210> 112 <211> 47 <212> DNA

<213> Artificial sequence <220>

<223> primer <4 00> 112

ggggaccact ttgtacaaga aagctgggtt tggccgacga cgacgac 47

<210> 113

<211> 2193

<212> DNA

<213> Oryza sativa

<4 00> 113

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300 <210> 114 <211> 42 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (2) .. (2) <223> / replace = "Going" <220>

<221> VARIANT

<222> (6) .. (6)

<223> / replace = "Asn"

<220>

<221> VARIANT <222> (10) .. (10) <223> / replace = "He" <220>

<221> VARIANT <222> (12) .. (12) <223> / replace = "Tyr" <220>

<221> VARIANT <222> (16) .. (16)

<223> / replace = "lie" / replace = "read" <220>

<221> VARIANT

<222> (17) .. (17)

<223> / replace = "Gly"

<220>

aaaaaatcttatgaagatatttgtgcattcttagtaattagtacttacgctcaactagcatgaattcaagaattttacagaaaaaaagaaacagagtggctccgcaacaaaaccaagcatggaggcatcccgaccgccttcacctcctcctgtagtacgggatttgggatggttttcaatgattttgtgaaataaaagtactatcctttgatgagattgaacagtagtcccctgttcttcctttctggttgctgtagttctttctgatctattatttttttgtcctcaatcctgtagaagagctgtaatctggtgtagct

tctagctgaatctgaacgtagtcatatcgcaagacaattgacacactttgacacatctctcactccaccaaatagcatgattttgctcgttgcccacagaccttttaacacctcctcctcaagccaagaacttcgatccatcacagggtagcgttgatgtagaggggttccgtctggagagtaccttttgcggttgtttgtttattccctatgattgattccatcacgaacgatttgcttcagttcaatgagttaataggccatttttaattattagctctttgttttcatttctttttggggatagttatgccactttc

ctcaatgggt

ttggcaaaga

acatcattaa

acttattttt

tgctcatgtg

aatatcactc

tcaccagacc

aaagtatgaa

gcgcgagcgc

acaacccaca

gcaggctttg

ccatctataa

gagggagagc

tatcttccgg

tgtgcccttc

taggaaaggg

ttgatgttgc

gctctatgga

tttgaggtaa

gtcctcgatt

attgaacaaa

cttaagcctg

attcatggaa

tagtcccaga

aattgattgc

taatacccct

ttatatgaaa.

tcaccccttc

aattcacatc

gttattcctt

tactgcttgt

accagcaaag

aaagagagag attttttttatttaaacata taattatata

ggacatgtctattatttatccatgtgtgaggcctatttaaacttttaataacgaactattcaatctcccaaaaaacgatgcggccaggagattcctccccaccaaggacatcgagttcttggttgttcttgatctgtatcatgttatcggaatgaaatggaatcagagcactggtagtgaaataatccaatccaaaatttacagttataaattttttttctacaaataatatagtttagttgaactgtagattattctgagattatctatgactgcttgatcttatgattttc

tactccatccttttttcgattgcacctccttacatttaggatatctaaaataggtttttctattgggcacatctaacggaagaggaggagccttttcccccgcgactagcggtcgatctcggatttattgtgtgatgattttcggtttgatttagggtacccggtgattttgcttctcgactttgaagaccgcagctggctcctcaggaaccaaatatctgcttttatagcaggagaagacataagcagtgctgaaagtcgcattatcctttacagaaagcatttccttt

aaaaaatagaattttatagtcaatttttattagatgcaagcaatacacgttagcaatatctacaaaaaatacatacaaaaacaggcaacaggacagcaagaggcaaagaatctctatataagaagccgagttccctcctcttctaggttgcctgttcttgttagtagtatggaatcttgctgcttggtgttttgacgaagggtcccgttgttgtttagatcaggggattctaaaaagtcacgttatcctaacttatccgaattcatttggtggcatgaaccttgtatctaaaatttatgagtgcagttct <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <223> <220> < 221> <222> <223> <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <223> <220> <221> <222><223><220><221><222><223><220><221><222><220><221><222> <223> <4 00> Read Leu1

Wing Leu Ile

VARIANT (18) .. (18) / replace

VARIANT

(19) .. (19) / replace

VARIANT

(20) .. (20) / replace

VARIANT (23) .. (23) / replace

VARIANT

(30) .. (30) / replace

VARIANT

(31) .. (31) / replace

VARIANT

(32). . (32) / replace

VARIANT

(33) .. (33) / replace

VARIANT (35) .. (35) / replace

VARIANT

(37) .. (37) / replace

VARIANT

(38). . (38) / replace

VARIANT (41). . (41) / replace114

= "Wing" / replace = "Go"

"Go"

= "Go"

= "Pro" / replace = "Wing" / replace = "Asn"

= "Phe"

= "Be"

"To be"

"Asn" / replace = "Glu"

= "Cys / replace =" Arg "/ replace =" Ser '

= "Glu"

= "lie" / replace = "Arg" / replace = "Lys"

= "Asp"

Lys Lys

Wing5

Phe

Ile20

Met asp

Asp Ser Lys35

<210> 115 <211> 19 <212> PRT

<213> Arti sequence <220>

<223> Arti sequence <220>

<221> VARIANT <222> (4) .. (4)

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

10 15

Being Thr Lys Gly Lys Read Tyr Glu Tyr Wing Thr

25 30

Asn Ile Leu Glu Arg40

<223> <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <222> <220> <221> <222> < 223> <220> <221> <222> <223> <220> <221> <222> <220> <221> <222> <223> <220> <221> <222> <223> <220><221><222> <223> <4 00>

/ replace = "Be"

VARIANT

(5) .. (5) / replace

VARIANT

(6) .. (6)

/replace

VARIANT (8) .. (8) / replace

VARIANT

(9) ■ · (9)

/replace

VARIANT

(10). . (10) / replace

VARIANT (H) · ■ (H) / replace

VARIANT (12). . (12) / replace

VARIANT

(13) .. (13) / replace

VARIANT

(14) .. (14) / replace115

= nArg "

= "lie"

"Thr" / replace = "Ser"

"lie"

= "Asn"

"Arg" / replace = "Asn"

= "Cys" / replace = "Arg"

= "His"

= "Lys"

Lys Leu Lys Wing Lys1 5

Met Gly Glu

Val Glu Wing Leu

Gln10

Lys Ser Gln Arg

His15

Read

<210> 116 <211> 11 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (2) .. (2)

<223> / replace = "Gln" / replace <220>

<221> VARIANT <222> (6) .. (6) <223> / replace = "Phe" <220>

<221> VARIANT

<222> (7). . (7)

<223> / replace = "Phe"

<220>

<221> VARIANT <222> (8) .. (8)

"Go" / replace = "Wing" "Phe"

<223> / replace = "Phe" <220>

<221> VARIANT <222> (9) .. (9) <223> / replace = "Phe" <220>

<221> VARIANT <222> (10) .. (10)

<223> / replace = "Cys" / replace <220>

<221> VARIANT <222> (11) .. (11) <223> / replace = "Met" <400> 116

Gln Pro Gln Thr Be Ser Ser Ser Ser Ser Phe1 5 10

<210> 117 <211> 7 <212> PRT

<213> Artificial sequence <220>

<223> Artificial sequence <220>

<221> VARIANT <222> (1) .. (1)

<223> / replace = "Wing" / replace = "Go" / replace = "Read" <220>

<221> VARIANT <222> (2) .. (2) <223> / replace = "Pro" <220>

<221> UNSAFE <222> (3) .. (3)

<223> Xaa can be any amino acid, preferably Leu, Phe or His <220>

<221> UNSAFE <222> (6) .. (6)

<223> Xaa can be any naturally occuring amino acid, preferably Val orLeu

<220>

<221> VARIANT <222> (7). . (7) <223> / replace = "His" <4 00> 117

Gly Leu Xaa Trp Met Xaa Ser1 5

<210> 118 <211> 735 <212> DNA

<213> Hordeum vulgare <4 00> 118

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg ccaggtcacc 60

ttctccaagc gccgctcggg gctgctcaag aaggcgcacg agatctccgt gctctgcgac 120gccgaggtcg gcctcatcat cttctccacc aagggaaagc tctacgagtt ctccaccgag 180tcatgtatgg acaaaattct tgaacggtat gagcgctact cttatgcaga aaaggttctc 240gtttcaagtg aatctgaaat tcagggaaac tggtgtcacg aatataggaa actgaaggcg 300aaggttgaga caatacagaa atgtcaaaag catctcatgg gagaggatct tgaatctttg 360aatctcaagg agttgcagca actggagcag cagctggaaa gctcactgaa acatatcaga 420gccaggaaga accaacttat gcacgaatcc atttctgagc ttcagaagaa ggagaggtca 480ctgcaggagg agaataaagt tctccagaag gaacttgtgg agaagcagaa ggcccaggcg 540gcgcagcaag atcaaactca catgatgagggggggggggggggggggggggggc

<210> 119 <211> 244 <212> PRT

<213> Hordeum vulgare <4 00> 119

Met Gly Arg Gly Lys1 5

Arg Gln Val Thr Phe20

His Glu Ile Ser Val35

Be Thr Lys Gly Lys50

Lys Ile Leu Glu Arg65

Val Ser Ser Glu Ser85

Lys Leu Lys Wing Lys100

Met Gly Glu Asp Leu115

Glu Gln Gln Leu Glu130

Gln Leu Met His Glu145

Read Gln Glu Glu Asn165

Lys Wing Gln Wing Wing180

Ser Ser Ser Ser Phe195

Ser Asn His Pro Ala210

Gln Pro Gln Val Pro225

His Ile Asn Gly

<210> 120 <211> 1196 <212> DNA <213> Hordeum vulgare <4 00> 120

ctctcccctc ccacttcacc caaccacctg acagccatgg ctccgccacc tcgcctccgc 60

ccgcgcctct gagagtagcc gtcgcggtcg ctcgctcgct cgctcgctgc tgccggtgtt 120

ggcccggtcc tcgagcggag atggggcgca ggaaggtgca gctgaagcgg atcgagaaca 180

agatcaaccg ccaggtcacc ttctccaagc gccgctcggg gctgctcaag aaggcgcacg 240

agatctccgt gctctacgac gccgaggtcg gcctcatcat cttctccacc aagggaaagc 300

tctacgagtt ctccaccgag tcatgtatgg acaaaattct tgaacggtat gagcgctact 360

cttatgcaga aaaggttctc gtttcaagtg aatctgaaat tcagggaaac tggtgtcacg 420

aatataggaa actgaaggcg aaggttgaga caatacagaa atgtcaaaag catctcatgg 480

gagaggatct tgaatctttg aatctcaagg agttgcagca actggagcag cagctggaaa 540

gctcactgaa acatatcaga gccaggaaga accaacttat gcacgaatcc atttctgagc 600

ttcagaagaa ggagaggtca ctgcaggagg agaataaagt tctccagaag gaacttgtgg 660

agaagcagaa ggcccaggcg gcgcagcaag atcaaactca gcctcaaacc agctcttctt 720

cttcttcctt catgatgagg gatgctcccc ctgtcgcaga taccagcaat cacccagcgg 780

cggcaggcga gagggcagag gatgtggcag tgcagcctca ggtcccactc cggacggcgc 840

ttccactgtg gatggtgagc cacatcaacg gctgaagggc ttccagccca tgtaagcgta 900

ctattcagta cgagtaacaa gttgcagcgg ccagcctggt gtatcatgcg gttgcgaaca 960

tgctaacccc atggagggga gaggaaaaga aatcagagta aagcagcaag ctgcaggaat 1020

gtgtatattt cacttcgtcc acctcagttt cctttccacc tgggctgaga tggctgtacg 1080

agtaatctac catgtaattt atatgtagca tgagtgacga attttcaact ttcgatgata 1140

tccgttgctc ctgggtgttg tttctgtgaa ttaacctatc gaatatgagc gttgtg 1196

Val Gln Read Lys Arg10

Ser Lys Arg Arg Ser25

Read Cys Asp Wing Glu40

Read Tyr Glu Phe Ser55

Tyr Glu Arg Tyr Ser70

Glu Ile Gln Gly Asn90

Val Glu Thr Ile Gln105

Glu Ser Leu Asn Leu120

Ser Ser Leu Lys His135

Ser Ile Ser Glu Leu150

Lys Val Leu Gln Lys170

Gln Gln Asp Gln Thr185

Met Met Arg Asp Ala200

Wing Wing Gly Glu Arg215

Leu Arg Thr Wing Leu230

Ile Glu Asn Lys Ile Asn15

Gly Leu Leu Lys Lys Ala30

Val Gly Leu Ile Ile Phe45

Thr Glu Ser Cys Met Asp60

Tyr Wing Glu Lys Val Leu75 80

Trp Cys His Glu Tyr Arg95

Lys Cys Gln Lys His Leu110

Lys Glu Leu Gln Gln Leu125

Ile Arg Wing Arg Lys Asn140

Gln Lys Lys Glu Arg Ser155 160

Glu Leu Val Glu Lys Gln175

Gln Pro Gln Thr Ser Ser

Pro Pro Val Wing Asp Thr205

Glu Wing Asp Val Wing Val220

Pro Read Trp Met Val Ser235 240 <210> 121 <211> 244 <212> PRT

<213> Hordeum vulgare <400> 121

Met Gly Arg Arg Lys Val Gln1 5

Arg Gln Val Thr Phe Ser Lys20

His Glu Ile Ser Val Leu Tyr35

Being Thr Lys Gly Lys Leu Tyr

50 55

Lys Ile Read Glu Arg Tyr Glu65 70

Val Ser Ser Glu Ser Glu Ile85

Lys Leu Lys Wing Lys Val Glu100

Met Gly Glu Asp Leu Glu Ser115

Glu Gln Gln Leu Glu Ser Ser130 135

Gln Read Met His Glu Ser Ile145 150

Read Gln Glu Glu Asn Lys Val165

Lys Wing Gln Wing Wing Gln Gln180

Ser Ser Ser Ser Phe Met Met195

Ser Asn His Pro Wing Ala Wing210 215

Gln Pro Gln Val Pro Read Arg225 230

His Ile Asn Gly

Read Lys Arg10

Arg Arg Ser25

Asp Wing Glu40

Glu Phe Ser

Arg Tyr Ser

Gln Gly Asn90

Thr Ile Gln

105Leu Asn Leu120

Read Lys His

Being Glu Leu

Read Gln Lys170

Asp Gln Thr185

Arg Asp Ala200

Gly Glu ArgThr Wing Leu

Ile Glu Asn LysGlyValThr

Tyr

75

Trp

Lys

Lys

Ile

Gln155Glu

Gln

Pro

Allah

Pro235

Leu Leu Lys30

Gly Leu Ile45

Glu Ser Cys60

Glu Lys Wing

Cys His Glu

Cys Gln Lys110

Glu Leu Gln125

Arg Wing Arg140

Lys Lys Glu

Read Val Glu

Pro Gln Thr190

Pro Val Wing

205Glu Asp Val220

Read Trp Met

Ile Asn15

Lys Wing

Ile Phe

Met asp

Val Leu80Tyr Arg95

His leu

Gln Leu

Lys asn

Arg Ser160Lys Gln175

To be to be

Asp Thr

Val Wing

Val ser240

<210> 122

<211> 1161

<212> DNA

<213> Trit

<4 00> 122

ggcacgagct

cggcacgagc

accggcaggt

ccgtgctctg

agttctccac

cagaaaaggt

ggaaactgaa

atcttgaatc

tgaaacatat

agaaggagag

agaaggccca

ccttcatgat

gcgagagggc

tgtggatggt

agtagagagt

gaccccttgg

atatttcact

atatgtagct

gtttatgtga

ataaatcagg

<210> 123

icum aestivum

ctctctctctggagatggggggaccttctcccgacgccgagcgagtcatgttctcgtttcaggcgaaggtttttgaatctccagatccagggtcactgcagggcgggggcggggtgtgtgtgtgtgt

ctctctctct ctctctctct

cgcgggaaggaagcgccgctgtcggcctcaatggacaaaaagtgaatctggagacaatacaaggagttgcaagaaccaacgaggagatacaagatcagaccccctgccggcagtgcagcaacggctgaaaccggccagtaaatatagtagtag

tgcagctgaacggggctgcttcatcttctcttcttgaacgaaattcagggagaaatgtcaagcaactggattatgcacgaaagttctccactcagcctcacagctaccagcgcaggccccgggcttccagctggtggtggtggtggtggtg

ctctctcctcgcggatcgagcaagaaggcgcaccaagggagtatgagcgcaaactggtgtaaagcatctggcagcagctgatccatttctgaaggaactcaacaagctctcattcatccaaccccggacgcccatatagggcgctctctctct

gtgccgaattaacaagatcacacgagatctaagctctacgtactcttatgcacgaatataatgggagagggaaagctcacgagcttcagagtcgagaagctccttcttcttgcggcggcaggggcttccaccgtgggggggggggggtg

601201802403003604204805406006607207808409009601020108011401161 <211> 244 <212> PRT

<213> Triticum aestivum <400> 123 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Ser Gly Leu Lys Lys Wing 20 25 30 His Glu Ile Ser Val Leu Cys Asp Glu Val Wing Val Gly Leu Ile Ile Phe 35 40 45 Ser Val Lu Gys Lys Leu Tyr Glu Phe Ser Val Glu Cyr Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Glu Lys Val Leu65 70 75 80Val Being Glu Being Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu 100 105 110 Met Gly Glu Asp Leu Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Leu Glu Be Ser Leu Lys His Ile Arg Be Arg Lys Asn 130 135 140 Gln Leu Met His Glu Ser Ile Be Glu Leu Glys Lys Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln 165 170 175 Lys Ala Gln Ala Ala Gln Ala Ala Gln Ala Ala Gln Ala Ala Gn Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Ala Gly Asp Wing Val Wing 210 215 220 Gln Pro Gln Wing Pro Pro Arg Thr Gly Leu Pro Read Trp Met Val Ser225 230 235 240His

<210> 124 <211> 1210 <212> DNA

<213> Triticum aestivum <4 00> 124

tccctctcct ccctctcttc cgcctcaccc aaccacctga cagccatggc tccgcccccc 60

cgcccccgcc tgcgcctgtc ggagtagccg tcgcggtctg ccggtgttgg aggcttgggg 120

tgtagggttg gccccgttct ccagcggaga tggggcgcgg gaaggtgcag ctgaagcgga 180

tcgagaacaa gatcaaccgg caggtgacct tctccaagcg ccgctcgggg ctgctcaaga 240

aggcgcacga gatctccgtg ctctgcgacg ccgaggtcgg cctcatcatc ttctccacca 300

agggaaagct ctacgagttc tccaccgagt catgtatgga caaaattctt gaacggtatg 360

agcgctactc ttatgcagaa aaggttctcg tttcaagtga atctgaaatt cagggaaact 420

ggtgtcacga atataggaaa ctgaaggcga aggttgagac aatacagaaa tgtcaaaagc 480

atctgatggg agaggatctt gaatctttga atctcaagga gttgcagcaa ctggagcagc 540

agctggaaag ctcactgaaa catatcagat ccaggaagaa ccaacttatg cacgaatcca 600

tttctgagct tcagaagaag gagaggtcac tgcaggagga gaataaagtt ctccagaagg 660

aactcgtcga gaagcagaag gcccaggcgg cgcaacaaga tcagactcag cctcaaacaa 720

gctcttcttc ttcttccttc atgatgaggg atgctccccc tgccgcaact accagcattc 780

atccagcggc atcaggagag agggcagagg atgcggcagt gcagccgcag gccccacccc 840

ggacggggct tccactgtgg atggttagcc acatcaacgg ctgaagggct tccagcccat 900

ataagcgtac tattcagtag agagtaacaa gttgcaccgg ccagcctggt gtatgttgcg 960

gttgctagca tgcctgaccc cttggagggg aaaggaaaag aaatcagagt aaagtagcaa 1020

gctgcagtga tgtgtatatt tcactttgtc cacctcagtt tccctcccag ctgggctcaa 1080

tttaccatgt aatctatatg tagcttgagt gatgaatttt caagtttcca tgatacccgt 1140

ctcgagcggg tgttgtttat gtgaattaac ctatcaaata tgagcattgt gtaaaaaaaa 1200

aaaaaaaaaa 1210 <210> 125 <211> 244 <212> PRT

<213> Triticum aestivum <400> 125

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile Glu Asn

1 5 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Read

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Be Thr Glu Ser

50 55 60

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu65 70 75

Val Being Glu Being Glu Ile Gln Gly Asn Trp Cys His

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln

100 105

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys145 150 155

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val

165 170

Lys Wing Gln Wing Wing Gln Gln Asp Gln Thr Gln Pro Gln

180 185

Be Ser Be Ser Phe Met Met Arg Asp Ala Pro Pro Ala195 200 205

Ser Ile His Pro Ala Wing Ser Gly Glu Arg Wing Glu Asp

210 215 220

Gln Pro Gln Pro Pro Arg Thr Gly Leu Pro Leu Trp225 230 235

His Ile Asn Gly

Lys Ile Asn15

Lys Lys Ala30

Ile Ile Phe

Cys Met Asp

Lys Val Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Asn

Glu Arg Ser160

Glu Lys Gln

175Thr Ser Ser190

Thr Thr Wing

Wing Val Wing

Met Val Ser240

<210> 126 <211> 735 <212> DNA <213> Triti <4 00> 126atggggcgcgttctccaagcgccgaggtcgtcatgtatgggtttcaagtgaaggttgagaaatctcaaggtccaggaagactgcaggagggcgcagcaaggatgctccccgatgcggcagcacatcaacg <210> 127 <211> 244 <212> PRT <213> Triti <4 00> 127Met Gly Arg1

Arg Gln Val

cum monococcum

ggaaggtgcagccgctcggggcctcatcatacaaaattctaatctgaaatcaatacagaaagttgcagcaaccaacttatagaataaagtatcaaactcactgccgcaaatgcagccgcaggtga

gctgaagcgggcttctcaagcttctccacctgaacggtattcagggaaacatgtcaaaaaactggagcaggcacgaatcctctccagaaggcctcaaacctaccagcattggccccaccc

atcgagaacaaaggcgcacgaagggaaagcgagcgctatttggtgtcacgcatctcatggcagctggaaaatttctgagcgaactcgtggagctcttcttcatccagcggcggacgggggc

cum monococcum

Gly Lys Val Gln Read Lys Arg Ile

5 10

Thr Phe Be Lys Arg Arg Be Gly20 25

agatcaaccgagatctccgttctacgagttcttatgcagaaatataggaagagaggatctgctcactgaatgcagaagaaagaagcagaacttcttccttcggcaggcgattccaccgtg

gcaggtgaccgctctgcgacctccaccgagaaaggttctcactgaaggcgtgaatctttgacatatagaggagaggtcaggcccatgcgcatgctgagggagggcagaggatggtgagc

Glu Asn Lys Ile Asn15

Leu Leu Lys Lys Ala30

60120180240300360420480540600660720735His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Be Thr Glu Ser

50 55 60

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu65 70 75

Val Being Glu Being Glu Ile Gln Gly Asn Trp Cys His

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln

100 105

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys145 150 155

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val

165 170

Lys Wing His Wing Wing Gln Gln Asp Gln Thr Gln Pro Gln

180 185

Ser Ser Ser Ser Phe Met Leu Arg Asp Ala Pro Ala195 200 205

Ser Ile His Pro Wing Wing Wing Gly Glu Arg Wing Wing Glu Asp

210 215 220

Gln Pro Pro Gln Pro Pro Arg Thr Gly Leu Pro Pro Trp225 230 235

His Ile Asn Gly

Ile Ile Phe

Cys Met Asp

Lys Val Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Asn

Glu Arg Ser160

Glu Lys Gln

175Thr Ser Ser190

Asn Thr Wing

Wing Val Wing

Met Val Ser240

<210> 128 <211> 735 <212> DNA

<213> Triticum aestivum <4 00> 128

atggggcggg ggaaggtgca gctgaagcgg atcgagaacattctccaagc gccgctcggg gcttctcaag aaggcgcacggccgaggtcg gcctcatcat cttctccacc aagggaaagctcatgtatgg acaaaattct tgaacggtat gagcgctattgtttcaagtg aatctgaaat tcagggaaac tggtgtcacgaaggttgaga caatacagaa atgtcaaaag catctcatggaatctcaagg agttgcagca actggagcag cagctggaaatccaggaaga accaacttat gcacgaatcc atttctgagcctgcaggagg agaataaagt tctccagaag gaactcgtgggcgcagcaag atcaaactca gcctcaaacc agctcttcatgatgctcccc ctgccgcaaa taccagcatt catccagcgggatgcggcag tgcagccgca ggccccaccc cggacggggccacatcaacg ggtga <210> 129 <211> 244 <212> PRT

<213> Triticum aestivum <400> 129

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Be Thr Lys Gly Lys Read Tyr Glu Phe

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Val Be Glu Be Glu Ile Gln Gly Asn Trp85 90

agatcaaccgagatctccgttctacgagttcttatgcagaaatataggaagagaggatctgcctcactgaattcagaagaaagaagcagaacttcttccttcaacaggcgattccaccgtg

gcaggtgaccgctctgcgacctccaccgagaaaggttctcactgaaggcgtgaatctttgacatatagaggagaggtcaggcccatgcgcatgctgagggagggcagaggatggtgagc

Glu Asn Lys Ile Asn15

Leu Leu Lys Lys Ala30

Gly Leu Ile Ile Phe45

Glu Ser Cys Met Asp60

Glu Wing Lys Val Leu80

Cys His Glu Tyr Arg95

60120180240300360420480540600660720735Lys Read Lys

Met Gly Glu115

Glu Gln Gln

130Gln Read Met145

Read Gln Glu

Lys Ala His

Ser Ser Ser195

Ser Ile His210Gln Pro Gln225

His Ile Asn

Lys100 wing

Asp Leu

Read Glu

His glu

Glu Asn165Ala Ala180

Be PhePro AlaAla ProGly

Val Glu Thr

Glu Ser Leu120

Ser Ser Leu

135Ser Ile Ser150

Lys Val Leu

Gln Gln Asp

Met Leu Arg200

Thr Gly Wing

215Pro Arg Thr230

Ile Gln Lys105

Asn leu lys

Lys His Ile

Glu Leu Gln155

Gln Lys Glu

170Gln Thr Gln185

Asp Wing Pro

Glu Arg Wing

Gly Leu Pro235

Cys Gln

Glu Leu125Arg Ser140

Lys Lys

Read val

Pro Gln

Pro Ala205Glu Asp220

Pro Trp

Lys His Leu110

Gln Gln Leu

Arg Lys Asn

Glu Arg Ser160

Glu Lys Gln

175Thr Ser Ser190

Asn Thr Wing

Wing Val Wing

Met Val Ser240

<210> 130 <211> 738 <212> DNA

<213> Lolium perenne <4 00> 130

atggggcgcg gcaaggtgca gctcaagcgg atcgagaacattctccaagc gccgctcggg gctgctcaag aaggcgcacggccgaggtcg ggctcatcat cttctccacc aagggaaagctcatgtatgg acaaaattct tgagcggtat gagcgctactatttcaaccg aatctgaaat tcagggaaac tggtgtcatgaaggttgaga caatacagag atgtcaaaag catctaatggaatctcaagg agttgcagca actagagcag cagctggaaagccagaaaga accagcttat gcacgaatcc atatctgagcctgcaggagg agaataaaat tctccagaag gaactcatagcagcaagcgc agtgggagca aactcagccc caaaccagctatgggggaag ctaccccagc aacaaattgc agtaatccccgcagaggatg cgacggggca gccttcagct cgcacggtgccacatcaaca atggctga <210> 131 <211> 245 <212> PRT

<213> Lolium perenne <400> 131

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile1 5 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Being Thr Lys Gly Lys Read Tyr Glu Phe Wing Thr

50 55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Ile Be Thr Glu Ile Be Glu Ile Gln Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Arg

100 105

Met Gly Glu Asp Leu Glu Ser Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130 135

Gln Read Met His Glu Ser Ile Be Glu Leu Gln145 150 155

agatcaaccgagatctccgttctacgagttcctatgcagaaatataggaagagaggatctgttcactgaattcaaaagaaagaagcagaacttcctcctccagcagcggcttccaccatg

ccaggtcaccgctctgcgaccgcaaccgacgaaagtgctcactgaaggcgtgaatcattgacatattagaggagaggtcaggcccacacgctcctttatgcagcgacagagatggtgagt

Glu Asn

Leu Leu

Gly Leu

45Asp Ser60

Glu Wing

Cys his

Cys Gln

Glu Leu125Arg Ala140

Lys Lys

Lys ile

15Lys Lys30

Ile Ile

Cys met

Lys val

Glu Tyr

95Lys His110

Gln GlnArg LysGlu Arg

Asn

Allah

Phe

Asp

Leu80Arg

Read

Read

Asn

Ser160

60120180240300360420480540600660720738Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Ile Glu Lys Gln

165 170 175

Lys Wing His Thr Gln Gln Wing Gln Trp Glu Gln Thr Gln Pro Gln Thr

180 185 190

Be Ser Ser Ser Ser Ser Phe Met Met Gly Glu

195 200 205

Asn Cys Be Asn Pro Pro Wing Wing Wing Be Asp Arg Wing Glu Asp Wing

210 215 220

Thr Gly Gln Pro Ser Wing Arg Thr Val Leu Pro Pro Trp Met Val Ser225 230 235 240

His Ile Asn Asn Gly245

<210> 132 <211> 738 <212> DNA

<213> Lolium temulentum <4 00> 132

atggggcgcg gcaaggtgca gctcaagcgg atcgagaaca agatcaaccg ccaggtcacc 60

ttctccaagc gccgctcagg cctgctcaag aaggcgcacg agatctccgt gctctgcgac 120gcagaggtcg ggctcatcat cttctçcacc aagggaaagc tctacgagtt cgccaccgac 180tcatgtatgg acaaaattct tgagcggtat gagcgctact cctatgcaga gaaagtgctc 240atttcaactg aatctgaaat tcagggaaac tggtgtcatg aatataggaa actgaaggcg 300aaggttgaga caatacagag atgtcaaaag catctaatgg gagaggatct tgaatcattg 360aatctcaagg agttgcagca actagagcag cagctggaaa gttcactgaa acatattaga 420tccagaaaga gccagcttat gcacgaatcc atatctgagc ttcaaaagaa ggagaggtca 480ctgcaagagg agaataaaat tctccagaag gaactcatag agaagcagaa ggcccacacg 540cagcaagcgc agttggagca aactcagccc caaaccagct cttcctcctc ctcctttatg 600atgggggaag ctaccccagc aacaaatcgc agtaatcccc cagcagcggc cagcgacaga 660gcagaggatg cgacggggca gcctccagct cgcacggtgc ttccaccatg gatggtgagt 720cacctcaaca atggctga 738

<210> 133 <211> 245 <212> PRT

<213> Lolium temulentum <4 00> 133

Met Gly Arg Gly Lys Val Gln Read Le Lys Arg Ile Glu Asn Lys Ile Asn15 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe

35 40 45

Be Thr Lys Gly Lys Read Tyr Glu Phe Wing Thr Asp Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Ile Be Thr Glu Be Glu Ile Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Arg Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Be Ser Read Lys His Ile Arg Be Arg Lys Be

130 135 140

Gln Read Met His Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Ile Glu Lys Gln

165 170 175

Lys Wing His Thr Gln Gln Wing Gln Read Glu Gln Thr Gln Pro Gln Thr

180 185 190

Be Ser Ser Ser Ser Ser Phe Met Met Gly Glu

195 200 205

Asn Arg Be Asn Pro Pro Wing Ala Wing Be Asp Arg Wing Glu Asp Ala210 215 220Thr Gly Gln Pro Pro Wing Arg Thr Val Leu Pro225 230 235

His Leu Asn Asn Gly245

<210> 134 <211> 738 <212> DNA <213> Zea mays <400> 134

atggggcgcg ggaaggtgca gctgaagcgg atcgagaacattctccaagc gccgctcggg gctgctcaag aaggcgcacggccgaggtcg cgctcatcat cttctccacc aaagggaagctcatgtatgg acaaaattct tgaccggtac gagcgctactatttcagcag aatctgaaac tcagggcaat tggtgccacgaaggtcgaga caatacaaaa atgtcaaaag cacctcatggaatctcaaag agcttcagca actagagcag cagctggagaaccaggaaga accaacttat gctcgagtca atttcggagcctgcaggagg agaacaaggt tctgcagaag gagctcgcggaagcaagtgc aatggggcca aacccaacag cagaccagttataagggaag ctgccccaac aacaaatatc agcatttttcttggtggaag gtgcagcagc gcagccacag gctcgcgttgagccacctga gcagctga

<210> 135

<211> 245

<212> PRT

<213> Zea mays

<400> 135

Pro Trp Met Val Ser240

agatcaaccgagatctccgttctacgagtacctatgcagaagtatagaaagagaggatctgttcactgaatccaacggaaagaagcagaacgtcttcctcctgtggcagcgactaccacc

ccaggtgaccgctctgcgacttccaccgataaaggttcttactaaaggcgtgaaacgttgacatatcagaggagagagagccccccagcgggtgcttcgtgaggcgggggatggatgctt

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile1 5 10 Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly 20 25 His Glu Ile Be Val Leu Cys Asp Wing Glu Val 35 40 Ser Thr Lys Gly Lys Leu Tyr Glu Tyr Be Thr 50 55 Lys Ile Read Asp Arg Tyr Glu Arg Tyr Be Tyr65 70 75Ile Be Wing Glu Be Glu Thr Gln Gly Asn Trp 85 90 Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys 100 105 Met Gly Glu Asp Leu Glu Thr Leu Asn Leu Lys 115 120 Glu Gln Gln Leu Glu Be Ser Leu Lys His Ile 130 135 Gln Leu Met Leu Glu Ser Ile Be Glu Leu Gln145 150 155Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu 165 170 Lys Wing Gln Arg Lys Gln Val Gln Trp Gly Gln 180 185 Be Ser Ser Ser Ser Ser Ser Cys Phe Val Ile

195 200

Asn Ile Ser Ile Phe Pro Val Wing Gly Gly Wing

210 215

Wing Wing Wing Gln Pro Wing Gln Arg Wing Val Gly Leu225 230 235

Be His Leu Ser Ser245

<210> 136 <211> 738 <212> DNA <213> Zea mays

Glu Asn

Leu Leu

Leu45 Wing

Asp Ser60

Glu Wing

Cys his

Cys Gln

Glu Leu125Arg Thr140

Arg lys

Read Wing

Thr gln

Wing Wing205Arg Leu220

Pro pro

Lys Ile Asn15

Lys Lys Ala30

Ile Ile Phe

Cys Met Asp

Lys Val Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Asn

Glu Lys Ser160

Glu Lys Gln

175Gln Gln Thr190

Pro thr thr

Val Glu Gly

Trp Met Leu240

60120180240300360420480540600660720738 <400> 136

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccgttctccaagc gccgctcggg gctactcaag aaggcgcacg agatctccgtgccgaggtcg cgctcatcat cttctccacc aagggcaagc tctacgagtatcatgtatgg acaaaattct tgaacggtat gagcgctact cctatgcagaatttccgcag aatatgaaac tcagggcaat tggtgccatg aatatagaaaaaggtcgaga caatacagaa atgtcaaaag cacctcatgg gagaggatctaatctcaaag agcttcagca actagagcag cagctggaga gttcactgaaacaaggaaga gccagcttat ggtcgagtca atttcagcgc tccaacggaactgcaggagg agaacaaggt tctgcagaag gagctcgcgg agaagcagaacagcaagtgc aacgggacca aactcaacag cagaccagtt cgtcttccacttaagggaag ctgccccaac aacaaatgtc agcatcttcc ctgtggcagcgtggtggaag gggcagcagc gcagccgcag gctcgcgttg gactgccaccagccatctga gctgctga <210> 137 <211> 245 <212> PRT <213> Zea mays <400> 137

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile Glu Asn1 5 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Leu Read

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

40

ccaggtgacagctctgcgacctctaccgataaaggttctcactaaaggcgtgaaactttgacatatcagaggagagtcaagaccagcgggcccccatcatagaggcgggaggatggatgctt

35

Leu45 Wing

Lys ile

15Lys Lys30

Ile Ile

AsnAla

Phe

Be Thr Lys Gly Lys Read Tyr Glu Tyr Be Thr Asp Be Cys Met Asp

50

55

60

Leu80Arg

Read

Read

To be

Ser160Gln

Thr

Thr

Gly

Leu240

<211> 762 <212> DNA

<213> Oryza sativa <4 00> 138

atggggcggg gcaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaggtgaccttctccaagc gcaggtcggg gctgctcaag aaggcgaatg agatctccgt gctctgcgacgccgaggtcg cgctcatcat cttctccacc aagggcaagc tctacgagta cgccaccgactcatgtatgg acaaaatcct tgaacgttat gagcgctact cctatgcaga aaaggtccttatttcagctg aatctgacac tcagggcaac tggtgccacg aatataggaa actgaaggctaaggttgaga caatacagaa atgtcaaaag cacctcatgg gagaggatct tgaatctttgaatctcaaag agctgcagca gctggagcag cagctggaaa attcgttgaa acatatcaga

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Val65 70 75 Ile Be Wing Glu Tyr 85 Glu Thr Gln Gly Asn 90 Trp Cys His Glu Tyr 95Lys Leu Lys Wing 100 Lys Val Glu Thr Ile 105 Gln Lys Cys Gln Lys 110 HisMet Gly Glu 115 Asp Leu Glu Thr Leu 120 Asn Leu Lys Glu Leu 125 Gln GlnGlu Gln 130 Gln Leu Glu Be Ser 135 Leu Lys His Ile Arg 140 Thr Arg LysGln Leu Met Val Glu Ser Ile Ser Wing Ala Leu Gln Arg Lys Glu Lys145 150 155 Leu Gln Glu Glu Asn 165 Lys Val Leu Gln Lys 170 Glu Leu Wing Glu Lys 175Lys Asp Gln Arg 180 Gln Gln Val Gln Arg 185 Asp Gln Thr Gln Gln 190 GlnSer Ser Ser 195 Ser Thr Be Phe Met 200 Leu Arg Glu Wing Wing 205 Pro ThrAsn Val 210 Ser Ile Phe Pro Val 215 Wing Wing Gly Gly Arg 220 Val Val GluAla Wing Gln Pro Gln Wing Arg Val Gly Leu Pro Trp Met225 230 235 Ser His Leu Ser Cys 245 <210> 138

60120180240300360420480540600660720738

60120180240300360420tccagaaaga gccaactaat gctcgagtcc attaacgagc ttcaacggaa ggaaaagtca

ctgcaggagg agaataaggt cctacagaaa gaaaaccctt gctccttcct acagctggtg

gagaagcaga aagtccagaa gcaacaagtg caatgggacc agacacaacc tcaaacaagt

tcctcatcat cctccttcat gatgagggaa gcccttccaa caactaatat cagtaactac

cctgcagcag ctggcgaaag gatagaggat gtagcagcag ggcagccaca gcatgttcgc

attgggctgc caccatggat gctgagccac atcaacggct aa <210> 139 <211> 253 <212> PRT

<213> Oryza sativa <4 00> 139 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr 20 Phe Ser Lys Arg Arg 25 Ser Gly Leu Lys 30 Lys AlaAsn Glu Ile 35 Ser Val Leu Cys Asp 40 Glu Wing Val Wing Leu 45 Ile Ile PheSer Thr 50 Lys Gly Lys Leu Tyr 55 Glu Tyr Wing As Thr 60 Ser Cys Met AspLys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80Ile Be Wing Glu Ser 85 Asp Thr Gln Gly Wing 90 Trp Cys His Glu Tyr 95 ArgLys Leu Lys Wing 100 Lys Val Glu Thr Ile 105 Gln Lys Cys Gln Lys 110 His LeuMet Gly Glu 115 Asp Leu Glu Be Leu 120 Asn Leu Lys Glu Leu 125 Gln Gln LeuGlu Gln 130 Gln Leu Glu Asn Ser 135 Leu Lys His Ile Arg 140 Ser Arg Lys SerGln Leu Met Leu Glu Ser Ile Asn Glu Leu Gln Arg Lys Glu Lys Ser145 150 155 160Leu Gln Glu Glu Asn 165 Lys Val Leu Gln Lys 170 Glu Asn Pro Cys Ser 175 PheLeu Gln Leu Val 180 Glu Lys Gln Lys Val 185 Gln Lys Gln Gln Val 190 Pro Gln Thr Be 200 Ser Be Ser Be Ser 205 Ser Phe Met MetArg Glu 210 Wing Leu Pro Thr Thr 215 Asn Ile Be Asn Tyr 220 Pro Wing AlaGly Wing Glu Arg Ile Glu Asp Val Wing Gly Gln Wing Pro Gln His Val Arg225 230 235 240Ile Gly Leu Pro Pro 245 Trp Met Leu Being His 250 Ile Asn

<210> 140

<211> 741

<212> DNA

<213> Oryza sativa

<400> 140

atggggcggg gcaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaggtgaccttctccaagc gcaggtcaaa actgctcaag aaggcgaatg agatctccgt gctctgcgacgccgaggtcg cgctcatcat cttctccacc aagggcaagc tctacgagta cgccaccgactcatgtatgg acaaaatcct tgaacgttat gagcgctact cctatgcaga aaaggtccttatttcagctg aatctgacac tcagggcaac tggtgccacg aatataggaa actgaaggctaaggttgaga caatacagaa atgtcaaaag cacctcatgg gagaggatct tgaatctttgaatctcaaag agctgcagca gctggagcag cagctggaaa attcgttgaa acatatcagatccagaaaga gccaactaat gctcgagtcc attaacgagc ttcaacggaa ggaaaagtcactgcaggagg agaataaggt cctacagaaa gaactggtgg agaagcagaa agtccagaagcaacaagtgc aatgggacca gacacaacct caaacaagtt cctcatcatc ctccttcatgatgagggaag cccttccaac aactaatatc agtaactacc ctgcagcagc tggcgaaaggatagaggatg tagcagcagg gcagccacag catgaacgca ttgggctgcc accatggatgctgagccaca tcaac210 <2 <11 <11

480540600660720762

60120180240300360420480540600660720741 <212> PRT

<213> Oryza sativa <4 00> 141

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Lys Read Leu Lys Lys Wing

20 25 30

Asn Glu Ile Ser Val Leu Cys Asp Glu Val Wing Leu Ile Ile Phe

35 40 45

Be Thr Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Ile Be Wing Glu Be Asp Thr Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Leu Glu Be Leu Asn Leu Lys Glu Leu Gln Gln Leu

115 120 125

Glu Gln Gln Read Glu Asn Be Read Lys His Ile Arg Be Arg Lys Be

130 135 140

Gln Leu Met Leu Glu Ser Ile Asn Glu Leu Gln Arg Lys Glu Lys Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln

165 170 175

Lys Val Gln Lys Val Gln Val Val Gln Trp Asp Val Gln Thr

180 185 190

Ser Ser Ser Ser Ser Ser Ser Phe Met Met Arg Glu Wing Read Pro Thr Thr

195 200 205

Asn Ile Ser Asn Tyr Pro Wing Wing Wing Gly Glu Arg Ile Glu Asp Val

210 215 220

Wing Gly Wing Gln Pro Gln His Glu Arg Ile Gly Leu Pro Pro Trp Met225 230 235 240

Read His Ile Asn Gly245

<210> 142 <211> 735 <212> DNA

<213> Dendrocalamus Iatiflorus <4 00> 142

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaagtgact 60

ttctccaagc gccggtcggg gctgctcaag aaggcgcatg agatctccgt cctctgcgac 120gccgaggtcg gccttatcat cttctccacc aagggcaagc tctacgagta cgccaccgac 180tcatgtatgg acaaaattct tgaacggtac gagcgttact cctatgcaga aaaggttctt 240atttcagccg aatctgaaac tcagggcaac tggtgtcacg aatataggaa actgaaggcg 300aaggttgaga cgatacagaa atgtcaaaag cacctcatgg gagaggatcc tgaatctttg 360aatctcaagg agctgcagca actcgagcag cagctggaaa gttcagtgaa acatatcaga 420tccagaaaga gccagcttat gctcgagtcc atttccgagc ttcaaaagaa ggagaagtca 480ctgcaggagg agaacaaggt tctgcagaag gaactcgtgg agaagcagca ggtccataaa 540cggttagtgc aatgggacca aactcagccg caaactagtt cctcttcctc gtccttcatg 600atgagggaag ctctcccaac aacaaatatc agtatt

<210> 143 <211> 244 <212> PRT

<213> Dendrocalamus latiflorus <4 00> 143

Met Gly Arg Gly Lys Val Gln Read Le Lys Arg Ile Glu Asn Lys Ile Asn15 10 15

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly Read Leu Lys Lys Wing

20 25 30

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val Gly Leu Ile Ile Phe35 40 45

Be Thr Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Be Cys Met Asp

50 55 60

Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Val Leu65 70 75 80

Ile Be Glu Wing Be Glu Thr Gln Gly Asn Trp Cys His Glu Tyr Arg

85 90 95

Lys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His Leu

100 105 110

Met Gly Glu Asp Pro Glu Being Read Asn Read Lys Read Glu Read Read Gln Read Read

115 120 125

Glu Gln Gln Read Glu Be Ser Val Lys His Ile Arg Ser Arg Lys Ser

130 135 140

Gln Leu Met Leu Glu Ser Ile Ser Glu Leu Gln Lys Lys Glu Lys Ser145 150 155 160

Leu Gln Glu Glu Asn Lys Val Leu Gln Lys Glu Leu Val Glu Lys Gln

165 170 175

Gln Val His Lys Arg Read Val Gln Trp Asp Gln Thr Gln Pro Gln Thr

180 185 190

Ser Ser Ser Ser Ser Ser Ser Phe Met Met Arg Glu Wing Read Pro Thr Thr

195 200 205

Asn Ile Ser Ile Tyr Wing Wing Wing Wing Gly Wing Glu Arg Wing Wing Asp Wing

210 215 220

Gly Wing Gln Pro Gln Ile His Ile Gly Leu Pro Pro Trp Met Val Ser225 230 235 240

His Ile Asn Gly

<210> 144 <211> 735 <212> DNA

<213> Dendrocalamus latiflorus <4 00> 144

atggggcgcg ggaaggtgca gctgaagcgg atcgagaaca agatcaaccg gcaagtgact 60

ttctccaagc gccggtcggg gctgctcaag aaggcgcatg agatctccgt cctctgcgac 120gccgaggtcg gccttatcat cttctccacc aagggcaagc tctacgagta cgccaccgac 180tcatgtatgg acaaaattct tgaacggtac gagcgttact cctatgcaga aaaggttctt 240atttcagccg aatctgaaac tcagggcaac tggtgtcacg aatataggaa actgaaggcg 300aaggttgaga cgatacagaa atgtcaaaag cacctcatgg gagaggatct tgaatctttg 360aatctcaagg agctgcagca actcgagcag cagctggaaa gttcagtgaa acatatcaga 420tccagaaaga gccagcttat gctcgagtcc atttccgagc ttcaaaagaa ggagaagtca 480ctgcaggagg agaacaaggt tctgcagaag gaactcgtgg agaagcagca ggtccataaa 540cggttagtgc aatgggacca aactcagccg caaactagtt cctcttcctc gtccttcatg 600atgagggaag ctctcccaac aacaaatatc agtatt

