CN1821406A - Resistance gene Pi 36 of rice blast and its use - Google Patents
Resistance gene Pi 36 of rice blast and its use Download PDFInfo
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Abstract
The present invention discloses the nucleotide sequence, coded amino acid polypeptide sequence and application of one new rice resistance gene Pi36. The gene is member of nonTIR-NBS-LRR resistance gene family, and is one constitutive expression gene. The present invention also relates to the application of the gene in transforming rice or other plant to breed disease resisting variety and the application of the molecular marker generated based on the gene sequence in breeding variety.
Description
Technical field
The present invention relates to gene engineering technology field, be specifically related to a kind of separating clone and application of resistance gene Pi 36 of rice blast.
Background technology
Plant usually is subjected to the infringement of multiple pathogen in the process of growth, plant then takes multiple defence policies to protect self, avoids being subjected to it to attack.A most important defense mechanism is exactly the defence answering system that can discern the existence of obligate pathogenic microorganism and start self in the plant.Plant is mediated by disease-resistant gene the identification of pathogenic bacteria.Therefore, the analysis of disease-resistant gene product structure and research are the bases of understanding plant disease-resistant mechanism, also have important directive significance for the prevention and the control of Plant diseases.
So far, from 3 kinds of monocotyledonss and 5 kinds of dicotyledonss, separated more than 40 disease-resistant gene.To the structure of these disease-resistant genes and discovering of product, though host plant difference, the pathogen of institute's antagonism also has fungi, bacterium, differences such as virus and nematode, but the structure of disease-resistant gene and product have many common constitutional featuress, there is rich leucine tumor-necrosis factor glycoproteins (leucine-rich repeat as the C-end, LRR), there is nucleotide binding site (nucleotide binding site in the N-end, NBS), and leucine zipper (leucine zipper, LZ), coiled coil structural domain (coiled-coil, CC), and membrane spaning domain (transmembrane domain, TM), protein kinase (protein kinase, PK), and fruit bat Toll albumen and Mammals interleukin-1 receptor (Toll andinterleukin-1 receptor, TIR) etc.According to they coded proteic constitutional featuress, disease-resistant gene can be divided into 7 classes (Hammond-Kosack ﹠amp; Jones, 1997; Dangl ﹠amp; Jones 2001; Iyer ﹠amp; McCouch2004).
The first kind, toxin reduction enzyme disease-resistant gene.As corn disease-resistant gene Hm1, it is the 1st plant disease resistance genes that quilt is cloned, and it is responsible for the resistance to fungi Cochliobolus carbonum microspecies 1.Hm1 coding separate the HC toxin that toxenzyme can the passivation pathogenic fungi be produced, and the HC toxin is the virulence factor that fungi C.carbonum microspecies 1 produce, it determines this germ can only infect some genotype of corn (Johal etc., 1992).Second class, NBS-LRR class disease-resistant gene.The nearly N end of their encoded protein is NBS, and nearly C end then is made up of LRR.As RPS2 (Bent etc., 1994), RPM1 (Grant etc., 1995), I2 (Simon etc., 1998), RPP5 (Parker etc., 1997), N (Dodd etc., 2001), L6 (Lawrence etc., 1995), Mla1 (Zhou etc., 2001), Mla6 (Halterman etc., 2001); Disease-resistant gene of paddy rice such as Xa1 (Yoshimura etc., 1998), Pib (Wang etc., 1999), Pita (Bryan etc., 2000) etc.The 3rd class, PK class disease-resistant gene.As tomato Pto gene, its product is one and is positioned at intracellular serine-threonine protein kinase enzyme, do not have LRR structural domain (Martin etc., 1993).The 4th class, LRR-TM class disease-resistant gene.Gene C f-2 (Dixon etc., 1996), Cf-4 (Thomas etc., 1997), Cf-5 (Dixon etc., 1998), the Cf-9 (Jones etc., 1994) of the different physiological strains of the anti-leaf mold of tomato, and the gene Hs1 of the anti-Cyst nematode of beet
Ppro-1(Cai etc., 1997) etc.The 5th class, LRR-TM-PK class disease-resistant gene is representative with rice bacterial blight resistance gene Xa21 (Song etc., 1995).The 6th class is representative with the RPW8 of Arabidopis thaliana, and its encoded protein only contains complete CC and NBS structural domain (Xiao etc., 2001).The 7th class is representative with the xa5 gene of paddy rice, and its encoded protein is a transcription factor (TFIIA γ) (Iyer ﹠amp; McCouch, 2004).
The disease-resistant proteic disease-resistant gene of coding NBS-LRR class is a class disease-resistant gene maximum in the plant disease resistance genes, constructional feature according to the disease-resistant protein N terminal of NBS-LRR class, this genoid can be divided into two big class TIR-NBS-LRR (TNL) and CC-NBS-LRR (CNL) (Meyers etc., 1999 again; Pan etc., 2000; Cannon etc., 2002; Richly etc., 2002).TNL class disease-resistant gene is mainly found in dicotyledons, does not also find (Bai etc., 2002 so far in the monocotyledons genome; Meyers etc., 2002).The disease-resistant gene that in monocotyledons, identifies the at present disease-resistant albumen of CNL class of mainly encoding, also there is a large amount of CNL class disease-resistant genes in the dicotyledons, comparatively speaking, the CNL class disease-resistant gene in the monocotyledons is than more rich and varied (Cannon etc., 2002) in the dicotyledons.
Studies show that NBS, CC, TIR structural domain may participate in signal conduction (Hammond-Kosack ﹠amp; Jones 1997), although this class R albumen does not have the intrinsic kinase activity, NBS can activate kinases or G albumen, NBS has in conjunction with ATP or GTP and hydrolytic enzyme activities (Traut, 1994).The signal conduction (Jones, 1994) in the disease-resistant defense response of TIR structure possibility involved in plant downstream.Nearest evidence shows that the L family diversity of flax selects to also occur in the TIR zone, and this zone forms specificity (Luck etc., 2000) with corresponding LRR zone coevolution.The function of LRR structural domain be mainly concerned with protein-protein and with interaction (the Jones ﹠amp of part; Jones, 1996; Kajava, 1998), be product or the indirect interactional position of product (Bent, 1996 that the disease-resistant gene product is direct and the pathogenic bacteria nontoxic gene is encoded by inference; Baker etc., 1997).Jia etc. (2000) are processed as one the 176 specific exciton of amino acid whose activated protein AVR-Pita176 microspecies by yeast two-hybrid proof AVR-Pita encoded protein, be delivered to plant cytoplasm and combine with the LRD zone of Pita acceptor specifically, thus the defense response of Pita mediation in the activating cells.Av-rPita176 can not combine with LRD after the Ala among the Pita becomes Ser, thereby shows susceptible.This result has proved that directly the LRR structural domain may be exactly the zone of pathogenic bacteria identification; Mutual " gene pairs gene " relation of making to have verified from molecular level for the first time paddy rice and Pyricularia oryzae of Pita and AvrPita.
Paddy rice is one of most important food crop in the world, and it is staple food with rice that population over half is arranged approximately.(no condition: Pyricularia grisea Sacc.) rice blast that causes is that Rice Production is endangered one of severe diseases, annually all causes serious grain loss by pathogenic fungi Magnapothe grisea Barr..From the viewpoint of Sustainable development of environment protection with agricultural, breed and the utilization of disease-resistant variety is the safe and effective procedure of control rice blast.But, because the diversity and the volatility of rice blast fungus population, people lack effective utilization of antagonism gene in addition, and antagonism mechanism lacks understanding fully, so that the susceptibleization problem of disease-resistant variety not only is not resolved, and becomes thremmatology man stubborn problem the most because of the shortage of effective anti-source gene and short-livedization of disease-resistant variety on the contrary.Therefore, excavate, identify and clone disease-resistant gene and reasonably be applied to the breeding for disease resistance plan and become the preferential major issue that solves in the agri-scientific research.
Along with molecular biological fast development, so far, have at least more than 80 rice blast master to imitate resistant gene and be in the news, wherein located for 60 by molecule.At present, also not identified the rice blast master on the 3rd karyomit(e) of paddy rice imitates the resistant gene, all identified the main resistant gene site of imitating on remaining 11 karyomit(e), and contain a plurality of rice blast resistances site on the karyomit(e) that has, and the resistant gene of some gene locus is that cluster exists.It is worthy of note in the localized resistant gene of numerous quilt successes, to have only two resistant genes of resistant gene Pib and Pita successfully to be cloned, they lay respectively on the 2nd karyomit(e) and the 12nd karyomit(e) of paddy rice.Before proposing, the present patent application also do not have the report that rice blast resistance gene is cloned on paddy rice the 8th karyomit(e).
Clone's disease-resistant gene is the prerequisite to the research of paddy rice resistance mechanism, discloses the molecule mechanism of rice anti-rice blast and can control and reduce the harm of Pyricularia oryzae to paddy rice better.Simultaneously,, can control and increase the disease resistance of plant artificially, widen the anti-spectrum of plant clone's the modification and the transformation of disease-resistant gene.These aspects are that employing conventional plant breeding and improving technology institute are inaccessiable.
Summary of the invention
A rice blast resistance gene that the objective of the invention is to carry among the separating clone rice varieties Q61 and the dna fragmentation that comprises the promotor of regulating and control this gene.
Another object of the present invention provides the coded protein of above-mentioned rice blast resistance gene.
Another object of the present invention provides the above-mentioned carrier that contains above-mentioned resistant gene.
Another object of the present invention provides above-mentioned carrier transgenic plant transformed.
Another object of the present invention provides the application of above-mentioned protein in preparation resisting rice blast bacteria medicine.
Further purpose of the present invention provides molecule marker that above-mentioned resistant gene produces and rice blast is had application in the paddy rice of disease resistance in seed selection.
The present invention relates to separate and use a kind of dna fragmentation of the Pi36 of comprising gene, this fragment is given plant produces specificity (specialization) to the caused disease of Pyricularia oryzae (Magnaporthe grisea) disease resistance response.This invention is applicable to all plants to this pathogenic bacteria sensitivity.These plants comprise monocotyledons and dicotyledons.Wherein, described fragment perhaps is equivalent to the dna sequence dna shown in the SEQ ID NO:1 basically shown in sequence table SEQ ID NO:1, and perhaps its function is equivalent to the subfragment of sequence shown in the SEQ ID NO:1.A kind of NBS-LRR proteinoid of this dna sequence encoding, its aminoacid sequence such as SEQ ID NO:2 are listed, and structure is as shown in Figure 6.Dna fragmentation shown in the present is constitutive expression at the leaf tissue of paddy rice.
The Pi36 resistant gene coding NBS-LRR albumen that separates, clones.This albumen comprises two main structural domain: NBS and LRR zone, and wherein the NBS structural domain contains conservative kinase 1a:GMGGLGKTT, kinase 2a:IVIDDIWDKP and kinase 3a:GSKILVTTRK.And this proteic C-end is 13 LRR repetitions, and its leucine content is 17.0%.The nucleotide fragments of coding NBS or LRR may have independently function in the Pi36 gene.Fragment and the reorganization of other nucleic acid fragments with coding different structure territory in the Pi36 gene can constitute mosaic gene or protein, make it to have new function.The Pi36 gene is modified or transformed, can change or increase certain function of gene.For example, the LRR zone of this gene is replaced with the structural domain of other resistant genes, maybe the NBS structural domain with this gene carries out rite-directed mutagenesis, may cause the change of the anti-spectrum of the forfeiture of gene resistance or gene.
The present invention comprises that equally the primary structure part with the Pi36 resistant gene connects the upward formed mosaic gene of proper regulation sequence effectively, and comprises the plant of this gene and the seed of this kind of plant in genome.As this gene can be natural or chimeric.For example, will comprise the fragment of this gene and the promotor of a constitutive expression and be connected, this promotor can and be expressed under any condition cytocerastic any period.The promotor of this constitutive expression comprises the promotor of cauliflower mosaic virus 35S etc.On the other hand, also promotor that can the promotor of this gene and a tissue specific expression or developmental stage is specific expressed or accurately the promotor of environmental induction be connected, these promotors are referred to as inducible promoter.Like this, the change of environment, the difference of developmental stage can change this expression of gene, and is same, also this expression of gene can be limited in some tissues, makes by this gene induced disease resistance response and obtains artificial control.Wherein envrionment conditions comprises attack, anaerobic condition and the light etc. of pathogenic bacteria, and tissue and developmental stage comprise leaf, fruit, seed and flower etc.
According to Pi36 gene order information provided by the invention (SEQ ID NO:1), those skilled in the art can easily obtain the gene that is equal to Pi36 by the following method: (1) obtains by database retrieval; (2) with the Pi36 gene fragment be genomic library or the acquisition of cDNA library of probe Screening of Rice or other plant; (3) according to Pi36 gene order information design oligonucleotides primer, from genome, mRNA and the cDNA of paddy rice or other plant, obtain with the method for pcr amplification; (4) on the basis of Pi36 gene order, obtain with the gene engineering method transformation; (5) method with chemosynthesis obtains this gene.
Rice blast resistance gene Pi36 provided by the invention has important use and is worth.One of application is that described Pi36 gene order is connected to any plant conversion carrier, with any method for transformation the Pi36 disease-resistant gene is imported paddy rice or other plant cell, can obtain the transgenosis disease-resistant variety of expressing said gene, thereby be applied to produce.Of the present invention gene constructed in plant conversion carrier, can suitably modify described gene or its regulating and controlling sequence, also can before its transcription initiation codon, replace the original promotor of described gene, thereby widen and strengthen the resistance of plant pathogenic bacteria with other promotor.
The Another application of resistant gene provided by the invention is to produce specific molecule marker according to described gene order information, includes but not limited to SNP (mononucleotide polymorphic), SSR (simple sequence repeats polymorphic), RFLP (restriction enzyme length is polymorphic), CAP (the cutting amplified fragments is polymorphic).Can identify the resistant gene type of paddy rice or other plant with these marks, be used for the molecular marker assisted selection breeding, thereby improve the efficiency of selection of breeding.
The present invention has following beneficial effect: change clone's disease-resistant gene over to susceptible plant, help to produce new disease-resistant plants.Particularly can be with transformation technology a plurality of disease-resistant genes that in plant, add up, and can not produce the chain problem of bad gene in the genome of following appearance in the traditional breeding technology, and can shorten breeding time.The clone of disease-resistant gene overcomes the prerequisite that can not shift the problem of disease-resistant gene in the traditional breeding method between plant species.In addition, disease-resistant transfer-gen plant that the present invention can further provide or the above-mentioned dna fragmentation of applications exploiting obtains and corresponding seed, and with gene of the present invention or based on the recombinant chou plants transformed of this gene or the seed that obtains by this class plant.Can gene of the present invention be changed over to other plant with the mode of sexual hybridization.
Description of drawings
Fig. 1 is the map based cloning synoptic diagram of resistance gene Pi 36 of rice blast;
Fig. 2. be the high-res genetic map and the electronics physical map of resistance gene Pi 36 of rice blast;
Fig. 3 is the PCR (3a) and Southern (3b) the detected result figure of selective marker hygromycin gene of the partial resistance transformant of resistance gene Pi 36 of rice blast;
Fig. 4 is that the resistance of resistance gene Pi 36 of rice blast transformed plant is identified figure;
Fig. 5 is the gene structure figure of resistance gene Pi 36 of rice blast;
Fig. 6 is a resistance gene Pi 36 of rice blast amino acids coding peptide sequence structure iron;
Fig. 7 is that the RT-PCR of resistance gene Pi 36 of rice blast and allelotrope pi36 expression characterization thereof detects figure;
The Pi36 loci gene type that Fig. 8 identifies parent and disease plant for molecule marker CRG3 is figure as a result;
Wherein, A is the high-res genetic map in Pi36 site among Fig. 2, and sea line is represented chromosomal region, and the top is mark/resistant gene, and the below numeral is the genetic distance (cM) between mark; B is the BAC/PAC clone electronics contig of Pi36 site areas, and sea line is represented the BAC/PAC clone, and vertical line is represented the landing position of linked marker; C is the electronics physical map in Pi36 site, and bracket inner digital is the recombinant chou number of each marker site, and sea line is represented genome area, and the below numeral is the physical distance (kb) between mark.