<210> 145 <211> 244 <212> PRT

<213> Dendrocalamus latiflorus <4 00> 145

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr 20 Phe Ser Lys Arg Arg 25 Ser Gly Leu Leys Lys 30 AlaHis Glu Ile 35 Ser Val Leu Cys Asp 40 Ala Glu Val Gly Leu 45 Ile Ile PheSer Thr 50 Lys Gly Lys Leu Tyr 55 Glu Tyr Ala Thr Asp 60 Ser Cys Met AspLys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Ala Glu Val Leu65 70 75 80Ile Ser Ala Glu Thr 85 Glu Thr Gln Gly Asn 90 Trp Cys His Glu Tyr 95 ArgLys Leu Lys Wing Lys Val Glu Thr Ile Gln Lys Cys Gln Lys His LeuMet

Glu

Gln145Leu

Gln

To be

Asn

Ala225His

Gly

Gln130Leu

Gln

Val

To be

Ile210Gly

Ile

100Glu Asp Leu115

Gln Leu Glu

Met Leu Glu

Glu Glu Asn165

His Lys Arg180Ser Ser Ser195

Ser Ile TyrGln Pro GlnAsn Gly

Glu Ser

Ser Ser135Ser Ile150

Lys val

Read val

To be Phe

Wing Ala215Ile His230

105

Leu Asn Leu120

Val Lys His

Being Glu Leu

Read Gln Lys170

Gln Trp Asp185

Met Met Arg200

Wing Wing GlyIle Gly Leu

Lys glu

Ile Arg140Gln Lys155

Glu Leu

Gln thr

Glu Wing

Glu Arg220Pro Pro235

110Leu Gln125

To be Arg

Lys glu

Val glu

Gln Pro190Leu Pro205

GluTrp Met Wing

Gln Leu

Lys Ser

Lys Ser160Lys Gln175

Gln thr

Thr thr

Asp Wing

Val ser240

<210> 146 <211> 813 <212> DNA <213> Zea mays <4 00> 146

atggggcgcg gcaaggtgca gctgaagcgg atagagaaca agataaaccg gcaggtgacc 60

ttctccaagc gccggaacgg gctgctgaag aaggcgcacg agatctccgt cctctgcgac 120gccgaggtcg ccgtcatcgt cttctccccc aagggcaagc tctacgagta cgcctccgac 180tcccgcatgg acaaaattct agaacgttat gagcgatatt cctatgctga aaaggctctt 240atttcagctg aatctgaaag tgagggaaat tggtgccacg aatacaggaa actgaaggcc 300aaaattgaga ccatacaaag atgccacaag cacctgatgg gagaggatct agagtctttg 360aatccaaaag agctccaaca actagagcag cagctggaga gctcactgaa gcacatcaga 420tcaagaaaga gccaccttat ggccgagtca atttctgagc tacagaagaa ggagaggtca 480ctgcaggagg agaacaagat tctacagaag gaactttcag agaggcagaa ggcggtcgct 540agccggcagc agcagcagca gcaggtgcag tgggaccagc agacacaggt ccaggtccag 600acaagctcat cgtcttcttc cttcatgatg aggcaggatc agcagggact gccacctcca 660caaaacatct gcttcccgcc gttgagcatc ggagagagag gcgaagaggt ggctgcggcg 720gcgcagcagc agctgcctcc tccggggcag gcgcaaccac agctccgcat cgcaggtctg 780ccgccatgga tgctgagcca cctcaatgca taa 813

<210> 147 <211> 270 <212> PRT <213> Zea mays <4 00> 147

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile

Glu Asn

Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu

10

20

25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35

40

Be Pro Lys Gly Lys Read Tyr Glu Tyr Wing Ser

50

55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr

65 70 75

Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Arg

100 105

Met Gly Glu Asp Read Glu Be Read Asn Pro Lys

115

120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130

135

His Leu Met Wing Glu Ser Ile Ser Glu Leu Gln

Val45 wing

Asp Ser60

Glu Wing

Cys his

Cys his

Glu Leu125Arg Ser140

Lys Lys

145

150

155

Lys Ile Asn15

Lys Lys Ala30

Ile Val Phe

Arg Met Asp

Lys Wing Leu80

Glu Tyr Arg95

Lys His Leu110

Gln Gln Leu

Arg Lys Ser

Glu Arg Ser160Leu Gln Glu Glu Asn Lys Ile Leu Gln Lys Glu Leu Ser Glu Arg Gln

165 170 175

Lys Wing Val Wing Be Arg Gln Gln Gln Gln Gln Gln Val Gln Trp Asp

180 185 190

Gln Gln Thr Gln Val Gln Val Gln Thr Be Ser Ser Ser Ser Ser Phe

195 200 205

Met Met Arg Gln Asp Gln Gln Gly Read Pro Pro Gln Asn Ile Cys

210 215 220

Phe Pro Pro Read It Ile Gly Glu Arg Gly Glu Glu Val Wing Wing Wing225 230 235 240

Gln Wing Gln Gln Leu Pro Pro Gly Gln Gln Wing Pro Gln Leu Arg

245 250 255

Ile Ala Gly Leu Pro Pro Trp Met Leu Be His Leu Asn Ala260 265 270

<210> 148 <211> 689 <212> DNA

<213> Sorghum bicolor <4 00> 148

gcaaggtgca gctcaagcgg atagagaaca agataaaccg gcaggtgacc ttctccaagc 60

gccgcaacgg gctgctcaag aaggcgcacg agatctccgt cctctgcgac gccgaggtcg 120ccgtcatcgt cttctccccc aagggcaagc tctatgagta cgccaccgac tcccgcatgg 180acaaaattct cgaacgttat gagcgatatt cctatgctga aaaggctctt atttcagctg 240aatctgaaag tgagggaaac tggtgccacg aatacaggaa actgaaggcc aaaattgaga 300ccattcaaaa atgccacaag cacctgatgg gagaggatct agagtctttg aatcccaaag 360agctccaaca actagagcag cagctggaga gctcactgaa gcacatcaga tcaagaaaga 420gccaccttat ggctgagtct atttctgaac tacagaagaa ggagaggtca ctgcaggagg 480agaacaaggc tctacagaag gaacttgcgg agaggcagaa ggcggccgcg agcaggcagc 540agcagcaagg tgcagtggga ccagcagaca cagacccagg cccagacaag ctcatcatcg 600tcctccttca tgatgaggca ggatcagcag ggtctgccgc ctccacaaaa catatgcttc 660ccgccgctga taatcggaga gagaggtga 689

<210> 149 <211> 228 <212> PRT

<213> Sorghum bicolor <4 00> 149

Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn Arg Gln Val Thr15 10 15

Phe Ser Lys Arg Asn Gly Leu Leu Read Lys Lys Wing His Glu Ile Ser

20 25 30

Val Leu Cys Asp Glu Wing Val Wing Val Ile Val Phe Ser Pro Lys Gly

35 40 45

Lys Leu Tyr Glu Tyr Wing As Thr Be Arg Met Asp Lys Ile Leu Glu

50 55 60

Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Wing Leu Ile Ser Wing Glu65 70 75 80

Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg Lys

85 90 95

Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu Met Gly Glu Asp

100 105 110

Leu Glu Be Leu Asn Pro Lys Glu Leu Gln Gln Leu Glu Gln Gln Leu

115 120 125

Glu Being Ser Liu His Ile Arg Being Arg Lys Being His Leu Met Wing

130 135 140

Glu Be Ile Be Glu Read Gln Lys Lys Glu Arg Be Read Gln Glu Glu145 150 155 160

Asn Lys Wing Leu Gln Lys Glu Leu Wing Glu Arg Gln Lys Wing Wing

165 170 175

Be Arg Gln Gln Gln Gln Gly Wing Val Gly Pro Wing Asp Thr Asp Pro

180 185 190

Gly Pro Asp Lys Leu Ile Ile Val Leu Read His Asp Glu Wing Gly Ser

195 200 205

Wing Gly Be Wing Wing Be Thr Lys His Met Leu Pro Wing Wing Asp Asn210 215 220

Arg Arg Glu Arg225

<210> 150 <211> 822 <212> DNA <213> Zea mays <400> 150

atggggcgcg gcaaggtaca gctgaagcgg atagagaaca agataaaccg gcaggtgacc 60

ttctccaagc gccggaacgg cctgctcaag aaggcgcacg agatctccgt cctctgcgat 120gccgaggtcg ccgtcatcgt cttctccccc aagggcaagc tctacgagta cgccaccgac 180tcccgcatgg acaaaattct tgaacgctat gagcgatatt cctatgctga aaaggctctt 240atttcagctg aatctgaaag tgagggaaat tggtgccacg aatacaggaa actgaaggcc 300aaaattgaga ccatacaaaa atgccacaag cacctgatgg gagaggatct agagtctttg 360aatcccaaag agctccagca actagagcag cagctggata gctcactgaa gcacatcaga 420tcaaggaaga gccaccttat ggccgagtct atttctgagc tacagaagaa ggagaggtca 480ctgcaggagg agaacaaggc tctgcagaag gaacttgcgg agaggcagaa ggccgtcgcg 54 0agccggcagc agcagcaaca gcagcaggtg cagtgggacc agcagacaca tgcccaggcc 600cagacaagct catcatcgtc ctccttcatg atgaggcagg atcagcaggg actgccgcct 660ccacacaaca tctgcttccc gccgttgaca atgggagata gaggtgaaga gctggctgcg 720gcggcggcgg cgcagcagca gcagccactg ccggggcagg cgcaaccgca gctccgcatc 780gcaggtctgc caccatggat gctgagccac ctcaatgcat 822 aa

<210> 151 <211> 273 <212> PRT

<213> Zea mays <4 00> 151 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Read Lys Lys Wing 20 25 30 His Glu Ile Ser Val Leu Cys Asp Glu Wing Val Val Val Ile Val Phe 35 40 45 Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Being Arg Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Ala Tyr Glu Lys Wing Leu65 70 75 80Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Be Leu Asn Pro Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Leu Asp Be Ser Leu Lys His Ile Arg Be Arg Lys Ser 130 135 140 His Leu Met Wing Glu Ser Ile Be Glu Leu Glys Lys Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Wing Glu Arg Gln 165 170 175 Ly s Wing Val Wing Wing Being Arg Gln Gln Gln Gln Gln Gln Gln Val Gln Trp 180 185 190 Asp Gln Gln Thr His Wing Gln Gln Thr Being Being Being Being Being 195 200 205 Phe Met Met Arg Gln Asp Gln Gln Gly Leu Pro Pro Pro His Asn Ile 210 215 220 Cys Phe Pro Pro Read Thr Met Gly Asp Arg Gly Glu Leu Wing Ala225 230 235 240Ala Wing Wing Wing Gln Gln Gln Pro Leu Pro Gly Gln Wing 245 250 255 Gln Leu Arg Ile Wing Gly Leu Pro Pro Trp Met Leu Be His Leu Asn 260 265 270 Ala <210> 152 <211> 786 <212> DNA

<213> Lolium temulentum <400> 152

atgggtcgcg gcaaggtgca gctgaagcgg atagagaaca agataaaccg tcaggtgaca 60

ttctccaagc gccgcaacgg gctactcaag aaggcgcacg agatctccgt cctctgcgac 120gccgaggtcg ccgtcgtcgt cttctccccg aaagggaagc tctatgagta cgccactgac 180tccagcatgg acaaaattct tgaacgttat gaacgctact cttatgctga aaaggctttg 240atttcagctg aatctgaaag tgagggaaat tggtgccatg aatacaggaa gctgaaggcg 300aagattgaga ctatacaaaa atgtcacaag cacctcatgg gggaggatct ggagtgtcta 360aacctgaaag agctccaaca actagagcag cagctggaga gttcattgaa gcacatcaga 420tcgagaaaga gccaccttat gatggagtcc atttctgagc tacagaagaa ggagcggtca 480ctccaggagg agaacaaggc tctacagaag gaactggtgg agaggcagaa ggcggccagg 54 0cagcagcagc aagagcagtg ggaccgtcag acccaaacac aacaagccca aaaccaacct 600caggcccaga cgagctcatc atcttcctcc ttcatgatga gggatcagca ggcccatgct 660caacaaaaca tctgttaccc gctggtgaca atgggtggag aggctgtggc cgcggcgcca 720gggcagcagg ggcagcttcg catcggaggc ctgccaccat ggatgctgag ccacctcaac 780gcttga 786

<210> 153 <211> 261 <212> PRT

<213> Lolium temulentum <4 00> 153 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu Lys Lys Wing 20 25 30 His Glu Ile Ser Val Valu Cys Asp Glu Val Wing Val Val Val Val Phe 35 40 45 Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Ala Thr Asp Being Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Lys Tyr Wing Leu65 70 75 80Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Cys His Lys His Leu 100 105 110 Met Gly Glu Asp Leu Glu Cys Leu Asn Leu Lys Glu Leu Gln Gln Leu 115 120 125 Glu Gln Gln Gln Leu Glu Be Ser Leu Lys His Ile Arg Be Arg Lys Ser 130 135 140 His Leu Met Met Glu Ser Ile Be Glu Leu Glys Lys Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val Glu Arg Gln 165 170 175 Lys Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Gln Gln Wing Asn Ile 210 215 220 Cys Tyr Pro Leu Val Thr Met Gly Gly Glu Val Wing Wing Pro225 230 235 240Gly Gln Gln Gly Gln Leu Arg Ile Gly Gly Leu Pro Trp Met Leu 245 250 255 Be His Leu Asn Ala 260 <210> 154

<211> 786 <212> DNA

<213> Lolium perenne <4 00> 154atgggtcgcg gcaaggtgca gctgaagcgg atagagaaca

ttctccaagc gccgcaacgg gctactcaag aaggcgcacg

gccgaggtcg ccgtcgtcgt cttctccccg aaagggaagc

tccagcatgg acaaaattct tgaacgttat gaacgctact

atttcagctg aatctgaaag tgagggaaat tggtgccatg

aagattgaga ctatacaaaa atgtcacaag cacctcatgg

aacctgaaag agctccaaca actagagcag cagctggaga

tcgagaaaga gccaccttat gatggagtcc atttctgagc

ctccaggagg agaacaaggc tctacagaag gaactggtgg

cagcagcagc aagagcagtg ggaccgtcag acccaaacac

caggcccaga cgagctcatc atcttcctcc ttcatgatga

caacaaaaca tctgttaccc gccggtgaca atgggtggag

gggcagcagg ggcagcttcg catcggaggc ctgccaccatgcttga <210> 155 <211> 261 <212> PRT

<213> Lolium perenne <4 00> 155

Met Gly Arg Gly Lys Val Gln Read Lys Arg Ile

15 10Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly

20 25His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40Ser Pro Lys Gly Lys Read Tyr Glu Tyr Wing Thr

agataaaccgagatctccgttctatgagtacttatgctgaaatacaggaagggaggatctgttcattgaatacagaagaaagaggcagaaaacaagcccagggatcagcaaggctgtggcggatgctgag

tcaggtgacccctctgcgaccgccactgacaaaggctttgactgaaggcgggagtgtctagcacatagaggagcggtcaggcggccaggaaaccaacctggcccatgctcgcggcgccaccacctcaac

50

55

Lys Ile Read Glu Arg Tyr Glu Arg Tyr Ser Tyr65 70 75

Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys

100 105

Met Gly Glu Asp Leu Glu Cys Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130 135

His Leu Met Met Glu Ser Ile Ser Glu Leu Gln145 150 155

Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu

165 170

Lys Wing Arg Wing Gln Gln Gln Gln Gln Glu Gln Trp

180 185

Thr Gln Gln Wing Gln Asn Gln Pro Gln Wing Gln

195 200

Ser Be Phe Met Met Asp Gln Gln Wing His

210 215

Cys Tyr Pro Val Thr Met Gly Gly Glu Ala225 230 235

Gly Gn Gln Gly Gnu Leu Arg Ile Gly Gly Gnu

245 250

Being His Leu Asn Ala260

<210> 156 <211> 831 <212> DNA

<213> Hordeum vulgare <4 00> 156

agggtggggg

Glu Asn

Leu Leu

Val45 wing

Asp Ser60

Glu Wing

Cys his

Cys his

Glu Leu125Arg Ser140

Lys Lys

Read val

Asp Arg

Thr Ser205Ala Gln220

Val AlaPro Pro

Lys ile

15Lys Lys30

Val Val

Be met

Lys Wing

Glu Tyr

95Lys His110

Gln Gln

Arg lys

Glu Arg

Glu Arg175Gln Thr190

To be to be

Gln asn

Wing wing

Trp Met255

Asn

Allah

Phe

Asp

Leu80Arg

Read

Read

To be

Ser160Gln

Gln

To be

Ile

Pro240Leu

60120180240300360420480540600660720780786

agataaatcgagatctccgttctatgagtacttatgctgaaatacaggaa

gcaggtgacccctctgtgaccgccaccgacaaaggctcttacttaaggcg

60120180240300aagattgaga ccatacagaa gtgtcacaag cacctcatgg gagaggatct ggattctctgaacctcaaag aactccaaca actggagcag cagctggaga gttcattgaa gcacatcagatcgagaaaga gccatcttat gatggagtcc atttctgagc tacagaagaa ggagaggtcactgcaggagg agaacaaggc cctacagaag gaactggtgg agaggcagaa ggcggccagcaggcagcagc agctgcagca gcagcaacaa caacaacaaa tgcaatggga gcaccaagcccagacccaaa cccataccca tactcaaaac cagccccaag cccagactag ctcatcatcttcctctttca tgatgaggga tcagcaggcc catgcccctc aacagaacat ttgtagctacccaccggtga cgatgggtgg ggaggcgacg gcggcggcgg cggcgccgga gcagcaggctcagcttcgca tatgcctacc gccatggatg ctgagccacc tcaacgcttg the <210> 15.7 <211> 276 <212> PRT

<213> Hordeum vulgare <400> 157

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Lys Ile Asn1 5 10 15 Arg Gln Val Thr Phe Ser Lys Arg Arg Asn Gly Leu Leu Lys Lys Wing 20 25 30 His Glu Ile Be Val Leu Cys Asp Wing Glu Val Wing Val Ile Val Phe 35 40 45 Be Pro Lys Gly Lys Leu Tyr Glu Tyr Wing Thr Asp Be Ser Met Asp 50 55 60 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Wing Leu65 70 75 80Ile Be Wing Glu Be Glu Be Glu Gly Asn Trp Cys His Glu Tyr Arg 85 90 95 Lys Leu Lys Ala Lys Ile Glu Thr Ile Gln Lys Cys His Lys His Leu 100 105 110 Met Gly Glu Asp Leu Asp Ser Leu Asn Leu Lys Glu Leu Gln Leu 115 120 125 Glu Gln Gln Glu Leu Glu Be Being Read Lys His Ile Arg Be Arg Lys Be 130 135 140 His Met Le Met Met Glu Be Ile Be Glu Leu Gln Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn Lys Wing Leu Gln Lys Glu Leu Val Glu Arg Gln 165 170 175 Lys Wing Wing Be Arg Gln Gln Gln Leu Gln Gln Gln Gln Gln Gln Gln 180 185 190 Gln Met Gln Trp Glu His Gln Wing Gln Thr Gln Thr His Thr His Thr 195 200 205 Gln Asn Gln Pro Gln Wing Gln Thr Being Ser Ser Ser Ser Ser 210 210 220 Met Arg Asp Gln Gln His Wing Pro Wing Gln Gln Asn Ile Cys Ser Tyr225 230 235 240Pro Pro Val Thr Met Gly Gly Glu Wing Thr Wing Wing Wing Pro Wing 245 250 255 Glu Gln Gln Wing Gln Leu Arg Ile Cys Leu Pro Trp Met Leu Ser 260 265 270 His Read Asn Wing

275 <210> 158 <211> 804 <212> DNA <213> Oryza sativa <400> 158

atggggcggg ggaaggtgca gctgaagcgg atagagaact cgatgaaccg gcaggtgacgttctccaaga ggaggaatgg attgctgaag aaggcgcacg agatctccgt cctctgcgacgccgaggtcg ccgccatcgt cttctccccc aagggcaagc tctacgagta cgccactgactccaggatgg acaaaatcct tgaacgttat gagcgctatt catatgctga aaaggctcttatttcagctg aatctgagag tgagggaaat tggtgccatg aatacaggaa gcttaaggcaaagattgaga ccatacaaaa atgtcacaaa cacctcatgg gagaggatct agaatccctgaatctcaaag aactccaaca gctagagcag cagctggaga gttcattgaa gcacataagatcaagaaaga gccaccttat gcttgagtcc atttccgagc tgcagaaaaa ggagaggtca

360420480540600660720780831

60120180240300360420480ctgcaggagg agaacaaggc tctgcagaag gaactggtggggccagcagc aagtagggca gtgggaccaa acccaggtccccccaagccc agacaagctt cttcttcttc ttcatgctgatcaccacaaa atatctgcta cccgccggtg atgatgggcccggcggcggt ggcggcccaa ggccaggtgc aacttccgcaatgctgagca ccttcaaggc ttaa <210> 159 <211> 267 <212> PRT

<213> Oryza sativa <400> 159

agaggcagaaaggcccaggcgggatcagcaagagaaatgattggaggctt

gaatgtgaggccaagcccaaggcacttctttgcggcggcgtccgccatgg

Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile Glu Asn Ser Met Asn1 5 10 15 Arg Gln Val Thr 20 Phe Ser Lys Arg Arg 25 Asn Gly Leu Leu Lys 30 Lys AlaHis Glu Ile 35 Ser Val Leu Cys Asp 40 Ala Glu Val Wing 45 Wing Ile Val PheSer Pro 50 Lys Gly Lys Leu Tyr 55 Glu Tyr Wing Thr Asp 60 Ser Arg Met AspLys Ile Leu Glu Arg Tyr Glu Arg Tyr Ser Tyr Wing Glu Lys Alu Leu65 70 75 80Ile Ser Wing Glu Ser 85 Glu Ser Glu Gly Asn 90 Trp Cys His Glu Tyr 95 ArgLys Leu Lys Wing 100 Lys Ile Glu Thr Ile 105 Gln Lys Cys His Lys 110 His LeuMet Gly Glu 115 Asp Leu Glu Being Leu 120 Asn Leu Lys Glu Leu 125 Gln LeuGlu Gln 130 Gln Leu Glu Being Ser 135 Leu Lys His Ile Arg 140 Being Arg Lys SerHis Leu Met Leu Glu Being Ile Being Glu Leu Gln Lys Glu Arg Ser145 150 155 160Leu Gln Glu Glu Asn 165 Lys Wing Leu Gln Lys 170 Glu Leu Val Glu Arg 175 GlnLys Asn Val Arg 180 Gly Gln Gln Gln Val 185 Gly Gln Trp Asp Gln 190 Thr GlnVal Gln Wing 195 Gln Wing Gln Wing Gln 200 Pro Gln Wing Gln Thr 205 Ser Phe PhePh and Phe 210 Phe Met Leu Arg Asp 215 Gln Gln Wing Leu Leu 220 Ser Pro Gln AsnIle Cys Tyr Pro Val Met Met Gly Gln Arg Asn Asa Wing Ala225 230 235 240Arg Arg Trp Arg 245 Pro Lys Wing Arg Cys 250 Asn Phe Arg Ile Gly 255 GlyPhe Pro Pro Trp 260 Met Read Ser Thr Phe 265 Lys Wing

<210> 160

<211> 804

<212> DNA

<213> Oryza sativa

<4 00> 160

atggggcggg ggaaggtgca gctgaagcgg atagagaaca agatcaacag gcaggtgacg

ttctccaaga ggaggaatgg attgctgaag aaggcgcacg agatctccgt cctctgcgac

gccgaggtcg ccgccatcgt cttctccccc aagggcaagc tctacgagta cgccactgac

tccaggatgg acaaaatcct tgaacgttat gagcgctatt catatgctga aaaggctctt

atttcagctg aatccgagag tgagggaaat tggtgccatg aatacaggaa acttaaggca

aagattgaga ccatacaaaa atgtcacaaa cacctcatgg gagaggatca tgaatccctg

aatctcaaag aactccaaca gctagagcag cagctggaga gttcattgaa gcacataata

tcaagaaaga gccaccttat gcttgagtcc atttccgagc tgcagaaaaa ggagaggtca

ctgcaggagg agaacaaggc tctgcagaag gaactggtgg agaggcagaa gaatgtgagg

ggccagcagc aagtagggca gtgggaccaa acccaggtcc aggcccaggc ccaagcccaa

ccccaagccc agacaagctc ctcctcctcc tccatgctga gggatcagca ggcacttctt

ccaccacaaa atatctgcta cccgccggtg atgatgggcg agagaaatga tgcggcggcg

gcggcggcgg tggcggcgca gggccaggtg caactccgca tcggaggtct tccgccatgg

540600660720780804

60120180240300360420480540600660720780atgctgagcc acctcaatgc ttaa <210> 161 <211> 267 <212> PRT

804

<213> Oryza sativa <400> 161 Met Gly Arg Gly Lys Val Gln Leu Lys Arg Ile1 5 10 Arg Gln Val Thr Phe Ser Lys Arg Asn Gly 20 25 His Glu Ile Ser Val Leu Cys Asp Wing Glu Val 35 40 Ser Pro Lys Gly Lys Leu Tyr Glu Tyr Wing Thr 50 55 Lys Ile Leu Glu Arg Tyr Glu Arg Tyr To Be Tyr65 70 75Ile Be Wing Glu Be Glu To Be Glu Gly Asn Trp 85 90 Lys Leu Lys Wing Lys Ile Glu Thr Ile Gln Lys 100 105 Met Gly Glu Asp His Glu Be Leu Asn Leu Lys 115 120 Glu Gln Gln Glu Leu Glu Be Ser Leu Lys His Ile 130 135 His Leu Met Leu Glu Be Ile Ser Glu Leu Gln145 150 155Leu Gln Glu Glu Asn Lys Ala Leu Gln Lys Glu 165 170 Lys Asn Val Arg Gly Gln Gln Gln Val Gly Gln 180 185 Val Gln Wing Gln Wing Gln Wing Gln Pro Gln Wing

Glu Asn LysLeuAla

Asp60Ala

Cys

Cys

Glu

Ile140Lys

Read

Trp

Gln

Read lys

30Ala Ile45

To be Arg

195

200

Ser Ser Ser Met Leu Arg Asp Gln Gln Wing Leu Leu210 215 220

Ile Cys Tyr Pro Pro Met Met Gly Glu Arg Asn225 230 235

Wing Wing Wing Val Wing Wing Gln Gly Gln Val Gln Leu

245 250

Leu Pro Pro Trp Met Leu Be His Leu Asn Ala260 265

<210> 162 <211> 668 <212> DNA

<213> Virgin Tradescantia <4 00> 162

gggctggtga agaaggctca tgagatctcd atkytrtgtgattttctcta ctaaaggcaa actatacgag tatgccactgcttgaacgct atgaacgtta ctcatatgct gagaaggcttttacagggaa attggtgcca agagtatgtt aaacttaaggaaaagccaaa ggcatcttat gggagagcaa ctagaagcgtcaactagagc atcaacttga aggttctttg aggcttgtcaatgttggact ccatttccga acttcagagg aaggaaaagtaacctagaga aggagatttt ggagaagcag aaagaaaaggtgggaacagc agaatcagcc actacaaagc actaattcgcgcagaaactc atccaacact aaacattgga aatttccaaggcagaagaaa gtctgcagcg tcagatgagg atcagcagcamttcacma <210> 163 <211> 222 <212> PRT

<213> Virgin Tradescantia <220>

<221> UNSAFE

Glu Lys

His glu

His Lys110Leu Gln125

To be Arg

Lys glu

Val glu

Asp Gln190Thr Ser205

Pro ProAsp AlaArg Ile

Ile Asn15

Lys Wing

Val phe

Met asp

Wing Leu80Tyr Arg95

His leu

Gln Leu

Lys Ser

Arg Ser160Arg Gln175

Thr gln

To be to be

Gln asn

Wing Ala240Gly Gly255

atgcygaggtattccaaaattaacttcatcctaaggttgatggatctcaaggtcaagaaactctggaagactctggcacactccaaggccgtagaacaaagcctactgcc

tgcgcttattggaaaatatcagatcctgaaggccttacatagaattgcaggactcaaatggcaaaacaagccaagctcaccttcgtgatttaccgtccatmycmtggatg

60120180240300360420480540600660668 <222> (11) .. (11)

<223> Xaa can be any naturally occurring amino acid <220>

<221> UNSAFE <222> (218) .. (218)

<223> Xaa can be any naturally occurring amino acid <220>

<221> UNSAFE <222> (221) .. (221)

<223> Xaa can be any naturally occurring amino acid <4 00> 163

Gly Leu Val Lys Lys Wing His Glu Ile Ser Xaa Leu Cys Asp Wing Glu15 10 15

Val Wing Leu Ile Ile Phe Ser Thr Lys Gly Lys Leu Tyr Glu Tyr Wing

20 25 30 Thr Asp Ser 35 Lys Met Glu Asn Ile 40 Leu Glu Arg Tyr Glu 45 Arg Tyr SerTyr Wing 50 Glu Lys Wing Leu Thr 55 Ser As Asp Pro Glu 60 Leu Gln Gly AsnTrp Cys Gln Glu Tyr Val Lys Leu Lys Wing Val Glu Wing Leu His65 70 75 80Lys Ser Gln Arg His 85 Leu Met Gly Glu Gln 90 Leu Glu Wing Leu Asp 95 LeuLys Glu Leu Gln 100 Gln Leu Glu His Gln 105 Leu Glu Gly Ser Leu 110 Arg LeuVal Arg Ser 115 Arg Lys Thr Gln Met 120 Met Leu Asp Ser Ile 125 Ser Glu LeuGln Arg 130 Lys Glu Lys Ser Leu 135 Glu Glu Gln Asn Lys 140 Asn Leu Glu LysGlu Ile Leu Glu Lys Gln Lys Ala Leu Wing His His Gln Wing His145 150 155 160Trp Glu Gln Gln Asn 165 Gln Pro Leu Gln Be 170 Thr Asn Ser Pro Pro 175 Arg Phe Val Ile 180 Wing Glu Thr His Pro 185 Thr Leu Asn Ile Gly 190 Asn PheGln Gly Arg 195 Thr Asn Thr Val His 200 Wing Glu Glu Ser Leu 205 Gln Arg Gln

Met Arg Ile Ser Ser Ser Leu Pro Xaa Trp Met Xaa His

215

220

210 <210> 164 <211> 723 <212> DNA

<213> Virgin Tradescantia <4 00> 164

gtgcagctga aacggatgga gaacaagatt aacaggcagg tgacgttttc taaacgtcga 60

ggagggctgc tgaagaaagc tcatgagatc tctattctat gtgatgctga gattgctctt 120attattttct ctactaaagg gaagctctat gagtatgcca ccaattccaa aatggacaat 180attcttgaac gctatgagcg ttactcatat gctgaaaagg ctctaacttc atcagatcct 240gatatacagg gaaattggtg ccaagagtat gctaaactta agtctaaggt tgaggcttta 300tgtaaaagcc aaaggcatct tatgggagag cagcttgaaa cattgaatct caaagaattg 360cagcaactag agcaacagct cgaaggttct ctaaagcatg tcaggtcaag aaagactcaa 420gttatgctgg actctatttc tgaacttcag aggaaggaaa agtcactaga ggagcaaaac 480aagaacctag agaaggagat tttggagaag cagaaaatca aggctcttgc acagcaggct 540cactgggaac accagaatca accagcacca aggggttcac ctcctaggcc atttgtgatt 600gcagagttctc atccgacact aaatattgga catttccaag gcaggacaaa tgcagtcgaa 660gcagaagaaa atcagcagcc tcakatgaga atttgcagta gcctccggcccccccc23gcccccccc

<210> 165 <211> 241 <212> PRT

<213> Virgin Tradescantia <220>

<221> UNSAFE <222> (228). . (228)

<223> Xaa can be any naturally occurring amino acid <4 00> 165

Val Gln Leu Lys Arg Met Glu Asn Lys Ile Asn Arg Gln Val Thr Phe1 5 10 15 Ser Lys Arg Arg Gly Gly Leu Leu Lys Lys Wing His Glu Ile Ser Ile 20 25 30 Leu Cys Asp Wing Glu Ile Wing Leu Ile Ile Phe Ser Thr Lys Gly Lys 35 40 45 Leu Tyr Glu Tyr Wing Thr Asn Be Lys Met Asp Asn Ile Leu Glu Arg 50 55 60 Tyr Glu Arg Tyr Be Tyr Wing Glu Lys Wing Leu Thr Be Ser Asp Pro65 70 75 80Asp Ile Gln Gly Asn Trp Cys Gln Glu Tyr Ala Lys Leu Lys Ser Lys 85 90 95 Val Glu Ala Leu Cys Lys Ser Gln Arg His Leu Met Gly Glu Leu 100 105 110 Glu Thr Leu Asn Leu Lys Glu Leu Gln Leu Gln Leu Gln Leu Glu 115 120 125 Gly Be Leu Lys His Val Arg Be Arg Lys Thr Gln Val Met Leu Asp 130 135 140 Be Ile Be Glu Leu Gln Arg Lys Glu Lys Be Leu Glu Glu Gln Asn145 150 155 160Lys Asn Leu Glu Lys Glu Ile Leu Glu Lys Gln Lys Ile Lys Wing Leu 165 170 175 Wing Gln Gln Wing His Trp Glu His Gln Asn Gln Pro Arg Gly 180 185 190 Ser Pro Pro Arg Pro Phe Val Ile Glu Wing Be His Pro Thr Leu Asn 195 200 205 Ile Gly His Phe Gln Gly Arg Thr Asn Wing Val Glu Glu Wing Asn 210 215 220 Gln Gln Pro Xaa Met Arg Ile Cys Ser Seru Leu Pro Pro Pro Met225 230 235 240

Read <210> 166 <211> 599 <212> DNA

<213> Virgin Tradescantia <4 00> 166

gggctkgtga agaaagctca tgagatctcg gtactttgtg atgctgagct tgctcttatt 60

atcttctctc ccaaaggcaa gctctatgag tatgccaccg attccaaaat ggaaattatt 120cttgaacgct atgaacgtta cacctacgct gaaaaagctt taattgcatc agatcctgat 180gtacagggaa actggtgtca tgagtacatt aagcttaaag ctaaatttga ggccttgaat 240aaaagccaga ggcatcttat gggagaacaa ctagatacgt tgaaccaaaa ggaattgctg 300caactagaga ctaagcttga aggttctctg aaaaacgtca ggtcaagaaa gactcaactt 360atgttggatt ccatttctga gcttcaagaa aagggaaagt cactccagga gcaaaacacc 420tgcctagaaa aggagatttt gggaaaacag aaagacaagg ctcccaaaca gcatgttcag 480tgggaaaaac agaatcaacc accacctacc tcttctgcgc caatgccatt cctcattggt 540gatattcacc caacccctaa tatcagaaat ttccaaggca gaacagtagc tgatgcaga 599

<210> 167 <211> 199 <212> PRT

<213> Virgin Tradescantia <4 00> 167

Gly 1 Leu Val Lys Lys 5 Wing His Glu Ile Ser 10 Val Leu Cys Asp Wing 15 GluLeu Wing Leu Ile 20 Ile Phe Ser Pro Lys 25 Gly Lys Leu Tyr Glu 30 Tyr AlaThr Asp Ser 35 Lys Met Glu Ile Ile 40 Leu Glu Arg Tyr Glu 45 Arg Tyr ThrTyr Wing 50 Glu Lys Wing Leu Ile 55 Wing Ser Asp Pro Asp 60 Val Gln Gly AsnTrp Cys His Glu Tyr Ile Lys Leu Lys Phe Glu Wing Leu Asn65 70 75

Lys Ser Gln Arg His Leu Met Gly Glu Gln Leu

85 90

Lys Glu Read Leu Gln Read Le Glu Thr Lys Read Le Glu

100 105

Val Arg Be Arg Lys Thr Gln Leu Met Leu Asp

115 120

Gln Glu Lys Gly Lys Being Read Gln Glu Gln Asn

130 135

Glu Ile Read Gly Lys Gln Lys Asp Lys Wing Pro145 150 155

Trp Glu Lys Gln Pro Gln Pro Pro

165 170

Phe Leu Ile Gly Asp Ile His Pro Thr Pro Asn

180 185

Gly Arg Thr Val Wing Asp Wing

195 <210> 168 <211> 753 <212> DNA

<213> Elaeis guineensis <400> 168

atggggagag ggagggtgca gctgaggcgg atcgagaacattctcgaagc gccggtcggg gctcctgaag aaagcccacggccgaggtcg ccctcatcat cttctcgacc aagggcaagctcctgcatgg aaaggattct tgaacgctat gaacgttacaatttcatctg gacccgaatt gcagggtaac tggtgccatgaaggttgagg ctttacaaaa aagccaaagg catctcatggaatctcaaag aactccagca actagagcaa cagcttgaaaaccagaaagt gccaactcat gtttgaatcc atctctgagcctgcaggagc agaacaagat gctggagaag gagctcatggctaaaccagc aggcaccttg ggagcagcaa ggcccgccgcacctccttcc tgatcggaga ctctctcccc accctgaataggaaatgaac atggggagga agcagcacaa ccccaggttcccaccttgga tgcttagcca tag cttgaacggg <210> 169 <211> 250 <212 > PRT

<213> Elaeis guineensis <4 00> 169

Met Gly Arg Gly Arg Val Gln Leu Arg Arg Ile15 10

Arg Gln Val Thr Phe Be Lys Arg Arg Be Gly

20 25

His Glu Ile Ser Val Leu Cys Asp Wing Glu Val

35 40

Being Thr Lys Gly Lys Read Tyr Glu Tyr Wing Thr

50 55

Arg Ile Read Glu Arg Tyr Glu Arg Tyr Thr Tyr65 70 75

Ile Be Ser Gly Pro Glu Read Gln Gly Asn Trp

85 90

Lys Leu Lys Wing Lys Val Glu Wing Leu Gln Lys

100 105

Met Gly Glu Gln Leu Glu Pro Leu Asn Leu Lys

115 120

Glu Gln Gln Read Glu Be Ser Read Lys His Ile

130 135

Gln Read Met Phe Glu Be Ile Be Glu Leu Gln145 150 155

Leu Gln Glu Gln Asn Lys Met

165 170

Lys Val Lys Wing Read Asn Gln Gln Wing Pro Trp

Asp Thr LeuGlySer

Thr140Lys

To be

Ile

Ser Leu110Ile Ser125

Cys leu

Gln his

Pro Wing

Arg Asn190

Asn

95

Lys

Glu

Glu

Val

Met175Phe

80Gln

Asn

Read

Lys

Gln160Pro

Gln

agataaaccgagatctccgttctacgagtacctatgcagaaatttggcaagtgagcaactgttctttaaattcaaaaaaaagaagcagaaagacaagctcttgggacatagtataggaaa

gcaggtgacgcctctgcgaccgccaccgacaaaagcactaactcaaagcttgagcccttggcatataagaggaaaagtcaggtgaaggcaatcat ccccaccaatgtagccagcctgtta

Glu Asn

Leu Leu

Leu45 Wing

Asp Ser60

Glu Wing

Cys his

Be gln

Glu Leu125Arg Thr140

Lys LysLeu MetGlu Gln

Lys Ile Asn15

Lys Lys Ala30

Ile Ile Phe

Cys Met Glu

Lys Wing Leu80

Glu Phe Gly95

Arg His Leu110

Gln Gln Leu

Arg Lys Cys

Glu Lys Ser160

Glu Lys Gln175

Gln Gly Pro

60120180240300360420480540600660720753180 185

Pro Gln Thr Be Be Ser Be Pro Thr Be Be

195 200

Read Pro Thr Read Asn Ile Gly Thr Tyr Gln Cys

210 215

Gly Glu Glu Wing Gln Pro Wing Gln Val Arg Ile225 230 235

Pro Pro Trp Met Leu Be His Leu Asn Gly245 250

<210> 170 <211> 699 <212> DNA <213> Allium sp. <400> 170

gtgcaattga agaggatgga aaacaagatt aatagacaagaatggtttgt tgaagaaagc tcatgagatt tcwgtgctttattgttttct ctgctaaagg aaaactctat gaatattcaaattctggaga ggtatgaacg ttattgcttt gcggagaaatgactcccagg aggattggag ccttgaatat cacaaactgaaacaacaggc aaaggcatct tatgggagag caacttgaatggacagcttg agcaacaact tgagaattct ctcaaaactggaattgttaa gttctatttc agagcttcag gacaaggagaaaagctttag aaaatgagct tatgaaaagg gccagggcaagcacgatgga agcatcataa tcataaacaa caggataatcattggaaatt accaaacaag gaacaatgag ggaggagttggtacgtgttg ttagaaattt gttgccccac tggatgctt <210> 171 <211> 233 <212> PRT < 213> Allium sp. <400> 171

Val Gln Read Lys Arg Met Glu Asn Lys Ile Asn1 5 10

Ser Lys Arg Arg Asn Gly Leu Read Lys Lys Wing

20 25

Leu Cys Asp Glu Wing Val Wing Leu Ile Val Phe

35 40

Leu Tyr Glu Tyr To Be Thr Asp To Be Met Glu

50 55

Tyr Glu Arg Tyr Cys Phe Ala Glu Lys Ser Ser 70 70 75

Asp Be Gln Glu Asp Trp Be Read Glu Tyr His

85 90

Val Glu Ser Leu Asn Asn Arg Gln Arg His Leu

100 105

Glu Be Read Le Be Arg Read Glu Ile Gly Gln Read Le

115 120

Asn Ser Read Lys Thr Val Arg Thr Arg Lys Ser

130 135

Ser Ile Ser Glu Read Gln Asp Lys Glu Lys Thr145 150 155

Lys Wing Leu Glu Asn Glu Leu Met Lys Arg Wing

165 170

Leu Glu Gln Gln Wing Arg Trp Lys His His Asn

180 185

Asn Read His Asn Pro Asn Ile Asn Ile Gly Asn

195 200

Asn Glu Gly Gly Val Glu Pro Wing Thr Asp Val

210 215

Arg Asn Leu Leu Pro His Trp Met Leu225 230

<210> 172 <211> 744

190

Read Ile Gly Asp Ser205

Ser Gly Asn Glu His220

Gly Asn Ser Leu Leu240

tgaccttctcgtgatgcagactgattcaagcatcaacaataggctaaggtctctgagtctttcggacgcgaaactttgcgaagctattctttcataatccagccagcaac

aaaaagaagaagttgcactttatggaaaaagagtgacatttgagagtttatcgagaaattcaagagccaaagatgagaacggaacaacaaaa atatcaacggatgttcaa

Arg

His

To be

Lys60Thr

Lys

Met

Glu

Gln140Leu

Arg

His

Tyr

Gln220

Gln

Glu

Allah

45

Ile

Met

Read

Gly

Gln125Glu

Arg

Allah

Lys

Gln205Val

Val

Ile

30

Lys

Read

To be

Lys

Glu110Gln

Read

Asp

Lys

Gln190Thr

Arg

Thr Phe15

To be val

Gly lys

Glu Arg

Asp Ile80Ala Lys95

Gln Leu

Read Glu

Read Ser

Glu Asn160Ala Ile175

Gln AspArg AsnVal Val

60120180240300360420480540600660699 <212> DNA

<213> Dendrobium grex Madame Thong-IN <400> 172

atgggtcgtg gcagggtgca gctgaagcga atcgagaatattctcgaagc ggagatctgg tttgcttaag aaggcgcacggctgaagttg ctctgatcgt tttttccaat aagggaaagctccagcatgg agaaaattct tgaacggtat gagcgttattttttccaatg aggccaaccc ccaggctgat tggcgccttgagggttgaaa gcttacagaa gagccaaagg caccttatggagcattaaag aactccaacg tctagagcaa cagcttgaaatccagaaaga cacagctcat actacattca atttccgagcttgctggagc aaaacaagac cttagagaag gagattatagttggtgcagc atgccccatg ggagaagcaa aaccagtccccctgtgattt cggattctgt cccaactccc accagcagaagaagaagaat cacctcagcc acagttaaga gtaagcaacactcagtcata tgaatggaca ATAA <210> 173 <211> 247 <212> PRT

<213> Dendrobium grex Madame Thong-IN <4 00> 173

Met Gly Arg Gly Arg Val Gln Read Lys Arg Ile

25

aaataaaccgagatctccgttttatgagtacatatgctgaaatataataagggagcaactgttccttgaatacaaaagatctaaagagaaaatatagctccgtttcaagcctctgctgcc

gcaggtgacggctctgtgacttccaccgataagagcattaactgaaggcatgactccttggtttatacgaggaaaaataagccaaagcttgcactcccgcagagccaatcccatggatg

1 5 Arg Gln Val Thr Phe Ser 20 His Glu Ile Ser Val Leu 35 Ser Asn Lys Gly Lys Leu 50 Lys Ile Leu Glu Arg Tyr65 70Phe Ser Asn Glu Ala Asn 85 Lys Leu Lys Ala Arg Val 100 Met Gly Glu Leu Asp 115 Glu Gln Gln Leu Glu Ser 130 Gln Leu Ile Leu His Ser145 150Leu Leu Glu Gln Asn Lys 165 Lys Wing Lys Wing Leu Val 180 Ser Gln Tyr Ser Ser Wing 195 Thr Pro Thr

55

105

120

135

185

200

210

215

Pro Gln Pro Gln Leu Arg Val Ser Asn Thr Leu

225

230

[N Arg Ile Glu Asn Lys Ile Asn10 15 Ser Gly Leu Leu Lys Lys Wing 30 Glu Val Wing Leu Ile Val Phe 45 Ser Thr Asp Being Met Glu 60 Ser Tyr Wing Glu Arg Wing Leu 75 80Asp Trp Arg Leu Glu Tyr Asn90 95 Gln Lys Being Gln Arg His Leu 110 Ile Lys Glu Leu Gln Arg Leu 125 Phe Ile Arg Being Arg Lys Thr 140 Leu Gln Lys Met Glu Lys Ile 155 160Lys Glu Ile Ile Wing Lys Glu170 175 Pro Trp Glu Lys Gln Asn Gln 190 Val Ile Ser Asp Ser Val Pro 205 Arg Wing Asn Glu Glu Glu Ser 220 Thr Leu Leu Pro Pro Trp Met 235 240