Swimming lane 1-8 is the pcr amplification product of hygromycin gene of the conversion seedling of resistance gene Pi 36 among Fig. 3; Swimming lane 9 is the PCR product of the hygromycin gene of carrier pCAMBIA1300 plasmid; Swimming lane 10 is the non-transformant of acceptor Q1063; M is molecular weight marker DL2000; The arrow indication is the part fragment 760bp of Hpt gene.
Green square box is represented the exon of resistance gene Pi 36 among Fig. 5, and the square box of band oblique line is respectively 5 ' and 3 ' non-translational region, and straight line is then represented intron.
Underlined letter is for forming 6 zones of inferring NBS among Fig. 6; The 590th amino-acid residue reflected the difference of protein sequence between the anti-sense of Pi36 allelotrope, is aspartic acid in susceptible allelotrope pi36; The amino-acid residue that independently list the lower section is the terminal LRR of C-zone.
A is that disease-resistant variety Q61 is before inoculation and the expression conditions of inoculation back 6,12,24,48,72h among Fig. 7; Actin is an internal reference; B is susceptible variety Q13, i.e. LTH is before inoculation and the expression conditions of inoculation back 6,12,24,48,72h; Actin is an internal reference.
P1 is disease-resistant parent Q61 among Fig. 8; P2 is that susceptible parent likes to know the rising sun; S1-S5, S6 and S7 are F
2Disease plant, gene pi36pi36; R1 is F
2Disease-resistant plant, genotype are Pi36pi36; Banding pattern before undigest:2 parent enzyme cut; M: molecular weight marker DL2000; The arrow indication is mark CRG3.
Embodiment
Embodiment 1: the genetic analysis of resistance gene Pi 36 of rice blast and Primary Location
As Fig. 1, the present invention utilizes the map based cloning method to clone resistance gene Pi 36.At first, in order to excavate and identify the rice blast new resistance gene, long-grained nonglutinous rice disease-resistant variety Q61 and japonica rice susceptible variety are liked to know the rising sun and Lijiang xintuanheigu (Lijiangxintuanheigu, LTH) F of 2 cross combination origin
2(the F of colony
2-1 and F
2-2), inoculation is to the bacterial strain CHL39 of the clearly demarcated non-affinity/compatible host response of parents' kind performance.The result shows, these 2 F
2Disease-resistant plant and disease plant separates than meeting 3: 1 in the colony.Infer that thus the resistance to inoculating strain CHL39 that Q61 showed is controlled by a pair of dominant gene.
In order to determine main chromosome position of imitating resistant gene apace, made up by 40 disease-resistant individualities and 580 F that susceptible individual is formed
2For mapping population, and utilize randomly amplified polymorphic DNA (randomamplified polymorphic DNA, RAPD) labeling technique, in conjunction with based on population mixture compartment analysis method (bulked-segregant analysis, BSA) (recessive-class analysis RCA), has screened 1200 random primers to recessive population analysis method altogether, 3 marks that polymorphism is good have been obtained, BA1126
550, BA253
450And OPP09
1800Because these 3 marks all are positioned at a side in this site, and BA1126
550Repeated best, so, this mark is cloned and is checked order.The reference sequences Japanese fine by flag sequence and order-checking kind carries out homology relatively, and this mark is landed on paddy rice the 8th karyomit(e).For the result who confirms that RAPD analyzes, we have quoted 4 sequence tagged site (sequencetagged site from public database, STS) mark, S5014, C30376, E60275 and C61344, and 3 little satellites (simple sequence repeats, SSR) mark, RM8018, RM1374 and RM5647 utilize the susceptible F of 580 plant height degree
2Individuality has carried out linkage analysis.The result shows that this resistant gene site is positioned on the 8th chromosomal galianconism, the zone of 5.8cM between the mark RM8018-RM5647.Because the position of oriented 4 rice blast resistance genes is different on this zone and the 8th karyomit(e), therefore, is Pi36 with this new unnamed gene.
Embodiment 2: the Fine Mapping of rice blast new resistance gene Pi36 and electronics physical mapping
In order to determine the position in Pi36 site subtly, we utilize the reference sequences in the public database, developed the mark of 5 polymorphisms, 1 STS mark R36STS2 and 4 candidate resistance gene (candidateresistance gene, CRG) mark, CRG1, CRG2, CRG3 and CRG4, and to the susceptible F of 580 plant height degree
2Individuality has carried out linkage analysis.The result shows, the Pi36 site is by the zone of Fine Mapping 0.6cM between CRG2-RM5647, and with mark CRG3 and CRG4 be divided into fully from.In order to make up the physical map in this site, we utilize fine bacterial artificial chromosome (the bacterialartificial chromosome with reference to kind Japan, BAC) and artificial chromosome (the P1-derived artificialchromosome of P1 mediation, PAC) clone, by bioinformatic analysis (bioinformatics analysis, BIA), made up the electronics physical map in this site.The result shows, the Pi36 site by physical positioning in the zone of about 17kb.
Embodiment 3: the prediction note and the sequential analysis of rice blast new resistance gene Pi36 candidate gene
In order to determine the candidate gene of Pi36, we utilize the fine reference sequences of Japan, by 3 kinds of predictive genes software RiceGAAS (http://ricegaas.dna.affrc.go.jp), Gramene (http: // 143.48.220.116/resources/) and the FGENESH (http://www.softberry.com) of Softberry predictive genes and note analysis have been carried out in the goal gene zone, the candidate resistance gene of tentatively having determined Pi36 is Pi36-1, Pi36-2 and Pi36-3, and wherein Pi36-3 has comprised Pi36-1 and Pi36-2.Therefore, with Pi36-3 by following its function of functional complementation experimental verification.
Embodiment 4: the conversion of rice blast new resistance gene Pi36 and the resistance of transformant are identified
Utilize long segment PCR (long-range PCR, LR-PCR) technology increased one the 16.0kb that contains Pi36 fragment and be cloned into binary vector transformation system pCAMBIA1300, imported agrobacterium strains EHA105.Will height susceptible variety Q1063 mature seed evoked callus on inducing culture.The EHA105 that contains the goal gene conversion carrier was cultivated 1 day for 28 ℃ at the YM nutrient agar, be collected in the MB liquid nutrient medium of the Syringylethanone that contains 100 μ mol/L.The rice callus tissue was immersed bacterium liquid 20 minutes, blot and transfer to the MB nutrient agar behind the bacterium liquid and cultivate.Shift callus to the culture medium culturing that contains the 50mg/L Totomycin, subculture was 1 time in per 14 days, subculture 2 times.Behind the resistance screening, change regeneration culture medium over to and differentiate the conversion seedling, obtained 274 strain transformant.
CRG3 has carried out the PCR-CAPS detection to transformant with the CRG mark.2 primers have been used in this experiment, and wherein, primer CRG3F is: gctagcaagcatggagttcgt-3 ', primer CRG3R is: 3 '-agcgggtaaggtagcataggt.The step of pcr amplification is: 94 ℃ of pre-sex change 4min; Next be 30 circulations, cycling program is as follows: 94 ℃ of sex change 30sec, and 60 ℃ of annealing 30sec, 72 ℃ are extended 2min; Last 72 ℃ are extended 7min, and temperature drops to 4 ℃ and promptly finishes amplification.Amplified production carries out enzyme with restriction enzyme HigfI to be cut, and system is 10 * enzyme cutting buffering liquid, 2 μ l, PCR product 8 μ l, and restriction enzyme HinfI10U adds two aqua sterilisa to 20 μ l that steam.
Simultaneously, the PCR and the Southern blot evaluation (Fig. 3) of hygromycin gene transformed plant have been carried out.Detected result proof order gene fragment has imported these transformant.The resistance qualification result shows that resistance gene Pi 36 has showed the identical resistance with resistant gene donor Q61 under susceptible variety Q1063 genetic background, shows that resistance gene Pi 36 successfully cloned (Fig. 4).
The structure of embodiment 5:Pi36 gene
The employing method of moving one's steps is measured the dna sequence dna of Pi36.Utilize 5 ' and 3 ' RACE reaction, obtained the full-length cDNA of Pi36 and it is checked order.Sequence 1 and 2 in the sequence table is respectively DNA and the cDNA sequence of Pi36.Pi36 gene DNA length is 16.0kb, and its full-length cDNA is 3963bp, contains the opening code-reading frame of a 3171bp, and 5 ' and 3 ' non-translational region is respectively 65bp and 726bp.By icp gene group DNA and cDNA, find that the opening code-reading frame of this gene contains 4 introns and 5 exons, and 3 ' non-translational region contains 1 intron (Fig. 5).
The structure of embodiment 6:Pi36 resistance protein
The protein sequence of Pi36 genes encoding is shown in SEQ ID NO:2 in the sequence table.1 protein polypeptide of forming by 1056 amino-acid residues of resistance gene Pi 36 coding, molecular weight is 120KD, iso-electric point is 6.61.Utilize 14-45 amino-acid residue of this protein polypeptide of COIL analysis revealed to have 95% possibility to be CC (coil-coil) structural domain.Pi36 albumen belongs to NBS-LRR albumen, and conservative kinase 1a (GMGGLGKTT) is positioned at the 206th amino-acid residue of this polypeptide in the NBS structural domain; Kinase2 (IVIDDIWDKP) is positioned at the 286th amino-acid residue of this polypeptide; Kinase 3a (GSKILVTTRK) is positioned at the 310th amino-acid residue of this polypeptide.And the 581-921 of this a proteic C-end amino-acid residue is 13 LRR repetitions, and its leucine content is 17.0%.
The database comparative analysis finds that coding proteic nucleotide sequence of Pi36 and barley powdery mildew resistant gene Mla1 and Mla6 have 40% and 37% homology respectively, have 30%, 27% and 24% homology respectively with resistance gene of rice blast Pib, Pita and paddy rice bacterial leaf spot resistant gene Xa1.
Embodiment 7:Pi36 expression of gene specificity analysis
With RT-PCR Pi36 expression of gene pattern is analyzed.From the blade of disease-resistant variety Q61 and susceptible variety LTH, extract total RNA, utilize reverse transcription test kit SuperScript
TMReverseTranscriptase II carries out the synthetic of reverse transcription cDNA article one chain.The RT-PCR primer is R36RT F:CGAGCTGCTGAAGGAGAAGT; R36RT R:AGCTCATCCCTTGGCTCATC.The PCR reaction is: 94 ℃ of pre-sex change 4min; Next be 35 circulations, cycling program is as follows: 94 ℃ of sex change 30sec, and 56 ℃ of annealing 30sec, 72 ℃ are extended 1.5min; Last 72 ℃ are extended 7min, and temperature drops to 4 ℃ and promptly finishes amplification.
Experimental result shows, the RNA reverse transcription template of the leaf tissue before and after the inoculation of resistant variety and susceptible variety all can amplify special fragment, illustrate that Pi36 and allelotrope pi36 thereof all can express in leaf tissue, promptly belong to the gene (Fig. 7) of constitutive expression.
Embodiment 8: transform the application of the resistant variety of Pi36 gene generation
The Pi36 gene clone to plant conversion carrier pCAMBIA1300, is imported agrobacterium strains EHA105, be used to transform susceptible rice varieties, obtained 274 strain transformed plants.T with the transformant generation
1Evaluation is sowed, sprouts and inoculated to seed, finds wherein to have 35 to transform (the T of system
1) the performance resistance separates, and the resistance of transformed plant and Pi36 gene be divided into from.This explanation can be used Pi36 gene transformation paddy rice susceptible variety, is used for producing.
The application of embodiment 9:Pi36 gene order in the molecular marker assisted selection breeding
Utilize Pi36 gene order information provided by the invention can produce molecule marker, be used to identify that the range gene type that has on the Pi36 site is the plant of Pi36Pi36, Pi36pi36 and pi36pi36, can in the molecular marker assisted selection breeding process, be applied.
Resistance gene Pi 36 of rice blast and application sequence table thereof
SEQUENCE?LISTING
<110〉Agricultural University Of South China
<120〉resistance gene Pi 36 of rice blast and application thereof
<130>
<160>2
<170>PatentIn?version?3.2
<210>1
<211>14450
<212>DNA
<213〉Oryza paddy rice (Orysa sativa L.)
<220>
<221>5’UTR
<222>(2489)..(2553)
<220>
<221>exon
<222>(2554)..(2972)
<220>
<221>gene
<222>(2554)..(11611)
<220>
<221>Intron
<222>(2973)..(3405)
<220>
<221>exon
<222>(3406)..(3834)
<220>
<221>Intron
<222>(3835)..(8904)
<220>
<221>exon
<222>(8905)..(10943)
<220>
<221>Intron
<222>(10944)..(11067)
<220>
<221>exon
<222>(11068)..(11334)
<220>
<221>Intron
<222>(11335)..(11593)
<220>
<221>exon
<222>(11594)..(11611)
<220>
<221>3’UTR