Leu Being His Met Asn Gly Gln245

<210> 174

<211> 2194

<212> DNA

<213> Oryza sativa

<4 00> 174

aatccgaaaa gtttctgcac cgttttcacc ccctaactaaaaatataaaa tgagacctta tatatgtagc gctgataactcatccaccta ctttagtggc aatcgggcta aataaaaaaagtttccttagtgtgtgtgtgtgtgtgt

caatatagggagaactatgcagtcgctacattagaatatacatcaaactc

aacgtgtgctaagaaaaactctagtttcgtcgttcacatcttcttgaata

60120180240300360420480540600660720744

60120180240300aaaaaatctt tctagctgaa ctcaatgggtatgaagatat tctgaacgta ttggcaaagattgtgcattc gtcatatcgc acatcattaattagtaatta aagacaattg acttatttttgtacttacgc acacactttg tgctcatgtgtcaactagca acacatctct aatatcactctgaattcaag cactccacca tcaccagaccaattttacag aatagcatga aaagtatgaaaaaaaaagaa ttttgctcgt gcgcgagcgcacagagtggc tgcccacaga acaacccacatccgcaacaa ccttttaaca gcaggctttgaaccaagcat cctccttctc ccatctataaggaggcatcc aagccaagaa gagggagagccgaccgcctt ctcgatccat atcttccggtacctcctcct cacagggtat gtgcctcccttgtgtagtac gggcgttgat gttaggaaagtggatttggg atagaggggt tcttgatgttatggttttca atcgtctgga gagctctatggcgattttgt gagtaccttt tgtttgaggtgtaataaagt acggttgttt ggtcctcgátgctatccttt gtttattccc tattgaacaagatgagattg aatgattgat tcttaagccttacagtagtc cccatcacga aattcatggaccctgttctt ccgatttgct ttagtcccagactttctggt tcagttcaat gaattgattgagctgtagtt cagttaatag gtaataccccatttctgatc tccattttta attatatgaagattattttt tttattagct ctcaccccttctgtcctcaa ttttgttttc aaattcacatacctgtagaa gtttcttttt gggggatta ctgcttgttggtgtagc ttgccacttt caccagcaaa <210> 175 <211> 1725 <212> DNA

<213> Arabidopsis thaliana <400> 175

atgaattcta acaactggct tggctttcctcatgaataca accttggctt ggtcagcgactggaatatga tcaatccaca cggtggaggagccgattttc tcggtgtgag caaaccggacaacgactcag actactactt ccataccaatgtcgttgtag cagcttgtga ctccaatactgagagtgctc acaatctaca gtcacttactgttgtagaca aagcttcacc atccgagaccgccgttgttg agacggccac gccaagacgtatctatcgtg gtgtcacaag acatcgatggaatagttgta gaagggaagg ccagtctagggataaagaag ataaagcagc aagatcatattcaactacta ctaatttccc cattacaaacatgacgaggc aagagttcgt ggctgccattgcttcgatgt atcgaggagt tacaaggcacggccgagtcg ccgggaacaa agacctctacgcagaagctt acgatatagc tgcaataaaggagatcaacc ggtacgacgt gaaagccattggcgcagcta aacggctcaa agaagctcaagagatgatag cccttggttc aagtttccagtccacctcat caagacttca gcttcaacctccttttctat ctcttcagaa caatgacatctcctcctctt ttaatcacca tagctatatcaacaattact tgcagcaaca gtcgagccagcatagcaatc cggctctgct tcatggacttaacaatggag gctctagtgg gagctacaac

aaagagagag attttttttatttaaacata taattatata

ggacatgtctattatttatccatgtgtgaggcctatttaaacttttaataacgaactattcaatctcccaaaaaacgatgcggccaggagattcctccccaccaaggacacgagttcttgtcggttgttcgggatctgtagcatgttatcgaaatgaaataaaatcagagtctggtagtgaaataatccagtccaaaattaacagttataaattttttttctacaaataatatagtttagatgaactgtacattattctgcgattatctatgactgcttgttcttatgatgttc

tactccatccttttttcgattgcacctccttacatttaggatatctaaaataggtttttctattgggcacatctaacggaagaggaggagccttttcccccgcgactagcgtcgatctctttggatttattctgtgatgaggttcggtttggtttagggacaccggtgatatgcttctcgactttgaagatcgcagctggatcctcaggacccaaatatctgcttttatatcaggagaaggcataagcagagctgaaagttgcattatccattacagaaatcatttcctt

aaaaaatagaattttatagtcaatttttattagatgcaagcaatacacgttagcaatatctacaaaaaatacatacaaaaacaggcaacaggacagcaagaggcaaagaatctctatataagaagccgagtccctcctcctgttctaggtttcctgttctgattagtagttcggaatctttttgcttggtatttgacgaacggtcccgttcttgtttagaacaggggattttaaaaagtcgcgttatcctaacttatccgtattcatttgctggcatgaatcttgtatctgaaatttatgtgtgcagttc

ctttcaccga acaactcttccatatggaca acccttttcaggagatgaag gaggagaggtgaaaaccaat ccaaccacctagcttgatgc ctagcgtccacctaacaaca gtagctatcattgtccatgg ggaccaccgcaccggggata acgctagcgggcattggaca ctttcggacaactggtcgat atgaggctcaaaaggaagac aagtttacttgatctagctg cacttaagtatacgagaaag aagtagaggaagaaggaaaa gtagtggattcaccaacatg gaagatggcattgggaactt ttagcactgatttagaggac ttaatgcagtctagagagta gcactcttccgctcttgagt cttcaaggaatacggtggtg gctcgagcactaccctctaa gcattcaacatctcattaca acaacaacaacagacacaac ttcatctccaaactctcagc agctctacaagtctctacct ctatcgttgaactgcagcat ttcttgggaa

tttgcctcctaacacaagagtccaaaagtgagtagcttacatcaaacgattgagcttcaatggtaataattggagcactaacgaacctcgtctatgggatgggtggatatctggggtcccaatgaagcacttcgagaggcagcaaggatcggaagaagcagaccaacttccatcggaggaacgcgaggcgaggctctggcaccattagagtgctcacgatccaacagacctgcgtatcttcaacaataatccacggtatt

360420480540600660720780840900960102010801140120012601320138014401500156016201680174018001860192019802040210021602194

601201802403003604204805406006607207808409009601020108011401200126013201380144015001560ggtattgggt ccagctcgac tgttggatcg accgaggagt ttccaaccgt taaaacagat 1620tacgatatgc cttccagtga tggaaccgga gggtatagtg gttggaccag tgagtctgtt 1680caggggtcaa accctggtgg tgttttcact atgtggaatg agtaa 1725

<210> 176 <211> 574 <212> PRT

<213> Arabidopsis thaliana <400> 176

Met Asn Be Asn Asn Trp Read Gly Phe Pro Read Be Pro Asn Asn Ser1 5 10 15 Be Read Pro Pro His Glu Tyr Asn Read Gly Leu Val Be Asp His Met 20 25 30 Asp Asn Pro Phe Gln Thr Glu Trp Asn Met Ile Asn Pro His Gly 35 40 45 Gly Gly Gly Asp Glu Gly Gly Glu Val Pro Lys Val Wing Asp Phe Leu 50 55 60 Gly Val Ser Lys Pro Asp Glu Asn Gln Ser Asn His Leu Val Wing Tyr65 70 75 80Asn Asp Ser Asp Tyr Tyr Tyr Phe His Thr Asn Be Leu Met Pro Be Val 85 90 95 Gln Be Asn Asp Val Val Val Wing Ala Cys Asp Be Asn Thr Pro Asn 100 105 110 Asn Be Tyr His Glu Leu Gln Be Wing His 125 Leu Thr Leu Be Met Gly Thr Thr Wing Gly Asn Asn Val Val Asp Lys 130 135 140 Wing Be Pro Be Glu Thr Thr Asn Asn Wing Be Gly Gly Wing Leu145 150 155 160Ala Val Val Glu Thr Ala Thr Pro Wing Ala Leu Asp Thr Phe Gly 165 170 175 Gln Arg Thr Being Ile Tyr rp Thr Gly 180 185 190 Arg Tyr Glu Wing His Leu Trp Asp Asn Be Cys Arg Arg Glu Gly Gln 195 200 205 Be Arg Lys Gly Arg Gln Val Tyr Leu Gly Gly Tyr Asp Lys Glu Asp 210 215 220 Lys Wing Ala Arg Be Tyr Asp Leu Wing Wing Leu Lys Tyr Trp Gly Pro225 230 235 240Ser Thr Thr Thr Asn Phe Pro Ile Thr Asn Tyr Glu Lys Glu Val Glu 245 250 255 Glu Met Lys His Met Thr Arg Gln Phe Val Ala Ile Arg Arg 260 265 270 Lys Be Be Gly Phe Be Arg Gly Wing Be Met Tyr Arg Gly Val Thr 275 280 285 Arg His His Gln His Gly Arg Trp Gln Wing Arg Ile Gly Arg Val Wing 290 295 300 Gly Asn Lys Asp Read Tyr Leu Gly Thr Phe Be Thr Glu Glu Glu Wing305 310 315 320Ala Glu Wing Tyr Asp Ile Wing Wing Ile Lys Phe Arg Gly Leu Wing Asn Wing 325 330 335 Val Thr Asn Phe Glu Ile Wing Tn Asp Val Lys Wing Ile Leu Glu 340 345 350 Ser Ser Thr Leu Pro Ile Gly Gly Gly Wing Wing Lys Arg Winged Lys Glu 355 360 365 Wing Gln Wing Leu Glu Be Ser Arg Lys Arg Glu Wing Glu Met Ile Wing 370 375 380 Leu Gly Ser Be Phe Gln Tyr Gly Gly Ser Be Thr Gly385 390 395 400Ser Thr Be Ser Arg Leu Gln Leu Gln Pro Tyr Pro Leu Ser Ile Gln 405 410 415 Gln Pro Leu Glu Pro Phe Leu Be Leu Gln Asn Asn Asp Ile Be His 420 425 430 Tyr Asn Asn Asn Asn Wing His Asp Be Ser Be Phe Asn His His Ser435

450

455

465

470

485

490

500

505

515

520

530

535

445 Gln Thr Asn Asn Tyr Leu 460 Leu Tyr Asn Wing Tyr Leu475 480Val Ser Thr Be Ile Val 495 Gly Ser Tyr Asn Thr Wing 510 Gly Ser Ser Ser Thr Val 525 Thr Asp Tyr Asp Met Pro 540 Trp Thr Ser Glu Ser Val

545

550

555

560

Gln Gly Being Asn Pro Gly Gly Val Phe Thr Met Trp Asn Glu

565

570

<210> 177 <211> 1707 <212> DNA

<213> Arabidopsis thaliana <400> 177

atgaattcta acaactggct cgcgttccct ctatcaccaa ctcactcttccacattcact cttcacaaaa ttctcatttc aatctaggttaacccttttc aaaaccaagg atggaatatg atcaatccacggagaggttc caaaagtggc tgatttctta ggagtgagcagatcacaacc tcgtacctta taacgacatt catcaaaccacaaaccaata gcttgttacc tacagtcgtc acttgtgcctgagcttcaag agagtgcaca caatttgcaa tctctcactcgctgccgctg cagaagtcgc cactgtgaaa gcctcgccggagtagcagca ctaccaacac aagcggagga gccatcgttg

ttggaaactt ttggacaacg aacctctatc tatcgtggagggtagatatg aagctcatct ttgggataat agctgtagaaggaagacaag tctacttagg tgggtatgac aaagaagagactagctgcac ttaaatattg gggtccctct actactaccagagaaggaag tagaggagat gaaaaacatg acgagacaagaggaagagta gcggattctc gcgtggtgca tccatgtatccaacatggaa gatggcaagc aaggatcggc cgagttgctgggaacattca gcacggagga agaagcagca gaagcttatgcgaggtctaa acgcggttac aaactttgag ataaatcggtgagagcaaca cacttcctat aggaggtggt gcggctaaacctagaatcat caagaaaacg agaggaaatg atagccctcgggtgcagcga gcggctcgag ctctgttgct tccagctctacctctaagca ttcaacaacc ttttgagcat cttcatcatccagaacaaca acgatatctc tcagtatcat gattcctttacatcttcacc aacaacaaac caacaattac ttgcagtctttacaatgctt atcttcagag taaccctggt ctgcttcatgaacacttcag ggtttcttgg aaacaatggg attggtattgtcatcggctg aggaagagtt tccagccgtg aaagtcgattggagctacag ggtatggagg atggaatagt ggagagtcttgg cgatgtggaa tgaataa <210> 178 <211>

<213> Arabidopsis thaliana <4 00> 178

Met Asn Ser Asn Asn Trp Leu Wing Phe Pro Leu15 10

Ser Leu Pro Pro Ile His Ser Ser Gln Asn

20 25

Gly Leu Val Asn Asp Asn Ile Asp Asn Pro Phe

35 40

Asn Met Ile Asn Pro Gly Gly Gly Gly Gly Glu

tggtcaacgaatggtggaggaatcgggggaacgcctccgactaatgctcctctctatgggctgagactagaggctacaccttacaagacagagaaggacaaagcagccagactttccgatagtttgtggcgtggagtaacgaaacaaagaacatagctgcatgatgtgaaggctcaaagagatcaaatttggcttcagctatcagcctttgttacattcactagtcacacgatttgtctcggtcaagctcacgatatgccctcaaggatc

tttgccgcctcaatatcgaccggcgaaggttcatcacaccctactacttttaataattataagtactggatgccgataatgagacggacttagatggaccatcaaggaaaagcatatgataactaactacttctataagaaaggcatcattctctacttggataaagtttagccatcctgagctcaagctccatcaatattcaaccttacacttactctagacgcagcttttcacagctctgataataactaccgttggatccttccggtgaatccagga

Ser Pro Thr His Ser15

Being His Phe Asn Leu30

Gln Asn Gln Gly Trp45

Gly Gly Glu Val Pro

60120180240 '300360420480540600660720780840900960102010801140120012601320138014401500156016201680170750 55 60

Lys Val Wing Asp Phe Leu Gly Val Ser Lys Ser Gly Asp His His Thr65 70 75 80

Asp His Asn Read Val Pro Tyr Asn Asp Ile His Gln Thr Asn Ala Ser

85 90 95

Asp Tyr Tyr Phe Gln Thr Asn Being Read Leu Pro Thr Val Val Thr Cys

100 105 110

Wing Be Asn Wing Pro Asn Asn Tyr Glu Read Gln Glu Be Wing His Asn

115 120 125

Read Gln Be Read Thr Read Read Be Met Gly Be Thr Gly Wing Wing Wing

130 135 140

Glu Val Wing Thr Val Lys Wing Ser Pro Wing Glu Thr Wing Wing Asp Asn145 150 155 160

Be Be Be Thr Thr Asn Thr Be Gly Gly Wing Ile Val Glu Wing Thr

165 170 175

Pro Arg Arg Thr Read Glu Thr Phe Gly Gln Arg Thr Be Ile Tyr Arg

180 185 190

Gly Val Thr Arg His Arg Trp Gly Arg Tyr Glu Wing His Leu Trp

195 200 205

Asp Asn Be Cys Arg Arg Glu Gly Gln Be Arg Lys Gly Arg Gln Val

210 215 220

Tyr Leu Gly Gly Tyr Asp Lys Glu Glu Lys Wing Arg Wing Wing Tyr Asp225 230 235 240

Read Wing Wing Read Lys Tyr Trp Gly Pro Be Thr Thr Thr Asn Phe Pro

245 250 255

Ile Thr Asn Tyr Glu Lys Glu

260 265 270

Gln Glu Phe Val Wing Be Ile Arg Arg Lys Be Gly Phe Be Arg

275 280 285

Gly Ala Be Met Tyr Arg Gly Val Thr His Arg His Gln His Gly Arg

290 295 300

Trp Gln Wing Arg Ile Gly Arg Val Wing Gly Asn Lys Asp Leu Tyr Leu305 310 315 320

Gly Thr Phe Be Thr Glu Glu Glu Wing Wing Glu Wing Tyr Asp Ile Wing

325 330 335

Wing Ile Lys Phe Arg Gly Leu Asn Wing Val Thr Asn Phe Glu Ile Asn

340 345 350

Arg Tyr Asp Val Lys Wing Ile Leu Glu Ser Asn Thr Leu Pro Ile Gly

355 360 365

Gly Gly Wing Wing Lys Arg Leu Lys Glu Wing Gln Wing Leu Glu Ser Ser

370 375 380

Arg Lys Arg Glu Glu Met Ile Wing Read Gly Ser Asn Phe His Gln Tyr385 390 395 400

Gly Ala Wing Be Gly Be Ser Be Val Wing Be Ser Be Arg Leu Gln

405 410 415

Read Gln Pro Tyr Pro Read Ser Ile Gln Gln Pro Phe Glu His Leu His

420 425 430

His His Gln Pro Read Leu Thr Read Le Gln Asn Asn Asn Asp Ile Ser Gln

435 440 445

Tyr His Asp Be Phe Ser Tyr Ile Gln Thr Gln Read His Read His Gln

450 455 460

Gln Gln Thr Asn Asn Tyr Leu Gln Be Ser Be His Thr Be Gln Leu465 470 475 480

Tyr Asn Ala Tyr Leu Gln Ser Asn Pro Gly Leu Read His Gly Phe Val

485 490 495

Be Asp Asn Asn Asn Thr Be Gly Phe Read Gly Asn Asn Gly Ile Gly

500 505 510

Ile Gly Be Ser Be Thr Val Gly Ser Be Wing Glu Glu Glu Phe Pro

515 520 525

Val Lys Wing Val Asp Tyr Asp Met Pro Pro Being Gly Gly

530 535 540

Tyr Gly Gly Trp Asn Be Gly Glu Be Wing Gln Gly Be Asn Pro Gly545 550 555 560Gly Val Phe Thr Met Trp Asn Glu565

<210> 179 <211> 1662 <212> DNA <213> Glycine max <400> 179

atgaacaaca actggctttc gttccctctt tctcctactcgatcttcaag caactcaata tcatcaattt tcccttgggtaaccctttcc aaaatcatga ttggaatctg attaacacccaaagtggctg attttctagg agtgagcaag tctgaaaatcaacgaaattc attcaaatga ttcagattat ctgttcacaacaaaaccctg tgttggacac acctagcaat gagtatcaagcaatcattga cattatccat gggaagtggt aaggattcaaaatagcacaa acactactgt tgaagttgca cctagaagaaagaacatcca tatatcgtgg agtaactcga catagatggactttgggata atagctgtag aagggaaggc caatcaagaaggtggatatg ataaagaaga gaaagcagct agagcttatgtgggggacat ccaccactac caactttcca attagcaactatgaaacaca tgacgagaca agaatttgtt gccgccattatccaggggtg catcaatgta tcgtggagtt acaaggcatcgcaaggattg gcagagttgc aggaaacaaa gatctttactgaagaggctg cagaagcata cgacatagca gcgataaagtacaaactttg acatgagccg ctacgacgtg aaagccattcataggaggag gcgctgcaaa gcgtctgaaa gaagctcaagcgcgaagaga tgattgcact aggctcatct tccacgttcctcttctaggc ttcacgctta ccctctaatg cagcaccacccctctgctaa ctcttcaaaa ccacgacata agttcttctccctttgcãtc atcagggtta catccaaacg cagcttcagttcttcttata gctttcagaa taatgctcag ttctacaatggcattgcttc agggaatgat gaacatgggg tcttcttcttaataataata gtaacaataa taataataat gttggtgggtatggcttcga atgcaacggc ggggaacacg gtggggacagaaggtggact atgacatgcc ggctggaggt tacggtggctcagacgtcaa atggtggggt gttcacaatg tggaatgatt <210> 180 <211> 553 <212> PRT <213> Glycine max <4 00> 180

Read Pro Wing His Asp Read Gln Wing Thr Thr Gln Tyr

attcttcctttagtgaacgaatagtagcaaagtcagacctacaacagtctaaaatgctaacatgtgaaacctttggatacctggaaggtaaaggacgccaatttagctgcatgagaaggagaaggaaaagaccaacacggtgggaacttttcagaggtctttgaaagcaactctagaatcaatacggaacaccagttcgaacttctctcatgcaccagcagttaccttcacttcctcatcttgtgggaagcggaggagttggtcggcggcaa

accagctcatgaacatggatcgaaattccatgcagccttaggtgcctatgtagtaatttgcagtggtgaaattcgggcagtgaagctcatagtttatttgactgaagtacattggatgaacagtggtttcaagatggcaacagtactgagcaacgctgtccactctcccattcgagaaaactcagcaagcgcaacctcaaccagcaagacgagtggcgctgaaccacccttgtgttggagtgggtttggtagggctggtgggactccatg

20

25

Gly Leu Val Asn Glu Asn Met Asp Asn Pro Phe

35

40

Asn Leu Ile Asn Thr His Being Ser Asn Glu Ile

50

55

Phe65

Read Gly Val Ser Lys Ser Glu Asn Gln Ser70

Asn Glu Ile His Ser Asn Asp Ser Asp Tyr Leu85

Read Val Val Met Gln Asn Read Val Val Asp Thr

100

105

Gln Glu Asn Wing Asn Ser Asn Leu Gln Ser Leu

115

120

Being Gly Lys Asp Being Thr Cys Glu Thr Being Gly

130

135

Thr Thr Val Glu Val Pro Wing Arg Arg Thr Leu

145

150

Arg Thr Be Ile Tyr Arg Gly Val Thr His165

Tyr Glu Wing His Leu Trp Asp Asn Ser Cys Arg

Leu Ser Pro Thr His Ser10 15 Gln Tyr His Gln Phe Ser Leu 30 Pro Phe Gln Asn His Asp Trp 45 Glu Ile Pro Lys Val Wing Asp 60 Gln Ser Asp Leu Wing Wing 75 75Tyr Leu Phe Thr Asn Asn Ser90 95 Asp Thr Pro Be Asn Glu Tyr 110 Be Read Thr Read Le Be Met Gly 125 Be Gly Glu Asn Be Thr Asn 140 Thr Read Asp Thr Phe Gly Gln 155 160Arg His Arg Trp Thr Gly Arg170 175 Cys Arg Arg Glu Gly Gln Ser

60120180240300360420480540600660720780840900960102010801140120012601320138014401500156016201662180 185 190 Arg Lys Gly Gly Tyr Asp Lys Glu Glu Lys 195 200 205 Ala Ala Ala Thr Ala Thr Ala Tyr Glu Lys Glu Read Asp Glu225 230 235 240Met Lys His Met Thr Arg Gln Phe Val Wing Wing Ile Arg Arg Lys 245 250 255 Be Ser Gly Phe Ser Arg Gly Wing Be Met Tyr Arg Gly Val Thr Arg 260 265 270 His His Gln His Gly Arg Trp Gln Wing Arg Ile Gly Arg Val Wing Gly 275 280 285 Asn Lys Asp Leu Tyr Leu Gly Thr Phe Be Thr Glu Glu Wing Ala 290 295 300 Glu Wing Tyr Asp Ile Wing Ile Lys Phe Arg Gly Leu Asn Wing Val305 310 315 320Thr Asn Phe Asp Met Be Arg Tyr Asp Val Lys Wing Ile Leu Glu Ser 325 330 335 Asn Thr Leu Pro Ile Gly Gly Gly Wing Lys Arg Leu Lys Glu Wing 340 345 350 Gln Wing Leu Glu Be Ser Arg Be Lys Arg Glu Met Ile Wing Leu Gly 355 360 365 Ser Be Ser Thr Be Phe Gln Tyr Gly Thr Ser Be Ser Be Be Arg Leu 370 375 380 His Wing Tyr Pro Leu Met Gln His His His Gln Phe Glu Gln Pro Gln385 390 395 400Pro Read Leu Thr Read Le Gln Asn His Asp Ile Be Ser Be His Phe Ser 405 410 415 His Gln Gln Asp Pro Read His His Gln Gly Tyr Ile Gln Thr Gln Leu 420 425 430 Gln Read His Gln Gln Ser Gly Ala Be Ser Tyr Ser Phe Gln Asn Asn 435 440 445 Ala Gln Phe Tyr Asn Gly Tyr Leu Gln Asn His Pro Ala Leu Leu Gln 450 455 460 Gly Met Met Asn Met Gly Ser Ser Ser Ser Ser Ser Val Glu465 470 475 480Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Val Gly Gly Phe Val Gly 485 490 495 Ser Gly Phe Gly Met Wing Being Asn Wing Thr Gly Wing Asn Thr Val Gly 500 505 510 Thr Wing Glu Glu Leu Gly Read Val Lys Val Asp Tyr Asp Met Pro Wing 515 520 525 Gly Gly Tyr Gly Gly Trp Ser Wing Asp Wing M et Gln Thr Ser Asn 530 535 540 Gly Gly Val Phe Thr Met Trp Asn Asp 545 550

<210> 181 <211> 1674 <212> DNA <213> Glycine max <4 00> 181

atgaacaaca actggctttc gttccctctt tctcctactc attcttcctt accagctcat 60

gatcttcaag caactcaata tcatcaattt tcccttgggt tagtgaacga gaacatggat 120

aaccctttcc aaaatcatga ttggaatctg attaacaccc atagtagcaa cgaaattcca 180

aaagtggctg attttctagg agtgagcaag tctgaaaatc agtcagacct tgcagcctta 240

aacgaaattc attcaaatga ttcagattat ctgttcacaa acaacagtct ggtgcctatg 300

caaaaccctg tgttggacac acctagcaat gagtatcaag aaaatgctaa tagtaatttg 360

caatcattga cattatccat gggaagtggt aaggattcaa catgtgaaac cagtggtgaa 420

aatagcacaa acactactgt tgaagttgca cctagaagaa ctttggatac attcgggcag 480

agaacatcca tatatcgtgg agtaactcga catagatgga ctggaaggta tgaagctcat 540

ctttgggata atagctgtag aagggaaggc caatcaagaa aaggacgcca agtttatttg 600

ggtggatatg ataaagaaga gaaagcagct agggcttatg atttagctgc actgaagtac 660tgggggacat ccaccactac caactttcca attagtaact atgagaagga attggatgaa 720

atgaaacaca tgacgcgaca agaatttgtt gctgccatta gaaggaaaag cagtggtttc 780

tccaggggtg catcaatgta tcgtggagtt acaaggcatc accaacacgg aagatggcaa 840

gcaagaattg gcagagttgc aggaaacaaa gatctttact tgggaacttt cagtactgaa 900

gaagaggctg ctgaagcata cgacatagct gcgataaagt tcagaggtct caacgctgtc 960

acaaactttg acatgagccg ctacgacgtg aaagccatcc ttgaaagcaa cactctccca 1020

ataggaggag gagctgcaaa gcgtctgaaa gaagctcaag ctctagaatc ttcgagaaag 1080

cgcgaagaga tgattgcact aggatcatcc acattccaat atggaaccac aagctctaat 1140

tctaggctac atgcttaccc tctaatgcag caccaccacc agtttgaaca acctcaacct 1200

ctgctaactt tgcaaaacca tgatatcagt tctcacttct ctcaccagca agaccctttg 1260

catcagggtt acatccaaac gcagcttcag ttgcaccagc agcagagtgg tggttcttct 1320

tcttatagct ttcagaataa taatataaat aatgctcagt tctataatgg ttataatctt 1380

cagaaccacc ctgcattgct tcagggaatg attaacatgg ggtcttcatc ttcttcatct 1440

gtgttggaga ataataatag taccaataat aatgttggtg ggtttgtggg aagtgggttt 1500

ggtatggctt ctaatgcaac gtcggggaac acggtgggga cggcggagga gctagggctg 1560

gtgaaggtgg actatgacat gccgactggt ggttacggtg gatggtcggc ggcggcggcg 1620

gcggagtcca tgcagacgtc gaatagtggg gtgttcacaa tgtggaatga ctga 1674 <210> 182 <211> 557 <212> PRT <213> Glycine max <4 00> 182

Met Asn Asn Asn Trp Read Be Phe Pro Read Be Pro Thr His Be Ser1 5 10 15 Read Pro Wing His Asp Read Gln Wing Thr Gln Tyr His Gln Phe Be Read 20 25 30 Gly Read Val Asn Glu Asn Met Asp Asn Pro Phe Gln Asn His Asp Trp 35 40 45 Asn Leu Ile Asn Thr His Being Ser Asn Glu Ile Pro Lys Val Wing Asp 50 55 60 Phe Leu Gly Val Being Lys Ser Glu Asn Gln Being Asp Leu Wing Ala Leu65 70 75 80Asn Glu Ile His Ser Asn Asp Ser Asp Tyr Leu Phe Thr Asn Asn Ser 85 90 95 Leu Val Pro Met Gln Asn Pro Val Leu Asp Thr Pro Ser Asn Glu Tyr 100 105 110 Gln Glu Asn Wing Asn Ser Asn Leu Gln Ser Leu Thr Leu Ser Met Gly 115 120 125 Be Gly Lys Asp Be Thr Cys Glu Thr Be Gly Glu Asn Be Thr Asn 130 135 140 Thr Thr Val Glu Val Wing Pro Arg Arg Thr Read Asp Thr Phe Gly Gln145 150 155 160Arg Thr Be Ile Tyr Arg Gly Val Thr Arg His Arg Trp Thr Gly Arg 165 170 175 Tyr Glu Wing His Leu Trp Asp Asn Be Cys Arg Arg Glu Gly Gln Ser 180 185 190 Arg Lys Gly Arg Gln Val Tyr Leu Gly Gly Tyr Asp Lys Glu Glu Lys 195 200 205 Wing Wing Arg Wing Wing Tyr Asp Leu Wing Wing Read Lys Tyr Trp Gly Thr Ser 210 215 220 Thr Thr Thr Asn Phe Pro Ile Ser Asn Tyr Glu Lys Glu Read Asp Glu225 230 235 240Met Lys His Met Thr Arg Gln Glu Phe Val Wing Ile Arg Arg Lys 245 250 255 Ser Be Gly Phe Ser Arg Gly Wing Be Met Tyr Arg Gly Val Thr Arg 260 265 270 His His Gln His Gly Arg Trp Gln Wing Arg Ile Gly Arg Val Wing Gly 275 280 285 Asn Lys Asp Leu Tyr Leu Gly Thr Phe Be Thr Glu Glu Wing Ala 290 295 300 Glu Wing Tyr Asp Ile Wing Ile Lys Phe Arg Gly Leu Asn Wing Val305 310 315 320Thr Asn Phe Asp Met Ser Arg Tyr Asp Val Lys Wing Ile Leu Glu Ser325 330

Asn Thr Read Pro Ile Gly Gly Gly Wing Wing Lys

340 345

Gln Wing Read Glu Ser Be Arg Lys Arg Glu Glu

355 360

Be Ser Thr Thr Phe Gln Tyr Gly Thr Thr Ser

370

375

Tyr Pro Wing Read Met Gln His His His Gln Phe385 390 395

Leu Leu Thr Leu Gln Asn His Asp Ile Ser Ser

405 410

Gln Asp Pro Read His Gln Gly Tyr Ile Gln Thr

420 425

Gln Gln Gln Be Gly Gly Be Ser Tyr Ser Ser

435 440

Ile Asn Asn Wing Gln Phe Tyr Asn

450 455

Wing Read Leu Gln Gly Met Ile Asn Met Gly Ser465 470 475

Val Leu Glu Asn Asn Asn Ser Thr Asn Asn Asn

485 490

Gly Ser Gly Phe Gly Met Wing Be Asn Wing Thr

500 505

Gly Thr Wing Glu Glu Leu Gly Leu Val Lys Val

515 520

Thr Gly Gly Tyr Gly Gly Trp

530 535

Gln Thr Be Asn Be Gly Val Phe Thr Met Trp545 550 555

<210> 183 <211> 1632 <212> DNA

<213> Medicago trunculata <400> 183

atgaacaata actggctttc attccctctc tcaccttctcgatcttcaag caactcaata tcatcacttt cctcttggataaccctttcc aaaatcatga ttggaatctg atgaacacacaaggttgcgg attttctcgg tgtatgcaag tctgaaaatcaacgaaattc aatctaatga ttcagattat ctgtttacaaatgcaaaacc aaatggttac aacatgcacc aatgagtatcaatttgcagt ctttgacatt atccatggga agtggtaaagggtgaaaata gtacaaacac tgttgaagtt gctgttcctaggacaaagaa cttcgatata tcgcggtgta acaaaacatagctcaccttt gggataacag ctgtagaagg gaaggtcagtggatatgata aagaagagaa agctgctagg tcttatgattgggacatcca ccactaccaa ctttccagtt agcaactatgaagcacatga caagacaaga atttgttgcc tctattagaaaggggtgcat caatgtaccg tggagttaca aggcatcaccaggattggca gagttgcagg aaataaagat ctatacttgggaggctgcag aagcatacga catagcagca ataaaattcaaactttgaca tgactcgtta cgacgtgaaa gccattctcgggaggaggag ctgcaaaaag actaaaagaa gcacaagctcgaagaaatgc ttgcacttaa ctcatcatct ttccaatatgactagactcc aaccctaccc tctcatgcaa tatcatcaccttgctaacat tacaaaacaa ccatgaaagc ttgaattctcggtggtggtt atttccaaac acagcttgag ttgtgtcaaccagaatagta acataggttc attctacaat ggttattatccagatgaata atataggatc ttcttcttca tcttcggtgatctagtggga tttatagtaa tagtggaggg ttaattagtagtgccggtta aggttgatta tgacatgcaa ggtgatggaagcggcaggag agaacatgca gactgctgagagtagtag10>

Arg

Met

Asn380Glu

His

Gln

Phe

Leu460Ser

Val

To be

Asp

Ala540Asn

Read Lys350Ile Ala365

To be Arg

Gln pro

Phe Ser

Read Gln430Gln Asn445

Gln asn

To be to be

Gly Gly

Gly Asn510Tyr Asp525

GluAsp Wing

335

Glu Wing

Read gly

Read his

Gln Pro400His Gln415

Read his

Asn asn

His pro

Ser Ser480Phe Val495

Thr ValMet ProSer Met

attcttcctttagtcaatgaacaacagcaaactcagatctataacaatacaagaaaaggcattcaacatgaaagaacttcgatggactggcgagaaaaggtagctgcactagaaggaaatggaaaagcagaacatggaaggaactttcaggaggactcaaaaagcaacactagaaacttcgaacatcaagaatttgaacaaacaattctcaacaaaaccaagaatcatcctgggaaataaataatgctgtgtggttttggtgtggaatga

accttctaatcaacatggaatgaagttccatgctacaccgtctcatgccatagtaatagttgaaactagtagagacatttaaggtatgaaccgccaaggttaagtactggagatgaaatgcggtttctctatggcaagcacactgaagaacgctgtaacaactgccaattgagaaaacgcctctagtaacacctcaaccatcaacaccaaacaaccatcttggtttgttttggtggtggttgaggaatttcggctggtcgctatgagaca

60120180240300360420480540600660720780840900960102010801140120012601320138014401500156016201632 <211> 557 <212> PRT

<213> Medicago trunculata <400> 184

Met Asn Asn Asn Trp Read Be Phe Pro Read Be Pro Thr His Be Ser1 5 10 15

Leu Pro Wing His Asp Leu Gln Wing Thr Gln Tyr His Gln Phe Ser Leu

20 25 30

Gly Leu Val Asn Glu Asn Met Asp Asn Pro Phe Gln Asn His Asp Trp

35 40 45

Asn Leu Ile Asn Thr His Being Ser Asn Glu Ile Pro Lys Val Wing Asp

50 55 60

Phe Leu Gly Val Ser Lys Ser Glu Asn Gln Ser Asp Leu Wing Wing Leu65 70 75 80

Asn Glu Ile His Ser Asn Asp Ser Asp Tyr Leu Phe Thr Asn Asn Ser

85 90 95

Leu Val Pro Met Gln Asn Pro Val Leu Asp Thr Pro Be Asn Glu Tyr

100 105 110

Gln Glu Asn Wing Asn Be Asn Read Gln Be Read Thr Read Read Be Met Gly

115 120 125

Being Gly Lys Asp Being Thr Cys Glu Thr Being Gly Lys Asp Being Thr Asn

130 135 140

Thr Thr Val Glu Val Wing Pro Arg Arg Thr Read Asp Thr Phe Gly Gln145 150 155 160

Arg Thr Being Ile Tyr Arg Gly Val Thr Arg His Arg Trp Thr Gly Arg

165 170 175

Tyr Glu Ala His Leu Trp Asp Asn Ser Cys Arg Arg Glu Gly Gln Ser

180 185 190

Gly Arg Lys Gly Arg Val Tyr Leu Gly Gly Tyr Asp Lys Glu Glu Lys

195 200 205

Wing Wing Arg Wing Wing Tyr Asp Read Wing Wing Wing Read Lys Tyr Trp Gly Thr Ser

210 215 220

Thr Thr Thr Asn Phe Pro Ile Ser Asn Tyr Glu Lys Glu Read Asp Glu225 230 235 240

Met Lys His Met Thr Arg Gln Glu Phe Val Wing Ile Arg Arg Lys

245 250 255

Be Ser Gly Phe Ser Arg Gly Ala Ser Met Tyr Arg Gly Val Thr Arg

260 265 270

His His Gn His Gly Arg Trp Gln Arg Wing Ile Gly Arg Val Gly Wing

275 280 285

Asn Lys Asp Leu Tyr Leu Gly Thr Phe Be Thr Glu Glu Glu Wing Wing

290 295 300

Glu Wing Tyr Asp Ile Wing Ile Wing Lys Phe Arg Gly Leu Asn Wing Val305 310 315 320

Thr Asn Phe Asp Met Ser Arg Tyr Asp Val Lys Ala Ile Leu Glu Ser

325 330 335

Asn Thr Leu Pro Ile Gly Gly Gly Ala Wing Lys Arg Leu Lys Glu Wing

340 345 350

Gln Wing Leu Glu Be Ser Arg Lys Arg Glu Glu Met Ile Wing Leu Gly

355 360 365

Be Thr Be Phe Gln Tyr Gly Thr Thr Be Asn Be Arg Read His

370 375 380

Tyr Pro Wing Read Met Gln His His His Gln Phe Glu Gln Pro Gln Pro385 390 395 400

Leu Leu Thr Leu Gln Asn His Asp Ile Being His Phe Being His Gln

405 410 415

Gln Asp Pro Read His Gln Gly Tyr Ile Gln Thr Gln Read Gln Read His

420 425 430

Gln Gln Gln Be Gly Gly Be Ser Tyr Ser Phe Gln Asn Asn Asn

435 440 445

Ile Asn Asn Wing Gln Phe Tyr Asn Gly Tyr Asn Read Gln Asn His Pro

450 455 460

Wing Read Leu Gln Gly Met Ile Asn Met Gly Be Ser Ser Ser Ser465 470 475

Val Leu Glu Asn Asn Asn Ser Thr Asn Asn Asn

485 490

Gly Ser Gly Phe Gly Met Wing Be Asn Wing Thr

500 505

Gly Thr Wing Glu Glu Leu Gly Leu Val Lys Val

515 520

Thr Gly Gly Tyr Gly Gly Trp

530 535

Gln Thr Be Asn Be Gly Val Phe Thr Met Trp545 550 555

<210> 185 <211> 2079 <212> DNA <213> Oryza sativa <400> 185

atggccacca tgaacaactg gctggccttc tccctctccctctcagacca actccactct catctccgcc gccgccaccatccaccggcg acgtctgctt caacatcccc caagattggatcggcgctcg tcgccgagcc gaagctggag gacttcctcgcagcagcatc atcacggcgg caagggcggc gtgatcccgagcgagctccg gcagcagcgt cggctacctg taccctcctcttcgccgact ccgtcatggt ggccacctcc tcgcccgtcgggcggcggca tggtgagcgc cgccgccgcc gcggcggccactgtccatga tcaagaactg gctccggagc cagccggcgctctctgtcca tgaacatggc ggggacgacg acggcgcaggctcgccggcg caggggagcg aggccggacg acgcccgcgtgcgcacggag cgacgacggc gacgatggct ggtggtcgcaagcggcagcg ccggcgccgt ggttgccgtc ggctcggagtgtggaggccg gcgcggcggc ggcggcggcg aggaagtccgacatcgatct accgcggcgt gacaaggcat agatggacagtgggacaaca gctgcagaag agagggccaa actcgcaaggaaagaggaaa aagctgctag agcttatgat ttggctgctcacgacgacaa attttccggt aaataactat gaaaaggagcacaaggcagg agttcgtagc ctctttgaga aggaagagcatccatttacc gtggagtaac taggcatcac cagcatgggaagagttgcag ggaacaagga cctctacttg ggcaccttcagaggcgtacg acatcgcggc gatcaagttc cgggggctcaatgagccgct acgacgtcaa gagcatcctc gacagcgctggccaagcgcc tcaaggacgc cgaggccgcc gccgcctacgcacctcggcg gcgacggcgc ctacgccgcg cattacggccgccgcctggc cgaccatcgc gttccaggcg gcggcggcgcctttaccacc cgtacgcgca gccgctgcgt gggtggtgcagtgatcgcgg cggcgcacag cctgcaggat ctccaccaccgccgcgcatg acttcttctc gcaggcgatg cagcagcagcaacgcgtcgc tcgagcacag caccggctcc aactccgtcgggcggaggcg gcggctacat catggcgccg atgagcgccggtggcgagca gccacgatca cggcggcgac ggcgggaagcagctacctcg tcggcgcaga cgcctacggc ggcggcggcggcgatgacgc cggcgtcggc gccggccgcc acgagcagcacatggcgcac agctcttcag cgtctggaac gacacataa <210> 186 <211> 692 <212> PRT

<213> Oryza sativa <4 00> 186

Met Wing Thr Met Asn Asn Trp Read Ala Phe Ser15 10

Gln Leu Pro Pro Being Gln Thr Asn Being Thr Leu

20 25

Thr Thr Thr Thr Wing Gly Asp Be Ser Thr Gly

35 40

Ile Pro Gln Asp Trp Being Met Arg Gly Being Glu

480

Val Gly Gly Phe Val495

Ser Gly Asn Thr Val510

Asp Tyr Asp Met Pro525

Wing Wing Glu Ser Met540

Asn asp

cgcaggatcaccaccaccgcgcatgagggggcggcatctcgcagcgccgccaagctcatctcgcccacgagtggcaacggcgcagccggcgcggcggcgccagagagcctaggagattaacaggcggcagtcgacacgttggaggtatgagtcgtcaaggtcaaatactgtggaggagatgtggtttctcgatggcaagcgcacgcaggaacgccgtcacccctccccgtacgtcggccgaccaccaccacgccgccgcaagcaggagcatcaacctcggacggcctcggtctacaacggtgtcggccacaggtgcagatccgggaggatgcgacatgac

gctcccgccgcggcgactccatcggagctccttctcggagcgcttgctacctcgctccagcggcgtcagccggcattggcgcaggcgctgcatggcgctcgtccacgtcgcgaggaaggccggcgccgtgcggccagagaggctcatctttggttatgacgggcccgacggaagcacatgcagaggtgcaaaggataggaggaggcggcgcaacttcgaccggcaccgcccatcgcctcgctcggccgcccgccgccgggggaccacgcccgccgccgcccagcatcgaccgacaatggcggccaccgcgggggtacgacgccatcctggcggagtctgc

601201802403003604204805406006607207808409009601020108011401200126013201380144015001560162016801740180018601920198020402079

Read Ser Pro Gln Asp15

Ile Ser Ala Wing Ala30

Asp Val Cys Phe Asn45

Leu Ser Wing Leu Val50 55 60

Glu Wing Pro Lys Read Glu Asp Phe Read Gly Gly Ile Be Phe Be Glu65 70 75 80

Gln Gln His Gly Gly Gly Lys Gly Gly Val Ile Pro Be Ser Ala

85 90 95

Wing Cys Wing Tyr Wing Be Ser Gly Ser Be Val Gly Tyr Leu Tyr Pro

100 105 110

Pro Pro Ser Ser Ser Ser Glu Phe Ala Asp Ser Val Val Val Wing

115 120 125

Thr Be Ser Pro Val Val Wing His Asp Gly Val Ser Gly Gly Gly Met

130 135 140

Val Ser Wing Ala Wing Ala Ala Ala Ala Ser Gly Asn Gly Gly Ile Gly145 150 155 160

Read Ser Met Ile Lys Asn Trp Read Arg Be Gln Pro Wing Pro Gln Pro

165 170 175

Gln Wing Wing Read Be Read Be Met Asn Met Wing Gly Thr Thr Thr Wing

180 185 190

Gln Gly Gly Gly Wing Met Wing Read Leu Read Gly Wing Gly Wing Glu Arg Gly

195 200 205

Arg Thr Thr Pro Wing Be Glu Be Read Be Thr Be Wing His Gly Wing

210 215 220

Thr Thr Wing Thr Met Wing Gly Gly Arg Lys Glu Ile Asn Glu Glu Gly225 230 235 240

Ser Gly Ser Wing Gly Val Wing Val Val Wing Val Gly Ser Glu Ser Gly Gly

245 250 255

Ser Gly Wing Val Val Glu Wing Gly Wing Wing Wing Wing Wing Wing Arg Lys

260 265 270

Being Val Asp Thr Phe Gly Gln Arg Thr Being Ile Tyr Arg Gly Val Thr

275 280 285

Arg His Arg Trp Thr Gly Arg Tyr Glu Wing His Leu Trp Asp Asn Ser

290 295 300

Cys Arg Arg Glu Gly Gly Thr Arg Lys Gly Gly Arg Glly Gly Gly Tyr Asp305 310 315 320

Lys Glu Glu Lys Wing Arg Wing Arg Wing Tyr Asp Leu Wing Wing Leu Lys Tyr

325 330 335

Trp Gly Pro Thr Thr Thr Asn Phe Pro Val Asn Asn Tyr Glu Lys

340 345 350

Glu Leu Glu Glu Met Lys His Met Thr Arg Gln Glu Phe Val Wing Ser

355 360 365

Read Arg Arg Lys Be Gly Phe Be Arg Gly Wing Be Ile Tyr Arg

370 375 380

Gly Val Thr Arg His His Gln His Gly Arg Trp Gln Wing Arg Ile Gly385 390 395 400

Arg Val Wing Gly Asn Lys Asp Leu Tyr Leu Gly Thr Phe Ser Thr Gln

405 410 415

Glu Glu Wing Wing Glu Wing Tyr Asp Ile Wing Ile Wing Lys Phe Arg Gly

420 425 430

Leu Asn Wing Val Thr Asn Phe Asp Met Ser Arg Tyr Asp Val Lys Ser

435 440 445

Ile Leu Asp Ser Wing Ward Leu Pro Val Gly Thr Wing Ward Lys Arg Leu

450 455 460

Lys Asp Wing Glu Wing Wing Wing Tyr Wing Wing Val Gly Arg Ile Wing Ser465 470 475 480

His Leu Gly Gly Asp Gly Wing Tyr Wing Wing His Tyr Gly His His

485 490 495

His Ser Wing Wing Wing Wing Trp Pro Thr Ile Wing Phe Gln Wing Wing Wing

500 505 510

Pro Wing Pro Pro His Wing Wing Gly Leu Tyr His Pro Tyr Wing Gln Pro

515 520 525

Read Arg Gly Trp Cys Lys Gln Glu Gln Asp His Wing Val Ile Wing Wing

530 535 540

Wing His Ser Read Le Gln Asp Read His His Leu Asn Leu Gly Wing Wing Ala545 550 555 560Ala Wing His Asp Phe Phe Ser Gln Wing Met Gln

565 570

Gly Ser Ile Asp Asn Wing Being Read Glu His Ser

580 585

Val Val Tyr Asn Gly Asp Asn Gly Gly Gly Gly

595 600

Pro Met Ward Be Val Ward Be Thr Thr Ward

610 615

His Glyph His Gly Gly Asly Gly Gly Lys Gln Val625 630 635

Ser Tyr Leu Val Gly Wing Asp Wing Tyr Gly Gly

645 650

Met Pro Be Trp Wing Met Thr Pro Wing Be Wing

660 665

Being Asp Met Thr Gly Val Cys His Gly Wing

675 680

Trp Asn Asp Thr

690 <210> 187 <211> 2133 <212> DNA <213> Zea mays <4 00> 187

atggccactg tgaacaactg gctcgctttc tccctctccccagacgacgg actccacact catctcggcc gccaccgccgtgcttcaaca tcccccaaga ttggagcatg aggggatcaggagccgaagc tggaggactt cctcggcggc atctccttctaactgcaaca tgatacccag cactagcagc acagtttgctaccggctacc atcaccagct gtaccaccag cccaccagcttccgtaatgg tggcctcctc ggccggtgtc cacgacggcggccgctaacg gtgtcgctgg cgctgccagt gccaacggcgattaagaact ggctgcggag ccaaccggcg cccatgcagcggcgcgcagg ggctctcttt gtccatgaac atggcggggaatgccacttc tcgctggaga gcgcgcacgg gcgcccgagaggtggagccg tcgtcgtcac ggcgccgaag gaggatagcggctctagtag ccgtgagcac ggacacgggt ggcagcggcggcaaggaaga cggtggacac gttcgggcag cgcacgtcgacatagatgga ctgggagata tgaggcacat ctttgggatacaaactcgta agggtcgtca agtctattta ggtggctatgagggcttatg atcttgctgc tctgaagtac tggggtgccagtgagtaact acgaaaagga gctcgaggac atgaagcacagcgtctctga gaaggaagag cagtggtttc tccagaggtgactaggcatc accaacatgg aagatggcaa gcacggattggatctttact tgggcacctt cagcacccag gaggaggcaggcgatcaagt tccgcggcct caacgccgtc accaacttcgaagagcatcc tggacagcag cgccctcccc atcggcagcggccgaggccg cagcgtccgc gcagcaccac cacgccggcgcgcatcgcct cgcagctcgg cgacggcgga gccctggcggcacggcgccg cctggccgac catcgcgttc cagccgggcgcacccgtacg cgcagcagcc aatgcgcggc ggcgggtggtgcggtgatcg cggccgcgca cagcctgcag gacctccaccggcgcgcacg actttttctc ggcagggcag caggccgccgggtagcatcg acagtgcgtc gctcgagcac agcaccggctggcggggtcg gcgacagcaa cggcgccagc gccgtcggcgatgccgatga gcgctgccgg agcaaccact acatcggcaacatgcacggg cctacgacga agccaagcag gctgctcagagtgaacgcgg agaacaatgg tggcggaagg atgtctgcatgccgcggcgg cagcagcaag cagcaacgac aacatggccggcgcagctct tcagtgtctg gaacgacact taa <210> 188 <211> 710 <212> PRT <213> Zea mays

Gln Gln His

Thr Gly Ser590

Gly Gly Tyr605

Wing Val Wing620

Gln Met Gly

Gly Gly Wing

Pro Wing Ala670

Gln Leu Phe685

Gly Leu575

Asn ser

Ile met

To be to be

Tyr Asp640Gly Arg655

Thr SerSer Val

cgcaggagct gccgccctcc 60accatgtctc cggcgatgtc 120agctttcggc gctcgtcgcg 180ccgagcagca tcacaaggcc 240acgcgagctc aggtgctagc 300cagcgctcca cttcgcggac 360gtgccatgct cagcgcggcc 420gcggcatcgg gctgtccatg 480cgagggtggc ggcggctgag 540cgacccaagg cgctgctggc 600gtgtatcgac gtcagcacag 660gtggcagcgg tgttgccggc 720gcgcgtcggc tgacaacacg 780tttaccgtgg cgtgacaagg 840acagttgcag aagggaaggg 900ataaagagga gaaagctgct 960caacaacaac aaattttcca 1020tgacaaggca ggagtttgta 1080catccattta caggggagtg 1140gacgagttgc agggaacaag 1200cggaggcgta cgacatcgcg 1260acatgagccg ctacgacgtg 1320ccgccaagcg cctcaaggag 1380tggtgagcta cgacgtcggc 1440cggcgtacgg cgcgcactac 1500ccgccagcac aggcctgtac 1560gcaagcagga gcaggaccac 1620acctgaacct gggcgcggcc 1680ccgctgcgat gcacggcctg 1740ccaactccgt cgtctacaac 1800gcagtggcgg tggctacatg 1860tggtgagcca cgagcaggtg 1920tggggtacga gagctacctg 1980gggggactgt cgtgtctgca 2040ccgacgtcgg ccatggcggc 2100 2133 <400> 188

Met Wing Thr Val Asn Asn Trp Read Ala Phe Be Read Ser Pro Gln Glu1 5 10 15

Leu Pro Pro Be Gln Thr Thr Asp Be Thr Read Leu Ile Be Wing Ward Thr

20 25 30

Asp Wing His Val Ser Gly Asp Val Cys Phe Asn Ile Pro Gln Asp Trp

35 40 45

Be Met Arg Gly Be Glu Leu Be Wing Leu Val Wing Glu Pro Lys Leu

50 55 60

Glu Asp Phe Read Gly Gly Ile Be Phe Be Glu Gln His His Lys Ala65 70 75 80

Asn Cys Asn Met Ile Pro Be Thr Be Be Thr Val Cys Tyr Wing Be

85 90 95

Be Gly Ala Be Thr Gly Tyr His His Gln Read Tyr His Gln Pro Thr

100 105 110

Ser Ser Ala Read His Phe Al Ser Asp Ser Val Val Val Ser Ser Ser Ala

115 120 125

Gly Val His Asp Gly Gly Gly Wing Met Read To Be Wing Wing Wing Wing Asn Gly

130 135 140

Val Wing Gly Wing Wing Be Wing Asn Gly Gly Gly Ile Gly Read Ser Met145 150 155 160

Ile .. Lys Asn Trp Read Arg Be Gln Pro Wing Pro Met Gln Pro Arg Val

165 170 175

Wing Wing Wing Glu Gly Wing Gln Gly Read Be Read Be Met Asn Met Wing

180 185 190

Gly Thr Thr Gln Gly Wing Gly Met Pro Pro Read Leu Wing Gly Glu Arg

195 200 205

Wing Arg Wing Pro Glu Be Val Be Thr Be Gln Wing Gly Gly Val Wing

210 215 220

Val Val Thr Wing Pro Lys Glu Asp Be Gly Gly Be Gly Val Wing Gly225 230 235 240

Wing Leu Val Wing Val Val Thr Thr Asp Thr Gly Gly Ser Gly Gly Wing Ser

245 250 255

Wing Asp Asn Thr Wing Arg Lys Thr Val Asp Thr Phe Gly Gln Arg Thr

260 265 270

Ser Ile Tyr Arg Gly Val Thr Arg His Arg Trp Thr Gly Arg Tyr Glu

275 280 285

Wing His Leu Trp Asp Asn Be Cys Arg Arg Glu Gly Gln Thr Arg Lys

290 295 300

Gly Arg Gln Val Tyr Leu Gly Gly Tyr Asp Lys Glu Glu Lys

Arg Wing Tyr Asp Leu Wing Wing Leu Lys Tyr Trp Gly Wing Thr Thr Thr

325 330 335

Thr Asn Phe Pro Val Ser Asn Tyr Glu Lys Glu Leu Glu Asp Met Lys

340 345 350

His Met Thr Arg Gln Glu Phe Val Wing Be Read Arg Arg Lys Be Ser

355 360 365

Gly Phe Be Arg Gly Wing Be Ile Tyr Arg Gly Val Thr Arg His

370 375 380

Gln His Gly Arg Trp Gln Arg Wing Ile Gly Arg Val Wing Gly Asn Lys385 390 395 400

Asp Leu Tyr Leu Gly Thr Phe Be Thr Gln Glu Glu Wing Wing Glu Wing

405 410 415

Tyr Asp Ile Wing Ile Wing Lys Phe Arg Gly Leu Asn Wing Val Thr Asn

420 425 430

Phe Asp Met Be Arg Tyr Asp Val Lys Be Ile Read Asp Be Ser Ala

435 440 445

Leu Pro Ile Gly Ser Wing Lys Arg Wing Leu Lys Glu Wing Glu Wing Wing

450 455 460

Wing Ser Wing Gln His His His Wing Gly Val Val Ser Tyr Asp Val Gly465 470 475 480

Arg Ile Wing Be Gln Leu Gly Asp Gly Gly Wing Leu Wing Wing Wing Wing Tyr485 490 495Gly Wing His Tyr His Gly Wing Trp Pro Thr

500 505

Gly Wing Wing Be Thr Gly Leu Tyr His Pro Tyr

515 520

Arg Gly Gly Gly Trp Cys Lys Gln Glu Gln Asp

530 535

Wing Wing His Ser Leu Gln Asp Leu His His Leu545 550 555

Gly Wing His Asp Phe Phe Ser Wing Gly Gln Gln

565 570

Met His Gly Read Gly Ser Ile Asp Ser Ala Ser

580 585

Gly Ser Asn Ser Ser Val Tyr Asn Gly Serly Val

595 600

Wing Be Wing Val Gly Gly Be Gly Gly Gly Tyr

610 615

Wing Wing Gly Wing Thr Thr Thr Be Wing Met Val625 630 635

His Wing Arg Wing Tyr Asp Glu Wing Lys Gln Wing

645 650

Glu Ser Tyr Leu Val Asn Wing Glu Asn Asn Gly

660 665

Trp Wing Gly Thr Val Val Ser Wing Wing Wing Wing

675 680

Asn Asp Asn Met Wing Asp Wing Val Gly His Gly

690 695

Ser Val Asp Asn Thr705 710

<210> 189 <211> 1374 <212> DNA

<213> Lotus corniculatus <4 00> 189

agatctctct ctgcagtgca caatccagtg gtaagcacatcaagaaaatg gtaatggtaa tttgcaatcg ttgacattattcaacatgtg aaaccagtgg tgacactagt accaacactaaccttggaga catttgggca aagaacatct atatatcgagaccggaaggt atgaagctca cctttgggac aatagctgtaaaaggtcgcc aaggtggata tgataaagaa gacaaagcaggcccttaagt actgggggac atccactact accaactttcgaagtggatg acatgaagca tatgacaaga caagaatttgagcagtggtt tctcgagggg tgcttcaatg tatcgtggaggggaggtggc aagcaaggat tggaagagtt gcaggaaacattcggtactg aggaagaggc ggcagaagct tacgacatagcttaacgcca tcaccaactt tgacatgaac cgttacgatgaacaccctcc caatcggagg aggagcttca aaaaggctaatcttcaagaa aacgtgaaga tcagatgatt gcactcggctattgcaaccc catcaagctc tacaaggcca caaccttacccacaatcagt ttgaacaaca gcctcaacca tttctaaccttctcaatact cccatcatca gcaggaccct tcgtttcagccttcagttgc agttgaatca acacggtggt ggtggttctacctcatcaga acagtgagtt ctataatggt ggaaattattatgatgatga acaatagtat ggggtcttgt tcttcatcgtaatgctgctg ctggtggggc ttcttttgtg ggtcccgcagaaggttgatt atgacatgga tgctgctgct ggcggtggttgagtccatgc acacgtcggc ggctggtggt ttgtttacta <210> 190 <2 11> 457 <212> PRT

<213> Lotus corniculatus <400> 190

Arg Be Read Sera Wing Val His Asn Pro Val Val1 5 10

Ile

Allah

His540Asn

Allah

Read

Gly

Met620Ser

Allah

Gly

Allah

Gly700

Phe510Gln Gln525 Wing

Val Wing

Read gly

Wing wing

Glu His590Asp Ser605

Met pro

His glu

Gln met

Gly Arg670Ala Ala685

Gln Wing

Gln pro

Pro met

Ile wing

Wing Ala560Ala Ala575

To be thr

Asn gly

Met ser

Gln Val640Gly Tyr655

Met SerSer SerLeu Phe

gtagtaatggccatgggaagtcgaagctgtgtgtaacaaggaagggaaggctagggcctacggttagcaatggcttccatttacaaggcaaagatctttactgcgataaatgaaagcataagaaactcacaacatttcacgctaaacggaggacgacgacgacgacgacgg

taatgagtcttggtaaggatgcctagaagaacatagatggacagtcaaggtgatttagctctatgagaagtagaaggaaatcatcaacatcttgggaactgttcagaggctctagagagcagctctggaatcaatacggaaatgcatcaggggaggggggggagggtagagt

601201802403003604204805406006607207808409009601020108011401200126013201374

Be Thr Cys Be Asn1 5Gly Asn Glu Be Gln Glu Asn Gly Asn Gly Asn Read Gln Be Read Thr

20 25 30

Read Being Met Gly Being Gly Lys Asp Being Thr Cys Glu Thr Being Gly Asp

35 40 45

Thr Be Thr Asn Thr Ile Glu Wing Val Pro Arg Arg Thr Leu Glu Thr

50 55 60

Phe Gly Gln Arg Thr Be Ile Tyr Arg Gly Val Thr Arg His Arg Trp65 70 75 80

Thr Gly Arg Tyr Glu Wing His Leu Trp Asp Asn Ser Cys Arg Arg Glu

85 90 95

Gly Gln Being Arg Lys Gly Arg Gln Gly Gly Tyr Asp Lys Glu Asp Lys100 105 110

Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Wing Ile Arg Arg Lys145 150 155 160Be Be Gly Phe Be Arg Gly Wing Be Met Tyr Arg Gly Val Thr Arg 165 170 175 His His Gln His Gly Arg Trp Gln Wing Arg Gly Thr Phe Gly Thr Glu Glu Glu Wing Wing 195 200 205 Glu Wing Tyr Asp Ile Wing Ile Lys Phe Arg Gly Leu Asn Wing Ile 210 215 220 Thr Asn Phe Asp Met Asn Arg Tyr Asp Val Lys Wing Ile Leu Glu Ser225 230 235 240Asn Thr Leu Pro Ile Gly Gly Gly Wing Be Lys Arg Leu Lys Glu Thr 245 250 255 Gln Wing Leu Glu Be Be Arg Lys Arg Glu Asp Gln Met Ile Wing Leu 260 265 270 Gly Be Thr Phe His Gln Tyr Gly Ile Wing Thr Pro Being Being Being Thr 275 280 285 Arg Pro et His His His Asn Gln Phe 290 295 300 Glu Gln Gln Pro Gln Pro Phe Leu Thr Read Le Gln Asn His Asp Ile Asn305 310 315 320Ser Gln Tyr Be His His Gln Gln Asp Pro Be Phe Gln Gln Ser Tyr 325 330 335 Ile Gln Thr Gln Leu Gln Leu Gln Leu Asn Gln His Gly Gly Gly Gly 340 345 350 Be Ser Tyr Wing Gln Glu Thr Wing Pro His Gln Asn Ser Glu Phe Tyr 355 360 365 Asn Gly Gly Asn Tyr Tyr Leu Gln Asn Leu Gln Gly Met Met Met Asn 370 375 380 Asn Be Met Gly Be Cys Be Be Ser Be Val Leu Glu Asn Asp His385 390 395 400Asn Wing Ward Wing Gly Wing Gly Wing Be Phe Val Gly Pro Wing Wing Glu Glu 405 410 415 Leu Gly Leu Val Lys Val Asp Tyr Asp Met Asp Wing Ward Wing Gly Gly 420 425 430 Gly Tyr Gly Gly Trp Being Wing Wing Glu Being Met His Thr Being Wing Wing 435 440 445 Gly Gly Leu Phe Thr Met Trp Asn Glu 450 455

<210> 191

<211> 202 <212> PRT

<213> Artificial sequence <220>

<223> Arath_PLTl and Arath_PLT2 AP2 domain <220> <221> VARIANT

<222> (4) .. (4)

<223> / replace = "Thr"

<220>

<221> VARIANT <222> (6) .. (6) <223> / replace = "Glu" <220>

<221> VARIANT

<222> (57) .. (57)

<223> / replace = "Glu"

<220>

<221> VARIANT

<222> (62) .. (62)

<223> / replace = "Wing"

<220>

<221> VARIANT

<222> (93) .. (93)

<223> / replace = "Asn"

<220>

<221> VARIANT <222> (102) .. (102) <223> / replace = "Be" <220>

<221> VARIANT <222> (187) .. (187) <223> / replace = "Asn" <4 00> 191

Pro Arg Arg Wing Read Asp Thr Phe Gly Gln Arg Thr Be Ile Tyr Arg15 10 15

Gly Val Thr Arg His Arg Trp Gly Arg Tyr Glu Wing His Leu Trp

20 25 30

Asp Asn Be Cys Arg Arg Glu Gly Gln Be Arg Lys Gly Arg Gln Val

35 40 45

Tyr Leu Gly Gly Tyr Asp Lys Glu Asp Lys Wing Wing Arg Be Tyr Asp

50 55 60

Leu Wing Ala Leu Lys Tyr Trp Gly Pro Be Thr Thr Thr Asn Phe Pro65 70 75 80

Ile Thr Asn Tyr Glu Lys Glu Val Glu Glu Met Lys His Met Thr Arg

85 90 95

Gln Glu Phe Val Wing Ward Ile Arg Arg Lys Be Ser Gly Phe Ser Arg

100 105 110

Gly Ala Be Met Tyr Arg Gly Val Thr His Arg His Gln His Gly Arg

115 120 125

Trp Gln Wing Arg Ile Gly Arg Val Wing Gly Asn Lys Asp Leu Tyr Leu

130 135 140

Gly Thr Phe Be Thr Glu Glu Glu Wing Wing Glu Wing Tyr Asp Ile Wing145 150 155 160

Wing Ile Lys Phe Arg Gly Leu Asn Wing Val Thr Asn Phe Glu Ile Asn

165 170 175

Arg Tyr Asp Val Lys Wing Ile Leu Glu Ser Be Thr Leu Pro Ile Gly

180 185 190

Gly Gly Wing Wing Lys Arg Winged Lys Glu Wing

195 200

<210> 192 <211> 8 <212> PRT

<213> Artificial sequence <220>

<223> reason 1 <220>

<221> VARIANT <222> (3) .. (3) <223> / replace = "Read" <220>

<221> VARIANT <222> (4). . (4) <223> / replace = "Glu" <4 00> 192

Pro Lys Val Wing Asp Phe Leu Gly

1 5

<210> 193

<211> 7

<212> PRT

<213> Artificial sequence <220>

<223> reason 2 <220>

<221> VARIANT

<222> (1) .. (1)

<223> / replace = "Read"

<220>

<221> VARIANT <222> (3) .. (3) <223> / replace = "Be" <220>

<221> VARIANT <222> (4) .. (4) <223> / replace = "Going" <220>

<221> VARIANT <222> (7) .. (7) <223> / replace = "Glu" <4 00> 193

Val Phe Thr Met Trp Asn Asp

1 5

<210> 194

<211> 2193

<212> DNA

<213> Oryza sativa

<4 00> 194

aatccgaaaa gtttctgcac cgttttcacc ccctaactaa caatataggg aacgtgtgct 60

aaatataaaa tgagacctta tatatgtagc gctgataact agaactatgc aagaaaaact 120catccaccta ctttagtggc aatcgggcta aataaaaaag agtcgctaca ctagtttcgt 180tttccttagt aattaagtgg gaaaatgaaa tcattattgc ttagaatata cgttcacatc 240tctgtcatga agttaaatta ttcgaggtag ccataattgt catcaaactc ttcttgaata 300aaaaaatctt tctagctgaa ctcaatgggt aaagagagag atttttttta aaaaaataga 360atgaagatat tctgaacgta ttggcaaaga tttaaacata taattatata attttatagt 420ttgtgcattc gtcatatcgc acatcattaa ggacatgtct tactccatcc caatttttat 480ttagtaatta aagacaattg acttattttt attatttatc ttttttcgat tagatgcaag 540gtacttacgc acacactttg tgctcatgtg catgtgtgag tgcacctcct caatacacgt 600tcaactagca acacatctct aatatcactc gcctatttaa tacatttagg tagcaatatc 660tgaattcaag cactccacca tcaccagacc acttttaata atatctaaaa tacaaaaaat 720aattttacag aatagcatga aaagtatgaa acgaactatt taggtttttc acatacaaaa 780aaaaaaagaa ttttgctcgt gcgcgagcgc caatctccca tattgggcac acaggcaaca 840acagagtggc tgcccacaga acaacccaca aaaaacgatg atctaacgga ggacagcaag 900tccgcaacaa ccttttaaca c gcaggctttg ggccaggag agaggaggag aggcaaagaa 960aaccaagcat cctcctcctc ccatctataa attcctcccc ccttttcccc tctctatata 1020ggaggcatcc aagccaagaa gagggagagc accaaggaca cgcgactagc agaagccgag 1080cgaccgcctt cttcgatcca tatcttccgg tcgagttctt ggtcgatctc ttccctcctc 1140cacctcctcc tcacagggta tgtgcccttc ggttgttctt ggatttattg ttctaggttg 1200tgtagtacgg gcgttgatgt taggaaaggg gatctgtatc tgtgatgatt cctgttcttg 1260gatttgggat agaggggttc ttgatgttgc atgttatcgg ttcggtttga ttagtagtat 1320ggttttcaat cgtctggaga gctctatgga aatgaaatgg tttagggtac ggaatcttgc 1380gattttgtga gtaccttttg tttgaggtaa aatcagagca ccggtgattt tgcttggtgt 1440aataaaagta cggttgtttg gtcctcgatt ctggtagtga tgcttctcga tttgacgaag 1500ctatcctttg tttattccct attgaacaaa aataatccaa ctttgaagac ggtcccgttg 1560atgagattga atgattgatt cttaagcctg tccaaaattt cgcagctggc ttgtttagat 1620acagtagtcc ccatcacgaa attcatggaa acagttataa tcctcaggaa caggggattc 1680cctgttcttc cgatttgctt tagtcccaga attttttttc ccaaatatct taaaaagtca 1740ctttctggtt cagttcaatg aattgattgc tacaaataat gcttttatag cgttatccta 1800gctgtagttc agttaatagg taatacccct atagtttagt caggagaaga acttatccga 1860tttctgatct ccatttttaa ttatatgaaa tgaactgtag cataagcagt attcatttgg 1920attatttttt ttattagctc tcaccccttc attattctga gctgaaagtc tggcatgaac 1980tgtcctcaat tttgttttca aattcacatc gattatctat gcattatcct cttgtatcta 2040cctgtagaag tttctttttg gttattcctt gactgcttga ttacagaaag aaatttatga 2100agctgtaatc gggatagtta tactgcttgt tcttatgatt catttccttt gtgcagttct 2160tggtgtagct tgccactttc accagcaaag ttc 2193

<210> 195 <211> 54 <212> DNA

<213> Artificial sequence <220>

<223> initiator: PRM08180 <4 00> 195

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgat caatccacac ggtg 54

<210> 196 <211> 50 <212> DNA

<213> Artificial sequence <220>

<223> initiator: PRM08181 <4 00> 196

ggggaccact ttgtacaaga aagctgggtt ccttgtttac tcattccaca 50

<210> 197 <211> 56 <212> DNA

<213> Artificial sequence <220>

<223> initiator: PRM08182 <400> 197

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgaa ttctaacaac tggctc 56

<210> 198 <211> 50 <212> DNA

<213> Artificial sequence <220>

<223> initiator: PRM08183 <4 00> 198

ggggaccact ttgtacaaga aagctgggtt catcttttat tcattccaca 50

<210> 199 <211> 1638 <212> DNA

<213> Populus tremuloides <4 00> 199

atgaattcta acaactggct ttcatttcct ctttctccta ctcatccttc cttgcctgct 60

catctacatg catcccaccc tcatcaattc tctctagggt tagtcaatga taatatggaa 120aacccatttc aaactcaaga gtggagtctt cttaacactc atcaaggcaa caatgaagtg 180ccaaaggttg cagactttct tggtgtgagc aaatctgaga atcaatcaga tcttgtagcc 240ttcaatgaaa ttcaagctaa tgaatctgac tatctctttt caaacaatag tctagtacca 300gtccaaaatg ctgttgtagg cgccaataat acctttgagt ttcaagaaaa tgctagcaat 360ttgcagtcat taacattgtc tatgggcagt gctagtggta aaggttctac atgtgaaccc 420agtggggata atagcactaa tactgttgaa gctgctgcac caagaagaac tttggataca 480tttgggcaaa gaacatccat atatcgtggt gtaacaaggc atcgatggac aggaaggtat 540gaagctcatt tatgggataa tagttgcaga agagaaggtc aatctaggaa aggaagacag 600ggtggctatg acaaagaaga aaaggcagct agggcttatg atcttgctgc acttaagtac 660tggggaacat ccaccactac caattttcca atcagcaact acgagaaaga aatagaggaa 720atgaagcaca tgaccaggca agaatttgtg gcctccatta gaaggaagag tagtggcttc 780tctaggggtg catccatgta tcgtggagtt acaaggcatc accagcatgg tagatggcaa 840gcaaggatag gcagagttgc aggaaacaaa gatctctact tgggaacttt tagcactgag 900gaggaggctg cagaagctta g tgacatagca caataaagt ttagagggct taatgcagtg 960actaactttg acatgaatcg atatgatgtg aagagcattc ttgaaagcaa tactttgcca 1020attggaggag gggcagccaa acggctaaag gaggctcaag caattgaatc atcacgaaaa 1080agagaagaaa tgattgctct tggctcaagt tttccatatg gatcaacttc aagctctagc 1140aggctacaag cttaccctct aatgcagaca ccatttgagc aacctcaacc tttacttact 1200ctacaaaatc aagacatttc tcagtacact caggattcct catcattcca ccaaaatttc 1260cttcaaacac agcttcattt gcaccagcaa tctacagggt ctaatttcct gcataaccaa 1320tcaaaccaaa accctcaata ttacaatagt tatatccaaa acaatccagc tttacttcat 1380ggattgtgga acatgggttc ttcatcatct gtaatggaga ataatggaag ttctagtggg 1440agctatagta ctggaggtta tctgggaaat gggctgggaa tggcttccaa ttcaacaggg 1500tctaatgcag .taggatcagc cgaggaactt gcacttgtca aagttgatta tgatatgcct 1560tctagtggct atggaagctg gtctggggac tcagtccagg gatccaatcc aggtgttttc 1620actatgtgga atgagtga 1638

<210> 200 <211> 545 <212> PRT

<213> Populus tremuloides <400> 200

Met Asn Be Asn Asn Trp Read Be Phe Pro Read Be Pro Thr His Pro15 10 15

Be Leu Pro His Wing Leu His Wing Be His Pro His Gln Phe Be Leu

20 25 30

Gly Leu Val Asn Asp Asn Met Glu Asn Pro Phe Gln Thr Gln Glu Trp

35 40 45

Ser Leu Read Asn Thr His Gln Gly Asn Asn Glu Val Pro Lys Val Wing

50 55 60

Asp Phe Leu Gly Val Be Lys Be Glu Asn Gln Be Asp Leu Val Ala65 70 75 80

Phe Asn Glu Ile Gln Wing. Asn Glu Ser Asp Tyr Leu Phe Ser Asn Asn

85 90 95

Ser Leu Val Pro Val Gln Asn Wing Val Val Gly Wing Asn Asn Thr Phe

100 105 110

Glu Phe Gln Glu Asn Wing Be Asn Read Gln Be Read Thr Read Be Met

115 120 125

Gly Being Ala Being Gly Lys Gly Being Thr Cys Glu Pro Being Gly Asp Asn

130 135 140

Ser Thr Asn Thr Val Glu Ward Wing Ward Pro Arg Arg Thr Read Asp Thr145 150 155 160

Phe Gly Gln Arg Thr Being Ile Tyr Arg Gly Val Thr Arg His Arg Trp

165 170 175

Thr Gly Arg Tyr Glu Wing His Leu Trp Asp Asn Ser Cys Arg Arg Glu

180 185 190

Gly Gln Being Arg Lys Gly Arg Gln Gly Gly Tyr Asp Lys Glu Glu Lys

195 200 205

Wing Wing Arg Wing Wing Tyr Asp Read Wing Wing Wing Read Lys Tyr Trp Gly Thr Ser

210 215 220

Thr Thr Thr Asn Phe Pro Ile Ser Asn Tyr Glu Lys Glu Ile Glu Glu225 230 235 240

Met Lys His Met Thr Arg Gln Glu Phe Val Wing Ser Ile Arg Arg Lys

245 250 255

Be Ser Gly Phe Ser Arg Gly Ala Ser Met Tyr Arg Gly Val Thr Arg

260 265 270

His His Gn His Gly Arg Trp Gln Arg Wing Ile Gly Arg Val Gly Wing

275 280 285

Asn Lys Asp Leu Tyr Leu Gly Thr Phe Be Thr Glu Glu Glu Wing Wing

290 295 300

Glu Wing Tyr Asp Ile Wing Ile Wing Lys Phe Arg Gly Leu Asn Wing Val305 310 315 320

Thr Asn Phe Asp Met Asn Arg Tyr Asp Val Lys Ser Ile Leu Glu Ser

325 330 335

Asn Thr Leu Pro Ile Gly Gly Gly Wing Ally Lys Arg Wing Leu Lys Glu Wing340 345 350Gln Wing Ile Glu Be Being Arg Lys Arg Glu Glu

355 360

Be Ser Phe Pro Tyr Gly Ser Thr Ser Ser

370 375

Tyr Pro Read Met Gln Thr Pro Phe Glu Gln Pro385 390 395

Read Gln Asn Gln Asp Ile Be Gln Tyr Thr Gln

405 410

His Gln Asn Phe Leu Gln Thr Gln Leu His Leu

425

Met

Ser380Gln

AspHis

Gly Ser Asn Phe Read His Asn Gln Ser Asn Gln Asn

420

435

440

Asn Ser Tyr Ile Gln Asn Asn Pro Wing Leu Leu

450

455

Met Gly Be Ser Be Ser Val Met Glu Asn Asn465 470 475

Being Tyr Being Thr Gly Gly Tyr Read Gly Asn Gly

485 490

Asn Be Thr Gly As Asn Wing Val Gly Be Wing

500 505

Val Lys Val Asp Tyr Asp Met Pro To Be Gly

515 520

Gly Asp Be Val Gln Gly Asp Pro Gly Val530 535

Glu545 <210> 201 <211> 1515 <212> DNA

<213> Populus tremuloides <400> 201

atgttttttt ttttgtttcc tttagagtgg agtcttcttagtgccaaagg ttgcagattt tcttggtgta agtaaatctggccttcaatg aaattcaagc cagtgattct gagtatctctccggtccaaa atgctgtggt agccgccagt actaactacgaatttgcagt cattaacatt gtctatgggc agtgctagtgaccagtggtg ataatagtac taattctgtc gaagctgctgacatttggtc aaagaacatc catctatcgt ggtgtaacaatatgaagctc atttatggga taatagttgc agaagagaagcaaggtggct atgacaaaga agacaaggct gctagggctttactggggaa catcgaccac taccaatttt cctatcagcagacatgaaga acatgaccag acaagaattt gtggcctccattctctaggg gtgcatccat gtatcgtgga gtcacaaggccaagcaagaa ttggtagagt tgcaggaaac aaagatctctgaggaggagg ctgcagaagc ttatgacata gcagcaataagtgactaatt ttgacatgaa tcgatatgat gtgaaaagcaccaattggag gaggggcagc caaaaggcta aaggaggctcaaacgagaag aaatgattgc tcttggatca agttatccatagtcgacaac aagcttactc tctaatgcag aaaccatttgactctacaaa atcaagacat ttctcagtac actcaagattcttcaaacac agcttcattt gcaccagcta tctgcagggtcaatcaagcc aaaatcctca gtattacaac agctatatcccatggattgt ggaacatggg ttcttcatca tctctaatgggggagttata gtactgtcgg ttatctggga aatgggttgggggtctaatg cagtagctga ggaacttcca cttgttaagaggtggctatg gaagttggtc tggggaatca gttcagggatatgtggaatg agtga <210> 202 <211> 504 <212> PRT

<213> Populus tremuloides <400> 202

Met Phe Phe Phe Leu Phe Pro Leu Glu Trp Ser

His460Gly

Read

Glu

Tyr

Phe540

Ile365Arg

Pro

To be

Gln

Pro445Gly

To be

Gly

Glu

Gly525Thr

Wing Leu

Read Gln

Leu Leu

Ser Ser415'Gln Ser430

Gln tyr

Read trp

To be to be

Met Ala495Leu Ala510

Be trp

Gly

Allah

Thr400Phe

Thr

Tyr

Asn

Gly480Ser

LeuSerMet Trp Asn

acactcaaggagaatcaatctttcaagcaaaatttcaagagtaagggttcctccaagaagggcatcgatggtcaatccagatgatcttgcactatgagaattagaaggaaatcaccaacaacttgggaacagtttagaggttcttgagagaagcaatcatgatagatcactacatacatcata

caacaatgaaagatctcgtatagtctgttgaaatcctagctaaatgtgaagactttggatgacaggaagggaaaggaagatgcacttaagagaactagaggagtagtggctggaagatggttttagcactgcttaatgcacaatagtttgatcgtcacaattcaagctctacctttacttgcaaaattacgcataataaccactttgcttcagttctagtcaattcaacatatgccttcttgtttttaca

601201802403003604204805406006607207808409009601020108011401200126013201380144015001515

Leu Leu Asn Thr Gln1 5 10 15

Gly Asn Asn Glu Val Pro Lys Val Wing Asp Phe Leu Gly Val Ser Lys

20 25 30

Be Glu Asn Gln Ser Asp Leu Val Wing Phe Asn Glu Ile Gln Wing Ser

35 40 45

Asp Ser Glu Tyr Leu Phe Ser Ser Asn Ser Leu Pro Val Gln Asn

50 55 60

Val Wing Val Wing Wing Ser Thr Thr Wing Tyr Glu Phe Gln Glu Asn Pro Ser65 70 75 80

Asn Read Gln Be Read Thr Read Be Met Gly Be Wing Be Gly Lys Gly

85 90 95

Be Lys Cys Glu Thr Be Gly Asp Asn Be Thr Asn Be Val Glu Wing

100 105 110

Ala Wing Pro Arg Arg Thr Read Asp Thr Phe Gly Gln Arg Thr Be Ile

115 120 125

Gyr Arg Tyr Arg His Tyr Arg Gly Arg Thr Tyr Glu His Wing

130 135 140

Leu Trp Asp Asn Be Cys Arg Arg Glu Gly Gln Be Arg Lys Gly Arg145 150 155 160

Gln Gly Gly Tyr Asp Lys Glu Asp Lys Wing Ala Arg Wing Tyr Asp Leu

165 170 175

Wing Wing Read Lys Tyr Trp Gly Thr Be Thr Thr Thr Asn Phe Pro Ile

180 185 190

Ser Asn Tyr Glu Lys Glu Read Glu Asp Met Lys Asn Met Thr Arg Gln

195 200 205

Glu Phe Val Wing Be Ile Arg Arg Lys Be Gly Phe Be Arg Gly

210 215 220

Ala Ser Met Tyr Arg Gly Val Thr His His Gln His Gly Arg Trp225 230 235 240

Gln Wing Arg Ile Gly Arg Val Wing Gly Asn Lys Asp Leu Tyr Leu Gly

245 250 255

Thr Phe Ser Thr Glu Glu Glu Wing Wing Glu Wing Tyr Asp Ile Wing Wing

260 265 270

Ile Lys Phe Arg Gly Read Asn Wing Val Thr Asn Phe Asp Met Asn Arg

275 280 285

Tyr Asp Val Lys Ser Ile Leu Glu Ser Asn Ser Leu Pro Ile Gly Gly

290 295 300

Gly Wing Wing Lys Arg Leu Lys Glu Wing Gln Wing Ile Glu Ser Ser Gln305 310 315 320

Lys Arg Glu Glu Met Ile Wing Read Gly Ser Be Tyr Pro Tyr Gly Ser

325 330 335

Thr Be Be Be Be Arg Gln Gln Wing Tyr Be Read Met Gln Lys Pro

340 345 350

Phe Glu Gln Pro Gln Pro Read Leu Thr Read Leu Gln Asn Gln Asp Ile Ser

355 360 365

Gln Tyr Thr Gln Asp Being Ser Phe Gln Gln Asn Tyr Read Gln Thr Gln

370 375 380

Leu His Leu His Gln Leu Be Ala Gly Ser Asn Phe Leu His Asn Asn385 390 395 400

Gln Be Ser Gln Asn Pro Gln Tyr Tyr Asn Ser Tyr Ile Gln Asn Asn

405 410 415

Pro Thr Leu Read His Gly Leu Trp Asn Met Gly Ser Ser Ser Ser Ser

420 425 430

Met Glu Asn Asn Gly Be Ser Be Ser Gly Ser Tyr Ser Thr Val Gly Tyr

435 440 445

Read Gly Asn Gly Read Gly Met Ala Thr Asn Be Thr Gly Be Asn Wing

450 455 460

Val Wing Glu Glu Leu Pro Leu Val Lys Ile Asp Tyr Asp Met Pro Ser465 470 475 480

Gly Gly Tyr Gly Ser Trp Ser Gly Glu Ser Val Gln Gly Ser Asn Pro

485 490 495

Gly Val Phe Thr Met Trp Asn Glu500 <210> 203 <211> 785 <212> DNA

<213> Vitis vinifera <400> 203

tcttgataaa tcccaaaact gactgtgtag gtactgagga ggaagctgca gaagcctatg 60

atattgcagc aataaagttc agaggcctta atgcagtgac caactttgac atgaatcgat 120acgatgtgaa gagcattctt gaaagcaaca ctcttccgat aggtggggga gcggccaagc 180ggctaaagga ggctcaagca attgaatcat caagaaaacg ggaagaaatg atagccctag 240gttcaagttt ccaatatggg agctcgagct ctagcaggtt acagacatat cctctaatgc 300agcagcagtt tgagcaacct cagcctttac taacattaca gaaccaagaa ccattactaa 360ctttgcaaaa ccctgaaatt tctcagtacc cccaagactc ccagtttcac caaaactaca 420tccaaactca gttgcagttg caccagcaat ctgggtcgta cctgaaccat tcaagccaaa 480gtcctcagtt ctacaacagt tacctccaca acaacccggc tcttcttcat gggctgatga 540gtatgggctc ttcttcatct gtcatggaga ataatgggag ttctagtggg agttacaatg 600gaggmtactt caataatgga cttggggttg cttcgaattc tacggtggct agtgcagtag 660gatcagcaga ggagcttccc ctcatcaagg ttgattacga tatgccggcc gcaggctatg 720gcagctggtc aggtgactca gttcagggac agaatgctgg agtttttaca atgtggaatg 780actga 785

<210> 204 <211> 260 <212> PRT

<213> Vitis vinifera <400> 204

Leu Ile Asn Pro Lys Thr Asp Cys Val Gly Thr Glu Glu Glu Wing Ala1 5 10 15 Glu Wing Tyr Asp Ile Wing Ala Ile Lys Phe Arg Gly Ile Leu Glu Ser 35 40 45 Asn Thr Leu Pro Ile Gly Gly Gly Wing Ala Lys Arg Leu Lys Glu Wing 50 55 60 Gln Wing Ile Glu Ser Be Arg Lys Arg Glu Met Ile Wing Leu Gly65 70 75 80Ser Ser Phe Gln Tyr Gly Being Being Being Being Being Arg Read Gln Thr Tyr 85 90 95 Pro Read Met Gln Gln Gln Phe Glu Pro Gln Pro Read Leu Thr Read Le 100 105 110 Gln Asn Gln Pro Read Leu Thr Read Leu Gln Asn Pro Glu Ile Be Gln 115 120 125 Tyr Pro Gln Asp Be Gln Phe His Gln Asn Tyr Ile Gln Thr Gln Leu 130 135 140 Gln Read His Gln Gln Ser Gly Be Tyr Leu Asn His Ser Be Gln Ser145 150 155 160Pro Gln Phe Tyr Asn Ser Tyr Leu His Asn Wing Leu Read His 165 170 175 Gly Leu Met Be Met Gly Be Ser Be Val Ser Glu Asn Asn Gly 180 185 190 Be Ser Be Ser Gly Ser Tyr Asn Gly Serly Tyr Phe Asn Asn Gly Leu Gly 195 200 205 Val Wing Ser Asn Ser Thr Val Wing Ser Wing Val Gly Ser Wing Glu Glu 210 215 220 Leu Pro Leu Ile Lys Val Asp Tyr Asp Met Pro Wing Gly Wing Tyr Gly225 230 235 240Ser Trp Ser Gly Asp Ser Val Gln Gly Gln Asn

245

Met Trp Asn Asp260

<210> 205 <211> 150 <212> DNA

<213> brassica napus <400> 205

250

255ggaacatcgg ccgaggaaga gtttcccacg gttaaagttg attacgatat gcctccttta 60

ggtggagcca cagggtgtga acgatggact aatggagaga atggtcaggg gtcaaatcca 120ggaggtgtct tcacaatgtg gaatgaataa 150

<210> 206 <211> 49 <212> PRT

<213> Brassica napus <400> 206

Gly Thr Be Wing Glu Glu Glu Phe Pro Thr Val Lys Val Asp Tyr Asp1 5 10 15 Met Pro Pro Read 20 Gly Gly Wing Thr Gly 25 Cys Glu Arg Trp Thr 30 Asn GlyGlu Asn Gly Gln Met Trp Asn

35 40 45

Glu

<210> 207 <211> 162 <212> DNA

<213> Phaseolus coccineus <400> 207

tggagggggggggggggggggggg

<213> Phaseolus coccineus <400> 208

Be Asn Wing Be Ser Gly Asn Wing Val Gly Thr Wing Glu Glu Leu Gly15 10 15

Read Val Lys Val Val Asp Tyr Asp Met Pro Wing Gly Gly Tyr Gly Gly Trp

20 25 30

Be Wing Ala Wing Wing Glu Be Met Gln Thr Be Asn Gly Gly Val Phe

35 40 45

Thr Met Trp Asn Glu50

<210> 209 <211> 7 <212> PRT

<213> Artificial sequence <220>

<223> Reason 2 <220>

<221> VARIANT

<222> (1) .. (1)

<223> / replace = "Read"

<220>

60120162

<221> <222>

UNSAFE (3). . (3)

<223> Xaa can be any naturally occurring amino acid <220>

<221> VARIANT

<222> (4) .. (4)

<223> / replace = "Go"

<220>

<221> VARIANT <222> (7). . (7) <223> / replace = "Glu" <400> 209

Val Phe Xaa Met Trp Asn Asp1 5

<210> 210 <211> 2194 <212> DNA <213> Oryza sativa <400> 210

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> 211 <211> 1245 <212> DNA <213> Oryza sativa <400> 211

cttgttgttg atctgtgccc ccaagaagaa taacactcta ctcttacttg ttggaaaaaa 60

atagtattag caaccacgca tatgcaaatt ttaatgcagt aataataaga gatggatcga 120

tcgttttcca gctcttgtat atgtgactgg ccctgcttta tgtgtgtagt gttaatttca 180

gctttagcag tacgtgatta gtgatggaca ataattgtcg cagacgtatc tatcaattgc 240

tcctgttgtg tgatgcttta actgttggaa tcaaagttgc gttgcctttg ttgttatgag 300

gaggaatata tatgttgggg caggaaaaga atggaggaga gatcgttctc catatcctta 360

tcatcggcct cgtcactgct cgcagtttaa ctttttggtg atgcgagcga tggtcagcca 420

tatatatact cccatgctgc atgctagtaa tcaatatacg ccttgtaaaa gtaaacgatc 480

gtctagtaat tgcaatatca taggggtagc cattgacaga gatctacata gatagagggg 540

gaacaagaat tgacactcca cagatgctcc actcattcac ctttactaat ttatatcttt 600

tgatgtttga tcgatcgatc gatccgtccg tcggtgtctc gacgaataaa aactgcaaat 660

cgaactgtat gtatataata tagcgtcgta aattaaatta aattaaatcg aactgaatac 720

tacatgtcga agcaagaatt agttcaacta aaagatttag tttttccggt tgcaatatct 780

gtgaaattaa ttgaagaaat taagaagaaa actggagaga tatatatatg gatgagacaa 840

aatgagataa gacgcatgat ggtccctcgg atgatgtcgt ccgttcctta tttccattcc 900

atggcagctg ctatcgctat ctagtgcgcg cggcatctcc aatcccatcc attctagtgg 960

tcgatctagc tactactgag tattgttttt tcttcttttt actactgttg attattctgc 1020aactgcagtt agatgcttgc tactcctaca tcgatctctc

cattcactac tgatgatccg tgggtgtagt gtgggtggct

ttgccattgc tcctcaggcc agcaactgag aagctccata

gccgacaagg ccagagaagg aagaagaagc tctcatcatc

<210> 212

<211> 675

<212> DNA

<213> Oryza sativa

<400> 212

atgaagagca ggaagaacag cacgacgagc acaaaagcagagcagcggag gaggaggagg cggcggcaac tgctatagcacgcaaggatg tggagaagaa tcggcgcctc cacatgaaggtccctcatcc ccgccgccgc tccccgccgc catcaccacctcatcgccgc cctcctccac caaggaggct gtgacgcagcgcggcgtaca tcaagcagct caaggggagg atcgacgagcgcggcggcac tcaccaccag caccagcaat ggcggcggcggtgcggtgcc aggatgggac gctggacgtg gtggtggtgaagggagaggg cggtgcggct gcacgaggtg atcggcgtgcgtggtgaacg ccagcttctc cgtcgtcggc gacaagatctgcgctctgct ccaggatcgg cctcgacgcc tccagggtctctcctccaat Attaa <210> 213 <211> 224 <212> PRT