<222>(11612)..(11780)
<220>
<221>Intron
<222>(11781)..(11903)
<220>
<221>3’UTR
<222>(11904)..(12459)
<400>1
Resistance gene Pi 36 of rice blast and application sequence table thereof
cttcgacacg?caaacgtgca?cacagccacc?tatcgtgacc?tcctcagcct?ccacgcatcc 60
gtacgtcctc?tagtgcagcc?actaaactgt?aagtccaagt?tctaaattgt?aaccgacttt 120
gtacatatat?tcgatacaaa?ctctaacaat?atcttcacta?accctatctt?agccgaataa 180
caactaaaat?tttttctatt?caaacatagc?taaaccatgg?ttaacaaata?caacaatcct 240
aacaaccaca?taaggtaaca?gttggaatcg?aacgctatat?tgttacaaca?ctgatacatt 300
tgtcgtgcac?aggtagcttg?aaatttgcac?aagtcacgaa?tattgtcgtc?aaacggagat 360
agaacaaatc?ctgcaagaag?ctctggtaca?ctttaaaatg?accgctctag?tcctcaaaat 420
ccagagatct?cctacaaagc?aattttagga?tacggctcgt?ttaaattgta?aaagatgtat 480
tgtgtaattt?caacagactt?gacgatgcac?atgctgattt?ttaagagctt?accattgtcc 540
actttaatgc?acctggacca?cggtcggttt?cacagttcgc?cgcaccaacc?ggttccggaa 600
ggtgagcgtg?ctggctgctg?agaagaaaaa?tagcttccca?acgtggcttc?aattaggggg 660
tgtttaggag?agagggactt?accaaacacc?cccttaatca?acgtgtggct?gtcttttcag 720
atgacccttt?tcctaagccc?tctctctcat?ttttcacaag?cacatttttc?aaattgttaa 780
acggtgtatt?tttttaaaaa?aaaagtttat?atactaaagt?tgcttaaaaa?attaaattaa 840
ttatttttta?aaaaaatagc?aaatgtttaa?ttaattacgt?tctattggac?cgtttcggtt 900
ttcgtaggta?ccatgcacct?ccccatccga?ctgtagcgaa?catcttccca?tccggctgta 960
gcgatcacac?ccagagggtc?tcatgtgacg?cgttcgaggt?ggagccaacg?gcgtcttaca 1020
agttgagtaa?cttgagtgat?aaacacagta?ttgcctacca?cgcgttttga?aacttaatta 1080
cacgtgtttt?cgccctcttg?ttatgttcat?gctccaatgg?aaacaaacaa?atcggatata 1140
acacttcaat?cctgaggcag?gattttgtat?gcaaaatggt?actatactca?gataaatggc 1200
tcatcaatta?taatgatggg?tttacaatta?caaatgaaga?tatttaagca?acttaattta 1260
ttcattctag?ttgcataggt?ctagaactcg?ttgcatttgg?agtgagatgg?agtaaatatt 1320
atttcacatt?ttgtcaattc?aaacatttac?acgttttaat?gttttttcga?gaatggatgg 1380
cgtgagctcc?gataagagta?ctataagatt?ataaaccagc?tataaacata?ttttaagaaa 1440
ataaaagagg?agagataaga?gcaacggatt?ataaatttat?agccagctgc?agcatagact 1500
ccaagataca?tgtgtgtatg?ataggtgaga?tcaaacatta?atcatatagt?acatgtttat 1560
agataactat?tgtatgaatt?gactttagaa?gatttggagc?caatagttga?atatactact 1620
cccttctttc?caaattaatc?atcatgtaag?aataggtaca?acagcagatt?attagccagc 1680
tataaacata?ttttaatgag?ataaaagatg?agagagaaga?gcagcaggct?atagatctgt 1740
agccagctat?aacacggact?ccaagacgca?atgggtgtat?gacatgtggg?accatatatt 1800
aatagtataa?taagcaacta?ttgtatgaat?tggctattag?atcggctata?gatgaattgg 1860
agctagtagt?gggctatact?agtaaacttg?ctctaataga?atttaaaatt?tttcaaattg 1920
gtcattatat?aactgcatgg?acacggattt?catcggaata?caattaatac?agtatgagag 1980
aatatgtgta?tgctagggtg?taattgacat?gaggttttaa?ttgagtatag?gagaatgtgt 2040
gtatgttgtc?ttgattgatg?tgatttaaat?tgtcattggt?cttggtgcat?aaaattatac 2100
tccctccgtt?ccataatata?aggcacaact?atttttttta?tgttccataa?tataaggcat 2160
acatgcatgt?atacaattaa?ttaggacatc?ttctccatta?aattattatt?tttttaaatc 2220
attcactctc?aagatctcta?aatttattgg?atgcatgcat?gtattatatt?tattaggatg 2280
Resistance gene Pi 36 of rice blast and application sequence table thereof
atccaaatta?gaaaataata?ataattgttt?cttgaaaaag?tggttgtacc?ttatattttg 2340
gaacatatgg?agtatgatga?ttaatttggg?aaggagggag?tattaaactt?gctctaaccc 2400
aattgtgaca?agtccattaa?ttaacctatc?agctagtagg?ctagtagttg?cacatatata 2460
caggagtgca?atgtatctgc?tcctacccag?ctaccaaccc?agcaatacca?tccactcaac 2520
tcaagtcaag?tcaaccacag?ctagctagca?agc?atg?gag?ttc?gtc?act?ggg?gcg 2574
Met?Glu?Phe?Val?Thr?Gly?Ala
1 5
atg?agt?agc?ctg?atc?ccc?aag?ctg?ggc?gag?ctg?ctg?aag?gag?aag?tac 2622
Met?Ser?Ser?Leu?Ile?Pro?Lys?Leu?Gly?Glu?Leu?Leu?Lys?Glu?Lys?Tyr
10 15 20
aat?ctg?cag?aag?aac?ata?agg?ggg?aag?atc?gag?tct?ctc?tcg?gcg?gag 2670
Asn?Leu?Gln?Lys?Asn?Ile?Arg?Gly?Lys?Ile?Glu?Ser?Leu?Ser?Ala?Glu
25 30 35
ctc?cgg?cga?gcg?caa?gcc?gtc?ctc?cgc?atg?gtc?ggc?gag?gtg?cca?caa 2718
Leu?Arg?Arg?Ala?Gln?Ala?Val?Leu?Arg?Met?Val?Gly?Glu?Val?Pro?Gln
40 45 50 55
gac?cag?cag?aac?gag?ctg?gtg?aag?ctc?tgg?gcg?agc?gac?ctc?agg?gag 2766
Asp?Gln?Gln?Ash?Glu?Leu?Val?Lys?Leu?Trp?Ala?Ser?Asp?Leu?Arg?Glu
60 65 70
gcg?tcc?tac?gac?atg?gag?gat?atc?atc?gac?gcc?ttc?ttg?gtt?cat?gtc 2814
Ala?Ser?Tyr?Asp?Met?Glu?Asp?Ile?Ile?Asp?Ala?Phe?Leu?Val?His?Val
75 80 85
gac?gac?gat?ggc?ccc?aag?cca?gcc?gac?cca?cat?gtg?ctc?agg?cgc?ctc 2862
Asp?Asp?Asp?Gly?Pro?Lys?Pro?Ala?Asp?Pro?His?Val?Leu?Arg?Arg?Leu
90 95 100
ggc?aag?cag?gtg?aag?aaa?ctg?ttc?aag?aag?acc?aag?cat?cgc?gtc?acc 2910
Gly?Lys?Gln?Val?Lys?Lys?Leu?Phe?Lys?Lys?Thr?Lys?His?Arg?Val?Thr
105 110 115
atc?gcg?gac?gca?atc?caa?gaa?atg?gaa?aag?aag?ctc?ctg?gag?att?gat 2958
Ile?Ala?Asp?Ala?Ile?Gln?Glu?Met?Glu?Lys?Lys?Leu?Leu?Glu?Ile?Asp
120 125 130 135
gca?agg?cat?ggc?ag gtatggaaaa?tatttactcc?ctccgtccca?aaaaaaattc 3012
Ala?Arg?His?Gly?Arg
aatttctaag?tttccgtatc?caacatttga?ttgtccgtct?tatatgaaat?ttttctatga 3072
ttagtatttt?tattgttgtt?agatgataaa?acataaataa?tactttatgc?gtgcgtgatt 3132
tatcttttta?ttttttttat?aattttttca?aataagacgg?acggtcaaac?gttgaacacg 3192