<213> Oryza sativa <400> 213

tcgcgcgggc gtatgcattgataaataggg cagggtgcggcaagtaagca gcagctagttatcac

caggcagctggcagcagtaggtctctgcctactactccactggatcaccttgaagaagaggcgggatgccgcgaggcgattggaggaagatctacactctcccacaggct

ccacaccagccaagatggagcaagctctccctcctcctccggagcaggcggaagcagcagggtggtggagcaggggagaggcgcggagccactcccaggcgcaacctc

Met Lys Be Arg Lys Asn Be Thr Thr Be Thr Lys Wing Wing Gly Ser1 5 10 15 Cys His Thr Be 20 Be Gly Gly Gly 25 Gly Gly Gly Gly Asn 30 Cys TyrSer Be 35 Be Ser Lys Met Glu 40 Arg Lys Asp Val Glu 45 Lys Asn ArgArg Leu 50 His Met Lys Gly Leu 55 Cys Leu Lys Leu Ser 60 Ser Leu Ile ProAla Wing Pro Arg Arg His His His Tyr Ser Thr Ser Ser 70 70 75 80Ser Ser Pro Pro Ser 85 Ser Thr Lys Glu Wing 90 Val Thr Gln Leu Asp 95 HisLeu Glu Gln Wing 100 Wing Wing Tyr Ile Lys 105 Gln Leu Lys Gly Arg 110 Ile AspGlu Leu Lys 115 Lys Arg Lys Gln 120 Wing Wing Wing Leu Thr 125 Thr Be ThrSer Asn 130 Gly Gly Gly Gly Gly 135 Met Pro Val Val Glu 140 Val Arg Cys GlnAsp Gly Thr Read Asp Val Val Val Val Ser Glu Wing Ile Arg Glu Glu

145

150

155

Arg Glu Arg Wing Val Arg Read His Glu Val Ile Gly Val

Val Val

165

170

Glu Gly Wing Glu Val Val Asn Wing Ser Phe Ser

180

185

Ile Phe Tyr Thr Read His Ser Gln Wing Read Cys

195

200

Asp Wing Be Arg Val Be His Arg Read Arg Asn

210 215

<210> 214 <211> 696 <212> DNA <213> Oryza sativa <400> 214

atggatcagg agaaggcgga cggcggcggc aggaggaggaagcggcagcg gcgcgagcag cacggcggcg gcggcggcggcgcaggaggc ggcaggacat gaagggcctc tgcgtcaagcaggagggcagggcgg

160

Read Glu Glu175

Gly Asp Lys190

Ser Arg Ile Gly Leu205

Leu Leu Leu Gln Tyr220

ggagcagggc gacgagtagccggagaggaa ggagatggagtcgcctctct catccccaaacccagctggg cagcctggac

1080114012001245

60120180240300360420480540600660675

60120180240gaagcggcgg cctacatcaa gaagctcaag gaaagggtggagcatgatga gtatcacatc atcgcgctgc cgctcaggaggctgctggcc agtcgacgag cggcggcggc ggcggggaagaggacgacgg cggcggcggc ggtggtggag gtgcggcagcatcagcttgg acgtggtgct gatctgcagc gcagcgaggcatcaccgtcc tcgaggaaga aggcgccgac atcatctccgcacaatttct actacaccat ctactccagg gcctttagcttcgaggattt ctgagagact acgggcattg gtatga <210> 215 <211> 231 <212> PRT

<213> Oryza sativa <400> 215

Met Asp Gln Glu Lys Wing Asp Gly Gly Gly Arg1 5 10

Wing Thr Be Be Gly Be Gly Wing Be Be

20 25

Wing Wing Glu Arg Lys Glu Met Glu Arg Arg Arg

acgagctgcagaggaggaggaagaagaagaacgtgcaggacggtcaagttccaacttctccaagaattgg

ccacaagaggaccagctgctagatatgacgggggtcgctgccacgacgtccctcgccgcccatagaggct

35

40

Gly Leu Cys Val Lys Leu Wing Ser Leu Ile Pro

50

55

Met Ser Lys Met Gln Wing Wing Be Arg Thr Gln

Arg Arg Arg

Thr Wing Ala30

Arg Gln Asp45

Lys Glu His60

Read gly ser

65

70

75

Glu Wing Wing Wing Tyr Ile Lys Lys Leu Lys Glu Arg Val Asp

85

90

His His Lys Arg Be Met Met Be Ile Thr Be

100

105

Gly Gly Gly Gly Gly Pro Wing Wing Wing Wing Gly

115 120

Gly Gly Gly Gly Glu Glu Glu Glu Glu Asp Met

130 135

Wing Wing Wing Val Val Glu Val Arg Gln His Val145 150 155

Ile Ser Leu Asp Val Val Leu Ile Cys Ser Ala

165 170

Phe His Asp Val Ile Thr Val Leu Glu Glu Glu

180 185

Be Wing Asn Phe Be Wing Ala Wing His Asn Phe

195 200

Being Arg Wing Phe Being Being Arg Ile Gly Ile Glu

210 215

Glu Arg Leu Arg Wing Leu Val225 230

<210> 216 <211> 633 <212> DNA <213> Oryza sativa <400> 216

atggagatcc cgccgccgtc agctggtggc ggaggcggcgacgacggagc gcatccgccg cgagcagatg aacaagctctgtccgctccg ctccccccac agttaattcc attccttcactaccatcaaa gaaaattacg gattctcggt ggggcggcggaggttggggg tggcggcgga gtacataagg cagacgcagggagaagaaga gggagctcac cggcggcggc ggcggcggctggggcggcca cggccgccgc gccggaggtg gaggtgcagcgccatcctct tcaccggcgc gccgcccacc gacggcgcctgccgtcgagg acgccggcgg ccaggtgcag aacgcgcactgccgtctaca ccatccacgc catgattgga gatggatatgcagagattaa aggaagcaat acggagcaac cup <210> 217 <211> 210 <212> PRT

<213> Oryza sativa

Be Arg Cys110

Gln Ser Thr

125Thr Arg Thr140

Gln Glu Gly

Arg Pro Wing

Gly Wing Asp190

Tyr Tyr Thr

205Ala To Be Arg220

To be Arg15

Wing wing

Met lys

Cys ser

Leu Asp80Glu Leu95

Arg Ser

Be gly

Thr wing

Ser Leu160Val Lys175

Ile IleIle TyrIle Ser

gcggcaagcc cgaccggaagactcccacct cgactccctcattcaaattc aaattcaaaa

cggcgacgacagagggtggacctcgtcgtcacctggggtcccttccaccgtctccgtcgcgaggcattga

gaggccggaccatgctgagggtccggcgcccggcctgcaccgccgtccgccggcgccaaggagggtggtg

300360420480540600660696

60120180240300360420480540600633 <400> 217

Met Glu Ile Pro Pro Be Wing Gly Gly Gly Gly Gly Gly Gly Gly Lys1 5 10 15

Pro Asp Arg Lys Thr Thr Glu Arg Ile Arg Arg Glu Gln Met Asn Lys

20 25 30

Leu Tyr Be His Leu Asp Ser Leu Val Arg Sera Pro Pro Thr Val

35 40 45

Asn Be Ile Pro Be His Be Asn Be Asn Be Lys Tyr His Gln Arg

50 55 60

Lys Leu Arg Ile Leu Gly Gly Wing Wing Wing Wing Thr Thr Arg Pro Asp65 70 75 80

Arg Leu Gly Val Wing Wing Glu Tyr Ile Arg Gln Thr Gln Glu Arg Val

85 90 95

Asp Met Leu Arg Glu Lys Lys Arg Glu Read Thr Gly Gly Gly Gly Gly

100 105 110

Gly Be Ser Be Ser Be Gly Wing Gly Wing Wing Thr Wing Wing Wing Pro Wing

115 120 125

Glu Val Glu Val Gln His Leu Gly Ser Gly Leu His Wing Ile Leu Phe

130 135 140

Thr Gly Wing Pro Pro Thr Asp Gly Wing Be Phe His Arg Wing Val Arg145 150 155 160

Val Glu Wing Asp Wing Gly Gly Gln Val Gln Asn Wing His Phe Ser Val

165 170 175

Wing Gly Wing Lys Wing Val Tyr Thr Ile His Wing Met Ile Gly Asp Gly

180 185 190

Tyr Gly Gly Ile Glu Arg Val Val Gln Arg Read Le Lys Glu Wing Ile Arg195 200 205

Ser Asn210 <210> 218 <211> 546 <212> DNA

<213> Arabidopsis thaliana <400> 218

atggggagag caagagaaat aggagaagga aactcatcgt cgttaaggga acaacgaaac 60

ctcagagaga aggatcgaag aaacatctct tctctatact ttcttctcat gatgcgcatg 120gtttctccca ctcgcaagtt accagtgcct caccttatag atcaagcgac atcatacatg 180atccaattga aagagaatgt aaattatttg aaagagaaga aaaggacatt gttacaagga 240gaactcggga atctctacga agggtcgttt cttctaccca aactcagtat tcgttcgcgg 300gattcgacca tagaaatgaa tctgatcatg gatctaaaca tgaaaagagt aatgttacac 360gagcttgtga gtatttttga agaagaagga gctcaagtaa tgagtgctaa tcttcagaac 420ttgaatgata ggaccactta cacaatcata gcccaggcta tcattagtcg gattggcatt 480gatccatcaa ggatagaaga gagagtacgg aaaatcatct atggatatat atattttgaa 540gcatga 54 6th

<210> 219 <211> 188 <212> PRT

<213> Arabidopsis thaliana <400> 219

Met Gly Arg Wing Arg Glu Ile Gly Glu Gly Asn Ser Ser Ser Ser Leu Arg15 10 15

Glu Gln Arg Asn Leu Arg Glu Lys Asp Arg Arg Met Arg Met Lys His

20 25 30

Leu Phe Ser Ile Leu Ser Be His Val Ser Pro Thr Arg Lys Leu Pro

35 40 45

Val Pro His Leu Ile Asp Gln Wing Thr Be Tyr Met Ile Gln Leu Lys

50 55 60

Glu Asn Val Asn Tyr Leu Lys Glu Lys Lys Arg Thr Leu Read Leu Gln Gly65 70 75 80

Glu Leu Gly Asn Leu Tyr Glu Gly Ser Phe Leu Pro Le Lys Leu Ser

85 90 95

Ile Arg Be Arg Asp Be Thr Ile Glu Met Asn Leu Ile Met Asp Leu100 105 110Asn Met Lys 115 Arg Val Met Leu His 120 Glu Leu Val Ser Ile 125 Phe Glu GluGlu Gly 130 Wing Gln Val Met Ser 135 Wing Asn Leu Gln Asn 140 Leu Asn Asp ArgThr Thr Tyr Thr Ile Ile Wing Gln Val Pro His Pro His Wing Tyr Ala145 150 155 160Ile Ile Being Arg Ile 165 Gly Ile Asp Pro Being 170 Arg Ile Glu Glu Arg 175 ValArg Lys Ile Ile 180 Tyr Gly Tyr Ile Tyr 185 Phe Glu Wing

<210> 220 <211> 525 <212> DNA

<213> Arabidopsis thaliana <400> 220

atggaaagag cgagagaaat aggagaagga agcgcatcgt cattacggga acaacgaaac 60

ctcagagaga aagaacgacg aatgcgcatg aaacatctct tctccatact ctcttctcat 120gtttctccca ctcgtaggtt gccagtgcct caacttatag atcaagcggt atcatacatg 180atccaactga aagagaaggt aaactatttg aatgagatga aaaggagaat gttaggagga 240gaagtcaaaa atcgctctga agggtcgtct cttctgccaa aactcagtat tcgttcactg 300gattcgatca tagaaatgaa tctggttatg gatctaaaca tgaaaggagt aatgttacac 360aagcttgtga gtgtttttga agaagaagga gctcaagtga tgagtgctaa tcttcagaac 420ttgaatgata ggacctttta tacaatcata gcccaggcta tcatatgtcg gatcgggatt 480gatccatcaa ggatagaaga gagattaagg gatataatct catga 525

<210> 221 <211> 174 <212> PRT

<213> Arabidopsis thaliana <400> 221

Met Glu Arg Wing Arg Glu Ile Gly Glu Gly Be Wing To Be To Be Leu Arg1 5 10 15 Glu Gln Arg To Asn 20 Leu Arg Glu Lys Glu 25 Arg Met Met 30 Met His Lys Phe To Be 35 Ile Leu To Be His 40 Val Ser Pro Thr Arg 45 Arg Leu ProVal Pro 50 Gln Leu Ile Asp Gln 55 Wing Val Ser Tyr Met 60 Ile Gln Leu LysGlu Lys Val Asn Tyr Leu Asn Glu Met Lys Arg Arg Met Leu Gly Gly65 70 75 80Glu Val Lys Asn Arg 85 Ser Glu Gly Ser Ser 90 Leu Le Pro Lys Le 95 95 SerIle Arg Ser Leu 100 Asp Ser Ile Ile Glu 105 Met Asn Leu Val Met 110 Asp LeuAsn Met Lys 115 Gly Val Met Leu His 120 Lys Leu Val Ser Val 125 Phe Glu GluGlu Gly 130 Ala Gln Val Met Ser 135 Wing Asn Leu Gln Asn 140 Leu Asn Asp ArgThr Phe Tyr Thr Ile Ile Wing Gln Ile Ile Cys Arg Ile Gly Ile145 150 155 160Asp Pro To Be Arg Ile 165 Glu Glu Arg Leu Arg 170 Asp Ile Ile Ser

<210> 222 <211> 369 <212> DNA

<213> Arabidopsis thaliana <400> 222

atgatccaat tgaaagagaa tgtaaattat ttgaaagaga agaaaaggac attgttacaa 60

ggagaactcg ggaatctcta cgaagggtcg tttcttctac ccaaactcag tattcgttcg 120cgggattcga ccatagaaat gaatctgatc atggatctaa acatgaaaag agtaatgtta 180cacgagcttg tgagtatttt tgaagaagaa ggagctcaag taatgagtgc taatcttcag 240aacttgaatg ataggaccac ttacacaatc atagcccagg ctatcattag tcggattggc 300attgatccat caaggataga agagagagta cggaaaatca tctatggata tatatatttt 360gaagcatga 369 <210> 223 <211> 122 <212> PRT

<213> Arabidopsis thaliana <400> 223

Met Ile Gln Leu Lys Glu Asn Val Asn Tyr Leu Lys Glu1 5 10

Thr Leu Leu Gln Gly Glu Leu Gly Asn Leu Tyr Glu Gly

20 25

Leu Pro Lys Leu Be Ile Arg Be Arg Asp Be Thr Ile

35 40 45

Leu Ile Met Asp Leu Asn Met Lys Arg Val Met Leu His

50 55 60

Ser Ile Phe Glu Glu Glu Gly Ala Gln Val Met Ser Ala65 70 75

Asn Read Asn Asp Arg Thr Thr Tyr Thr Ile Ile Wing Gln

85 90

Be Arg Ile Gly Ile Asp Pro Be Arg Ile Glu Glu Arg

100 105

Ile Tyr Gly Tyr Ile Tyr Phe Glu Wing

115 120

<210> 224 <211> 573 <212> DNA

<213> Arabidopsis thaliana <400> 224

atggagccga gccattcaaa cacaggtcaa tcaagatctg tagatcgcaaaaaaatagaa ggatgcaaat gaagtctctc tactcagaac tcatctctctcattcttcta cggagccttt aacactacct gatcagctag atgaagctgcaagaagctac aagtgaacgt ggagaaaaag agagaaagga aaaggaacctacaactttgg agaaactgaa ttccgtcgga tcttcatcgg tttcgtcgagtccgtgccaa gaaagctgcc aaaaatcgag attcaagaaa ctggttccattttcttgtga caagcttgga acacaagttt atgttttgtg agatcattcggaggaattag gagctgagat cactcatgct ggatactcca ttgttgatgacacacccttc actgcaaggt ggaagaacac gattatggag ctaggagtcaagactggaga agattgtgaa cagtgttcac taa <210> 225 <211> 190 <212 > PRT

<213> Arabidopsis thaliana <400> 225

Met Glu Pro Be His Ser Asn Thr Gly Gln Be Arg Ser15 10

Lys Thr Val Glu Lys Asn Arg Arg Met Gln Met Lys Ser

20 25

Glu Leu Ile Ser Leu Leu Pro His His Being Ser Thr Thr

35 40 45

Leu Pro Asp Gln Leu Asp Glu Wing Asn Wing Tyr Ile Lys

50 55 60

Val Asn Val Glu Lys Lys Arg Glu Arg Lys Arg Asn Leu65 70 75

Thr Thr Read Glu Lys Read Asn Be Val Gly Be Ser Ser

85 90

Ser Val Asp Val Ser Val Pro Arg Lys Leu Pro Lys Ile

100 105

Glu Thr Gly Ser Ile Phe His Ile Phe Read Val Thr Ser115 120 125

Lys Phe Met Phe Cys Glu Ile Ile Arg Val Leu Thr Glu

130 135 140

Glu Wing Ile Thr His Wing Gly Tyr Ser Ile Val Asp Asp145 150 155

His Thr Read His Cys Lys Val Glu Glu His Asp Tyr Gly165 170

Lys Lys Arg15

Ser Phe Leu30

Glu Met Asn

Glu Leu Val

Asn Leu Gln80

Ile Ile95 Wing

Val Arg Lys110

aacggttgagtcttcctcataaactacatccgttgcgactcgtcgatgtcttttcacatctgttctcacctgctgtcttcaattcctgaa

60120180240300360420480540573

Val Asp Arg15

Read Tyr Ser30

Pro Leu Thr

Lys Leu Gln

Val Wing Thr80

Val Ser Ser95

Glu Ile Gln110

Read Glu His

Glu Leu Gly

Val Phe160 Wing

Arg Wing Ser175Gln Ile Pro Glu Arg Read Le Glu Lys Ile Val Asn Ser Val His180 185 190

<210> 226 <211> 423 <212> DNA

<213> Medicago truncatula <400> 226

atgacatctc taacaaagga ggcgatttca gtgccggatc agctaaagga agcaacaaac 60

tacataaaga aattgcagat caacctggag aaaatgaagg aaaagaagaa ttttctacta 120ggaattcaaa ggccaaatgt gaatttgaat agaaaccaga agatgggatt aaagtctcca 180aaaattaaga tacaacaaat tggtttagtc ttagaggttg ttctaataac tggattggag 240tctcagtttt tgttcagcga aacctttcga gttcttcatg aagaaggagt tgatattgtt 300aatgctagtt ataaggtcaa tgaagattct gttttccatt caatacactg ccaggtagga 360gaatttggca atgaagctgc aagaatatct gagagattga agaagtttat gcaagactat 420tag 423

<210> 227 <211> 140 <212> PRT

<213> Medicago truncatula <400> 227

Met Thr Be Leu Thr Lys Glu Wing Ile Be Val Pro Asp Gln Leu Lys1 5 10 15

Glu Wing Thr Asn 20 Tyr Ile Lys Lys 25 Gln Ile Asn Leu Glu 30 Lys MetLys Glu Lys 35 Lys Asn Phe Leu Read 40 Gly Ile Gln Arg Pro 45 Asn Val AsnLeu Asn 50 Arg Asn Gln Lys Met 55 Gly Leu Lys Ser Pro 60 Lys Ile Lys IleGln Gln Ile Gly Leu Val Leu Glu Val Val Leu Ile Thr Gly Leu Glu65 70 75 80Ser Gln Phe Leu Phe 85 Ser Glu Thr Phe Arg 90 Val Leu His Glu Glu 95 GlyVal Asp Ile Val 100 Asn Ala Ser Tyr Lys 105 Val Asn Glu Asp Ser 110 Val PheHis Ser Ile 115 His Cys Gln Val Gly 120 Glu Phe Gly Asn Glu 125 Wing Wing ArgIle Ser 130 Glu Arg Leu Lys Lys 135 Phe Met Gln Asp Tyr 140 <210> 228

<211> 882 <212> DNA

<213> Hordeum vulgare <400> 228

atgaatgcct gcgctctgaa ctcgatggag ccggtgaagg cgaagccggc gaggggcggg 60

aagaggagca gggagagcgg cggcacggcg gtggtgctgc tggagaagaa ggagtcggag 120aaggagagga ggaagcgcat gaaggcgctc tgcgagaagc tcgcatccct catcccaagg 180gaacactgct gctccaccac tgatacaatg acccagctag gcagcctgga tgtgggggca 240tcgtatatca agaagctgaa ggagagggtc gatgagctac aacgtaggat gacctctgcg 300cagaccttgg ataccttcag aggagacact agcatcccaa cgcccactac caccactacc 360acgaaaagcg ttgtagggtc gctggaagaa gagaaagctc gggaggcatc ggcacccgta 420ttgcaggtgc ggcaacacga cgattcaagc atggaggtga gattgatatg ctgcatgaag 480aggccgatca agctccatga ggtgatcacc atccatgagg aagaaggtgc tgagatcatc 540aacgccaatc actctgttgc tggccaaaaa atgttctaca ctatacactc tcgggcctct 600agctcgagaa ttggcataga tgttccaagg gtttctgaac gactgcgagc attgctccaa 660ctttattcgc atgaaaatca ggcaccgtcg tgcatgcaca acaatcaagt cctcaggtac 720aatgatgggg ccaacgcgac tcctcccaag gagctcgtcg gcaacaatga tataggtgga 780accaacaagg tggcaaacca cgagtgtgag tcttgggttg agcaagacca agtgatgtgg 840agttccctgc ttgcgtcaat atcaccagag ttgctcgagt 882 g

<210> 229

<211> 293

<212> PRT

<213> Hordeum vulgare

<400> 229Met Asn Cys Wing Wing Read Asn Be Met Glu Pro Val Lys Pro Lys Wing1 1 10 15 Arg Wing Gly Gly Lys Arg Be Arg Glu Be Gly Thr Val Val 20 25 30 Leu Leu Glu Lys Lys Glu Be Glu Lys Glu Arg Arg Lys Arg Met Lys 35 40 45 Wing Leu Cys Glu Lys Leu Wing Wing Be Leu Ile Pro Arg Glu His Cys Cys 50 55 60 Be Thr Thr Asp Thr Met Thr Gln Leu Gly Be Leu Asp Val Gly Ala65 70 75 80Ser Tyr Ile Lys Lys Leu Lys Glu Arg Val Asp Glu Leu Gln Arg Arg 85 90 95 Met Thr Be Wing Gln Thr Read Asp Thr Phe Arg Gly Asp Thr Be Ile 100 105 110 Pro Thr Thr Thr Thr Thr Lys Be Val Val Gly Ser Leu 115 120 125 Glu Glu Glu Lys Arg Wing Glu Wing Ser Pro Pro Val Wing Leu Gln Val Arg 130 135 140 Gln His Asp Asp Being Met Glu Val Arg Leu Ile Cys Cys Met Lys145 150 155 160Arg Pro Ile Lys Leu His Glu Val Ile Thr Ile His Glu Glu Glu Gly 165 170 175 Wing Glu Ile Ile Asn Wing Asn His Ser Val Wing Gl y Gln Lys Met Phe 180 185 190 Tyr Thr Ile His Be Arg Wing Be Be Be Arg Ile Gly Ile Asp Val 195 200 205 Pro Arg Val Be Glu Arg Leu Arg Wing Read Leu Gln Leu Tyr Be His 210 215 220 Glu Asn Gln Wing Pro Be Cys Met His Asn Asn Gln Val Leu Arg Tyr225 230 235 240Asn Asp Gly Asn Wing Asa Thr Pro Pro Lys Glu Leu Val Gly Asn Asn 245 250 255 Asp Ile Gly Gly Asn Lys Val Wing Asn His Glu Cys Glu Be Trp 260 265 270 Val Glu Gln Asp Gln Val Met Trp Ser Be Leu Read Ala Ser Ile Ser

275 280

Pro Glu Leu Leu Glu

290 <210> 230 <211> 2193 <212> DNA

<213> Oryza sativa <400> 230

aatccgaaaa gtttctgcac cgttttcacc ccctaactaaaaatataaaa tgagacctta tatatgtagc gctgataactcatccaccta ctttagtggc aatcgggcta aataaaaaagtttccttagt aattaagtgg gaaaatgaaa tcattattgctctgtcatga agttaaatta ttcgaggtag ccataattgtaaaaaatctt tctagctgaa ctcaatgggt aaagagagagatgaagatat tctgaacgta ttggcaaaga tttaaacatattgtgcattc gtcatatcgc acatcattaa ggacatgtctttagtaatta aagacaattg acttattttt attatttatcgtacttacgc acacactttg tgctcatgtg catgtgtgagtcaactagca acacatctct aatatcactc gcctatttaatgaattcaag cactccacca tcaccagacc acttttaataaattttacag aatagcatga aaagtatgaa acgaactattaaaaaaagaa ttttgctcgt gcgcgagcgc caatctcccaacagagtggc tgcccacaga acaacccaca aaaaacgatgtccgcaacaa ccttttaaca gcaggctttg cggccaggagaaccaagcat cctcctcctc ccatctataa attcctccccggaggcatcc aagccaagaa gagggagagc accaaggacacgaccgcctt cttcgatcca tatcttccgg tcgagttcttcacctcctcc tctagcccttc ggttgttctt

285

caatatagggagaactatgcagtcgctacattagaatatacatcaaactcattttttttataattatatatactccatccttttttcgattgcacctccttacatttaggatatctaaaataggtttttctattgggacacatctaacggaagaggaggagcccggcgcgcggctg

aacgtgtgctaagaaaaactactttttttcgtcgttcacatcttcttgaataaaaaaaatagaattttatagtcaatttttattagatgcaagcaatacacgttagcaatatctacaaaaaatacatacaaaaacaggcaacaggacagcagcagtagcctagcccctagtcct

601201802403003604204805406006607207808409009601020108011401200aaacaatgaa gagcaggaag aacagc

cagagtgaaa gagtggtgtg

56

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> 231 <211> 56 <212> DNA

<213> Artificial sequence <220>

<223> initiator: prm06808 <400> 231

ggggacaagt ttgtacaaaa aagcaggctt <210> 232 <211> 50 <212> DNA

<213> Artificial sequence <220>

<223> initiator: prm06809 <400> 232

ggggaccact ttgtacaaga aagctgggtg <210> 233 <211> 2194 <212> DNA <213> Oryza sativa <400> 233

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

50gctatccttt 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> 234 <211> 1065 <212> DNA

<213> Arabidopsis thaliana <400> 234

atggagttgt taatgtgttc gggtcaggcc gagtcaggtg gttcctcttc caccgagtct 60

tcttcactca gtggtggact caggtttggt cagaagatct acttcgagga tggatccgga 120

tccagaagca agaaccgggt caataccgtt egtaagtcgt ctaccacggc gaggtgccaa 180

gtggaaggtt gtagaatgga tctaagcaat gttaaagctt attactcgag acacaaagtt 240

tgttgcattc actctaaatc atctaaagtc attgtctctg gtcttcatca aaggttttgt 300

caacaatgta gcaggtttca ccagctttct gagtttgact tggagaaaag aagttgtcgc 360

agaagactcg cttgtcataa cgaacgacga agaaaaccac aacccacaac ggctcttttc 420

acttctcatt actctcgaat cgctccatct ctttacggaa accccaatgc tgcaatgatt 480

aaaagcgttt tgggagatcc tactgcgtgg tcaaccgcaa gatcagtgat gcagcggcct 540

ggaccgtggc agattaatcc agttagggaa acccatccac acatgaatgt tttatcacat 600

ggaagctcaa gctttactac atgtccagag atgataaaca acaatagcac agattcaagc 660

tgtgctctct ctcttctgtc aaactcatac ccaattcatc agcagcaact tcagacacca 720

acaaatacat ggcgaccatc ttctggtttc gactcgatga tctcattctc cgataaggtt 780

acaatggctc agccaccgcc catttcaacc catcagccgc ccatctcaac acatcagcag 840

tacctcagcc aaacttggga agtcatcgcg ggcgaaaaga gcaattcaca ttatatgtct 900

cctgtgagtc aaatctcgga gccagcagat ttccagataa gcaatggcac cacaatgggt 960

ggatttgagc tgtatcttca tcagcaggtt ctgaagcaat acatggaacc cgagaacaca 1020

agagcttatg actcctctcc tcaacatttc aattggtctc tttga 1065 <210> 235 <211> 354 <212> PRT

<213> Arabidopsis thaliana <400> 235

Met Glu Read Leu Met Cys Be Gly Gln Wing Glu Be Gly Gly Be Ser1 5 10 15 Be Thr Glu Be Be Read Le Be Gly Gly Leu Arg Phe Gly Gln Lys 20 25 30 Ile Tyr Phe Glu Asp Gly Be Gly Be Arg Be Lys Asn Arg Val Asn 35 40 45 Thr Val Arg Lys Be Be Thr Thr Wing Arg Cys Gln Val Glu Gly Cys 50 55 60 Arg Met Asp Leu Be Asn Val Lys Wing Tyr Tyr Be Arg His Lys Val65 70 75 80Cys Cys Ile His Be Lys Being Ser Lys Val Ile Val Ser Gly Leu His 85 90 95 Gln Arg Phe Cys Gln Gln Cys Being Arg Phe His Gln Leu Being Glu Phe 100 105 110 Asp 125 Arg Arg Arg Lys Pro Gln Pro Thr Thr Wing Leu Phe Thr Be His Tyr 130 135 140 Ser Arg Ile Pro Wing Be Tyr Gly Asn Pro Asn Wing Ala Met Ile145 150 155 160Lys Ser Val Leu Gly Asp Pro Wing Trp Be Thr Wing Arg Ser Val 165 170 175 Met Gln Arg Pro Gly Pro Trp Thr His180 185 190

Pro His Met Asn Val Leu Being His Gly Being Being Phe Thr Thr Cys

195 200 205

Pro Glu Met Ile Asn Asn Asn Be Thr Asp Be Ser Cys Ala Read Ser

210 215 220

Leu Leu Be Asn Ser Tyr Pro Ile His Gln Gln Gln Read Le Gln Thr Pro225 230 235 240

Thr Asn Thr Trp Arg Pro Be Gly Phe Asp Be Met Ile Be Phe

245 250 255

Be Asp Lys Val Thr Met Wing Pro Gln Pro Ile Be Thr His Gln

260 265 270

Pro Pro Ile Be Thr His Gln Gln Tyr Read Be Gln Thr Trp Glu Val

275 280 285

Ile Wing Gly Glu Lys Be Asn Be His Tyr Met Be Pro Val Be Gln

290 295 300

Ile Ser As Glu Pro Asp Phe Gln Ile Ser As Asn Gly Thr Thr Met Gly305 310 315 320

Gly Phe Glu Leu Tyr Leu His Gln Gln Val Leu Lys Gln Tyr Met Glu

325 330 335

Pro Glu Asn Thr Arg Wing Tyr Asp Being Being Pro Gln His Phe Asn Trp340 345 350

To be read

<210> 236 <211> 1128 <212> DNA

<213> Arabidopsis thaliana <400> 236

atggagatgg gttccaactctcctccactg agtcatcctcgaggacggtg gtggtggatccgtggaggcg ggtcgggtcaatggatctaa ccaatgcaaaaaaacaccta aagtcactgttttcatcagc ttccggaattcataatgagc gacgaaggaagggaggatcg caccttcgctgggaaccaag agataggatgccagtgtcgt caccgtcatgggtggaggag ggacaagcttaagggaattg gcgactcaaagacaacaaca acaacaacaatcaggttttg gcccgatgaccagtatctga acccgccttgaatttaggtc gatacaccgagagttcgagt tatctgatcaacaagggctt atgactcttc <210> 237 <211> 375 <212> PRT <213> Arabidopsis thaliana <400> 237

Met Glu Met Gly Being Asn Being Gly Pro Gly His Gly Pro Gly Gln Ala1 5 10 15 Glu Being Gly Gly 20 Being Being Thr Glu Being 25 Being Being Phe Being Gly 30 Gly LeuMet Phe Gly 35 Gln Lys Ile Tyr Phe 40 Glu Asp Gly Gly Gly 45 Gly Being GlySer Being 50 Being Being Gly Gly Arg 55 Being Asn Arg Arg Val 60 Arg Gly Gly GlySer Gly Gln Being Gly Gln Ile Pro Cys Gln Val Glu Gly Cys Gly65 70 75 80Met Asp Read Thr Asn Wing Lys Gly Tyr Tyr Ser Arg His Arg Val Cys

gggtccgggttttcagtggacgggtcttctgtcgggtcagaggttattacggctggtatctgacctagaggccacagcctttacgaaaatgccaagttcagcagatcaatctcatctccactgtgctctccacacacatcatcataccaacgaccata

catggtccgggggctcatgttcctcaggtgataccaaggttcgagacaccgaacagaggtaaaaggagttgcgtctctctggtgatgctgagaacattggccaatgaatggagattatggtctcttctgtaacaacaacgagacagagagacagagacagagacagagcagaccag

gtcaggcagattggccagaagtcgttcaaagccaagtggagagtttgtggtttgtcaacagccgcaggagctgtgttagcgaatgaatggatacaagagttatttagtcaacactaa atcaaatccacaacaatacatgcacctgcacggggtctg

gtcgggtggtgatctacttccagacgtgtcaggttgtgggagtgcactctgtgcagcaggactcgctggtttctcgttacaagctttcttgatgaggcggggggctagagagggggggggggggggggg

6012018024030036042048054060066072078084090096010201080112885 90 95

Gly Val His Ser Lys Thr Pro Lys Val Thr Val Wing Gly Ile Glu Gln

100 105 110

Arg Phe Cys Gln Gln Cys Be Arg Phe His Gln Read Pro Glu Phe Asp

115 120 125

Read Glu Lys Arg Be Cys Arg Arg Arg Read Leu Wing Gly His Asn Glu Arg

130 135 140

Arg Arg Lys Pro Gln Pro Wing To Be Read To Be Val Read To Wing To Be Arg Tyr145 150 155 160

Gly Arg Ile Wing Pro Being Read Tyr Glu Asn Gly Asp Wing Gly Met Asn

165 170 175

Gly Be Phe Read Gly Asn Gln Glu Ile Gly Trp Pro Be Ser Arg Thr

180 185 190

Read Asp Thr Arg Val Met Arg Arg Pro Val Ser Be Pro Ser Trp Gln

195

200

205

Ile Asn Pro Met Asn Val Phe Being Gln Gly Being Val Gly Gly Gly Gly

210

215

220

Thr Be Phe Be Be Pro Glu Ile Met Asp Thr Lys Read Glu Be Tyr

225

230

235 240

Lys Gly Ile Gly Asp Be Asn Cys Wing Read Be Read Read Be Be Pro

245 250 255

His Gln Pro His Asp Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn Asn

260 265 270

Asn Asn Asn Thr Trp Arg Wing Be Ser Gly Phe Gly Pro Met Thr Val

275 280 285

Thr Met Wing Pro Gln Pro Pro Pro Wing Be Gln His Gln Tyr Leu Asn

290

295

300

Pro Pro Trp Val Phe Lys Asp Asn Asp Asn Asp Met Ser Pro Val Leu

305

310

315

320

Asn Read Gly Arg Tyr Thr Glu Pro Asp Asn Cys Gln Ile Be Ser Gly

325

330

335

Thr Ala Met Gly Glu Phe Glu Read Asp His His His Gln Be Arg

340 345 350

Gln Arg Tyr Met Glu Asp Glu Asn Thr Arg Wing Tyr Asp Ser Ser Ser

355 360 365His His Thr Asn Trp Ser Leu

370 375 <210> 238 <211> 1149 <212> DNA

<213> Aquilegia formosa χ Aquilegia pubescens <400> 238

atggaaatgg gttccagctc tttcgctggt ggtgggggta agggtgcttc tggttcatct 60

gattcttcac tgaatggttt gaaatttggg aagaaaatct actttgaaga tgtgggtatt 120

ggagttttag gcaagtcaag tgttgggtct ccgtcaattt tggtttctga agctcggttg 180

ccaccggctt cgtcggtgaa gaagggtaga ggggttttgc aaggacaacc acctagatgt 240

tctgttgaag gttgtaagct tgatcttact gatgctaagc cttattactc aaggcacaaa 300

gtctgtggta tgcactctaa atctcctaaa gtaattgttg gtggtttgga gcagaggttt 360

tgccagcagt gtagcagatt tcatctactc tgtgaatttg accaaggcaa gcgaagctgt 420

cgtagacgtc tagctggcca caatgagcgt cgaagaaaac cacaacctgg atcaatattt 480

tcaccgcgct atggtcgtgt gtcaccatct ttccaagaaa atagcaccag aggaggaggt 540

tttctaatgg acttcacagc gtacccaagg ctggcaggaa gggatgcatg gcaaacagta 600

aaagccggca attgggcaga tggaaaccaa acctctccta ttaagaagct tctcccacat 660

caatggcaag gcaattcaga gaatcctcct cctattgtct attctcaggc acctcacccg 720

tatctgcaag gtgtggctag tggatcaatt ttttccagta caagaatacc ttcagccgag 780

tgttttgctg gtgtctctga ctccagctgt gctctctctc ttctgtcaaa tcaaccatgg 840

aaccccagaa accagacttc cgatttggag gtgaataaca tagtgaacgg tgaaggggta 900

tccatggcaa gatctatagc acctcatagt gctgtagtca ccaacttcac gaacaacaca 960

tggaatttta agggcaacag tgaagttagt ggcagttccc atgaaattcc acgtcaggtg 1020

tcgcagccag tcaccaatca tttctccagt gagctcgatc tagctcagca ggggaacagg 1080

cagtacatgg agatccggca ttcaagggat tttggttctt ccactcacca gatgcactgg 1140tctctttga 1149 <210> 239 <211> 382 <212> PRT

<213> Aquilegia formosa χ Aquilegia pubescens <400> 239

Met Glu Met Gly Being Being Being Phe Ala Gly Gly Gly Gly Lys Gly Ala1 5 10 15

Being Gly Being Being Asp Being Being Read Asn Gly Read Lys Phe Gly Lys Lys

20 25 30

Ile Tyr Phe Glu Asp Val Gly Ile Gly Val Leu Gly Lys Ser Ser Val

35 40 45

Gly Ser Pro Ser Ile Leu Val Ser Glu Wing Arg Leu Pro Pro Wing Ser

50 55 60

Ser Val Lys Lys Gly Arg Gly Val Leu Gln Gly Gln Pro Pro Cys65 70 75 80

Ser Val Glu Gly Cys Lys Leu Asp Leu Thr Asp Wing Lys Pro Tyr Tyr

85 90 95

Being Arg His Lys Val Cys Gly Met His Being Lys Being Pro Lys Val Ile

100 105 110

Val Gly Gly Read Glu Gln Arg Phe Cys Gln Gln Cys Be Arg Phe His

115 120 125

Leu Leu Cys Glu Phe Asp Gln Gly Lys Arg Be Cys Arg Arg Arg Leu

130 135 140

Wing Gly His Asn Glu Arg Arg Arg Lys Pro Gln Pro Gly Ser Ile Phe145

Be Pro Arg Tyr Gly Arg Val Be Pro Be Phe Gln Glu Asn Be Thr

165 170 175

Arg Gly Gly Gly Phe Leu Met Asp Phe Thr Wing Tyr Pro Arg Leu Ala

180 185 190

Gly Arg Asp Trp Wing Gln Thr Val Lys Wing Gly Asn Trp Wing Asp Gly

195 200 205

Asn Gln Thr Be Pro Ile Lys Lys Read Leu Pro His Gln Trp Gln Gly

210 215 220

Asn Be Glu Asn Pro Pro Ile Val Tyr Be Gln Pro Wing His Pro225 230 235 240

Tyr Read Gln Gly Val Wing Be Gly Be Ile Phe Be Be Thr Arg Ile

245 250 255

Pro Be Wing Glu Cys Phe Wing Gly Val Be Asp Be Wing Cys Wing

260 265 270

Be Read Leu Be Asn Gln Pro Trn Asn Pro Arg Asn Gln Thr Be Asp

275 280 285

Leu Glu Val Asn Asn Ile Val Asn Gly Glu Gly Val Ser Met Wing Arg

290 295 300

Ser Ile Wing Pro His Ser Wing Val Val Thr Asn Phe Thr Asn Asn Thr305 310 315 320

Trp Asn Phe Lys Gly Asn Be Glu Val Be Gly Be Be His Glu Ile

325 330 335

Pro Arg Gln Val Ser Gln Pro Val Thr Asn His Phe Ser Ser Glu Leu

340 345 350

Asp Leu Wing Gln Gln Gly Asn Arg Gln Tyr Met Glu Ile Arg His Ser

355 360 365

Arg Asp Phe Gly Be Being Thr His Gln Met His Trp Being Read

370 375 380

<210> 240 <211> 1170 <212> DNA

<213> Gossypium hirsutum <400> 240

atggaaatgg gttcgggctc tttgactgag tcgggcggtt ctcccaccaa ctcttccgcc 60

gagtcactca acggcttgaa gttcggtaaa aagatctact ttgaggatac agccgctgtc 120gccgccgccg ctggtggtaa aagtgttggt ggtggtgcaa acatagggac tccatccaag 180tccggtccag catcttcaag cttagccggg tcgtgtagga aagccagggt tgatggaatg 240gcgcaagggg ttctgccttc taggtgtcaa gtagaagggt gtaaagtgga tctgagtgat 300gctaaggctt actattcaag gcataaggtt tgttgtatgc actctaagtc atctaaagtc 360Gly Gly Ser

Phe Gly Lys30

Gly Gly45 Wing

Be gly

attgttgctg gtctcgagca aagattttgt cagcagtgta gcagatttca tcagctttctgaatttgaca aagggaaacg gagttgtcgt agacggcttg caggtcacaa tgagcgacgcaggaaaccac cacctggatc attattttcc tctccttatg gccggctttc ttcctctattattgaaaatg gcagtagagg tggaagcttt atagtggatt tctcagcata cccaaggctttcaggaaggg atgcatggcc agcagctcga tcgttagaat gcataactgg aaatcgaagcacagccactg gaagctcatt ttcacatcca cggcaaaaca actccagcaa acctcctcatgaccatttct tgcaaggttc accatgtggg actagtttct ccagcactgg aatttctccaggagaatgct tcacagggtc tggtgactca agctgtgctc tctctcttct gtcaaatcaaccgtggggct ccaggaacca ggctttgaat ttttctgtaa atggcgtgat aagtactgaagggtcctctg cggcacaacc aacaacgctt catggtgcag ttgtgaaccc ttattcaaatgcctctttgg atttcaatgg cagtgacact gttcgcagtt ctcacaagat gctgccacacctagatttgg gtcaaatccc agaccctgtt aactgtcaat tctctagtga ccttgagttgtctcaacaaa gctggaggtc atatatcgaa catgagcagt ccggggcagc ctatgacgactccatgcagc atatccactg gacgctctaa <210> 241 <211> 389 <212> PRT

<213> Gossypium hirsutum <400> 241

Met Glu Met Gly Ser Gly Ser Leu Thr Glu Ser1 5 10

Asn Be Ser Ala Glu Be Read Asn Gly Leu Lys

20 25

Tyr Phe Glu Asp Thr Wing Wing Val Wing Wing Wing

35 40

Val Gly Gly Gly Wing Asn Ile Gly Thr Pro Ser Lys

50 55 60

Be Be Be Read Leu Wing Gly Be Cys Arg Lys Wing Arg Val Asp65 70 75

Gln Wing Gly Val Leu Pro To Be Arg Cys Gln Val Glu Gly Cys

85 90

Asp Read Be Asp Wing Lys Wing Tyr Tyr Be Arg His Lys Val100 105 110

Met His Ser Lys Ser Ser Lys Val Ile Val Wing Gly Leu Glu

115 120 125

Phe Cys Gln Gln Cys Be Arg Phe His Gln Read Be Glu Phe

130 135 140

Gly Lys Arg Be Cys Arg Arg Arg Read Wing Gly His Asn Glu145 150 155

Arg Lys Pro Pro Gly Be Read Phe Ser Be Pro Tyr Gly

165 170

Being Being Being Ile Ile Glu Asn Gly Being Arg Gly Gly Being Phe180 185 190

Asp Phe Be Wing Tyr Pro Arg Read Gly Arg Wing Asp Wing Trp

195 200 205

Wing Arg Be Read Glu Cys Ile Thr Gly Asn Arg Be Thr Wing

210 215 220

Be Be Phe Be His Pro Gln Asn Asn Be Be Lys Pro225 230 235

Asp His Phe Read Gln Gly Ser Pro Cys Gly Thr Ser Phe Ser

245 250

Gly Ile Being Pro Gly Glu Cys Phe Thr Gly Being Gly Asp Ser260 265 270

Wing Read Be Read Read Be Asn Gln Pro Trp Gly Be Arg Asn

275 280 285

Read Asn Phe Ser Val Asn Gly Val Ile Ser Thr Glu Gly Ser

290 295 300

Wing Gln Pro Thr Thr Read His Gly Wing Val Val Asn Pro Tyr305 310 315

Wing Be Read Asp Phe Asn Gly Be Asp Thr Val Arg Be Ser

325 330

Met Leu Pro His Leu Asp Leu Gly Gln Ile Pro Asp Pro Val340 345 350

Pro Thr1 5

Lys ile

Lys Ser

To the wing

Gly Met80Lys Val95

Cys cys

Gln arg

Asp Lys

Arg Arg160Arg Leu175

Ile val

To the wing

Thr gly

Pro His240Ser Thr255

Be cys

Gln Wing

To be a wing

Ser Asn320His Lys335

Asn cys

4204805406006607207808409009601020108011401170Gln Phe Ser Ser Asp Leu355

Ile Glu His Glu Gln Ser370

Ile His Trp Thr Leu385

<210> 242 <211> 1017 <212> DNA <213> Ipomoea nil <400> 242

atggaactgg gttcttcttc ttcttctacc tctacctccg ccgactcctc atccgacggc 60ttgaagttcg gcaaaaaagt ctactttgaa gatgcgggtg gtgggagtgg tgggtcgtcg 120ctgccggcca agagagggag gagcgcggtg gcgcaaggcg gacagccacc caggtgccag 180gtggaagggt gcaaggcaga tctgagtgaa tttaaggctt attactcaaa gcataaagtt 240tgtggtatgc actccaagtc tcctaaggtc attgtttctg ggcttgaaca gagattctgc 300cagcagtgca gcaggtttca tcaattgagt gaatttgatc aagtaaaaag gagctgccgt 360aggcgtttgg ctggtcataa tgagcgtcgt aggaagcccc cacttggatc catattgtcc 420acacattatg ggactctttc ttcttcaatg tttggaaaca atggccactt tgtgatggat 480ttcgcctcat acccatatcc gggtggtaag ggctcatggc ggccaagtac caatggcgtg 540gggaactttc ttcaacaacc atggcagaga aactctgaag atcccccacc caagcttctt 600ttgctaggtt cggatgctgc tgctagggct acttatccca gtccttgtgg agaatacttc 660aatggggtct cggattccac ccgtgctctc tctcttctgt caaactcaaa tcagccctgg 720ggctcgagaa accaacaacc ctctggtctc ggggttaata gcttacttaa cactgatgga 780acgcttgctg ttcatccatc cggttcccat gctgccgtta tcaatgaatt ttcttcaagt 840ccatggggtt ttaaaggcaa c tcaagccact cagctcag ataagatcct tcctgatagt 900cactattctg gtgagctcga gatgatggct catcaacaaa ctggacgagc atacatggga 960atggagtact cgacgggtta tgattcttct gtccagaatg tgcactggga 17c