gaaacccaaa?aattgagtta?ttttaagacg?aagggagtaa?tatggaagaa?atcttaataa 3252
tatgttgtat?tatattgcat?tatattgatc?tgtcatgagg?tacatacata?tatagtacga 3312
gttatagctg?atttatctca?agaattctat?ttcttgtagt?cttacattcg?tcatctcttg 3372
atgttgtctc?ttgactctca?tctattacta?cag?a?tac?cca?gtt?gat?aac?ata 3424
Tyr?Pro?Val?Asp?Asn?Ile
145
gtt?atc?cct?gca?cgt?ccg?acg?cct?att?gat?cct?cgc?atc?cta?aat?att 3472
Val?Ile?Pro?Ala?Arg?Pro?Thr?Pro?Ile?Asp?Pro?Arg?Ile?Leu?Ash?Ile
150 155 160
gag?aaa?atg?gca?aac?aat?ctt?gtt?ggc?att?gat?gag?cca?agg?gat?gag 3520
Glu?Lys?Met?Ala?Asn?Asn?Leu?Val?Gly?Ile?Asp?Glu?Pro?Arg?Asp?Glu
165 170 175
ctt?att?aag?atg?ttg?tca?caa?cat?gag?cgt?gac?aac?aat?ctt?aat?acg 3568
Leu?Ile?Lys?Met?Leu?Ser?Gln?His?Glu?Arg?Asp?Asn?Asn?Leu?Asn?Thr
Resistance gene Pi 36 of rice blast and application sequence table thereof
180 185 190
tcc?aat?agg?aag?aca?aag?ata?gta?tac?gtc?gtt?gga?atg?ggt?ggt?ctg 3616
Ser?Asn?Arg?Lys?Thr?Lys?Ile?Val?Tyr?Val?Val?Gly?Met?Gly?Gly?Leu
195 200 205 210
ggc?aaa?acc?acc?ctt?gcc?aca?gca?gta?tat?gag?aag?att?aaa?gtg?ggt 3664
Gly?Lys?Thr?Thr?Leu?Ala?Thr?Ala?Val?Tyr?Glu?Lys?Ile?Lys?Val?Gly
215 220 225
ttt?cct?ttg?aat?gct?ttt?gtt?cca?atc?ggt?caa?aat?cct?aat?atg?aag 3712
Phe?Pro?Leu?Asn?Ala?Phe?Val?Pro?Ile?Gly?Gln?Asn?Pro?Asn?Met?Lys
230 235 240
gca?atc?ttg?tgg?aac?att?ctt?cat?cgc?ctt?ggc?tcg?gag?aag?tac?ctg 3760
Ala?Ile?Leu?Trp?Asn?Ile?Leu?His?Arg?Leu?Gly?Ser?Glu?Lys?Tyr?Leu
245 250 255
aat?tgt?cca?aat?att?gag?atg?ttg?act?gta?cag?gag?ctc?atc?ggt?gaa 3808
Asn?Cys?Pro?Asn?Ile?Glu?Met?Leu?Thr?Val?Gln?Glu?Leu?Ile?Gly?Glu
260 265 270
ctc?aag?caa?ttc?atc?aag?gga?aag?ag?gtatgaatat?gacgacccac 3854
Leu?Lys?Gln?Phe?Ile?Lys?Gly?Lys?Ser
275 280
caaatttgtt?tttatatact?acctcttttt?cttaataaat?cacgccattg?acttttgaaa 3914
taacgtttga?ccttcaaaaa?tttagtgtaa?atatgctcaa?gtataagtca?taccttaaag 3974
tttctttaat?aataaaacaa?gtcacaacaa?aataaatagc?atttatataa?tttgtttgaa 4034
tatgactaat?ggttaaacgt?catatcaaaa?gtcaacatct?tctgttagaa?tacagaggga 4094
gtatatgtcc?ataacctatg?ctaccttacc?cgcttatgaa?tgaatatttg?cgctacacac 4154
aatcatagtc?ttaaagctat?ttaatttgtc?tttgcgcttg?ctatttgaaa?aaaaaatgat 4214
gtacctcttt?ttaatcgatc?acagtaatag?tccgggttta?cccgttcttt?aggctccact 4274
tacatcatga?gcaagctaaa?aatcgtgatc?aagatattaa?acctaaaatg?accttgatgt 4334
cacctagagg?ggtgtccatg?gagcgactcc?tcggacagcg?aggagtggag?acccctcccc 4394
gagtttgaag?attcagccag?ggggttagca?gtgcaagacg?atagaaaaag?ccaaacagag 4454
aaaagatttg?tggggcctga?tctccccgcc?aagcaaaagt?caccagagtt?tatacaggtt 4514
caagccaccc?ggaggtgtaa?caccctactc?ccgtgttggt?tttgttgttt?ttcttatatc 4574
agcttagaac?aagggatgcc?tttttgggct?acagacaaga?agttctgatc?caaaaaatcc 4634
aactcttcat?ccgtagcttg?tccccttcct?tttatagatc?aaggatgggc?tcacaattct 4694
acaaatctac?ttccttgtct?tgtctatcag?ctgtcttgtc?aatgcctgtt?ggttccctca 4754
cacagtcttc?gagtaaccca?ctgacatttg?aagcttgcat?tgcccccaag?cacccgggct 4814
tctgtccgag?acagctgggt?cgtgccagca?tgttcaccat?gtggcgagtg?gtgggcctgg 4874
tgagacccga?cctgcctgaa?acagcaccac?ggtttcgttt?ctacccgttt?ggctaaatca 4934
ggcagcatgg?ggcatgtgcg?acacgtgtcc?aagatgccac?tggttctaat?gactcagtga 4994
tggtgccttg?tccacatcct?ttcagctcca?gactagctgg?ggctccccca?taaacctgtt 5054
taatttatgg?cggagaggac?tacttcacaa?cagcccgtgg?agcacgccca?acaccccagc 5114
tggtcctgag?gtcgcgccct?tgcacattta?tggagatagt?ttagctattc?ttacacaagc 5174
ttcttgaaga?tatcttagat?aatatctaat?gtttcgacag?agacattcct?ccttaatcaa 5234
gggacgtaac?ctacatgcag?acaaaggtga?gcaagctaac?aaggcaagca?aaaggggata 5294
tcggcaaaag?tagttaatgg?gttttcacaa?acaacaacta?aactactagt?tgcaccaaat 5354
Resistance gene Pi 36 of rice blast and application sequence table thereof
agcaaaactg?gaggagtgag?agagagagat?accaccggga?ttcttgcaaa?ctgatatgtt 5414
actgaatcta?ctctccgttg?aggagctcac?aatgatatat?atatatacat?atatatatat 5474
atatatacat?atatatacat?atacatatat?atatacatac?atatacatac?atacatacat 5534
acatacatac?atacatacat?atatacatac?atacatacat?acatacatac?atacatacat 5594
acatacatac?atacatacat?acatacatac?atacatacat?acatacatac?atacatacat 5654
acatacacac?acacatacat?acatacatac?atacatacat?acatacatat?atatatacat 5714
acatacatac?atacatacat?acatacatac?atacatacat?acatacatac?atatatatac 5774
atacatacat?acatatatat?atatacatac?atacatatat?gtatacatac?atatatatat 5834
aatgtttcga?cactcttcgt?tgtagaggaa?tgtaaaactc?acaaggagta?aggtgtgccc 5894
gggtcctagg?acgaggccgg?acataggcat?acctcagttt?atctcttaat?catatgtctc 5954
caccctccac?cccttgtaga?aagtgtgacc?tatgaagatt?aaggtaagcc?caggacccag 6014
gtcaaggccg?tgtctgaggc?aaaccttagc?ttattcttct?taagatgtat?atcttgtagg 6074
tctctacgct?cgtctacgta?ggacttacaa?ggactaaggt?gagcccgaga?cccaggacaa 6134
gacagggtcc?gaggcatacc?ttaggctaca?ttcttaagat?gtagatttta?tacgtctcca 6194
cactcctctt?ctcagagcaa?tgtgtgactt?gtgcggacta?aggtgagccc?gagacccagg 6254
ataaggccga?gtctgaggca?taccttagtt?tattcccttt?aagatgtaga?tcttgtaggt 6314
ctctacgctc?ctctcaccga?gtctgaggca?cacgttacat?tettaagatg?tacttcagtg 6374
gacctgggtc?ccgaagcgga?actaggccta?ggaccaacct?tattactcag?agttttaagt 6434
cgttgtactt?ttgacatgca?tcattccccc?ttaatcaagg?ggagtaacct?acatgcagac 6494
aaaggtgggc?ccgggaccaa?ccaaggtggg?tccctagtga?cataacccta?tttttaagga 6554
tggtttaagc?atgccattta?tcttatttat?aattgttttg?ctgggggaaa?tgggtagcct 6614
aattcccatt?tcaccgacaa?ggggggtaaa?taagcgttta?taacttttat?gtaaatgtgg 6674
aggctaaaca?tttcagatat?tccaaaatat?ttttggatat?tctaggatat?tttcagatat 6734
tccgaaaatc?tcaggttttc?gagcaagcta?acaaggcaag?caaaggggat?atcggcaaaa 6794
gtagttaatg?ggtttgcaca?aacaacgact?aaactactag?ctgcaccaaa?tagcaaaact 6854
agaggagtga?gagagagaga?taccaccgag?attgttgcaa?actgatatgt?tactgaatct 6914
acactccgtt?gaggagttca?caatgagcca?agtcttttcc?aaccctatcc?tcgtgaagtt 6974
gcataaatgc?actcatcttt?tcattaaggg?tatctcttcc?ctttcagagg?tgagatctga 7034
ccttcacaaa?ctccctcggc?acaccaccta?caaatcgatg?tctcgaggtc?aactcctagc 7094
cgtccaggta?tcctcaacca?ccaagagtta?caaatgcaac?aaagatttct?tggggatgaa 7154
tcgactgctc?gacaaatctt?cacgaaatca?acaccaagca?ctcaaataaa?ctcaaatcaa 7214
ccactcacac?atacacgaaa?tccgagatgg?aagatcggtg?gagagtaggg?atggcaattt 7274
gacccgattg?tccgtttacc?catgggtaaa?aaccctaata?gagttgggtt?tgttctacat 7334
ttagtatcca?tgggtttctt?attgggtcaa?aacctgtacc?caatgggcaa?gcgggcatgg 7394
gcatgggtac?ccatgcccgc?ttcgcccatt?tacccgcgaa?aaacaactga?catgtgggcc 7454
ctcaatcaac?ttaatataaa?aagcaccatc?taattctagg?gttttcttcc?tttcctccac 7514
agccgctact?ccttctcttc?ccctttcctc?tacagctacc?actcctcttc?ctccacagac 7574
Resistance gene Pi 36 of rice blast and application sequence table thereof
catagccacc?actccaggct?gccaaggcgc?caggctgaca?cgggacagcc?ggacgagagg 7634
ggcgcgcaat?tcgccaccac?ctcccacctc?cgctcgtcgc?tggcggcccc?accctccagc 7694
cgacgacttt?catagcggcc?acgggggagg?gggggcaacc?aaggaagctg?cgaccgccgt 7754
cgccttctac?cccgtctgcg?ccgagctcct?ctccaaccag?tcgtcgatgt?cctctagcca 7814
tcggccgcca?gcccgccaca?ccttcctcca?gtcgcgcctc?ccacctccgg?tcgccgccgc 7874
ttcctccaga?cgcgcctccc?acctctggtt?cccagatgtg?acgggtaaac?gggcatgcct 7934
gcgggcacat?ggcgcctacg?ggggacgggc?attggaagtg?ctccttgccc?atcatgtagt 7994
aacgggcatg?cccgcggaca?tgaattacct?cgcggacacg?ggatgggtag?tcactacccg 8054
tgcccgttgt?gcccgactgc?catccctagt?ggaaagtaag?aagtgagagg?gcaaggctca 8114
aagtgaatct?caaatcaaca?caagcaaggg?gcacaagaaa?gcaatggagc?gagcaatctt 8174
gacttggggt?ggagggggtt?atttataacc?acaacctcaa?atctgattgt?tggaggcaaa 8234
tcaagattca?actttttgaa?aaattttgaa?aaccattttt?ggacaagtct?gaaaaacaca 8294
agaaagcaat?ttcatttgag?caaccatttt?ccgaatttcc?aaaaaaattt?cggaattatt 8354
ttcggaagtt?gctacaatac?ttttggaaat?tccaaaaagt?ataaattaca?gcacaaactc 8414
agcaaaaaaa?aggaaaaata?acatttcaga?aattctgaaa?atatttcgaa?aaaatctgaa 8474
atcaccagag?agctttttca?caaattttat?cattttggaa?attccaaaaa?ccattttcgg 8534
aaaattctag?aaaacaccaa?aagatgtgtt?ggccaccgcc?gtcacgtgag?catgtcacat 8594
ttgaactcat?gacggaggag?cacactgccc?acttgtcagc?gttatccctg?tcactattct 8654
agtgcaccac?tggttcctag?ctacaagacc?gtcgtgccca?cacattatcc?aaaagcacat 8714
gccaatgcat?aggctaatga?ataattaatt?gcaatatctc?acttttaatg?catgtttatc 8774
tctcttatct?agattttctt?cggcactaaa?aattattgct?ccttcacata?taaaaagaaa 8834
accacatctt?attgatgact?cgtaattttc?agtgattggg?gtgcttatac?agttcttcta 8894
ctattctagg?t?ttt?tta?tcg?tta?tag?atg?aca?tat?ggg?ata?agc?cat?cat 8944
Phe?Leu?Ser?Leu Met?Thr?Tyr?Gly?Ile?Ser?His?His
285 290 295
ggc?aaa?tac?tcg?aaa?gtg?gtt?tac?aag?ata?atg?att?acg?gaa?gta?aaa 8992
Gly?Lys?Tyr?Ser?Lys?Val?Val?Tyr?Lys?Ile?Met?Ile?Thr?Glu?Val?Lys
300 305 310
tac?ttg?taa?cta?ctc?gta?aat?ctg?agg?ttg?caa?caa?tta?tca?gcg?atg 9040
Tyr?Leu Leu?Leu?Val?Asn?Leu?Arg?Leu?Gln?Gln?Leu?Ser?Ala?Met
315 320 325
tgt?aca?ata?tga?aac?cac?ttt?ctc?atg?ata?act?cta?aag?aat?tat?tgt 9088
Cys?Thr?Ile Asn?His?Phe?Leu?Met?Ile?Thr?Leu?Lys?Asn?Tyr?Cys
330 335 340
ata?caa?gaa?ctg?gta?gtg?aag?gca?aat?ctc?ttg?aca?gtt?cat?caa?ctg 9136
Ile?Gln?Glu?Leu?Val?Val?Lys?Ala?Asn?Leu?Leu?Thr?Val?His?Gln?Leu
345 350 355
agg?cat?gtg?aca?aga?ttt?tga?aga?aat?gtg?ctg?gtg?tgc?cat?tag?cta 9184
Arg?His?Val?Thr?Arg?Phe Arg?Asn?Val?Leu?Val?Cys?His Leu
360 365 370
tca?tta?cga?tag?cta?gct?tgt?tgg?cta?gta?gat?cag?gtc?tgg?act?ggt 9232
Ser?Leu?Arg Leu?Ala?Cys?Trp?Leu?Val?Asp?Gln?Val?Trp?Thr?Gly
375 380 385
ctg?agg?tgt?aca?gag?cta?ttg?att?ttg?gag?aag?aag?aca?act?atg?aaa 9280
Leu?Arg?Cys?Thr?Glu?Leu?Leu?Ile?Leu?Glu?Lys?Lys?Thr?Thr?Met?Lys
390 395 400
Resistance gene Pi 36 of rice blast and application sequence table thereof
tgg?caa?ata?cta?aaa?gga?tat?tgt?ctt?tta?gct?act?acg?atc?tgc?ctt 9328
Trp?Gln?Ile?Leu?Lys?Gly?Tyr?Cys?Leu?Leu?Ala?Thr?Thr?Ile?Cys?Leu
405 410 415
cac?att?taa?aga?att?gtt?tgc?tct?atc?taa?gca?tgt?ttc?cag?aag?att 9376
His?Ile Arg?Ile?Val?Cys?Set?Ile Ala?Cys?Phe?Gln?Lys?Ile
420 425 430
ata?aaa?ttg?aca?aaa?atc?att?tga?tat?gga?tgt?gga?tag?ctg?aag?gtt 9424
Ile?Lys?Leu?Thr?Lys?Ile?Ile Tyr?Gly?Cys?Gly Leu?Lys?Val
435 440 445
ttg?tcc?ctg?aga?aac?aaa?aca?caa?att?tag?gat?tat?atg?agc?ttg?ggg 9472
Leu?Ser?Leu?Arg?Asn?Lys?Thr?Gln?Ile Asp?Tyr?Met?Ser?Leu?Gly
450 455 460