<210> 243 <211> 338 <212> PRT <213> Ipomoea nil <400> 243

Met Glu Read Gly Ser Ser 5

Ser Ser Asp Gly Leu Lys20

Gly Gly Gly Ser Gly Gly35

Val Wing Gln Wing Gly Gly50

Lys Wing Asp Leu Ser Glu65 70

Cys Gly Met His Ser Lys85

Gln Arg Phe Cys Gln Gln100

Glp Asp Val Lys Arg Ser115

Arg Arg Arg Lys Pro Pro130

Thr Read Ser Ser Ser Met145 150

Phe Ala Ser Tyr Pro Tyr165

Thr Asn Gly Val Gly Asn180

Glu Asp Pro Pro Pro Lys195

Arg Wing Thr Tyr Pro Ser210

Asp Ser Thr Arg Wing Leu225 230

Glu Read Be Gln Gln Be Trp Arg Be Tyr

360 365

Gly Wing Tyr Asp Wing Asp Ser Met Gln His375 380

Ser Ser Ser Ser Thr

Phe Gly Lys Lys Val25

Ser Ser Leu Pro Ala40

Gln Pro Pro Arg Cys55

Phe Lys Wing Tyr Tyr75

Ser Pro Lys Val Ile90

Cys Ser Arg Phe His105

Cys Arg Arg Arg Leu120

Leu Gly Ser Ile Leu135

Phe Gly Asn Asn Gly155

Pro Gly Gly Lys Gly170

Phe Leu Gln Gln Pro185

Leu Leu Leu Leu Gly200

Pro Cys Gly Glu Tyr215

Seru Leu Seru Asn235

Thr Ser Wing Asp Ser15

Tyr Phe Glu Asp Ala30

Lys Arg Gly Arg Ser45

Gln Val Glu Gly Cys60

Ser Lys His Lys Val80

Val Ser Gly Leu Glu95

Gln Read Ser Glu Phe110

Gly Wing His Asn Glu125

Ser Thr His Tyr Gly140

His Phe Val Met Asp160

Ser Trp Arg Pro Ser175

Trp Gln Arg Asn Ser190

Be Asp Wing Wing Wing205

Phe Asn Gly Val Ser220

Ser Asn Gln Pro Trp240Gly Ser Arg Asn Gln Gln Pro Ser Gly Leu Gly Val Asn Ser Leu

245 250 255

Asn Thr Asp Gly Thr Read Wing Val His Pro Be Gly Ser His Wing

260 265 270

Val Ile Asn Glu Phe Ser Ser Ser Ser Pro Trp Gly Phe Lys

275 280 285

Wing Thr Thr Be Asp Lys Ile Leu Pro Asp Be His Tyr Be Gly

290 295 300

Glu Leu Glu Met Met Wing His Gln Gln Thr Gly Arg Wing Tyr Met Gly305 310 315 320

Met Glu Tyr To Be Thr Gly Tyr Asp To Be Val Gln Asn Val His Trp325 330 335

Thr leu

<210> 244 <211> 1035 <212> DNA

<213> Lactuca sativa <400> 244

atggagatgg gtggttcgag tggttcttcg gagtcgcaacttcgggaaag aaatctattt tgaggatgtg ggagttggaggggttgtctc cggcgagcgg cggcgatgct gctggagggcgctggtgggg tggtgagtgg ttttggacaa cagcaaccactgtaatctgg atctgagtga tgctaaaagt tactattcaacattcgaaaa cggctaaggt cattgttaat ggccttgaacagcaggttcc atcaactacc agagtttgac cagggaaaaagctgggcaca atgaacgtag aagaaagcca tctctgctattcctcctcaa tctttgaaaa caatgggaat tctggaggcttgctcaagag gaaggattca gtggccagga acaagggcggatcgacctcc caattgccgg agaaaagttc ccaccgcttcaatccacctc cttatgttcc accaggaggg tgttttaatgtgtgctctct ctcttctgtc aaatcactca tctggctcaagagtactata tcaaccctga agctggtgca tatgtgcaccggaaccggag ctgggttcgg aaccatggtt gaggtcaccgacccatgatg cccatttggg tttgggtcac gtcccacagtgaggttggac ttggtccaca tggtggggga aggcgatatggactggtcac tttga <210> 245 <211> 344 <212> PRT

<213> Lactuca sativa <400> 245

Met Glu Met Gly Gly Be Ser Gly Be Ser Glu1 5 10

Ile Gly Read Gln Phe Gly Lys Glu Ile Tyr Phe

20 25

Gly Wing Gln Val Lys Ser Asp Asp Gly Leu Ser

35 40

Asp Wing Wing Gly Gly Pro Gln Lys Lys Gly Arg

50 55

Val Ser Gly Phe Gly Gln Gln Gln Pro Pro Arg

tgctcaaaatctcaggttaacgcagaagaacgaggtgtcaggcacaaagtagagattctggaagctgcagccactcgctattctaatggacaccaccgcccatggcaaaggagtccatgaggaaccaatcagcttgcgcgggtggtggtg

tggtttgcaaatccgatgatagggagaactggtggaaggtttgtggtgctccaacagtgcgagacgattgtggaactgtccttttcgtcatcgagccgcccaacctggatggactccaaccctgagccataccaacaaccgggtactg

To be

Glu

Pro

Thr60Cys

65

70

Cys Asn Leu Asp Leu Ser Asp Asa Wing Lys Ser

75Tyr

85

90

Tyr

Val Cys Gly Wing His Ser Lys Thr Wing Lys Val Ile

100 105

Glu Gln Arg Phe Cys Gln Gln Cys Ser Arg Phe His

115

120

Phe Asp Gln Gly Arg Lys Be Cys Arg Arg Arg

130 135

Glu Arg Arg Arg Lys Pro To Be Read To Be Thr145 150 155

Being Being Being Ile Phe Glu Asn Asn Gly Asn Being

Leu140Arg

Gly

Gln Leu Leu Lys 15 Asp Val Gly Val 30 Wing Ser Gly Gly45 Wing Gly Gly ValGln Val Glu Gly 80Ser Arg His Lys 95 Val Asn Gly Leu 110 Gln Leu Pro Glu125 Wing Gly His AsnTyr Gly Thr Val 160Gly Phe Leu Met

6012018024030036042048054060066072078084090096010201035165 170

Asp Phe Be Cys Be Arg Gly Arg Ile Gln

180 185

Wing Pro Wing Pro Pro Wing Wing Wing Ile Asp Leu

195 200

Lys Phe Pro Pro Leu Pro Trp Gln Ser Asn Leu

210 215

Tyr Val Pro Gly Gly Cys Phe Asn Gly Val225 230 235

Cys Wing Read Be Read Read Be Asn His Be

245 250

To Be Read To Be His Glu Tyr Tyr Ile Asn Pro Glu

260 265

His Gln Read His Gln Pro Thr Thr Gly Thr Gly275 280

Val Glu Val Thr Wing Gly Trp Val Tyr290 295

Read Gly Read Gly His Val Pro Gln Ser Gly310

Met

His305

Trp pro

Pro Ile205Asp Asn220

His glu

Gly ser

Gly wing

Gly285Glu Thr300 Wing

Gly Gly

315

Glu Val Gly Leu Gly Pro His Gly Gly Gly Arg Arg Tyr

325

330

175Gly Thr190

Gly wing

Pro pro

Asp Ser

Arg Asn255Ala Tyr270

Phe Gly

His asp

Tyr ser

Asp Ser335

Arg

Glu

Pro

Asn240Gln

Val

Thr

Allah

Gly320Ser

Val Asp His Ile Asp Trp Ser Leu340

<210> 246 <211> 1137 <212> DNA

<213> Malus domestic <400> 246

atggaaatgg gctcgagttc taagaccgag tcagcgagctaactcctccg ctgagtcact caacggcttg aaattcggccgggggttttg gagctctgca caaatcatca tgcgggtctgggggctacgc cgccaaagaa gcaaaggggc ggcggaaattcaggtggagg gctgcgaggt agatctgagt ggtgccaaaggtctgtggct tgcactctaa aactcccact gtcattgttgtgccaacagt gtagcaggtt tcatttactt, cctgaatttgcgtagacgct tggctgggca taatgagcgt cgtagaaaactctacgcgtg gcagactttc ttcgtctctc tacgaaaacactgatggact tcactgcata cccaaggttt tctgggagggtctgagcgag cacctgttaa tcaaaatgcc aatgacgcagtggcagagca actctgatat ttctacatcc ggcttttaccactagttatc ctggtcctgg aattcctcca ggagaatgtgagctgtgctc tctctcttct gtcaaatcag ccatggggctgctgggatga attccttgat gaacactcaa ggggtacctgtctgcaacct ccaatcactt tccgaccact tcgtggggttagcagctcac acgggatgct tccagatctg ggtctcggtcagtcagtact ctggtgtgct ggagctgtct caacagggtactcggacaca ccaggggcta tgactccacc agtcagcaga <210> 247 <211> 378 <212> PR

<213> Malus domestic <400> 247

Met Glu Met Gly Ser Be Ser Lys Thr Glu Ser1 5 10

Be Ser Pro Pro Asn Ser Sera Wing Glu Ser Leu

cttcctcctcggaaaatctactgctgggtctgggtcagcccttactattcctggtcttgaatcaaggaaacacctccagggcagcagaatatacatggacggaagtttctcacagtagtctcgaaaccgggcgcgggcgggcgggcggcgggg

ttcgccgcccctttgaggatttcctccgccgccgcggtgtcaggcacaaaacagaggtttacgtagttgtatccatactgtggaagctttaacaagaaccacaacagccgagcaggcgggcactgactcaagtattgggtagaccctgcgaa

20

25

Gly Arg Lys Ile Tyr Phe Glu Asp 35 40Being Cys Gly Being Wing Gly 50 55 Pro Lys Lys Gln Arg Gly Gly65 70 Gln Val Glu Gly Cys Glu Val Asp

75

Wing Ser Ser Ser Ser15

Asn Gly Leu Lys Phe30

Gly Wing Read His Lys45

Wing Gly Wing Thr Pro60

Gln Pro Pro Arg Cys80

Wing Lys Wing Tyr Tyr

6012018024030036042048054060066072078084090096010201080113785 90 95 Ser Arg His Lys 100 Val Cys Gly Leu His 105 Ser Lys Thr Pro Thr 110 Val IleVal Ala Gly 115 Leu Glu Gln Arg Phe 120 Cys Gln Cn Gln Cys Ser 125 Arg Phe His Ln Phe Arg Ser Cys 140 Arg Arg Arg LeuAla Gly His Asn Glu Arg Arg Arg Arg Lys Pro Pro Gly Ser Ile Leu145 150 155 160Ser Thr Arg Gly Arg 165 Leu Be Ser Leu 170 Tyr Glu Asn Ser Be 175 ArgIle Gly Ser Phe 180 Leu Met Asp Phe Thr 185 Wing Tyr Pro Arg Phe 190 Ser GlyArg Asp Thr 195 Trp Thr Thr Thr Thr 200 Ser Glu Arg Wing Pro 205 Val Asn GlnAsn Wing 210 Asn Asp Wing Gly Lys 215 Phe Leu Gln Gln Pro 220 Trp Gln Ser AsnSer Asp Ile Ser Thr Be Gly Phe Tyr Leu Gln Gly Ser Wing Gly Gly225 230 235 240Thr Ser Tyr Pro Gly 245 Pro Gly Ile Pro 250 Gly Glu Cys Val Thr 255 ValVal Thr Asp Ser 260 Ser Cys Wing Leu Ser 265 Leu Pro TrpGly Ser Arg 275 Asn Arg Val Leu Gly 280 Wing Gly Met Asn Ser 285 Leu M et AsnThr Gln 290 Gly Val Pro Val Ala 295 Gln Pro Val Pro His 300 Ser Ala Thr SerAsn His Phe Pro Al Thr Thr Gly Phe Lys Gly Asn Glu Asn Gly305 310 315 320Ser Ser Ser His Gly 325 Met Leu Pro Asp Leu 330 Gly Leu Gly Gln Ile 335 SerGln Pro Leu Ser 340 Ser Gln Tyr Ser Gly 345 Val Leu Glu Leu Ser 350 Gln GlnGly Arg Arg 355 Gln Gln His Met Glu 360 Leu Gly His Thr Arg 365 Gly Tyr Asp

Be Thr Be Gln Gln Met His Trp Be Read

370 375 <210> 248 <211> 1014 <212> DNA

<213> Medicago trunculata <400> 248

atggattcag gaggcaactc ttcttcagaa gagtcctcac ttaatggctt gaaatttggc 60

caacgaatct atttcgaaga tacagctctt actgctgctt ctgctgctgc tgctagtact 120

accattgctg ctggttctcc ttcttcttct ggttcaaaga aaggaagagg tgggtcagtt 180caacattctc aaccacctag gtgtcaagtt gaaggatgta aactagatct gactgatgct 240

aaagcttact attctagaca caaagtttgt agcatgcact ctaaatcccc tactgttact 300

gtttctggtc ttcaacaaag gttttgtcaa caatgtagca gatttcatca gcttgctgag 360

tttgatcaag gaaaaagaag ttgtcggaga cgactagctg gtcataatga gcgtcgcaga 420

aagcccccac ccagctctct cttaacctca cgttttgcca ggctttcttc gtctgttttt 480

ggtaacagcg acagaggtgg cagctttttg atggaatttg cttcaaatcc gaaacatagt 540

ctgaggaatt cacccggaaa tcaaaccaca gcaatcggtt ggccttggcc ggggaacacg 600

gagtcgccat ctagcaacct tttcttgcaa ggttcggtgg gtgggacaag cttccctggt 660

gccaggcatc ctcccgagga aacttacact ggagtcacag attcaaactg tgctctctct 720

cttctgtcaa atcaaacatg gggttctcaa aacacagaac caagtcctgg attgaataac 780

atgctgaatt tcaacgggac acccatgaca caacttggta catcttctca tggtgtagcc 840

atgcatcaaa ttccaaacaa ttacgaggtt gtccctgatc ttggtcgggg tcacattttg 900

catcctcttg gtagccaaca ctctggcgag cttgatctgt tgcagcaggg aaggaggcat 960

tatatggatg tagaacattc cagggcctat gaatcttctc agtggtcact gtaa 1014 <210> 249 <211> 337 <212> PRT

<213> Medicago trunculata <400> 249 Met Asp Be Gly Gly Asn Be Be Glu Glu Be Be Read Asn Gly1 5 10 15 Read Lys Phe Gly Gln Arg Ile Tyr Phe Glu Asp Thr Wing Read Thr Wing 20 25 30 Wing Be Wing Wing Wing Wing Wing Thr Thr Ile Wing Wing Gly Be Pro Be 35 40 45 Be Be Gly Be Lys Gly Arg Gly Gly Be Val Gln His Be Gln 50 55 60 Pro Pro Arg Cys Gln Gly Cys Lys Leu Asp Leu Thr Asp Ala65 70 75 80Lys Ala Tyr Tyr Be Arg His Lys Val Cys Ser Met His Be Lys Ser 85 90 95 Pro Thr Val Thr Val Ser Gly Leu Gln Arg Phe Cys Gln Gln Cys 100 105 110 Ser Arg Phe His Gln Leu Ala Glu Phe Asp Gln Gly Lys Arg Be Cys 115 120 125 Arg Arg Arg Read Leu Gly His Wing Asn Glu Arg Arg Read Lys Pro Pro 130 135 140 Ser Be Read Leu Thr Be Arg Phe Wing Arg Read Le Be Ser Be Val Phe145 150 155 160Gly Asn Ser Asp Arg Gly Gly Ser Phe Leu Met Glu Phe Ala Ser Asn 165 170 175 Pro Lys His Be Read Arg Asn Be Pro Gly Asn Gln Thr Thr Wing Ile 180 185 190 Gly Trp Pro Trp Pro Gly Asn Thr Glu Be Pro Be As As Leu Phe 195 200 205 Leu Gln Gly Be Val Gly Gly Thr Be Phe Pro Gly Wing His Pro 210 215 220 Pro Glu Glu Thr Tyr Gly Val Thr Asp Ser Asn Cys Ala Leu Ser225 230 235 240Leu Read Asn Gln Thr Trp Gly As Gln Asn Thr Pro Glu 245 250 255 Gly Leu Asn Asn Met Leu Asn Phe Asn Gly Thr Pro Met Thr Gln Leu 260 265 270 Gly Thr Be Ser His Gly Val Val Wing Met His Gln Ile Pro Asn Asn Tyr 275 280 285 Glu Val Val Pro Asp Leu Gly Arg Gly His Ile Leu His Pro Leu Gly 290 295 300 Ser Gln His Ser Gly Glu Read Asp Leu Read Gln Gln Gly Arg Arg His305 310 315 320Tyr Met Asp Val Glu His Ser Arg Wing Tyr Glu Ser Be Gln Trp Ser 325 330 335 Leu

<210> 250 <211> 1143 <212> DNA

<213> Bentamian Nicotiana <400> 250

atggaacttc tggcttctgc ttcttcttct acttctacta attctacttc ccctgactct 60

cctcccaaca ctttaaaatt tggtcaaaaa atctactttg agcatgttgg acttcagcac 120

ctcaaatcag caactgggtc gtcgtcgtca tcgccgccgg tcatcggaac tccggcgccg 180

gcgatgtcca agaaaggaag agggggtggt gtagttcaag gtcggcaacc acctaggtgt 240

caagttgaag ggtgtgaagc agatctgagt gatgttaagg cttactattc aaggcacaaa 300

gtctgtgcta cacattctaa gtctcctgtg gtcattgttg ctggtcttga acaaagattt 360

tgtcaacagt gtagcaggtt tcatcggttg ccagaatttg accaagggaa acgcagttgc 420

cgcaggcgcc tagcaggcca taatgagcgt cggaggaaac ctccacctgg atctcttttg 480

tctaatcgct atggaagtct ttcttcatca atatttgaaa acaatggcag atctggaagt 540

tttctggttg acttcactgc atatccgaat ctcactggag gtgcatggcc aaatactaga 600

tcatctgatc gaggatggga taatcaatcc actgcgtcag ggaagcttct ccaaagtcat 660

tggctgaaca gttctgaaaa tccgacatcc gaccttgttc tgcaaggttc agttgctagg 720

ggtgccaatt attctggtcc tggtattatt ccttccggaa actgcttctc tggagtctca 780gattccaatg gtgctctctc tcttctgtca aatgagccat ggggctcgag gaaccaatcctctagcctcg gggttaacgg cttggtcaac actgatggcg gacataccgt tcacccatcgggttcccatg ctgctcctgt caatcactac tcaggccctc tatggggatt taaaggaaatgaagctagta gcagttcaca tgcaatacct cctgatctcg ggctgggtca catttctcaacatgctgtca atcagtactc tggtgagcct gggatggctc agcacagtgg aagacagtacatgggactgg agcattcaaa gggttataat tcttctgttc agaatgtgca ctggacactttga

<210> 251 <211> 380 <212> PRT

<213> Bentamian Nicotiana <400> 251

Met Glu Leu Leu Wing Ala Be Wing Be Ser Be Thr Be Thr Asn Be Thr1 5 10 15 Be Pro Asp Be Pro Pro Asn Thr Read Lys Phe Gly Gln Lys Ile Tyr 20 25 30 Phe Glu His Val Gly Leu Gln His Leu Lys Be Ala Thr Gly Be Ser 35 40 45 Be Ser Be Pro Pro Val Ile Gly Thr Pro Pro Wing Pro Met Wing Be Ser Lys 50 55 60 Lys Gly Gly Gly Val Val Gln Gly Arg Gly Pro Gl Cys65 70 75 80Gln Val Glu Gly Cys Glu Wing Asp Leu Be Asp Val Lys Wing Tyr Tyr 85 90 95 Be Arg His Lys Val Cys Wing Thr His Be Lys Ser Pro Val Val Ile 100 105 110 Val Wing Gly Leu Glu Arg Phe Cys Gln Cn Gln Cys Be Arg Phe His 115 120 125 Arg Leu Pro Glu Phe Asp Gln Gly Lys Arg Be Cys Arg Arg Arg Leu 130 135 140 Wing Gly His Asn Glu Arg Arg Lys Pro Pro Gly Be Leu145 150 155 160Ser Asn Arg Tyr Gly Be Read Be Ser Ile Phe Glu Asn Asn Gly 165 170 175 Arg Be Gly Be Phe Leu Val Asp Phe Thr Wing Tyr Pro Asn Leu Thr 180 185 190 Gly Gly Wing Trp Pro Asn Thr Arg Be Be Asp Arg Gly Trp Asp Asn 195 200 205 Gln Be Thr Wing Be Gly Lys Leu Read Gln Be His Trp Leu Asn Be 210 215 220 Be Glu Asn Pro Thr Be Asp Leu Val Leu Gln Gly Ser Val Wing Arg225 230 235 240Gly Wing Asn Tyr Ser Gly Pro Gly Ile Ile Pro Ser Gly Asn Cys Phe 245 250 255 Ser Gly Val Ser Asp Ser Asn Gly Wing Leu Ser Leu Le As Ser Gln 260 265 270 Pro Trp Gly Be Arg Asn Gln Be Ser Be Leu Gly Val Asn Gly Leu 275 280 285 Val Asn Thr Asp Gly Gly His Thr Val His Pro Be Gly Be His Wing 290 295 300 Pro Wing Val Asn His Tyr Be Gly Pro Read Trp Gly Phe Lys Gly Asn305 310 315 320Glu Wing Be Being Be Being His Wing Ile Pro Pro Asp Read Gly Leu Gly 325 330 335 His Ile Be Gln His Wing Val Asn Gln Tyr Be Gly Glu Pro Gly Met 340 345 350 Wing Gln His Be Gly Arg Gln Tyr Met Gly Read Glu His Ser Lys Gly 355 360 365 Tyr As n Ser Ser Val Gln Asn Val His Trp Thr Leu 370 375 380 <210> 252 <211> 1254

<212> DNA <213> Oryza sativa <400> 252

atggagatgg ccagtggagg aggcgccgcc gccgccgccg gcggcggagt aggcggcagc 60

ggcggcggtg gtggtggagg ggacgagcac cgccagctgc acggtctcaa gttcggcaag 120

aagatctact tcgaggacgc cgccgcggca gcaggcggcg gcggcactgg cagtggcagt 180

ggcagcgcga gcgccgcgcc gccgtcctcg tcttccaagg cggcgggtgg tggacgcggc 24 0

ggagggggca agaacaaggg gaagggcgtg gccgcggcgg cgccaccgcc gccgccgccg 300

ccgccgcggt gccaggtgga ggggtgcggc gcggatctga gcgggatcaa gaactactac 360

tgccgccaca aggtgtgctt catgcattcc aaggctcccc gcgtcgtcgt cgccggcctc 420

gagcagcgct tctgccagca gtgcagcagg ttccacctgc tgcctgaatt tgaccaagga 480

aaacgcagct gccgcagacg ccttgcaggt cataatgagc gccggaggag gccgcaaacc 540

cctttggcat cacgctacgg tcgactagct gcatctgttg gtgagcatcg caggttcaga 600

agctttacgt tggatttctc ctacccaagg gttccaagca gcgtaaggaa tgcatggcca 660

gcaattcaac caggcgatcg gatctccggt ggtatccagt ggcacaggaa cgtagctcct 720

catggtcact ctagtgcagt ggcgggatat ggtgccaaca catacagcgg ccaaggtagc 780

tcttcttcag ggccaccggt gttcgctggc ccaaatctcc ctccaggtgg atgtctcgca 840

ggggtcggtg ccgccaccga ctcgagctgt gctctctctc ttctgtcaac ccagccatgg 900

gatactacta cccacagtgc cgctgccagc cacaaccagg ctgcagccat gtccactacc 960

accagctttg atggcaatcc tgtggcaccc tccgccatgg cgggtagcta catggcacca 1020

agcccctgga caggttctcg gggccàtgag ggtggtggtc ggagcgtggc gcaccagcta 1080

ccacatgaag tctcacttga tgaggtgcac cctggtccta gccatcatgc ccacttctcc 1140

ggtgagcttg agcttgctct gcaggggaac ggtccagccc cagcaccacg catcgatcct 1200

gggtccggca gcaccttcga ccaaaccagc aacacgatgg attggtctct gtag 1254 <210> 253 <211> 417 <212> PRT

<213> Oryza sativa <400> 253

Met Glu Met Wing Be Gly Gly Gly Wing Wing Wing Wing Wing Gly Gly Gly1 5 10 15 Val Gly Gly Gly Gly Gly Gly Gly Gly Asp Glu His Arg Gln 20 25 30 Leu His Gly Leu Lys Phe Gly Lys Lys Ile Tyr Phe Glu Asp Wing Wing 35 40 45 Wing Wing Wing Gly Gly Gly Gly Gly Thr Gly Be Gly Ser Gly Ser 50 55 60 Wing Pro Pro Be Ser Be Lys Wing Gly Gly Gly Gly65 70 75 80Gly Gly Gly Lys Asn Lys Gly Lys Gly Val Wing Ala Wing Ala Wing Pro Pro 90 90 95 Pro Pro Pro Pro Pro Pro Cys Gln Val Glu Gly Cys Gly Cys Asp 100 105 110 Read Ser Gly Ile Lys Asn Tyr Cys Arg His Lys Val Cys Phe Met 115 120 125 His Ser Lys Wing Pro Arg Val Val Val Wing Gly Leu Glu Gln Arg Phe 130 135 140 Cys Gln Gln Cys Be Arg Phe His Leu Pro Glu Phe Asp Gln Gly145 150 155 160Lys Arg Be Cys Arg Arg Leu Wing Gly His Asn Glu Arg Arg Arg 165 170 175 Arg Pro Gln Thr Pro Read Wing Ala Be Arg Tyr Gly Arg Read Wing Wing Be 180 185 190 Val Gly Glu Arg His Arg Phe Arg Be Phe Thr Read Asp Phe Be Tyr 195 200 205 Pro Arg Val Pro Be Ser Val Arg Asn Wing Trp Pro Wing Ile Gln Pro 210 215 220 Gly Asp Arg Ile Be Gly Gly Ile Gln Trp His Arg Asn Val Wing Pro225 230 235 240His Gly His Being Ser Wing Val Wing Gly Tyr Gly Wing Asn Thr Tyr Being 245 250 255 Gly Gln Gly Being Ser Being Being Gly Pro Pro Val Phe Wing Gly Pro Asn 260 265 270 Leu Pro Pro Gly Gly Cys Read Wing Gly Val Gly Wing Wing Thr Asp Ser275

280

290

295

305

310

325

330

Tyr Met Pro Wing Pro Trp Thr Gly Ser Arg

340

345

Gly Arg Ser Val Wing His Gln Leu Pro His Glu

355

360

370

375

385

390

285Pro Trp Asp 300 Wing Met315 Wing Ser MetArg Wing Gly HisGlu Val Ser 365Phe Ser Gly 380 Wing Pro Arg395 Asn Thr Met

410

405Leu

<210> 254 <211> 1182 <212> DNA <213> Oryza sativa <400> 254

atggcgaccg gcggcagcgg cggcggcggc ggaggtggaggggctcaagt tcggcaagaa gatctacttc gagcaggacggcggcggtgg agtcgtcgtc gacgtcgtcg ggcggaggcggcggcggcgg cggcgccccc gccgccgctg ccgccgaggtgtggatctga gcggcgtcaa gccgtactac tgccgccacaaaggagccca tcgtcgtcgt cgccggcctc gagcagcgctttccaccaat tacctgaatt tgatcaagaa aaaaaaagctcacaatgaac gccggaggaa gccgacacct ggacctctttgctgcatcct ttcatgaaga gccaggcagg tccagaagctccaagggttc caagcagtgt gagggatgcg tggcctgctatccggttcaa tccagtggca agggggccat gaactccatcggatacagtg atcaccatgc gttcagcagc catggtggctctccaccacc cagcctttga gctcacctca ggtggatgtctccagctgtg ctctctctct tctgtcaact cagccatgggagccacaacc ggtccccgcc aatgtcgtca acggccagcgccggtgtcgc cctcggtcat ggcaagcaac tacatggcggtccagccggg gccatgacgg cgccaggaac gtgcacctgcctgaacgagg tccctccggg ctctgtccac cacggccattgcactgcagg gaggtgcccc gtccaaccgg ccggaagccggccttcagcc actccaccaa tgccatgaac tggtc2102 <2> <12> <12> <12>

<213> Oryza sativa <400> 255

Met Wing Thr Gly Gly Gly Gly Gly Gly Gly Gly1 5

Thr thr thr

Be Thr Thr320

Gly Ser Wing

335Glu Gly Gly350

Read Asp Glu

Glu Leu Glu

Ile Asp Pro400

Asp Trp Ser415

gtggtggtgacggcggcgtcgcaagaaggggccaggtggaaggtgtgctatctgccaacagccgcagacgcttctcgctattgtggtagattcagcccgcagcagcggctgggcgcgcgcgcgggcgggg

cgatgtccacggcgtcggcggaagggcgtggggttgcggccatgcacgccgtgcagcaggccttgcaggttggccggctttttctcataccgatcgcatgcgcgggcggcgggcgggcggggcggg

10

Gly

Asp Asp Val His Gly Leu Lys Phe Gly Lys Lys Ile

20 25

Asp Wing Wing Wing Be Wing Be Wing Wing Wing Val Glu

35

40

Gly

Tyr

Ser45

Gly

Phe

30

To be

Gly Gly15

Glu GlnSer Thr

Be Gly Gly Gly Gly Gly Lys Gly Lys Gly Lys Gly Val Wing Wing Wing Wing Wing 55 55 60 Wing Pro Pro Pro Leu Pro Pro Cys Gln Val Glu Gly Cys Gly65 70 75 80Val Asp Leu Being Gly Val Lys Pro Lys Val Cys 85 90 95 Tyr Met His Wing Lys Glu Pro Ile Val Val Val Wing Gly Leu Glu Gln 100 105 110 Arg Phe Cys Gln Cys Ser Arg Phe His Gln Leu Pro Glu Phe Asp

601201802403003604204805406006607207808409009601020108011401182115

120

125

Gln Glu Lys Lys Be Cys Arg Arg Arg Read Ala Gly His Asn Glu Arg 130 135 140 Arg Arg Lys Pro Thr Gly Pro Read Be Ser Arg Tyr Gly Arg Leu145 150 155 160Ala Wing Be Phe His Glu Pro Glu Gly Arg Be Arg Phe Val Val 165 170 175 Asp Phe Be Tyr Pro Arg Val Pro Be Ser Val Arg Asp Wing Trp Pro 180 185 190 Wing Ile Gln Pro Be Asp Arg Met Be Gly Be Ile Gln Trp Gly 195 200 205 His Arg Be Wing Val Wing Gly Tyr Be Asp 210 215 220 His His Wing Phe Be Being His Gly Gly Wing Wing Gly Wing Pro Met225 230 235 240Leu His His Wing Wing Phe Glu Leu Thr Be Gly Gly Cys Leu Wing Gly 245 250 255 Val Ala Thr Asp Be Ser Cys Ala Read Le Be Ser Leu Read Ser Gln Pro 260 265 270 Trp Asp Thr Thr Gln Ser Thr Be His Asn Arg Ser Pro Pro 275 280 285 Ser Ser Thr Be Wing Phe Gly Gly Gly Asn Asn Pro Val Ser Pro 290 295 300 Ser Val Met Wing Ser Asn Tyr Met Wing la Ser Pro Gly Trp Asn Ser305 310 315 320Ser Ser Arg Gly His Asp Gly Arg Wing Asn Val His Leu Pro Pro 325 330 335 His Gly Val Valu Leu Asn Glu Val Pro Pro Gly Glu Leu Glu Leu Wing Leu Gln Gly Gly Wing Pro Ser 355 360 365 Asn Arg Pro Glu Wing Glu His Gly Ser Gly Ser Gly Wing Phe Ser His

370

375

380

Be Thr Asn Ala Met Asn Trp Be Read

385 390

<210> 256

<211> 1119

<212> DNA

<213> Solanum tuberosum <400> 256

atggaactgg gttcagtttc ttcttctggtttgaagtttg gtaagaaaat ctactttggaggaagtgggt cgtcgccggt gaccggagataagaggggga ggggtgggtt ggtgcagggttgtcaggcag atctgagtga tgctaaggctcactctaagt ctcctactgt tgttgttgctagcaggttcc atcaattaac tgaattcgacgcatgccata atgagcgtcg taggaagcctgggaatcttt cttcatcaat atttgaaaatgacttcagct cacaccaaaa tgtcaatgatcaaggatggg atcatcaatc atcagggaaggaaaatgctg ccagtgagct tgttttgcaagttccttctg gagactattt tcctggagtctcaaatcggt cctggggatc aaggaatcgacacattgatg gggtacacac cattcaacctttctcaagcc cttcattgag ttttaaaggacctcctgatc tcggtttggg tcaaatgttacttgggatgg ctcagcatgg tgatggacgatatcatcctt ctgttcagaa tgtgcactgg <210> 257 <211> 372 <212> PRT

<213> Solanum tuberosum

aattcaagctaatgtgggtggggaacatgcggtcatccactactattcaaggtcttgaaccaggggaaaaccatctggataatagcagcaagttcatggcttccttcaacggttcagctatcagattcaatctccaagtctcagggcctcaatgaatctgaatctga

catctgattcttggagttcacaccggctccctaggtgtcaggcataaagtagagggttttgggagttgccgctcttttctcgatccggaagcaaatactcggtccttggctccaggaccaggggcggcggcgggcggcgcgcg

tttgaatggtggtcaagaatggcgacgactagttgaaggtttgtggtatgccaacagtgtcaggagactgtacacactacctttctggtcagcatctgaagaataactctttattctctcttctgcagccaagttacggggggggtg

60120180240300360420480540600660720780840900960102010801119 <400> 257

Met Glu Read Gly Be Val Be Ser Be Gly Asn Be Ser Be Ser Asp15 10 15

Be Leu Asn Gly Leu Lys Phe Gly Lys Lys Ile Tyr Phe Gly Asn Val

20 25 30

Gly Val Gly Val Gln Val Lys Asn Gly Ser Gly Ser Ser Val Pro

35 40 45

Gly Asp Gly Asn Met Pro Pro Pro Wing Pro Thr Thr Lys Arg Gly Arg

50 55 60

Gly Gly Read Val Gln Gly Gly His Pro Pro Arg Cys Gln Val Glu Gly65 70 75 80

Cys Gln Wing Asp Read Be Asp Wing Lys Wing Tyr Tyr Be Arg His Lys

85 90 95

Val Cys Gly Met His Be Lys Ser Pro Thr Val Val Val Wing Gly Leu

100 105 110

Glu Gln Arg Phe Cys Gln Gln Cys Be Arg Phe His Gln Leu Thr Glu

115 120 125

Phe Asp Gly Gly Lys Arg Be Cys Arg Arg Arg Read Leu Wing Cys His Asn

130 135 140

Glu Arg Arg Arg Pro Lys Pro Be Gly Be Read Phe Be Thr His Tyr145 150 155 160

Gly Asn Read Be Ser Be Ile Phe Glu Asn Asn Be Ser Be Arg Be Gly

165 170 175

Be Phe Leu Val Asp Be Phe Be His Gln Asn Val Asn Asp Be Ser

180 185 190

Trp Pro Asn Thr Arg Wing Be Glu Gly Trp Pro Asn His Gln Be Ser

195 200 205

Gly Lys Phe Read Gln Arg Pro Trp Read Asn Asn Be Glu Asn Wing Ward

210 215 220

Ser Glu Leu Val Leu Gln Gly Ser Wing Thr Arg Thr Ser Tyr Pro Ser225 230 235 240

Val Pro Gly Asp Tyr Phe Pro Gly Val Ser Asp Be Gly Wing

245 250 255

Read Be Read Read Be Asn Arg Be Trp Gly Be Arg Asn Arg Be Pro

260 265 270

Be Leu Gly Val Asn Be Gln Val His Ile Asp Gly Val His Thr

275 280 285

Gln Pro Being Gly Being His Gly Pro Wing Thr Asn His Phe Being Being Pro

290 295 300

Be Read Be Phe Lys Gly Asn Glu Wing Be Be Be His Glu Met305 310 315 320

Pro Pro Asp Leu Gly Leu Gly Gln Met Leu Gln Wing Be Asp Asn Pro

325 330 335

Tyr Cys Gly Glu Read Gly Met Wing Gln His Gly Asp Gly Arg Gln Tyr

340 345 350

Met Glu Read Asp Gln Be Lys Gly Tyr His Pro Be Val Gln Asn Val

355 360 365

His Trp Thr Leu

370 <210> 258 <211> 1140 <212> DNA

<213> Vitis vinifera <400> 258

atggaaaggg gttcgagctc tttgaccgtt tccagctctt cggccaactc gtctgagtcg 60ctcaacgggt tgaaatttgg gcagaagata tattttgaag atttgggcgt tggagctccg 120gccaaatcgg gaaccggctc ctcctcctcc tcctctgccg ccggctccgg tggtcgccca 180cctccggcgc cgccaaagaa ggtaagaggt agtggggttg ttcagggagg ccaaccaccg 240aggtgtcaag ttgaagggtg taaactagat ctgagtgatg ccaaagctta ctattcaagg 300cataaagtgt gtggtatgca ttcgaagtct ccaacggtca ttgttgcggg ccttgagcag 360aggttttgcc agcagtgtag cagatttcat cagcttgccg aatttgacca aggaaaacga 420agttgtcgta ggcgcctggc tggtcataat gagcgtcgca ggaagccacc acctggatct 480ttattgtcct cacgctatgg gcgactttct tcatccattt ttgaaaacag cagcagggtg 540ggaggaggct ttctgatgga ctttgctgca tacccaaggc atcccgagag ggatacttgg 600ccaactacaa gagcatctga tcgggtacct ggaaatcaaa ccactgcgat gggaaggttt 660cttccacatc catggcagag caactctgag aatcctctct ttctgcaagg ttcagccggc 720gggaccagct ttcatggtcc tggaattcct tcaggagaat gtttcacagg ggcctccgac 780tcaagctgtg ctctctctct tctgtcaaat cagccatgga gctccaggaa tcgagcatct 840ggtcttggag caaacagctt g catgaatcct aggggcat ccatggcgca acccacagct 900cctcatagtg cagctatcaa tcacttccca agcacctcgt gggatttcaa gggcaatgaa 960ggtagtagca gttcgcagga gatgccacct gatcttggtc ttggtcaaat ttcacagcct 1020attaatagcc agttctcagg tgggggcgag ttgccccaac agagtggaag gcaatacatg 1080gaactcgagc actccagggc ttatgacact tccactcagc agatgcactg gtcactttag 1140 <210> 259 <211> 379 <212> PRT <213> Vitis vinifera <400> 259

Met Glu Arg Gly Ser Ser Ser Leu Thr Val Ser Ser Ser Ala Asn15 10 15

Being Glu Being Glu Asn Gly Leu Lys Phe Gly Gln Lys Ile Tyr Phe

20 25 30

Glu Asp Leu Gly Val Gly Pro Wing Lys Ser Gly Thr Gly Ser Ser

35 40 45

Be Be Be Be Wing Gly Wing Gly Wing Gly Arg Pro Wing Pro

50 55 60

Pro Lys Lys Val Arg Gly Ser Gly Val Val Gly Gly Gly Gly Pro Pro65 70 75 80

Arg Cys Gln Val Glu Gly Cys Lys Read Asp Read Be Asp Wing Lys Wing

85 90 95

Tyr Tyr Be Arg His Lys Val Cys Gly Met His Be Lys Ser Pro Thr

100 105 110

Val Ile Val Wing Gly Leu Glu Gln Arg Phe Cys Gln Gln Cys Ser Arg

115 120 125

Phe His Gln Read Glu Wing Phe Asp Gln Gly Lys Arg Be Cys Arg Arg

130 135 140

Arg Leu Wing Gly His Asn Glu Arg Arg Arg Lys Pro Pro Gly Ser145 150 155 160

Read Read Be Ser Arg Tyr Gly Arg Read Be Ser Ser Ile Phe Glu Asn

165 170 175

Be Ser Arg Val Gly Gly Gly Phe Read Met Asp Phe Ala Wing Tyr Pro

180 185 190

Arg His Pro Glu Arg Asp Thr Thr Pro Thr Thr Arg Wing Be Asp Arg

195 200 205

Val Pro Gly Asn Gln Thr Thr Wing Met Gly Arg Phe Read Pro His

210 215 220

Trp Gln Be Asn Be Glu Asn Pro Read Phe Read Gln Gly Be Wing Gly225 230 235 240

Gly Thr Be Phe His Gly Pro Gly Ile Pro Be Gly Glu Cys Phe Thr

245 250 255

Gly Wing Be Asp Be Be Cys Wing Read Be Read Read Be Asn Gln Pro

260 265 270

Trp Ser Be Arg Asn Arg Wing Be Gly Read Gly Wing Asn Be Phe Met

275 280 285

Asn Pro Glu Gly Wing Be Met Wing Gln Pro Thr Wing Pro His Be Wing

290 295 300

Alle Ile Asn His Phe Pro To Be Thr Be Trp Asp Phe Lys Gly Asn Glu305 310 315 320

Gly Be Ser Be Ser Gln Glu Met Pro Pro Asp Read Gly Read Gly Gln

325 330 335

Ile Be Gln Pro Ile Asn Be Gln Phe Be Gly Gly Gly Gly Glu Leu Pro

340 345 350

Gln Gln Be Gly Arg Gln Tyr Met Glu Read Glu His Be Arg Wing Tyr

355 360 365

Asp Thr Be Thr Gln Gln Met His Trp Ser Leu370 375 <210> 260 <211> 1209 <212> DNA <213> Zea mays <400> 260

atggagtccg gcggtggcgg ggacgaccagtacttcgagg acgccgccgg ctccagcagcgcgcctccag cgacgcagca gccgtcgccgggcggcagga ggggcagggc cgcggccggcgcgcgctgcc aggtcgacgg ctgcaacgtttgccgccaca aggtgtgcaa aatgcactccgagcagcgct tctgccagca gtgcagcaggaagaagagct gccgcaaacg cctcgcaggcggaccccttg cgtcacgata cggccgtcacagaagcttca tgctggattt ctcgtacccgcccgcggtga gggccggtgg tgaaagggtggatcctcgtc accaccaagg cgcggtcgcgggtagctcgt cgtcggcgag gccgccggtgtgccttgcag gagtccccgc ggactccagctgggatgctg cccacagcca cagccacagcggcagccctg tggcgccctc cctcatggcgaccgagaccg actcctgggg ccacgaaggcgacgtccccc tcggcgaggt gcactccggcgagctcgagc tcgccctgca gggaaacaggccgcgcaata atcagggctc cgcgggcacgtcgctctag <210> 261 <211> 402 < 212> PRT <213> Zea mays <400> 261

Met Glu Gly Gly Gly Gly Gly Asp Asp Gln Read His Gly Read Lys Phe

1 5 10 15 Gly Lys Lys Ile 20 Tyr Phe Glu Asp Wing 25 Wing Gly Ser Be Ser 30 Gly SerSer Be Gly 35 Gly Gly Ser Wing Pro 40 Wing Pro Thr Wing 45 Gln ProSer Pro Wing Wing Ser Pro Arg Wing Pro Pro Gly Wing Gly Gly Arg Arg 50 55 60 Gly Arg Wing Wing Gly Wing Gly Wing Gly Pro Wing Being Thr Pro Wing Wing Pro65 70 75 80Ala Arg Cys Gln Val 85 Asp Gly Cys Asn Val 90 Asp Leu Thr Asp Val 95 LysPro Wing Tyr Tyr 100 Cys Arg His Lys Val 105 Cys Lys Met His Ser 110 Lys GluPro Arg Val 115 Leu Val Asn Gly Leu 120 Glu Gln Arg Phe Cys 125 Gln Glys CysSer Arg 130 Phe His Gln Leu Pro 135 Glu Phe Asp Gln Leu 140 Lys Lys Ser CysArg Lys Arg Leu Wing Gly His Asn Glu Arg Arg Arg Arg Pro Pro145 150 155 160Gly Pro Leu Wing To Be 165 Arg Tyr Gly Arg His 170 Wing Wing To Be Read Gly 175 GluPro Gly Arg Leu 180 Arg Be Phe Met Leu 185 Asp Phe Be Tyr Pro 190 Arg ValSer Ser Wing 195 Met Arg Gly Gly Phe 200 Pro Wing Val Arg Wing 205 Gly Gly GluArg Val 210 Pro Gly Gly Ile Gln 215 Trp Gln Wing Gly Leu 220 Asp Pro Arg HisHis Gln Gly Wing Val Wing Gly Tyr Gly Wing His Tyr Gly Being Glu Gly225 230 235 240Gly Being Being Being 245 Arg Wing Pro Pro Val 250 Phe Pro Gly Pro Glu 255 Leu

ctgcacggcc tcaagttcgg caagaagatc 60ggcagcagca gcggcggtgg cagcgcgccc 120ccggccgctt cgcctagggc cccggccggc 180ggcgcgggcc cctcgacggc gcccgcgccc 240gacctcaccg acgtcaagcc cgcctactac 300aaggagcccc gcgtcctcgt caacggcctc 360ttccaccagc tgcctgaatt cgaccagcta 420cacaacgagc gccggaggag gccgccgcct 480gctgcgtcgc tcggcgagcc cggcaggctc 540agggtctcaa gcgccatgag gggtgggttt 600cctggcggga tccagtggca agcgggcttg 660ggatacggcg cccactatgg gagcgagggt 720ttccctggcc cggagctgcc cccaggtgga 780tgtgctctct ctcttctgtc aactcagcca 840cacgctgcgc caacagcggg tttcgacggc 900gcgagtagct acatcgcgcc gagcccctgg 960gggcggagcg tgcctcagct gccacctgac 1020tcgagcagcc accacggcca gttctcaggt 1080ccagcgccag ggtcggcggc accgccagcg 1140ttcgaccagg ctggcaacac gatggactgg 1200 1209Pro Gly Gly Cys Leu Ala Gly Val Ser Ala

260 265 270

Read Be Read Read Be Gln Pro Trp Asp Wing Ward His Be His Be

275 280 285

His Ser His Wing Ala Pro Thr Ala Gly Phe Asp Gly Gly Ser Pro Val

290 295 300

Pro Wing Be Met Le Ward Wing Be Ser Tyr Ile Wing Pro Be Pro Trp305 310 315 320

Thr Glu Thr Asp Be Trp Gly His Glu Gly Gly Arg Be Val Pro Gln

325 330 335

Leu Pro Pro Asp Asp Val Pro Leu Gly Glu Val His Ser Gly Ser Ser

340 345 350

Be His His Gly Gln Phe Be Gly Glu Leu Glu Leu Wing Leu Gln Gly

355 360 365

Asn Arg Pro Pro Wing Gly Be Ala Pro Wing Pro Pro Asn Wing Asn Asn

370 375 380

Gln Gly Being Wing Gly Thr Phe Asp Gln Wing Gly Asn Thr Met Asp Trp385 390 395 400

To be read

<210> 262 <211> 1137 <212> DNA <213> Zea mays <400> 262

atggcgaccg gcggcggcag cagcaggagc gacgacgtgc gcgggctcaa gtttggcaag 60aagatctact tcgagcagga cggcgggagc gggagcgggg cgggggcggt gggcggcagg 120aaggggaagg gcgtggccac cggtggcgcg aggccggcgt ccgccgcctc cgcagcccag 180ccgccgaggt gccaggtgga cgggtgcggc gtggatctga gcgccgtcaa gcagtactac 240tgccggcaca aggtgtgcaa catgcactcc aaggagccgc gcgtcttcgt cgccggcatc 300gagcagcgct tctgccaaca gtgcagcagg ttccaccagc tacatgaatt tgaccaaggg 360aaacgtagct gccgccgccg cctcatcggt cacaacgagc gccggaggaa gccaccacct 420ggacctctca cttcacgata tggccggctc gctgcatcac ttcaagagcc tggcaggttc 480agaagcttcc tgctcgactt ctcgtaccca agggttccaa gcagcgtgag ggatgcgtgg 54 0ccaggaatcc agcacggtgg cgacaggatg ctgggcaccg tccagtggca tgggcaccaa 600gaacctcctc acccacaccg cagtgcagct gctggctatg gcaaccatgc tgcatacaac 660tgccatggcg gcttggtagc aggcggggcc ccaatgctct cctctgccgc ctttgagctc 720ccgcctggcg gatgtgtcgc gggagttgcc gccgactcca gctgtgctct ctctcttctg 780tcaactcagc catgggacac gacctcccac gaccaccggt ccccagcaat gcccgcggcc 840ggcgccttcg acggcacccc t ggtggcgccg ccgtcatgg cgagcagcta cgcggcgtcg 900agcgcctgga cgggctcgcg ggaccccgct gccgacggcg ccaggaacgc gcagcgtctc 960gacgatgctc tgcacctggt ccacccaggc tccgcggcgg tccacttctc cggcgagctc 1020gagctcgccc tgcagggaag cggcgggccg ccacacctgc cgcgcgtcga ccatggcggc 1080tccggcggcg gcaccttcaa ccattccacc accagcgcga tgaactggtc cctgtag 1137

<210> 263 <211> 378 <212> PRT <213> Zea mays <400> 263

Met Wing Thr Gly Gly Gly Ser Ser 5

Lys Phe Gly Lys Lys Ile Tyr Phe20

Gly Wing Gly Wing Val Gly Gly Arg

35 40

Gly Wing Arg Pro Wing Be Wing Wing

50 55

Gln Val Asp Gly Cys Gly Val Asp65 70

Cys Arg His Lys Val Cys Asn Met85

Val Wing Gly Ile Glu Gln Arg Phe100

Arg Ser Asp Asp Val Arg Gly Leu

10 15

Glu Gln Asp Gly Gly Ser Gly Ser25 30

Lys Gly Lys Gly Val Wing Thr Gly45

Be Wing Gln Pro Pro Arg Cys60 Wing

Read Ser Ala Val Lys Gln Tyr Tyr

75 80

His Ser Lys Glu Pro Arg Val Phe

90 95

Cys Gln Gln Cys Be Arg Phe His105 110Gln Read His Glu Phe Asp Gln Gly Lys Arg Be Cys Arg Arg Arg Read

115 120 125

Ile Gly His Asn Glu Arg Arg Arg Lys Pro Pro Gly Pro Read Thr

130 135 140

Be Arg Tyr Gly Arg Read Wing Ward Be Read Gln Glu Pro Gly Arg Phe145 150 155 160

Arg Be Phe Read Le Asp Phe Be Tyr Pro Arg Val Pro Be Ser Val

165 170 175

Arg Asp Trp Pro Wing Gly Ile Gln His Gly Gly Asp Arg Met Leu Gly

180 185 190

Thr Val Gln Trp His Gly His Gln Glu Pro His Pro His Arg

195 200 205

Wing Wing Wing Wing Gly Tyr Gly Asn His Wing Wing Wing Tyr Asn Cys His Gly Gly

210 215 220

Leu Val Wing Gly Gly Wing Pro Met Leu Being Ser Wing Wing Phe Glu Leu225 230 235 240

Pro Gly Gly Cys Val Wing Gly Val Gly Wing Wing Asp Ser Ser Cys Wing

245 250 255

Read Be Read Read Be Gln Pro Thr Asp Thr Thr Be His Asp His

260 265 270

Arg Be Pro Wing Met Pro Wing Wing Gly Wing Phe Asp Gly Thr Pro Val

275 280 285

Pro Wing To Be Val Met Wing To Be Tyr Wing Wing To Be Wing Trp Thr

290 295 300

Gly Ser Arg Asp Pro Wing Asp Wing Gly Wing Arg Asn Wing Wing Gln Arg Leu305 310 315 320

Asp Asp Wing Read His Leu Val His Pro Gly Ser Wing Wing Val His Phe

325 330 335

Ser Gly Glu Leu Glu Leu Wing Wing Leu Gln Gly Ser Gly Gly Pro Pro His

340 345 350

Read Pro Arg Val Asp His Gly Gly Gly Gly Gly Thr Phe Asn His

355 360 365

Be Thr Thr Be Ala Met Asn Trp Be Read

370 375

<210> 264 <211> 403 <212> DNA

<213> Sorghum propinquum <400> 264

atggagtccg gtggcgccag tggcggcggc ggcggggacg accagctgca cggcctcaag 60ttcggcaaga agatctactt cgaggacgcc gccgcggtcg gctccagcag cggtggcggt 120ggcggtggca gtggcagtgc tagcgcgacc cccgcgcctc cagcgacgca gcagccgtcc 180ccgccgcagg ccgcttcgcc cagggcagcc agcggcggcg gcggcaggag gggcagggcc 240gcgggcggcc cctccccggc gcccgcgccc gcgcgctgcc aggtcgacgg ctgcaacgtg 300gacctcaccg acgtcaagcc ctactactgc cgccacaagg tctgcaaaat gcactccaag 360gagccccgcg tcgtcgtcaa cggcctcgag cagcgcttct gcc 403

<210> 265 <211> 134 <212> PRT

<213> Sorghum propinquum <400> 265

Met Glu Be Gly Gly Wing Be Gly Gly Gly Gly Gly Asp Asp Gln Leu15 10 15

His Gly Leu Lys Phe Gly Lys Lys Ile Tyr Phe Glu Asp Wing Wing Wing

20 25 30

Val Gly Be Ser Be Gly Gly Gly Gly Gly Gly Be Gly Be Wing

35 40 45

Wing Pro Thr Wing Pro Pro Wing Thr Gln Gln Pro Be Pro Gln Pro Wing

50 55 60

Wing Be Pro Arg Wing Wing Be Gly Gly Gly Gly Arg Arg Gly Arg Ala65 70 75 80

Gly Gly Wing Pro Pro Pro Wing Pro Wing Pro Arg Wing Cys Gln Val Asp85 90 95Gly Cys Asn Val Asp Read Thr Asp Val Lys Pro

100 105

Lys Val Cys Lys Met His Be Lys Glu Pro Arg

115 120

Read Glu Gln Arg Phe Cys

130 <210> 266 <211> 732 <212> DNA <213> Allium strains <400> 266

atggaaatgg gttcgagctc tgagccaatt tctgactctcgaaaaaattt actttgaaga ttccactacc agtactgcaagctgctcctc ccaaaaaggg aaaatcttta gcttcttcgtcagcaacaaa taagcatacc tagatgccag gttgaaggatgctaaagcct attactgcag gcataaggtt tgcggagttcgtggttgctg gaattgaaca gaggttttgt cagcagtgcagaatttgacc aaggaaaaag aagctgccgc agacgcctagagaaagccgc ctccaggccc ttcccgttat ggaaggatgtagtaacagat tcagaggatt cctgatggac ttcagttaccccaggagatc atctatggcc tagactgtcc aacattccatactgcatcta acattcatgt tgctcaaccg tacatgcatggagcttccac acaggaatac atggtctggc gtctgcgactctgtcactca c <210> 267 <211> 244 <212> PRT <213> Allium strain <400> 267

Met Glu Met Gly Ser Ser Ser Ser Glu Pro Ile Ser1 5 10

Tyr Tyr Cys Arg His110

Val Val Val Asn Gly125

tcgatgggct cacattcggcgtactactag caatacaattccactaatat caactctaat

gcaaagttgaattcaaaatcgcaggttccactggacacaacatcatctgtcgaggcccatcatacccagccaatgaatcatgagctgtgc

tttgactggaaccaaaagttccagttgcaatgagcgtcgtctatgatggcacagcctcctaaacccaaacctacagttcctctctctctt

Asp Ser

Read Thr Phe Gly Glu Lys Ile Tyr Phe Glu Asp Ser Thr

20

25

Wing Be Thr Thr Be Asn Thr Ile Wing Pro

35

40

Be Read Ala Be Be Be Thr Asn Ile Asn Be

50

55

Ser Ile Pro Arg Cys Gln Val Glu Gly Cys Lys

Pro Lys45

Asn gln60

Val asp

65

70

75

Lys Wing Tyr Wing Tyr Cys Arg His Lys Val Cys Gly Val

85 90

Ser Pro Lys Val Val Val Wing Gly Ile Glu Gln Arg Phe

100

105

Cys Ser Arg Phe His Gln Read Gln Glu Phe Asp

115

120

Cys Arg Arg Arg Leu Wing Gly His Asn Glu Arg

130

135

Pro Gly Pro Be Arg Tyr Gly Arg Met Be

145

150

155

Ser Asn Arg Phe Arg Gly Phe Read Met Asp Phe

165 170

Glle Ile Pro Pro Gly Asp His Leu Trp Pro

180 185

Pro Ser Tyr Pro Wing Asn Pro Wing Asn Thr Wing

195 200

Gln Pro Tyr Met His Wing Met Asn His Tyr Ser

210 215

Arg Asn Thr Trp Ser Gly Val Cys Asp Leu Ser225 230 235

Leu Ser Leu Asn

Gln Gly125Arg Arg140

To be val

To be tyr

Arg Leu

Asn Ile205Ser Glu220

Cys Wing

Read Asp Gly15

Thr Ser Thr30

Lys Gly Lys

Gln Gln Ile

Read Thr Gly80

His Ser Lys95

Cys Gln Gln110

Lys Arg Ser

Lys Pro Pro

Tyr Asp Gly160

Pro Arg Pro175

Ser Asn Ile190

His Val Wing

Read pro his

Leu Ser Leu240

60120180240300360420480540600660720732

<210> 268 <211> 969 <212> DNA

<213> Antirrhinum majus <400> 268

tcagctgctg gtggagcaga ggaatctctc aatgggttga agtttggcaa gaaaatatac 60

tttgaggagg ctaaggcaaa gaaagggaag agtaccggtg gggtggttag gtgccaggtg 120

gaggggtgtg aggtagatct gagtgatgct aaggcttact atttgagaca caaagtttgt 180

agtatgcatt caaagtctcc aaaggtcatt gttgctggaa tagaacaaag gttttgccag 240

cagtgcagca ggttccatca attgcctgaa tttgaccaag gaaaacggag ttgccgcaga 300

cgccttgcag gccacaacga acgtcggagg aagccatctc caggatctat gatgtctcct 360

tactatggaa gtctttctcc aaccttattt gataaccaaa atagaactgg aggctttttg 420

atggacttca gcacttaccc aaatctcgct gggaaagatt catggccaaa tacaataccc 480

gaacgaggat tgggaggtcc agcaagtcca tggcagagcg acatgcaaaa tcctgtacct 540

gagtttttgc gaggtacaac aaataggcca agtttttctg gtcttggagt atcttccgaa 600

gaatgtttta gcggagtctc taattccagc actgctctct ctcttctgtc aaatcagtcc 660

tggggctcca gaaactcgaa caattttctt ggtaccaatg gaaacgggcc aaccatagtt 720

cagccgtcta ttaaccctgg tgccacaatt ggacagttta cctgtccctc ttggggtttt 780

ggaggcaacc cagctgataa cacctcccat gatatgcctc ccaatctgaa tttaggacaa 840

ttttctcact ccagtaacag tcactatact ggagagcctg gggtagtcca actgagccac 900

ggacaattcc aggacctcga tcactcaaga ggctatgatt cttccgttca ggacatgcat 960

tggtcactt 969 <210> 269 <211> 323 <212> PRT

<213> Antirrhinum majus <400> 269

20

25

35

40

50

55

65

70

85

100

105

115

120

130

135

145

150

165

Asn Pro Val Pro Glu Phe Read Arg Gly Thr Thr

180

185

Be Gly Leu Gly Val Be Glu Glu Cys Phe

195

200

Be Ser Thr Wing Read Be Read Read Asn Gln

210

215

Asn Ser Asn Asn Phe Read Gly Thr Asn Gly Asn

225

230

Gln Pro Ser Ile Asn Pro Gly Wing Thr Ile Gly245

260

265

275

280

290

295

Leu Asn Gly Leu Lys Phe Gly10 15 Wing Lys Lys Gly Lys Ser Thr 30 Gly Cys Glu Val â R Asp 95 Asn Glu Arg Arg Arg Lys Pro 110 Tyr Gly Be Read Be Pro Thr 125 Gly Phe Read Met Asp Phe Be 140 Be Trp Pro Asn Thr Ile Pro 155 160Pro Trp Gln Be Asp Met Gln170 175 Thr Asn Arg Pro Be Phe 190 Cys Phe Be Gly Val Be Asn 205 Asn Gln Be Trp Gly Be Arg 220 Gly Asn Gly Pro Thr Ile Val 235 240Ile Gly Gln Phe Thr Cys Pro250 255 Asp Asn Thr Be His Asp Met 270 Be His Be Asn Be His 285 Gly Gln Phe Gln 300Asp Read Asp His Be Arg Gly Tyr Asp Be Be Val Gln Asp Met His305 310 315 320

Trp Ser Leu

<210> 270 <211> 633

<212> DNA

<213> Brassica napus <400> 270

gacttggaga aaagaagttg tagaagacgt ctagcttgtcccccaagcaa caacagcagc tcttttggct tctggttacttacggaagcg ttttgggaga tcctacaacg tggtcaaccgtccgcaccgt gggatagcca tcaactgatg aacgttttgtagtataacat acccagagat ggtgaacaat aatagcacagcttctgtcaa actcaaacac aactcagcag cagcagcagaatggacgcag aaagggttac aatggctaag tcaccgcctgtcgaaacaaa cctgggagtt catgtcaggc gaaaagagcacctgttttgg gactgagaca aatctctgag ccagatgatgaatggcacca caatgggtgg attcgagctg aacctacagctactcttcta ctcaaaattt tacttggcct ctt

<210> 271 <211> 211 <212> PRT

<213> Brassica napus <400> 271

ataacgaaagctagaatcgccaagatctgtcacagggaagactcaagctgcatcaaccaatttcagtacaattggccttgacctccagttaggagcaggt

aagaagaaagtccatctcttgatgggacggttcaaggttttgctctctcttgcttacttgcaatcagtactgtgtcgtcccctgatgagctctgaggcaa

Asp Leu Glu Lys Arg Be Cys Arg Arg Arg Leu Wing Cys His Asn Glu1 5 10 15 Arg Arg Arg Lys 20 Pro Gln Wing Thr Thr 25 Wing Wing Leu Leu Wing 30 Ser GlyTyr Ser Arg 35 Ile Wing Pro Ser Leu 40 Tyr Gly Ser Val Leu 45 Gly Asp Pro Thr Thr 50 Trp Being Thr Wing Arg 55 Ser Val Met Gly Arg 60 Being Wing Pro TrpAsp Being His Gln Leu Met Asn Val Leu Being Gln Gly Being Ser Arg Phe65 70 75 80Ser Ile Thr Tyr Pro 85 Glu Met Val Asn Asn 90 Asn Ser Thr Asp Ser 95 SerCys Ala Leu Ser 100 Leu Read Asn Ser 105 Asn Thr Thr Gln Gln 110 Gln GlnGln Thr Ser 115 Thr Asn Ala Tyr Leu 120 Met Asp Ala Glu Arg 125 Val MetAla Lys 130 Ser Pro Pro Val Ser 135 Val His Asn Gln Tyr 140 Ser Lys Gln ThrTrp Glu Phe Met Ser Gly Glu Lys Ser Asn Trp Pro Cys Val Ser Ser145 150 155 160Pro Val Leu Gly Leu 165 Arg Gln Ile Ser Glu 170 Pro Asp Asp Leu 175 GlnPhe Leu Met Ser 180 Asn Gly Thr Thr Met 185 Gly Gly Phe Glu Leu 190 Asn LeuGln Gln Glu 195 Gln Val Leu Arg Gln 200 Tyr

Trp Pro Leu210 <210> 272 <211> 852 <212> DNA

<213> Saccharum officinarum <400> 272

ctcgtcgtca acggcctcga gcagcgcttc tgccagcagtcctgaatttg accaactaaa gaaaagctgc cgcagacgtccggaggaggc cacctcctgg acctcttgca tcacgatatgggtgagcccg gcaggttccg aagctttatg ttggatttctaccatgaggg atgggtttcc ggcagttcga cctggcgaaa

gcagcaggttttgcaggccagtcgccttgccatacccaaggggtgcctgg

ccaccagctgcaatgaacgctgcatcatttggttccaggctagtatccag

60120180240300360420480540600633

60120180240300tggcaagcgg gcttagatcc tcatcatcat caaagcgcggtcatatggga gccagggtag ctcgtcgtcg tcaaggccacctccccccag gtggatgtct tgcaggagtc ccctcggactctgtcaactc agccatggga tactacccac agcgccggcccctgcaacag caggttttga cggcaaccct gtggcaccctattgcgccaa gcccctggac tgactcccgg ggccatgaagttgccacctg acgtccccct cagcgaggtg cactctggctttctcaggtg agctcgagct tgccctgcag ggaaacaggcgcgccgcgca atgatcaggg ctccacgggc acgttcgacctggtcgctct g <210> 273 <211> 283 <212> PRT

<213> Saccharum officinarum <400> 273

Leu Val Val Asn Gly Read Glu Gln Arg Phe Cys15 10

Phe His Gln Leu Pro Glu Phe Asp Gln Leu Lys

20 25

Arg Read Leu Gly His Asn Glu Arg Arg Arg Arg Arg

35 40

Read Wing Ala Arg Tyr Gly Arg Read Wing Ala Ser

50 55

Arg Phe Arg Ser Phe Met Read Asp Phe Ser Tyr65 70 75

Thr Met Arg Asp Gly Phe Pro Wing Val Arg Pro

tcgcaggatacggtgttcccctagctgtgcacagccatgcccctcatggcgcgggcggaacaagcagccacagcaccaggagtccggcaa

cggtgcccactggtccagagtctctctctttggatcaatggagtagctaccgtgcctcagtcacggccaggtcagcgccacacaatggac

60

85

90

Gly Ser Ile Gln Trp Gln Wing Gly Read Asp Pro

100

105

Val Wing Gly Wing Tyr Gly Wing His Ser Tyr Gly

115

120

140

Cys Wing

Be Ser Be Arg Pro Pro Val Phe Pro Gly Pro

130 135

Gly Cys Read Ala Gly Val Pro Ser Asp Ser Ser145 150 155

Read Ser Thr Gln Pro Asp Thr Thr His Be Ala Gly

Gly asn

Gln Cys Ser Arg 15 Ser Cys Arg 30 Pro Gly Pro45 Gly Glu Pro GlyArg Val Pro Gly 80Glu Arg Val Pro 95 His His Gln Ser 110 Gln Gly Ser125 Leu Pro Pro Gly

165

170

Gly Wing Ser Met Pro Wing Thr Gly Phe Asp Wing

180 185

Pro Be Read Met Wing Be Ser Tyr Ile Wing Pro Be Pro195 200 205

Be Arg Gly His Glu Gly Gly Arg Asn Val Pro Gln Leu

210 215 220

Val Pro Read Being Glu Val His Being Gly Being Being His225 230 235

Phe Ser Gly Glu Leu Glu Leu Wing Leu Gln Gly Asn Arg

245 250

Gly Ser Wing Pro Wing Pro Wing Asn Asp Gln

260 265

Asp Gln Be Gly Asn Thr Met Asp Trp Be Read

275 280

<210> 274 <211> 647 <212> DNA

<213> Festuca arundinacea <400> 274

ggcacgaggg tggatctgag cggctccaag acctactact gccgccacaaatgcactcca aggcgccccg cgtcgtcgtc gccggcctcg agcagcgctttgcagcaggt tccaccagtt gcctgaattt gacaatggaa aacgcagctgctcgcaggtc acaatgaacg ccgtaggaag ccgcctcctg gccctctggcggccgactcg ctgcatcctt tgaagaaccg ggcaggtaca gaagctttcttcctacccaa gggttccgag cagcgtgcgg gatgcttggc ctgcagttcgcgtatgccca gtgaaatcca gtggcaaggg aacctagacc tgcgtcctca

Leu Ser Leu160

His being his

175Pro Val Ala190

Trp Thr Asp

Pro Pro Asp

His Gly Gln240

Pro Wing Pro

255Gly Thr Phe270

ggtctgctccctgccagcagccgcagacgtgtcacgctatgttagatttcaccaggctaccacgggttat

360420480540600660720780840852

60120180240300360420ggcccacatg catatggcag ccacggcttc cccggtccag agctccctcc aggcgggtgt 4 80ctcacagggg tcgccaccga ctccagctgt gctctctctc ttctgtcaac tcagccatgg 540gataccacca cccacggtgc cagccacgac catcggtctg cggccatgtc cgcggccgcg 600ggcttcgacg gcagccctgc ggcagtgtca ccctccatca 647 tggcgag

<210> 275 <211> 215 <212> PRT

<213> Festuca arundinacea <400> 275

Gly Thr Arg Val Asp Read Gly Be Lys Thr Tyr Tyr Cys Arg His1 5 10 15 Lys Val Cys Be 20 Met His Be Lys Wing 25 Pro Arg Val Val Val 30 Wing GlyLeu Glu Gln 35 Arg Phe Cys Gln 40 Cys Ser Arg Phe His 45 Gln Leu ProGlu Phe 50 Asp Asn Gly Lys Arg 55 Ser Cys Arg Arg Arg 60 Leu Wing Gly HisAsn Glu Arg Arg Lys Pro Pro Gly Pro Leu Wing To Be Arg Tyr65 70 75 80Gly Arg Leu Wing Ala Ser Phe Glu Glu Pro Gly Arg Tyr Arg Be Phe 85 90 95 Leu Read Asp Phe 100 Be Tyr Pro Arg Val 105 Pro Be Ser Val Arg 110 Asp AlaTrp Pro Wing 115 Val Arg Pro Gly Tyr 120 Arg Met Pro Glu 125 Ile Gln TrpGln Gly 130 Asn Leu Asp Leu Arg 135 Pro His Gly Tyr 140 Gly Pro His AlaTyr Gly Be His Gly Phe Pro Gly Pro Glu Leu Pro Pro Gly Gly Cys145 150 155 160Leu Thr Gly Val Ala 165 Thr Asp Ser Ser Cys 170 Ala Leu Ser Leu Leu 175 SerThr Gln Pro Trp Asp Thr Thr Thr His Gly Wing Be His Asp His Arg 180 185 190 Ser Wing Ala 195 Met Ser Wing Ala Wing 200 Gly Phe As p Gly Ser 205 Pro Wing AlaVal Ser 210 Pro Ser Ile Met Wing 215 <210> 276

<211> 11 <212> PRT

<213> Artificial sequence <220>

<223> reason 1 <220>

<221> VARIANT <222> (1) .. (1) <223> / replace = "Thr" <220>

<221> VARIANT <222> (3) .. (3)

<223> / replace = "Arg" / replace = "Met" / replace = "Gln" <220>

<221> VARIANT <222> (6) .. (6)

<223> / replace = "Gln" / replace = "Arg" <220>

<221> VARIANT <222> (7) .. (7)

<223> / replace = "Arg" / replace = "Glu" <220>

<221> VARIANT <222> (8). . (8) <223> / replace = "Go" "Gly"

<220>

<221> VARIANT <222> (11) .. (11) <223> / replace <400> 276

Gly Leu Lys Phe Gly Lys Lys Ile Tyr Phe Glu15 10

<210><211><212><213> <220>

<223> DomainArath_SPL15 <400> 277

Thr Wing Arg Cys Gln Val Glu Gly Cys

277

78

PRT

Sequence

artificial

SPL of DNA transcription factor (DBD) transcription factor

1

Lys

Tyr wing

Tyr Ser Arg20

Ile Val Ser

Ser Lys Val35

Being Arg Phe His Gln Leu50

Arg Arg Arg Read Ala Cys

70

His Lys Val25

Gly Leu His40

Arg Met Asp Leu10

Cys Cys Ile His

Ser Asn Val15

Ser Lys Ser30

Gln Gln Cys

Gln Arg Phe Cys45

Be Glu Phe Asp Read Glu Lys Arg Be Cys55 60

His Asn Glu Arg Arg Arg Lys Pro75

65

<210> 278 <211> 11 <212> PRT

<213> Artificial sequence <220>

<223> reason 2 <220>

VARIANT (3). . (3)

/ replace = "Asn" / replace = "Thr"

VARIANT (4). . (4)

/ replace = "Gly" / replace = "Arg" VARIANT

(H) · · (H)

/ replace = "Thr" 278

Cys Wing Read Leu Read Leu Be Asn5 10

To be

<221><222><223><220><221><222><223><220><221><222><223> <400> Asp Ser1

<210> 279

<211> 1130

<212> DNA

<213> Oryza sativa

<400> 279

catgcggcta atgtagatgc tcactgcgct agtagtaaggtatacaaagc tgtaatactc gtatcagcaa gagagaggcacaggattaga aaaacgggac gacaaatagt aatggaaaaacatggcaata taaatggaga aatcacaaga ggaacagaatgtactcgtac gtaaaaaaaa gaggcgcatt catgtgtggagatttgaaac cactcaaatc caccactgca aaccttcaaatagaaagcac aggtaagaag cacaacgccc tcgctctccagcacctcgcg gatcggtgac gtggcctcgc cccccaaaaaacaccccggg cccacccacc tgtcacgttg gcacatgttgcaaaatatca acgcggcgcg gcccaaaatt tccaaaatccgccgctcttc cacccaggtc cctctcgtaa tccataatggtgtacgtggg cgggttaccc tgggggtgtg ggtggatgac

tactccagta cattatggaa 60cacaagttgt agcagtagca 120caaaaaaaaa caaggaaaca 180ccgggcaata cgctgcgaaa 240cagcgtgcag cagaagcagg 300cgaggccatg gtttgaagca 360ccctcccacc caatcgcgac 420tatcccgcgg cgtgaagctg 480gttatggttc ccggccgcac 540cgcccaagcc cctggcgcgt 600cgtgtgtacc ctcggctggt 660gggtgggccc ggaggaggtc 720cggccccgcg cgtcatcgcg gggcggggtg tagcgggtgcgaaaattcaa attaggaggt ggggggcggg gcccttggagtgcccctgac ttggcttggc tcctcttctt cttatcccttttctctcctc tcctcttctc ttctcttctc tggtggtgtgctccttcctc ctctcctttc ccctcctctc ttcccccctcgactctcccc aggtgaggtg agaccagtct ttttgctcgagcctcgcgcg gatctgaccg cttccctcgc ccttctcgca <210> 280 <211> 56 <212> DNA

<213> Artificial sequence <220>

<223> initiator: prm07277 <400> 280

ggggacaagt ttgtacaaaa aagcaggctt aaacaatgga <210> 281 <211> 51 <212> DNA

<213> Artificial sequence <220>

<223> initiator: prm07278 <400> 281

ggggaccact ttgtacaaga aagctgggtt gatgaagatc <210> 282 <211> 1110 <212> DNA

<213> Brassica rapa <400> 282

atggataggg gttccaactc gggtcttggt cttggtccagtcatccactg agtcatcctc tttaagtgga gggctcatgtgaggacgctg gaggtggaac cgggtcttct tcctccggcgggaagcgggt cgggtccttc gggtcagata ccaaggtgtcgatctaacca atgcaaaggg ttattacacg aggcatagagacaccaaaag tcattgtcgc tggtatagaa caaaggttttcatcagcttc cggaatttga cctagagaaa aggagttgccaatgagagac gaaggaagcc acagcctgcg tctctatctgaggatcactc cttctctata cggaaatggt gaaactacaatcccaagaaa tgggttggaa tagtgcaaga acgttggataccgtcgtggc agatcaatcc tatgaatgtg tttagtcatgggaggaggga taagcttctc atctccagag attatggacaggaattggca gcgactcaaa ctgtgctctc tctcttctgtgacaacaaca acagcaacaa cacatggaga acttcttcggacaatggctc agccaccacc tgcacctagc cagcagcatcgtattcaagg acgatgataa tagctgtccg aatgatatgtcggttcaccg agacagagat aagcggtgga acgactttggcatcatcatc agaataggag gcagtacatg gaaagtgagatcttcacacc ataacaactg gtctctctga <210> 283 <211> 369 <212> PRT

<213> Brassica rapa <400> 283

Met Asp Arg Gly Ser Asn Ser Gly Leu Gly Leu15 10

Glu Be Gly Gly Be Ser Thr

20 25

Met Phe Gly Gln Arg Ile Tyr Phe Glu Asp Wing

35 40

Be Ser Be Ser Gly Gly Ser Asn Arg Arg Val

50 55

Gly Pro Ser Gly Gln Ile Pro Arg Cys Gln Val65 70 75

Asp Leu Thr Asn Wing Lys Gly Tyr Tyr Thr Arg

gaaaaggaggaataagcggagtcctcgcaaggtgtgtccctcacaagagattcgacgcgcggattcagcc

cgatcggtacatcgcagataccccgcttcctgtctcccctgagagcgccactttcacgcc

gttgttaatg tgttcg

ttaaaaggtg a

gccagacagattggccagagggtcaaacagaagtggaaggtttgtggaatgtcaacagtggtaggagacttgttgtcttctgaatgggagcaagagtgatgatcagtaagctaaaccagacaaacccacagttttggtccagtatgggctgaact

gtcgggtggtgatctacttcaagggtacgtttgtggaatggcactctaaacagcaggtttcgctggtcattcgttatgggctttcttggtgagacggccatggggggagagctacaagtcggggcggggtgctgtg

Gly Pro Gly Gln Thr15

Leu Ser Gly Gly Leu30

Gly Gly Gly Thr Gly45

Arg Gly Ser Gly Ser60

Glu Gly Cys Gly Met80

His Arg Val Cys Gly

780840900960102010801130

56

51

60120180240300360420480540600660720780840900960102010801110Met His Ser Lys100

Phe Cys Gln Gln115

Glu Lys Arg Ser130

Arg Lys Pro Gln145

Arg Ile Thr Pro

Ser Phe Leu Gly180

Asp Thr Arg Val195

Asn Val Phe Ser210

Be Phe Ser Ser225

Gly Ile Gly Ser

His Gln Pro His260

Be Gly Phe Gly275

Pro Ser Gln Gln290

Asp Asp Asn Ser305

Arg Phe Thr Glu

Glu Leu Ser Asp340

Glu Asn Thr Arg355

Read

85

Thr Pro Lys Val

Cys Ser Arg Phe120

Cys Arg Arg Arg135

Pro Wing Ser Leu150

Ser Leu Tyr Gly165

Be Gln Glu Met

Met Arg Arg Pro200

His Gly Ser Val215

Pro Glu Ile Met230

Asp Ser Asn Cys245

Asp Asn Asn Asn

Pro Met Thr Val280

His Gln Tyr Leu295

Cys Pro Asn Asp310

Thr Glu Ile Ser325

His His His Gln

Tyr Gly Ser360 Wing

90

Ile Val Wing Gly105

His Gln Leu Pro

Read Wing Gly His140

Ser Val Leu Ser155

Asn Gly Glu Thr170

Gly Trp Asn Ser185

Pro Ser Trp Gln

To be Gly Gly Gly220

Asp Thr Lys Pro235

Wing Leu Ser Leu250

Be Asn Asn Thr265

Thr Met Wing Gln

Asn Pro Pro Trp300

Met Ser Pro Val315

Gly Gly Thr Thr330

Asn Arg Arg Gln345

To be his his

95

Ile Glu Gln Arg110

Glu Phe Asp Leu125

Asn Glu Arg Arg

To be Arg Tyr Gly160

Thr Met Asn Gly175

Wing Arg Thr Leu190

Ile Asn Pro Met205

Gly Gly Gly Ile

Glu Ser Tyr Lys240

Read Ser Asn Pro255

Trp Arg Thr Ser270

Pro Pro Pro Ala285

Val Phe Lys Asp

Leu Asn Leu Gly320

Read Gly Glu Phe335

Tyr Met Glu Ser350

Asn Asn Trp Ser365

<210> 284

<211> 1080

<212> DNA

<213> Glyc

<400> 284

atggcttcag

aaaatctatt

tcttcttctt

tctgttcaac

gatgcaaaag

gtcattgttg

cctgagtttg

cggagaaagc

gcttttgata

cttagtctga

acctccacac

tcggtgggtg

gtcaccgaca

acagcaccaa

cttgctgcat

aagggcattg

atttcacagc

aggcattata

<210> 285

<211> 359

<212> PRT

me max

acgccaaactttgaggatgtctgctgctaccagctcaacccttactattcctggtcttcaatcaaggaaaccccaacaagatagtggcaggaaattcactttagctggaaggacacctgtgcgtcttgggttcatctggagtgctcg

ctcctcttcttggtcttgcttgttacttcttcccaggtgttagacacaagacaaaggtttaagaagttgcctccctcttaaggtagcaactccaactccttggcaactcacgccagcccgtctctctcttgagtaacatgtgcatcagcccccccccccctcct

gagtccctcaactccagccatcttcttcttcaagttgaaggtctgtggcatgtcaacagtcgtaggcgacacctctcgcttttctgatggagatcatctggagacatcatggacatcctcctgtcaaatcataaatttcacatcaact

acggtttgaacctctcttaccaaggaaaggggtgcaaagttgcactctaagtagcaggttggccaatgccagactaattgacttcagctagctccctgaccttttcaggggaaagaaacatggggacgggacaccccaa

gaggtggtcc ctgatctaggggtgagctgg acctgtcccagcatatgaat ctgctcattg

attcggccaattcttcttctaagaggtgggagatctgagtatccccttcatcatcagctttaatgaacgtttcttcgtctatacccaaagtggaaatcaacttgcaaggtttacattgggttctagaaaccttgacacaagtggttcgtgtctctgtgtctg

6012018024030036042048054060066072078084090096010201080 <213> Glycine max <400> 285

Met Wing Be Asp Wing Lys Read Be Be Glu Be Read Asn Gly Leu1 5 10 15

Lys Phe Gly Gln Lys Ile Tyr Phe Glu Asp Val Gly

20 25 30

Ala Thr Be Leu Thr Be Ser Ser Ser Ser Ser Ser Ala Thr Val

35 40 45

Thr Be Be Be Be Be Arg Lys Gly Arg Gly Gly Be Val Gln Pro

50 55 60

Gln Pro Pro Wing Arg Cys Gln Val Glu Gly Cys Lys Val Asp Leu Ser65 70 75 80

Asp Wing Lys Wing Tyr Tyr Be Arg His Lys Val Cys Gly Met His Be

85 90 95

Lys Ser Pro Ser Val Ile Val Wing Gly Read Gln Gln Arg Phe Cys Gln

100 105 110

Gln Cys Being Arg Phe His Gln Read Pro Glu Phe Asp Gln Gly Lys Arg

115 120 125

Be Cys Arg Arg Arg Read Ala Wing Gly His Asn Glu Arg Arg Arg Lys Pro

130 135 140

Pro Thr Be Ser Read Leu Thr Be Arg Tyr Wing Arg Read Le Be Ser145 150 155 160

Ala Phe Asp Asn Be Gly Arg Gly Be Asn Phe Leu Met Glu Leu Thr

165 170 175

Be Tyr Pro Lys Read Be Read Arg Asn Be Read Pro Thr Pro Be

180 185 190

Be Glu Read Ala Pro Gly Asn Gln Thr Be Thr Read Leu Ser Trp Asn Gly

195 200 205

Asn Be Glu Thr Be Be Asp Leu Phe Leu Gln Gly Be Val Gly Gly

210 215 220

Thr Be Phe Ala Be Pro Gly His Pro Pro Gly Glu Be Tyr Ile Gly225 230 235 240

Val Thr Asp Thr Be Cys Wing Read Be Read Read Be Asn Gln Thr Trp

245 250 255

Gly Be Arg Asn Thr Wing Pro Be Read Gly Read Asn Met Ile Asn

260 265 270

Phe Asn Gly Thr Pro Read Thr Gln Read Wing Ala Ser Be His Gly Wing

275 280 285

Ser Ile His Gln Leu Pro Asn Thr Ser Trp Phe Phe Lys Gly Ile Asp

290 295 300

Ser Gly Asn Cys Ser Pro Glu Val Val Pro Asp Leu Gly Leu Gly Gln305 310 315 320

Ile Ser Gln Pro Read Asn Be Gln Read His Gly Glu Read Asp Read Le Ser

325 330 335

Gln Gln Gly Arg Arg His Tyr Met Asp Read Glu Gln Be Arg Wing Tyr

340 345 350

Glu Ser Ala His Trp Ser Leu

355 <210> 286 <211> 1128 <212> DNA

<213> Populus tremuloides <400> 286

atggaaatgg attcaggctc cctaaccgag tcagctactt ccaatgcaac ttctccgcca 60

gctgagtctg ttaatggatt gaaatttggt aagaagattt actttgagga tcacgtgggg 120gtcggtgctc cggctaagag cggaactggg tcatcctcat ccggttccgg gtcagggtca 180tctaggaagg ctcaaggtgg acagcaccag cagccaccaa ggtgtcaagt tgaagggtgc 240aaagtagatc tgagtgatgc taagacttac tattcaaggc acaaagtttg tagtatgcac 300tccaagtctc ctagagttat tgttgctggt ttggtgcaaa gattttgcca gcaatgtagc 360agatttcatc tacttcctga atttgaccaa ggaaaacgaa gttgccgcag gcgcctagct 420ggccataatg agcgacggag gaagccacca tctggatcgg tgttgtccgc tcgccatggc 480cgattctctc cctctttgtt tgataatagc agcagagctg gaggccttct tgtggacttt 540agtgcatatc caaggcatac tgggagagat ggatggcctg cagcaaggtc ttctgagctt 600acccccggga atgatactgc tgccacagga aggtctatat ctcatatgtg gcagataagc 660tcccagaatc ctccatccaa cctttgcttg caaggctcaa ctggcgggac tggccttttc 720agttcaggaa ttcctccggg agaatgcttc acaggagttg ctgtttcaga ctcgagctgt 780gctctctctc ttctgtcaaa tcaaccatgg ggctccacaa accgagcatc aagtcttgcg 840gtgaatgact tgtttagtgc cgaagaggca cccgtggttc aatcaacagc tcaccatggt 900gcggctgtca atcagtatcc the aatcccttgg gcttcaaga gcaatgaagg aagtaacagt 960tcacatgaga tgtgccctga tctaggtctg ggtcaaattt caatgcctct caacagtcaa 1020cttgctggtc agctcgagca gtctcaacag aataggaggc aatacatgga cctcgagcat 1080tccagggctt atgactcttc aacccagcac atccactggt cactttaa 1128

<210> 287 <211> 375 <212> PRT

<213> Populus tremuloides <400> 287

Met Glu Met Asp Be Gly Be Read Thr Glu Be Wing Thr Be Asn Wing 1 5 10 15

Thr Be Pro Pro Glu Wing Be Val Asn Gly Leu Lys Phe Gly Lys Lys

20 25 30

Ile Tyr Phe Glu Asp His Val Gly Val Gly

35 40 45

Thr Gly Be Ser Be Ser Gly Be Gly Ser Gly Ser Be Arg Lys Wing

50 55 60

Gln Gly Gly Gln His Gln Gln Pro Pro Arg Cys Gln Val Glu Gly Cys65 70 75 80

Lys Val Asp Read Be Asp Wing Lys Thr Tyr Tyr Be Arg His Lys Val

85 90 95

Cys Be Met His Be Lys Be Pro Arg Val Ile Val Wing Gly Leu Val

100 105 110

Gln Arg Phe Cys Gln Gln Cys Be Arg Phe His Read Leu Pro Glu Phe

115 120 125

Asp Gln Gly Lys Arg Be Cys Arg Arg Arg Read Leu Wing Gly His Asn Glu

130 135 140

Arg Arg Arg Lys Pro To Be Gly To Be Val Leu To Be Wing Arg His Gly145 150 155 160

Arg Phe Be Pro Be Leu Phe Asp Asn Be Be Arg Wing Gly Gly Leu

165 170 175

Read Val Asp Phe Be Wing Tyr Pro Arg His Thr Gly Arg Asp Gly Trp

180 185 190

Pro Wing Ala Arg Be Being Glu Leu Thr Pro Gly Asn Asp Thr Wing Ala

195 200 205

Thr Gly Arg Be Ile Be His Met Trp Gln Ile Be Be Gln Asn Pro

210 215 220

Pro Be Asn Read Cys Read Gln Gly Be Thr Gly Gly Thr Gly Leu Phe225 230 235 240

Be Ser Gly Ile Pro Pro Gly Glu Cys Phe Thr Gly Val Wing Val Ser

245 250 255

Asp Ser Ser Cys Ala Le Ser Ser Leu Ser Ser Asn Gln Pro Trp Gly Ser

260 265 270

Thr Asn Arg Wing Be Ser Leu Wing Val Asn Asp Leu Phe Ser Wing Glu

275 280 285

Glu Wing Pro Val Val Gln Ser Thr Wing His His Gly Wing Val Val Wing

290 295 300

Gln Tyr Pro Ile Pro Trp Being Phe Lys Being Asn Glu Gly Being Asn Ser305 310 315 320

Be His Glu Met Cys Pro Asp Read Gly Read Gly Gln Ile Be Met Pro

325 330 335

Leu Asn Be Gln Leu Wing Gly Gln Leu Glu Gln Be Gln Gln Asn Arg

340 345 350

Arg Gln Tyr Met Asp Read Glu His Be Arg Wing Tyr Asp Be Ser Thr

355 360 365

Gln His Ile His Trp Ser Leu370 375

<210> 288 <211> 1059 <212> DNA

<213> Citrus Clementine <400> 288

atggaactgg gttcagacta tttggctgaa tcaggtggtg gctccggctc cggctcaggc 60

tccggcttat catccgctga gccatcactt aatggtttga agtttggcaa aaaaatctat 120

tttgaggatg tcggtactgc cggagctcca tttccaggat ctgggtcatc atctgggtcc 180

gggtcagggt caggttcagg gtcagggagg aaggtgaggg gtgttggcgg tggtatggtt 240

actagtgggc agcagccacc aaggtgccaa gtggaggggt gtaaagttga tctgagtgat 300

gccaaagctt actattcaag gcacaaagtt tgtggcatgc attcaaagtc tcctgttgtc 360

actgttgccg gccttgagca gaggttttgc cagcaatgta gcagatttca tcagcttccg 420

gagtttgacc aaggaaaacg aagttgccgc aggcgcctgg caggccataa tgagcgccgg 480

aggaagccaa cttctggacc atttttgggc actcgttatg gcaggctctc ttcctctgtc 540

attgagaaca gcagccaagg tggaggattt ctgatcgact tcagtgcata tcagatggtt 600

ggtgggaggg atggatggcc agtgacaagt gtctccaagc aggtatctgg aaatcaaacc 660

actgtcacag caaggcatct tcctcagcca ctatggcaaa accactctca ggatcctcca 720

cctgatcgtt accttcagtg ttcaacagct gggactggtt tctctggtcc tggaattcct 780

tgtggaggat gcttcacagg agttgctgac tcaaactgtg ctctctctct tctgtcaaat 840

caaccatggg gctctaagaa cccgacaccg ggtcatggag tgggtgactt aatgcatgcc 900

cataccaaat ccgtcactca accagtatcg ccccatggag cagctattaa tcaatatcca 960

aacatgtcat ggggggttca agggcaatgc aacctggtag cagttccaca ccaaatggcc 1020

ccccaaatgg ggtttgggtt caacattccg gcccattaa 1059

<210> 289 <211> 352 <212> PRT

<213> Citrus Clementine <400> 289

Met Glu Leu Gly Ser Asp Tyr Leu Wing Glu Ser1 5

Gly Gly

Ser Gly Ser Gly Ser Gly Read Ser Sera Glu Pro Ser Ser

10

20

25

Read Lys Phe Gly Lys Lys Ile Tyr Phe Glu Asp

35 40 Pro Phe Pro Wing Gly Be Gly Ser Be Ser Gly 50 55 Gly Be Gly Ser Gly Arg Lys Val Arg Gly Val65 70 75Thr Be Gly Gln Pro Gl Cn Gln Val 85 90 Asp Leu Be Asp Wing Lys Wing Tyr Tyr Ser Arg 100 105 Met His Be Lys Be Pro Val Val Thr Val Wing 115 120 Phe Cys Gln Gln Cys Be Arg Phe His Gln Leu 130 135 Gly Lys Arg Be Cys Arg Arg Read Leu Wing Gly145 150 155Arg Lys Pro Thr Be Gly Pro Phe Leu Gly Thr 165 170 Be Ser Be Val Ile Glu Asn Be Ser Gln Gly 180 185 Asp Phe Ser Ala Tyr Gln Met Val Gly Gly Arg 195 200 Thr Be Val Ser Lys Gln Val Ser Gly Asn Gln 210 215 Arg Trp Gln Asn His225 230 235Pro Asp Arg Tyr Leu Gln Cys Be Thr Wing Gly 245 250 Pro Gly Ile Pro Cys Gly Gly Cys Phe Thr Gly 260 265 Cys Wing Read Leu Be Asn Gln Pro Trp 275 280

Val gly

45Being Gly60

Gly Gly

Glu Gly

His lys

Gly Leu125Pro Glu140

His asn

Arg tyr

Gly Gly

Asp Gly205Thr Thr220

Be gln

Thr gly

Val Wing

Gly ser285

Gly Ser Gly15

Read Asn Gly30

Thr Wing Gly

Be gly be

Gly Met Val80

Cys Lys Val95

Val Cys Gly110

Glu Gln Arg

Phe Asp Gln

Glu Arg Arg160

Gly Arg Leu

175Phe Leu Ile190

Trp Pro Val

Val Thr Wing

Asp Pro Pro240

Phe Ser Gly

255Asp Ser Asn270

Lys Asn ProThr Gly His Gly Val Gly Asp Read Met His

290 295

Val Thr Gln Pro Val Ser Pro His Gly Wing Ala305 310 315

Asn Met Being Trp Gly Val Gln Gly Gln Cys Asn

325 330

His Gln Met Pro Wing Gln Met Gly Phe Gly Phe340 345

<210> 290 <211> 565 <212> DNA

<213> Beta vulgaris <400> 290

atggatacgg gttcaaatta tccgaccgtt aaggggtcatgggttgtcgg attctttaaa tgggctgaaa tttgggcaaaggtggtgttg gaacttccgg caagtcttcc gtcgccggaaagggccggaa aaggggtggt gcaaagtggg caaccaccaaaagatagatc ttagtgatgc taaaacttat tattctaggctctaaatctt ctgttgttat tgttgctggt cttgagcaacagatttcatc ggcttcctga gtttgaccaa gggaaacgaaggtcataatg agcgtcgaag aaaaccacca cctgggtcttcgtctctctt catccctttt tggtgataac accggcggaattctcttcgt atccacggca ttctg <210> 291 <211> 188 <212> PRT

<213> Beta vulgaris <400> 291

Met Asp Thr Gly Ser Asn Tyr Pro Thr Val Lys15 10

Ser Ser Ser Ser Gly Read Asp Be Asp Read Asn

20 25

Gln Lys Ile Tyr Phe Glu Asp Val

35 40

Ser Ser Val Wing Gly Ser Gly Wing Gly Pro Wing

50 55

Gly Val Val Gln Ser Gly Gln Pro Pro Arg Cys65 70 75

Lys Ile Asp Read To Be Asp Wing Lys Thr Tyr Tyr

His Thr Lys Ser300 Wing

Ile Asn Gln Tyr Pro320

Leu Val Val Wing Pro335

Asn Ile Pro Wing His350

catcaacctcaaatatacttgtggtggtgcggtgtcaagtataaagtttggtttttgccagttgtcgcagtgttatcatcgtggtggatt

ttcatcatcttgaagatgtgtccggcgaaaagaagggtgttggtatgcacgcagtgcagcacgccttgctacgtttgggacttattggac

85

90

Cys Gly Met His Ser Lys Ser Ser Val Val Ile

100

105

Gln Arg Phe Cys Gln Gln Cys Be Arg Phe His

115 120

Asp Gln Gly Lys Arg Be Cys Arg Arg Arg Read

130 135

Arg Arg Arg Lys Pro Pro Gly Ser Leu Leu145 150 155

Arg Read Be Be Be Read Phe Gly Asp Asn Thr

165 170

Phe Leu Read Asp Phe Be Ser Tyr Pro Arg His180 185

<210> 292 <211> 536 <212> DNA

<213> Hevea brasiliensis <400> 292

atggaaatgg gttcgggctc tttgacagag tcaggtacctgctgagtcaa taaatgggtt gaaatttggc caaaaaatctaagactccgg ccaaatctgc acccgggtct tcatcttccgaaggttcacg gtgggcagca gcagcagcca cccaggtgtcgatctgagtg atgctaaggc ttattattcg aggcacaaagtctcctaagg tcattgttgc tggtttggag caaagatttt

Gly Ser Be Being Thr 15 Gly Leu Lys Phe Gly 30 Gly Thr Be Gly Lys 45 Lys Arg Wing Gly Lys60 Gln Val Glu Gly Cys 80Ser Arg His Lys Val 95 Val Wing Gly Leu Glu 110 Arg Leu Pro Glu Phe 125 Wing Gly His Asn Glu140 Ser Be Arg Read Le Gly 160Gly Gly Ser Gly Gly 175 Ser

60120180240300360420480540565

ccaacgccac ttctccacct 60attttgagaa tgcgggggct 120ggtccggggc cccgtccagg 180aagttgaggg atgtaaagtg 240tttgtggtat gcactctaag 300gccagcagtg tagtagattt 360catcagcttc ctgaatttga ccaaggaaaa cgaagttgccaatgaacggc ggaggaagcc accaactgga tcagtgctgtttttcaacaa tttttgataa cagcagccga gctgggggca <210> 293 <211> 177 <212> PRT

<213> Hevea brasiliensis <400> 293

gcagacgcct agctggtcat 420catctcgcta taacagactt 480ttcttgtgga tttcag 536

20

25

35

40

50

55

65

70

85

100

105

115

120

130

135

145

Asp

150

165

Glu Be Gly Thr Be Asn Ala10 15 Gly Leu Lys Phe Gly Gln Lys 30 Thr Pro Wing Lys Be Ala Pro 45 Pro Be Arg Lys Val His Gly 60 Gln Val Glu Gly Cys Lys Val 75 80Ser Arg His Lys Val Cys Gly90 95 Val Ala Gly Leu Glu Gln Arg 110 Gln Leu Pro Glu Phe Asp Gln 125 Wing Gly His Asn Glu Arg Arg 140 Be Ser Arg Tyr Asn Arg Leu 155 160Arg Wing Gly Gly Ile Leu Val170 175

<210> 294

<211> 1130

<212> DNA

<213> Oryza sativa

<400> 294

catgcggcta atgtagatgc tcactgcgcttatacaaagc tgtaatactc gtatcagcaacaggattaga aaaacgggac gacaaatagtcatggcaata taaatggaga aatcacaagagtactcgtac gtaaaaaaaa gaggcgcattgatttgaaac cactcaaatc caccactgcatagaaagcac aggtaagaag cacaacgcccgcacctcgcg gatcggtgac gtggcctcgcacaccccggg cccacccacc tgtcacgttgcaaaatatca acgcggcgcg gcccaaaattgccgctcttc cacccaggtc cctctcgtaatgtacgtggg cgggttaccc tgggggtgtgcggccccgcg cgtcatcgcg gggcggggtggaaaattcaa attaggaggt ggggggcgggtgcccctgac ttggcttggc tcctcttcttttctctcctc tcctcttctc ttctcttctcctccttcctc ctctcctttc ccctcctctcgactctcccc aggtgaggtg agaccagtctgcctcgcgcg gatctgaccg cttccctcgg <210> 295 <211> 2194 <212> DNA

<213> Oryza sativa <400> 295

aatccgaaaa gtttctgcac cgttttcaccaaatataaaa tgagacctta tatatgtagc

agtagtaagggagagaggcaaatggaaaaaggaacagaatcatgtgtggaaaccttcaaatcgctctccacccccaaaaagcacatgttgtccaaaatcctccataatggggtggatgactagcgggtgcggccggctctctctgt

tactccagtacacaagttgtcaaaaaaaaaccgggcaatacagcgtgcagcgaggccatgccctcccacctatcccgcggggttatggttccgcccaagcccgtgtgtaccgggtggggcccgaaagggagaggaggaggaggaggag

cattatggaaagcagtagcacaaggaaacacgctgcgaaacagaagcagggtttgaagcacaatcgcgaccgtgaagctgccggccgcaccctggcgcgtctcggctggtggaggaggtccgatcggacaccggcccctccccctgcccct

60120180240300360420480540600660720780840900960102010801130

ccctaactaa caatataggg aacgtgtgctgctgataact agaactatgc aagaaaaact

60120catccaccta 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> 296 <211> 1149 <212> DNA <213> Zea mays <220>

<221> misc_feature <222> (157) .. (157) <223> η is a, c, g, or t <220>

<221> misc_feature <222> (894) .. (894) <223> η is a, c, g, or t <220>

<221> misc_feature <222> (954) .. (954) <223> η is a, c, g, or t <400> 296

atgcacaact gcaaagaaac cagcaagaat gcgccggcgg tttatagttc cccgctactg 60

caccgctctc tgctgcaggc tgcagctttg actaccaact catacagtac tagtactata 120

ctagagtact actacgccat tagcacggtg gttgcantag gccgtggtct ggaggtctct 180

ctcagctcag acgaggatcg tcggagcaag cagcagcgcg tgcagcctct gggagaccct 240

ggcggcgtcg acgccgatcc tggggctgag cgcctgcgag tggacggtgt agaatatctt 300

gtccccgatg acggagaagc tggcgctgac cacctcggcg ccctcctgct ccagcacggt 360

gatgacctcg tgcagcttga agggcctggc ggcctcgctg atgagcacca cgtccagcgt 420

cccgtcctgg caccgcacct cgatgaccgg catgcgcacg cctccgccgc cgccggtgga 480

cccggccgcc gccgttgcag cctccgtggc cgcgccgccg ccgttgcagc agcccgcctt 540

ccgctgcttc agcgcctcga tccgctcctt gagctgcttg atgtacgccg ccgcgctgtc 600

cagctggtcc agctgcgtca ccgcgtcctg cttgttgccg ggattggagg acaccgccgc 660cgacgcggcg tcggagagga gggaggcgtg cgtggcggcg gcggcggcgg cgggagggat 720gagggaggag agcttgaggc agaggccctt catgtgcagc ctccggttct tctccacgtc 780cttcctctcc agcttgcacc ccccgccgct gctgtgcgcg ttcctctccc ccgcggcgca 840ggcgcccgag ctgcccccgc tgctctgcct ccggctcttc atctcgatcg accngaccgg 900tctgcagatc tctccttggc tgttctcgtc gctgctcctg ccgggcgtga agcngcgcag 960cctttggttg ggacttggga gggacaagtt gcaacagcac ccacgggcca cggcacggag 1020ggggcgaaag gcaaggaggc caggacaaga cgagagaaat acaggccggg ggagatggct 1080ccgtggcgcg tacgtgtgtc tacctgcatg ttggttgatc cgattgcatc tgctgtaacc 1140atatattaa 1149

<210> 297 <211> 382 <212> PRT <213> Zea mays <220>

<221> UNSAFE <222> (53) .. (53)

<223> Xaa can be any naturally occurring amino acid <400> 297

Met His Asn Cys Lys Glu Thr Ser Lys Asn Pro Wing Pro Val Tyr Ser15 10 15

Be Pro Read Leu Read His Arg Be Read Leu Gln Wing Wing Wing Read Thr Thr

20 25 30

Asn Be Tyr Be Thr Be Thr Ile Read Glu Tyr Tyr Tyr Wing Ile Be

35 40 45

Thr Val Val Wing Xaa Gly Arg Gly Leu Glu Val Ser Leu Ser Ser Asp

50 55 60

Glu Asp Arg Arg Be Lys Gln Gln Arg Val Gln Pro Read Gly Asp Pro65 70 75 80

Gly Gly Val Asp Wing Asp Pro Gly Wing Glu Arg Leu Arg Val Asp Gly

85 90 95

Val Glu Tyr Leu Val Pro Asp Asp Gly Glu Wing Gly Wing Asp His Leu

100 105 110

Gly Wing Leu Leu Leu Gln His Gly Asp Asp Leu Val Gln Leu Glu Gly

115 120 125

Pro Gly Gly Leu Wing Asp Glu His His Val Gln Arg Pro Val Leu Wing

130 135 140

Pro His Leu Asp Asp Arg His Wing His Wing Be Wing Wing Wing Gly Gly145 150 155 160

Pro Gly Arg Arg Arg Cys Be Read Arg Gly Arg Wing Wing Wing Wing Val Wing

165 170 175

Wing Wing Arg Leu Pro Leu Leu Gln Arg Leu Asp Pro Leu Leu Glu Leu

180 185 190

Leu Asp Val Arg Arg Arg Wing Val Gln Leu Val Gln Leu Arg His Arg

195 200 205

Val Leu Leu Val Wing Gly Ile Gly Gly His Arg Arg Arg Arg Gly Val

210 215 220

Gly Gly Gly Gly Gly Gly Gly Val Arg Gly Gly Gly Gly Gly Gly Gly Gly Arg Asp225 230 235 240

Glu Gly Gly Glu Leu Glu Wing Glu Wing Read His Val Gln Pro Pro Val

245 250 255

Leu Leu His Val Leu Pro Leu Gln Leu Pro Pro Wing Val Wing Wing

260 265 270

Arg Val Pro Leu Pro Arg Gly Wing Gly Wing Arg Wing Wing Pro Wing Wing

275 280 285

Leu Pro Pro Wing Leu His Leu Asp Arg Pro Asp Arg Be Wing Asp Leu

290 295 300

Ser Leu Val Wing Leu Val Wing Pro Wing Gly Arg Wing Glu Wing Wing Gln305 310 315 320

Pro Read Val Gly Thr Trp Glu Gly Gln Val Wing Thr Wing Pro Thr Gly

325 330 335

His Gly Thr Glu Gly Wing Lys Gly Lys Glu Wing Arg Thr Arg Arg Glu

340 345 350

Lys Tyr Arg Pro Gly Glu Met Pro Wing Trp Arg Val Arg Val Val Ser Thr355 360 365

Cys Met Read Val Asp Pro Ile Wing Be Val Val Wing Ile Tyr

370 375 380

<210> 298 <211> 660 <212> DNA <213> Zea mays <400> 298

atggagatga gagcgaagaa gagcagcaga agcagcacga gcaacaccagacgaccacgg cggtggagag gaaggagatc gagaggagga ggaggcagcactctgcgcca agctcgcctc cctcatcccg aaagaacact actcctccaaacccagctgg gtagcctaga cgaggcagcc acgtacataa agagactcaagaggagctgc ggcacaagag cgcctctgca cggctcttgg ccgctggcagcgaggcggag gaggaggagg cgcctccaca tcgtcggcag cgacgaccacggcgcaggat catctgaaga agccggccgg cgggaggacg acatgccgccgaggttcggc agcacaatga cgggtcaagc ctggacgtgg tgctcatcagcgacccttca agctgcacga ggtggtcacc gtgctggagg aagaaggcgcaacgccaacc tctccgtcgc cggcaccaaa atcttctaca ccatccactgtgcccaagaa tcggtataga tgtttcaaga gtttctgaaa gattaagagc <210> 299 < 211> 219 <212> PRT <213> Zea mays <400> 299

Met Glu Met Arg Wing Lys Lys Ser Ser Arg Ser

cggcagcacggatgaagagcggatgctatgggagagggtgtggccggtggcggtggtacagcgcgcgcgagaccgaccaaggcctttattgggatga

To be thr

Be Gly Be Thr Thr Thr Thr Wing Val Glu Arg Lys Glu

10

20

25

Arg Arg Arg Gln Gln Met Lys Being Read Cys Wing

35

40

Ile Pro Lys Glu His Tyr Being Ser Lys Asp Wing

50

55

Be Read Asp Glu Wing Wing Thr Tyr Ile Lys Arg

Lys Leu

45Met Thr60

Read lys

65

70

75

Glu Glu Leu Arg His Lys Be Wing Be Wing Arg Leu Leu

Ala Wing

85

90

Be Gly Thr Arg Arg Gly Gly Gly Gly

100 105

Wing Wing Thr Thr Wing Wing Ser Gly Gly Wing Gly Ser Ser115 120 125

Gly Arg Arg Glu Asp Asp Pro Pro Val Wing Val Glu

130 135 140

His Asn Asp Gly Ser Ser Leu Asp Val Val Le Le Ile Ser145 150 155

Arg Phe Lys Pro Leu His Glu Val Val Thr Val Leu Glu

165 170

Glu Thr Wing Asp Asn Wing Asn Leu Ser Val Val Gly Thr Wing

180 185

Tyr Thr Ile His Cys Lys Phe Cys Pro Wing Arg Ile Gly195 200 205

Ser Arg Val Ser Glu Arg Leu Arg Wing Leu Gly

210 215

<210> 300 <211> 669 <212> DNA <213> Glycine max <400> 300

atgggtcaac aacaacaggg aagtcaacct tctccaacca aagtcgaaaggagaaaaaca ggagaagcca gatgaagaac ctctattccg aactcaactcacccgtaatc ccaaggaagc gatgtcactg cctgatcaaa tagatgaagcatcaaaagcc tagagacaaa agtgaagctg gagcaggaga agaaagaaagaggaagagaa ctcgtggtgg ctgttcgagt tcttctgaag cacaaggaagccaaatatcc agattcacga aacgggaaat ttgcttgaag tcattctaac

Be Asn Thr15

Ile Glu Arg30

Ala Ser Leu

Gln Leu Gly

Glu Arg Val80

Wing Wing Gly95

Thr Ser Ser110

Glu Glu Wing

Val Arg Gln

Be Wing Wing160

Glu Glu Gly175

Lys Ile Phe190

Ile Asp Val

aaagattgtatcttctccctaatcaactatgttaaaggaacctgaaatcgatgtggggtc

60120180240300360420480540600660

60120180240300360gatagccagt tcatgttctg tgaaattatt cgaattttgc atgaagagaa cgtcgaggtc 420

atcaatgcca attcttcaat ggtcggagat ttagtgattc atgttgtgca cggggaggtt 480

gagccatcta tctatcaatt cggagcgacc aaagtgagtg agaagctgaa atggtttatg 540

aacggatcct tcagtgatgt ggaaatggag cctgaattaa tgtggaattt taaaattgat 600

gctactgagc cgtgggggct tctagatgat cttacactgg acaatgtctt accaccaaat 660

actttgtaa 669

<210> 301

<211> 222

<212> PRT

<213> Glycine max

<400> 301

Met Gly Gln Gln Gln Gln Gly Be Gln Pro Be Pro Thr Lys Val Glu1 5 10 15

Arg Lys Ile Val Glu Lys Asn Arg Arg Be Gln Met Lys Asn Leu Tyr

20 25 30

Be Glu Read Asn Be Read Leu Pro Thr Arg Asn Pro Lys Glu Ala Met

35 40 45

Ser Leu Pro Asp Gln Ile Asp Glu Wing Ile Asn Tyr Ile Lys Ser Leu

50 55 60

Glu Thr Lys Val Lys Leu Glu Gln Glu Lys Lys Glu Arg Leu Lys Glu65 70 75 80

Arg Lys Arg Thr Arg Gly Gly Cys Be Be Be Be Glu Wing Gln Gly

85 90 95

Be Leu Lys Be Pro Asn Ile Gln Ile His Glu Thr Gly Asn Leu Leu

100 105 110

Glu Val Ile Leu Thr Cys Gly Val Asp Being Gln Phe Met Phe Cys Glu

115 120 125

Ile Ile Arg Ile Read His Glu Glu Asn Val Glu Val Ile Asn Wing Asn

130 135 140

Ser Ser Met Val Gly Asp Leu Val Ile His Val Val His Gly Glu Val145 150 155 160

Glu Pro Ser Ile Tyr Gln Phe Gly Wing Thr Lys Val Ser Glu Lys Leu

165 170 175

Lys Trp Phe Met Asn Gly Be Phe As Asp Val Glu Met Glu Pro Glu

180 185 190

Leu Met Trp Asn Phe Lys Ile Asp Wing Thr Glu Pro Trp Gly Leu

195 200 205

Asp Asp Leu Thr Leu Asp Asn Val Leu Pro Pro Asn Thr Leu210 215 220

Claims (139)

Method for enhancing plant-related traits relative to control plants, characterized in that it preferably comprises increasing expression of a nucleic acid encoding an HAL3 polypeptide in a plant bud, and optionally selecting plants having increased yield. Method according to claim 1, characterized in that said increased expression is effected by introducing genetic modification, preferably at the locus of a gene encoding an HAL3 polypeptide. Method according to claim 2, characterized in that said genetic modification is effected by any one or more of: T-DNA5 TILLING activation and homologous recombination. Method according to claim 2, characterized in that said genetic modification is effected by any one or more of: T-DNA activation, TILLING and homologous recombination. A method according to claim 1, characterized in that said increased expression is effected by introducing and expressing a plant a nucleic acid encoding an HAL3 polypeptide. A method according to claim 5, wherein said nucleic acid encoding a HAL3 protein is represented by any of the nucleic acids SEQ ID NOs given in Table A, or a portion thereof, or a sequence capable of hybridizing to any of the acids. SEQ ID NOs given in Table A. Method according to claim 5, characterized in that said nucleic acid encoding a HAL3 protein encodes aortologist or paralog of any of the nucleic acids SEQ ID NOs given in Table A. Method according to any one of claims 1 to 7, characterized in that said improved performance-related characteristics comprise increased yield and / or early vigor. Method according to claim 8, characterized in that said increased yield is increased seed yield. A method according to claim 9, characterized in that said increased seed yield is one or more of: a) increased seed biomass; b) increased harvest index; and c) increased number of seeds (floods). A method according to claim 9, characterized in that said increase in seed yield is at least 10% relative to control plants. A method according to any one of claims 5 to 11, characterized in that said nucleic acid encoding a HAL3 protein is operably linked to a bud-specific promoter, preferably to a beta expansin promoter. A method according to any one of claims 5 to 12, characterized in that said nucleic acid encoding a HAL3 protein is of plant origin, preferably from a dicotyledonous plant, still preferably from the Brassicaceae family, more preferably Arabidopsis, more preferably from Arabidopsis. Thaliana. Plant or part thereof, including seeds, characterized in that it is obtainable by a method as defined in any preceding claim, wherein said plant or part thereof comprises a recombinant nucleic acid encoding a HAL3 polypeptide. 15. A construct, characterized in that it comprises: (a) a nucleic acid encoding an HAL3 polypeptide, (b) one or more control sequences capable of directing the nucleic acid sequence of (a) preferentially budded bud of a plant; and optionally (c) a transcription termination sequence. Construction according to claim 15, characterized in that said one or more control sequences is at least one shoot-specific promoter, preferably an expansin debeta promoter. Use of a construct as defined in claim 15 or 16 for enhancing plant yield characteristics, preferably for increasing yield and / or hearing, relative to control plants. Use according to claim 17, characterized in that said yield is increased seed yield. Use according to claim 17, characterized in that said vigor is early vigor. 20. Plant, plant part or plant cell, characterized in that it is transformed with the construction as defined in claims 15 or 16. 21. A method for producing a transgenic plant with improved yield-related characteristics compared to control plants, characterized in that it comprises: (i) preferentially introducing and enhancing bud expression of a plant of a nucleic acid encoding a HAL3 polypeptide; and (ii) cultivate the plant cell under conditions that promote plant growth and development. Transgenic plant, characterized by the fact that it has improved yield-related characteristics compared to control plants, said improved yield-related characteristics resulting from increased expression of a HAL3 polypeptide preferably in the bud. Transgenic plant according to claim 14, 20 or -22, characterized in that said plant is a cultivation plant or a monocotyledonea or a cereal, such as rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats. Harvestable parts of a plant as defined in any one of claims 14, 20, 22 or 23, characterized in that said harvestable parts are preferably seeds. Products, characterized in that they are derived from a plant as defined in claim 23 and / or harvestable parts of a plant as defined in claim 24. Use of a nucleic acid encoding a HAL3 polypeptide, whose nucleic acid is operably linked to a bud-specific promoter, characterized in that it is for improving plant yield traits. Use according to claim 26, characterized in that said yield-related characteristics include seed yield and / or early vigor. A method for improving plant yield characteristics, comprising comprising modulating, preferably increasing, expression in a plant of a nucleic acid encoding a MADS15 protein. A method according to claim 28, characterized in that said MADS15 protein comprises any one or more of the following reasons: SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQID NO: 51. A method according to claims 28 to 29, characterized in that said modulated expression is effected by any one or more identification by T-DNA activation, TILLING, site directed mutagenesis, directed evolution or homologous recombination. A method according to any one of claims 28 to 29, characterized in that said modulated expression is effected by introducing and expressing in a plant a nucleic acid encoding protein MADS15, and wherein the amino acid sequence of said protein MADS15 comprises any one or more for the following reasons: SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51. The method of claim 31, wherein said nucleic acid encoding a MADS15 protein is any of the SEQ LD NO nucleic acids given in Table D or a portion thereof or a sequence capable of hybridizing to any of the SEQ ID NO nucleic acids. data in Table D. A method according to claim 31, characterized in that said nucleic acid encoding a MADS15 protein encodes an ortholog or paralog of any of the SEQ IDNOs nucleic acids given in Table D. A method according to any one of claims 28 to 33, characterized in that said improved yield related characteristic is root biomass. A method according to any one of claims 31 to 34, characterized in that said nucleic acid encoding a MADS15 protein is operably linked to a constitutive promoter, preferably to a GOS2 promoter. A method according to any one of claims 31 to 35, characterized in that said nucleic acid encoding a MADS15 protein is of plant origin, preferably from a plantamonocotyledon, still preferably from the Poaceae family, preferably from the genus Oryza, more preferably from Oryza. sativa. Plant or part thereof, including seeds, characterized in that it is obtainable by a method as defined in any one of claims 28 to 36, wherein said plant or part thereof comprises a recombinant nucleic acid encoding a MADS15 protein. 38. Construction, characterized in that it comprises: (a) nucleic acid encoding a MADS15 protein, wherein the amino acid sequence of said MADS15 protein comprises any or more of the following reasons: SEQ ID NO: 48, SEQ ID NO: 49, SEQ (B) one or more control sequences capable of directing nucleic acid sequence expression of (a); and optionally (c) a transcription termination sequence. Construction according to Claim 38, characterized in that said one or more control sequences is at least one constitutive promoter, preferably a GOS2 promoter. Use of a construction as defined in claim 38 or 39, characterized in that it is for making plants having improved yield-related characteristics, particularly increased root yields, relative to control plants. 41. Plant, plant part or plant cell, characterized in that it is transformed with a construction as defined in claim 38 or 39. 42. A method for producing a transgenic plant with improved yield-related traits compared to control plants, characterized in that it comprises: (i) introducing and expressing into a plant a nucleic acid encoding a MADS15 protein, in which the amino acid sequence deduces The MADS15 protein comprises any one or more of the following motives: SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, and (ii) growing the plant cell under conditions that promote plant growth and development. . 43. Transgenic plant, characterized by the fact that increased yield over control plants, said increased yield resulting from increased expression of a nucleic acid encoding a MADS15 protein, wherein the amino acid sequence of the MADS15 protein comprises any one or more of the following: SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, or a transgenic plant cell derived from said transgenic plant. Transgenic plant according to claim 37, 41 or 43, characterized in that said plant is a cultivation plant or a monocotyledonea or a cereal such as rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats, or a transgenic plant cell derived from said transgenic plant. 45. Harvestable parts of a plant as defined in claim 44, characterized in that said harvestable parts are preferably roots. Products, characterized in that they are derived from a plant as defined in claim 44 and / or harvestable parts of a plant as defined in claim 45. 47. Use of a nucleic acid encoding a MADS15 protein, characterized in that it is for improving plant yield characteristics, wherein the amino acid sequence of the MADS15 protein comprises any one or more of the following motives: SEQ ID NO: 48, SEQ ID NO : 49, SEQ ID NO: 50, SEQ ID NO: 51. Use according to claim 47, characterized in that said improved yield-related trait is increased root biomass relative to control plants. 49. A method for improving yield-related characteristics in plants over suitable control plants, characterized in that it preferably comprises reducing expression of an endogenous MADS15 gene in said plant. A method according to claim 49, characterized in that said MADS15 protein comprises any one or more of the following reasons: SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ED NO: 117. A method according to claim 49 or 50, wherein said reduced expression is effected by RNA-mediated downregulation of gene expression. The method of claim 51, characterized in that said RNA-mediated down-regulation is effected by co-suppression. A method according to claim 51, characterized in that said RNA-mediated down-regulation is effected by using MADS15 antisense nucleic acid sequences. A method according to claim 49 or 50, characterized in that said reduced expression is effected using an inverted repeat of a MADS15 gene or fragment thereof, preferably capable of forming a staple structure. A method according to claim 49 or 50, characterized in that said reduced expression is performed using nucleotide specificity MADS15. A method according to claim 49 or 50, characterized in that said reduced expression is effected by insertion mutagenesis. A method according to any one of claims 49 to 56, characterized in that said endogenous MADS15 gene is an MADS15 gene as found in a plant in its natural form or is an isolated MADS15 nucleic acid subsequently introduced into a plant. A method according to claim 57, characterized in that said introduced MADS15 nucleic acid is operably linked to a constitutive promoter, preferably a GOS2 promoter. A method according to claim 57 or 58, wherein said introduced MADS15 nucleic acid is from a plant source or artificial source, preferably wherein plants are transformed with an MADS15 nucleic acid homologous to the endogenous MADS15 gene, more preferably in which sequences. MADS15 nucleic acid from monocotyledonous plants are used for transformation of monocotyledonous plants, and preferably in which Poaceae family MADS15 sequences are used for transformation into Poaceae family plants, more preferably in which MADS15 rice nucleic acid sequences are used to transform rice plants. rice. A method according to claim 59, characterized in that said rice MADS15 nucleic acid sequence comprises a sufficiently substantially contiguous nucleotide length of SEQ ID NO: 109 (OsMADS 15) or comprises a sufficient length of substantially contiguous nucleotides of a sequence of. nucleic acids encoding an ortholog or analog of OsMADS 15 (SEQ ID NO: 110). 61. The method according to claim 60, characterized in that said OsMADS 15 orthologs or paralogues are represented by SEQ ID NO: 147, SEQ ID NO: 149, SEQ ID NO: 151, SEQ ID NO: 153, SEQ ID NO. : 155, SEQ ID NO: 157, SEQ ID NO: 159 and SEQ ID NO: 161. A method according to claim 60 or 61, characterized in that said substantially contiguous nucleotides of the nucleic acid sequence encoding an OsMADS 15 ortholog or parallel (SEQ LD NO: 110) are substantially contiguous nucleotides of nucleic acid sequences represented by SEQ. ID NO: 146, SEQ ID NO: 148, SEQ ID NO: 150, SEQ ID NO: 152, SEQ ID NO: 154, SEQ ID NO: 156, SEQ ID NO: 158 and SEQ ID NO: 160. A method according to any one of claims 49 to 62, characterized in that said improved yield related characteristics comprise increased yield, preferably increased seed yield and / or increased biomass. A method according to claim 63, characterized in that said increased seed yield is selected from one or more of the following: (a) increased seed biomass (seed weight); b) increased number of flowers per plant; and c) increased number of seeds (floods). Plant or part thereof, including seeds, characterized in that it is obtainable by a method as defined in any one of claims 49 to 64, wherein said plant or part of plant comprises a recombinant MADS15 nucleic acid. 66. Construction, characterized in that it comprises: (i) an MADS15 nucleic acid capable of silencing an endogenous MADS15 gene, (ii) one or more control sequences capable of directing (a) nucleic acid sequence expression; and optionally (iii) a transcription termination sequence. Construction according to claim 66, characterized in that said one or more control sequences is at least one constitutive promoter, preferably a GOS 2 promoter. Use of a construct as defined in claim 66 or 67, characterized in that it is for making plants having improved yield characteristics, particularly increased seed yield and / or biomass, relative to control plants. 69. Plant, plant part or plant cell characterized by being transformed with the construction as defined in claim 66 or 67. 70. A method for producing a transgenic plant with improved yield-related traits relative to control plants, comprising: (i) introducing and expressing in a plant a MADS15 nucleic acid capable of silencing an endogenous MADS15 gene; and (ii) cultivate the plant cell under conditions that promote plant growth and development. 71. Transgenic plant, characterized by the fact that it has improved yield-related characteristics compared to control plants, said improved yield-related characteristics resulting from decreased expression of a MADS15 protein-encoding nucleic acid, wherein the amino acid sequence of said MADS15 protein comprises either or more of the following reasons: SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117; or a transgenic plant cell derived from said transgenic plant. Transgenic plant according to claim 65, 69 or 71, characterized in that said plant is a cultivated plant or a monocotyledonea or a cereal, such as rice, maize, wheat, barley, millet, rye, triticale, sorghum. and oats, or a transgenic plant cell derived from said transgenic plant. 73. Harvestable parts of a plant as defined in claim 72, characterized in that said harvestable parts are preferably seeds. Products, characterized in that they are derived from a plant as defined in claim 72 and / or harvestable parts of a plant as defined in claim 73. 75. Use of a MADS15-encoding nucleic acid to improve plant yield characteristics, characterized in that the amino acid sequence of said MADS15 protein comprises any or more of the following reasons: SEQ ID NO: 114, SEQ ID NO: 115, SEQ ID NO: 116, SEQ ID NO: 117. Use according to claim 75, characterized in that said improved yield characteristics comprise increased seed yield. 77. A method for increasing plant yield over suitable control plants, comprising increasing expression in a plant of a nucleic acid encoding a PLT transcription factor polypeptide, and optionally selecting increased plant yield. 78. Method for increasing abiotic stress resistance in relation to control plants, characterized in that it comprises increasing expression in a plant of a nucleic acid by encoding a PLT transcription factor polypeptide, and optionally selecting plants having increased abiotic stress resistance. 79. The method of claim 78, characterized in that said increased resistance to abiotic stress is increased nutrient absorption efficiency relative to control plants. A method according to any one of claims 77 to 79, characterized in that said increased expression is effected by introducing and expressing into a plant, plant part or plant cell a nucleic acid encoding a PLT transcription factor polypeptide. 81. The method of claim 80, wherein said nucleic acid encoding a transcription factor PLT is any of the SEQ ID NO nucleic acids given in Table I or a portion thereof or a sequence capable of hybridizing to any of the SEQ nucleic acids. ED NOs given in Table I. A method according to claim 80, characterized in that said nucleic acid encoding a transcription factor PLT encodes an ortholog or paralog of any of the nucleic acids SEQ IDNOs given in Table D. A method according to any of claims 77, 80, 81 and 82, characterized in that said increased yield is increased seed yield. A method according to claim 83, characterized in that said seed yield is selected from one or more of: a) increased seed biomass; b) increased number of flowers per plant; c) increased number of seeds (floods); d) increased grain filling rate; e) increased harvest index; f) Increased Mill Seed Weight (TKW) g) increased seed area; h) increased seed length; and i) increased seed width. A method according to any one of claims 80 to 84, characterized in that said nucleic acid is operably linked to a constitutive promoter, preferably a GOS2 promoter. A method according to any one of claims 80 to 84, characterized in that said nucleic acid encoding a PLT transcription factor polypeptide is of plant origin, preferably from a dicotyledonous plant, still preferably from the Brassicaceae family, more preferably from the genus Arabidopsis. more preferably Arabidopsis thaliana. A plant or part thereof, including seeds, characterized in that it is obtainable by a method as defined in any one of claims 77 to 86, wherein said plant or part thereof comprises a nucleic acid encoding a PLT transcription factor polypeptide whose Nucleic acid is operably linked to a constitutive engine. 88. Construction comprising: (i) a nucleic acid encoding a PLT transcription factor polypeptide (ii) one or more control sequences capable of directing the nucleic acid sequence of (i); and optionally (iii) a transcription termination sequence. Construction according to claim 88, characterized in that said constitutive promoter is a GOS2 promoter. Use of a construct as defined in claim 88 or 89 characterized in that it is for making plants having increased yield, particularly increased seed yield and / or increased resistance to abiotic stress relative to control plants. 91. Plant, plant part or plant cell, characterized in that it is transformed with the construction as defined in claim 88 or 89. 92. A method for producing a transgenic plant with increased bias and / or increased resistance to abiotic stress, comprising: (i) introducing and expressing a nucleic acid encoding a PLT transcription factor polypeptide in a plant cell; and (ii) cultivate the plant cell under conditions that promote plant growth and development. 93. Transgenic plant, characterized by the fact that increased terrestrial and / or increased resistance to abiotic stress relative to control plants, resulting from increased expression of a nucleic acid encoding a transcription factor polypeptide PLT5 or plant cell derived from said transgenic plant. Transgenic plant according to claim 87, 91 or-93, characterized in that said plant is a crop plant or a monocotyledonea or a cereal, such as rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats, or a transgenic plant cell derived from said transgenic plant. 95. Harvestable parts of a plant as defined in claim 94, characterized in that said harvestable parts are preferably seeds. Products, characterized in that they are derived from a plant as defined in claim 94 and / or harvestable parts of a plant as defined in claim 93. 97. Use of a nucleic acid encoding a PLT transcription defactor polypeptide, or use of a PLT transcription factor polypeptide, characterized in that it is to increase resistance to biotic stress and / or to improve plant yield, especially seed yield, relative to to control plants. 98. A method for improving plant yield characteristics, comprising modular plant expression of a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ SEQ ID NO: 225, SEQ ID NO: -297, SEQ ID NO: 299, SEQ ID NO: 301, or orthologs or paralogs of any of those mentioned above. 99. A method according to claim 98, characterized in that said modulated expression is effected by any one or more identification by T-DNA activation, TILLING, or homologous recombination. 100. The method of claim 98, characterized in that said modulated expression is effected by introducing and expressing into a plant a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, or orthologs or dialogues of any of the above mentioned SEQ ID NOs. Method according to any one of claims 98 to 100, characterized in that said improved yield related characteristic is early vigor. 102. The method of claim 100, wherein said nucleic acid is operably linked to a conductive promoter, preferably to a GOS2 promoter. A method according to any one of claims 100 to 102, characterized in that said nucleic acid is a portion of or an allelic variant of or a joining variant of or a sequence capable of hybridizing to a sequence of nucleic acids according to any one of SEQ ID NO: 212, SEQ ID NO: 214, SEQ ID NO: 216, SEQ ID NO: 218, SEQ ID NO: 220, SEQ ID NO: 222, SEQ ID NO: 224, SEQ ID NO : 226 and SEQ ID NO: 228, SEQ ID NO: 296, SEQ ID NO: 298, SEQ ID NO: 300, wherein said portion, allelic variant, splice variant, or hybridization sequence encodes a bHLH transcription factor. A method according to any one of claims 98 to 103, characterized in that said nucleic acid encoding a bHLH transcription factor is of plant origin, preferably from a plantamonocotyledon, still preferably from the Poaceae family, most preferably from the genus Oryza, more preferably from the genus Oryza. preferably from Oryza sativa. Plant or part thereof including seeds, characterized in that it is obtainable by a method as defined in any one of claims 98 to 104, wherein said plant or part thereof comprises a nucleic acid encoding a transcription factor bHLH. 106. Isolated polypeptide characterized in that it comprises: (i) an amino acid sequence represented by any one of SEQ ED NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301, (ii) an amino acid sequence having (a) preferably ascending at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity with any of the amino acid sequences represented by SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301, and (b) a bHLH domain; (iii) derived from any of the amino acid sequences given in ( i) or (ii) above. 107. Isolated nucleic acid molecule, characterized in that it comprises: (i) a nucleic acid represented by any one of SEQID NO: 296, SEQ ID NO: 298 and SEQ ID NO: 300 (ii) the complement of a nucleic acid represented by any one of SEQ ID NO: 296, SEQ ID NO: 298 and SEQ ID NO: 300 (iii) a nucleic acid encoding a transcription factor bHLH having (a) in ascending order preferably at least 50%, 55% , 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or more sequence identity with the amino acid sequence represented by any one of SEQ ID NO: 297, SEQ ID NO: 299 and SEQ ID NO: 301, and (b) a bHLH domain. 108. Construction comprising: (a) nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225 , SEQ ID NO: 297, SEQ ID NO: - 299, SEQ ID NO: 301, or orthologs or paralogues of any of the above-mentioned SEQ IDNOs: (b) one or more control sequences capable of directing the nucleic acid sequence expression of (The); and optionally (c) a transcription termination sequence. 109. Construction according to claim 108, characterized in that said one or more control sequences is at least one constitutive promoter, preferably a GOS2 promoter. 110. Use of a construct as defined in claim 108 or 109, characterized in that it is for making plants with improved yield-related characteristics, particularly early vigor, relative to control plants. 111. Plant, plant part or plant cell, characterized in that it is transformed with a construction as defined in claim 108 or 109. 112. A method for producing a transgenic plant with improved yield-related characteristics compared to control plants, characterized in that it comprises: (d) introducing and expressing in a plant a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID SEQ ID NO 215, SEQ ID NO 217, SEQ ID NO 219, SEQ ID NO: 219, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301, orortologists or paralogues of any of SEQ ID NOs mentioned above; and (e) cultivate the plant cell under conditions that promote plant growth and development. 113. Transgenic plant, characterized by the fact that terrification is increased over control plants, said increased yield resulting from increased expression of a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 215, SEQ ID NO 215, SEQ ID NO : 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: 297, SEQ ID NO: 299, SEQ ID NO: 301 orortologists or paralogues of any of the aforementioned SEQ ID NOs, or a transgenic plant cell derived from said transgenic plant. Transgenic plant according to claim 105, 111 or 113, characterized in that said plant is a crop plant or a monocot or cereal, such as rice, maize, wheat, barley, millet, rye, triticale, sorghum. and oats, or a transgenic plant cell derived from said transgenic plant. 115. Harvestable parts of a plant as defined in claim 114, characterized in that said harvestable parts are preferably seeds. Products, characterized in that they are derived from a plant as defined in claim 114 and / or harvestable parts of a plant as defined in claim 115. 117. Use of a nucleic acid encoding a bHLH transcription factor represented by any of SEQ ID NO 213, SEQ ID NO 215, SEQ ID NO: 217, SEQ ID NO: 219, SEQ ID NO 225, SEQ ID NO: - 297, SEQ ID NO: 299, SEQ ID NO: 301 or orthologs or paralogs of any of the above, characterized in that it is for improving plant yield characteristics. 118. Use according to claim 117, characterized in that said improved yield-related trait is early in force. 119. A method for increasing plant yield over appropriate control plants, characterized in that it comprises increasing expression in a plant of a nucleic acid encoding a SPL15 transcription factor polypeptide, and optionally selecting increased yield plantast. 120. A method according to claim 119, characterized in that said increased expression is effected by any one or more of: T-DNA activation, TILLING, site-directed mutagenesis, directed evolution, and homologous recombination. 121. The method of claim 119, wherein said increased expression is effected by introducing and expressing into a plant a nucleic acid encoding a transcription factor polypeptide SPL15. 122. The method of claim 121, wherein said nucleic acid encoding a transcription factor SP15 is any of the SEQ ID NO nucleic acids given in Table Nou a portion thereof or a sequence capable of hybridizing to any of the SEQ nucleic acids. ID NOs given in Table N. 123. The method of claim 121, characterized in that said nucleic acid encoding a transcription factor SP15 encodes an ortholog or paralog of any of the nucleic acids SEQ ID NOs given in Table N. A method according to any one of claims 119 to 123, characterized in that said increased yield is increased seed yield. 125. The method of claim 124, characterized in that said increased seed yield comprises one or more of: a) increased above-ground biomass b) increased number of flowers per plant; c) increased seed yield; d) increased number of seeds (floods)); e) Increased Thousand Seed Weight (TKW); and f) increased harvest index. 126. A method according to any one of claims 121 to 125, characterized in that said nucleic acid is operably linked to a constitutive promoter, preferably to a HMGB (high mobility group B) promoter or a GOS2 promoter, still preferably to a higher. HMGB rice promoter or a GOS2 rice promoter. A method according to any one of claims 121 to 126, characterized in that said nucleic acid encoding a SPL15 transcription factor polypeptide is of plant origin, preferably from a dicotyledonous plant, still preferably from the Brassicaceae family, more preferably from Arabidopsis thaliana. Plant or part thereof, including seeds, characterized in that it is obtainable by a method as defined in any one of claims 119 to 127, wherein said plant or part thereof comprises a recombinant nucleic acid encoding a SPL15 transcription factor. 129. Construction, characterized in that it comprises: (a) a nucleic acid encoding a SPL15 transcription factor polypeptide, (b) one or more control sequences capable of directing (a) nucleic acid sequence expression; and optionally (c) a transcription termination sequence. Construction according to claim 129, characterized in that said one or more control sequences is at least one constitutive promoter, preferably an HMGB promoter or a GOS2 promoter. Use of a construction as defined in claim 129 or 130, characterized in that it is for making plants having increased yields relative to control plants. 132. Plant, plant part or plant cell, characterized in that it is transformed with a construction as defined in claim 129 or 130. 133. A method for producing a transgenic plant which is enhanced relative to control plants, characterized in that it comprises: (f) introducing and expressing a nucleic acid encoding a SPL15 transcription factor polypeptide in a plant cell; and (g) cultivate the plant cell under conditions that promote plant growth and development. 134. A transgenic plant, characterized by the fact that terrification is increased compared to control plants, said increased yield resulting from increased expression of a nucleic acid encoding a transcription factor SPL1 5. Transgenic plant according to claim 128,132 or 134, characterized in that said plant is a crop plant or a monocot or cereal, such as rice, maize, wheat, barley, millet, rye, triticale, sorghum and oats. , or a transgenic plant cell derived from said transgenic plant. 136. Harvestable parts of a plant as defined in claim 135, characterized in that said harvestable parts are preferably seeds. Products, characterized in that they are derived from a plant as defined in claim 135 and / or harvestable parts of a plant as defined in claim 136. 138. Use of a nucleic acid encoding a transcription factor polypeptide SPL1 5, characterized in that it is for increase in plant yield over control plants. Use according to claim 138, characterized in that said increased yield is increased seed yield.