aga?gtt?act?tca?atg?agc?tga?taa?ata?gaa?gca?tga?tcc?agc?caa?ttg 9520
Arg?Val?Thr?Ser?Met?Ser Ile?Glu?Ala Ser?Ser?Gln?Leu
465 470
aga?atg?aat?ttt?ttg?ggt?gta?tag?aag?gtt?tgt?cgt?gtt?cca?tga?tat 9568
Arg?Met?Asn?Phe?Leu?Gly?Val Lys?Val?Cys?Arg?Val?Pro Tyr
475 480 485
ggt?gct?cga?tct?agc?tcg?ttc?att?gac?aag?tga?aca?aaa?ctt?tgt?cac 9616
Gly?Ala?Arg?Ser?Ser?Ser?Phe?Ile?Asp?Lys Thr?Lys?Leu?Cys?His
490 495 500
tgt?att?gga?taa?cga?tga?gca?acg?gaa?acc?tga?aag?cac?caa?tgc?tcg 9664
Cys?Ile?Gly Arg Ala?Thr?Glu?Thr Lys?His?Gln?Cys?Ser
505 510 515
tag?gtt?ggc?cct?gca?ccg?tac?gag?tat?cac?gag?cta?tcg?att?tgt?taa 9712
Val?Gly?Pro?Ala?Pro?Tyr?Glu?Tyr?His?Glu?Leu?Ser?Ile?Cys
520 525 530
tat?gga?tat?gaa?gaa?agt?gag?gtc?att?tgt?tgc?cac?tga?gtg?caa?taa 9760
Tyr?Gly?Tyr?Glu?Glu?Ser?Glu?Val?Ile?Cys?Cys?His Val?Gln
535 540
tgg?caa?taa?tag?tgt?ggc?acc?acc?aag?gtt?cca?agt?ttt?acg?tgt?gtt 9808
Trp?Gln Cys?Gly?Thr?Thr?Lys?Val?Pro?Ser?Phe?Thr?Cys?Val
545 550 555
gtc?ctt?gga?caa?atg?taa?tgg?tat?gga?gga?tta?tta?cat?aga?aag?tat 9856
Val?Leu?Gly?Gln?Met Trp?Tyr?Gly?Gly?Leu?Leu?His?Arg?Lys?Tyr
560 565 570
ttt?gca?gta?tgc?tgg?aag?gct?ggg?tca?cct?gag?gtg?cct?cca?gct?ttc 9904
Phe?Ala?Val?Cys?Trp?Lys?Ala?Gly?Ser?Pro?Glu?Val?Pro?Pro?Ala?Phe
575 580 585
ctc?aca?cac?aga?gtt?tca?tag?gct?ccc?taa?aga?att?agg?aga?tct?caa 9952
Leu?Thr?His?Arg?Val?Ser Ala?Pro Arg?Ile?Arg?Arg?Ser?Gln
590 595 600
gtt?tct?aa?aaat?act?gga?ctt?ggg?tga?ttg?tgg?cgg?tac?aat?aga?aga 10000
Val?Ser?Lys?Asn?Thr?Gly?Leu?Gly Leu?Trp?Arg?Tyr?Asn?Arg?Arg
605 610 615
gct?gcc?gga?gga?gtt?ggg?tct?cct?gac?aca?ttt?gtt?gtg?tct?acg?tat 10048
Ala?Ala?Gly?Gly?Val?Gly?Ser?Pro?Asp?Thr?Phe?Val?Val?Ser?Thr?Tyr
620 625 630
tcc?tcg?cag?att?gga?gat?gtt?gcc?agc?tgg?ttt?gat?cgg?aaa?act?gac 10096
Ser?Ser?Gln?Ile?Gly?Asp?Val?Ala?Ser?Trp?Phe?Asp?Arg?Lys?Thr?Asp
635 640 645 650
atc?gct?gca?aga?gct?atg?cat?aag?tct?tat?cga?aga?ggt?cgc?tgt?aag 10144
Ile?Ala?Ala?Arg?Ala?Met?His?Lys?Ser?Tyr?Arg?Arg?Gly?Arg?Cys?Lys
655 660 665
gca?ctt?tgt?gca?gga?gtt?ggg?caa?cct?cag?aga?act?gag?ggt?gct?cta 10192
Ala?Leu?Cys?Ala?Gly?Val?Gly?Gln?Pro?Gln?Arg?Thr?Glu?Gly?Ala?Leu
Resistance gene Pi 36 of rice blast and application sequence table thereof
670 675 680
tgc?caa?aat?tta?tat?agg?gct?gaa?aga?tga?gag?cat?gca?gag?aga?ttt 10240
Cys?Gln?Asn?Leu?Tyr?Arg?Ala?Glu?Arg Glu?His?Ala?Glu?Arg?Phe
685 690 695
tct?aca?gtc?ctt?agg?ctg?tct?aca?taa?ggt?cca?cac?tat?gaa?tat?ata 10288
Ser?Thr?Val?Leu?Arg?Leu?Ser?Thr Gly?Pro?His?Tyr?Glu?Tyr?Ile
700 705 710
tag?cat?tga?agt?aag?gga?atg?cac?cag?acc?aga?tgc?agc?agg?ttc?tgt 10336
His Ser?Lys?Gly?Met?His?Gln?Thr?Arg?Cys?Ser?Arg?Phe?Cys
715 720 725
gtc?ctg?tcc?tcg?gct?ctg?gca?gct?gtc?att?gcc?atg?cat?caa?gtt?ttt 10384
Val?Leu?Ser?Ser?Ala?Leu?Ala?Ala?Val?Ile?Ala?Met?His?Gln?Val?Phe
730 735 740
tag?cct?acc?tgt?gtg?gat?taa?ctc?atc?aac?tct?tgg?ttg?cct?ctc?gca 10432
Pro?Thr?Cys?Val?Asp Leu?Ile?Asn?Ser?Trp?Leu?Pro?Leu?Ala
745 750 755
tct?aga?cgt?gaa?agt?aca?ggt?tgt?gag?aga?gca?gga?cat?gga?aac?cct 10480
Ser?Arg?Arg?Glu?Ser?Thr?Gly?Cys?Glu?Arg?Ala?Gly?His?Gly?Asn?Pro
760 765 770
tgg?aag?gct?ccc?aat?gct?ctg?tta?cct?caa?att?gga?ttc?cca?ata?tac 10528
Trp?Lys?Ala?Pro?Asn?Ala?Leu?Leu?Pro?Gln?Ile?Gly?Phe?Pro?Ile?Tyr
775 780 785
cag?gct?aat?aag?cat?taa?gaa?gcc?tgc?tga?tga?tgg?cta?ctt?cca?aaa 10576
Gln?Ala?Asn?Lys?His Glu?Ala?Cys Trp?Leu?Leu?Pro?Lys
790 795 800
att?gag?att?ctt?taa?aac?tcc?tca?ttc?att?tgt?ccg?gtt?tga?tct?tca 10624
Ile?Glu?Ile?Leu Asn?Ser?Ser?Phe?Ile?Cys?Pro?Val Ser?Ser
805 810 815
tgg?ttg?cga?gtc?cag?cag?tgg?agc?atc?tag?ctt?cat?gcc?aag?act?tga 10672
Trp?Leu?Arg?Val?Gln?Gln?Trp?Ser?Ile Leu?His?Ala?Lys?Thr
820 825
aca?cat?tga?att?cag?tgt?cga?tgt?tcg?gtt?cct?aaa?aga?tgc?agc?aga 10720
Thr?His Ile?Gln?Cys?Arg?Cys?Ser?Val?Pro?Lys?Arg?Cys?Ser?Arg
830 835 840
cct?gca?tgc?tgg?tct?taa?caa?cct?gct?tgc?agg?ctt?caa?tga?ctt?tgg 10768
Pro?Ala?Cys?Trp?Ser Gln?Pro?Ala?Cys?Arg?Leu?Gln Leu?Trp
845 850 855
aag?aac?ttc?gct?caa?gag?tgt?cga?cgc?tag?gat?cca?ctg?caa?gga?tgc 10816
Lys?Asn?Phe?Ala?Gln?Glu?Cys?Arg?Arg Asp?Pro?Leu?Gln?Gly?Cys
860 865 870
cct?tgc?tgc?gga?ggt?gca?gga?agc?cga?ggt?ggc?att?ggc?aaa?tgc?agc 10864
Pro?Cys?Cys?Gly?Gly?Ala?Gly?Ser?Arg?Gly?Gly?Ile?Gly?Lys?Cys?Ser
875 880 885
aca?cgg?cca?tcc?caa?ccg?tcc?agt?cct?tcg?aac?tga?tat?ggc?ata?tga 10912
Thr?Arg?Pro?Ser?Gln?Pro?Ser?Ser?Pro?Ser?Asn Tyr?Gly?Ile
890 895 900
gag?gat?gat?gcg?ctc?acc?tga?tga?cga?taa?g?gtacgtgtat?gcacagtacg 10963
Glu?Asp?Asp?Ala?Leu?Thr Arg
905 910
taatgtactt?agatcatttc?aatcgatttt?ccttatgctg?gttttctgcc?tacatatata 11023
aaatctgatc?ttaattattt?cttgtgctta?tatatatttt?tcag?cc aac?aag?ctg 11078
Ala?Asn?Lys?Leu
gtc?cga?tca?aaa?att?ttt?tga?gat?aca?tgt?cta?tcc?gtc?gtc?gaa?tga 11126
Val?Arg?Ser?Lys?Ile?Phe Asp?Thr?Cys?Leu?Ser?Val?Val?Glu
915 920 925
Resistance gene Pi 36 of rice blast and application sequence table thereof
cca?tca?tcg?tta?cct?tag?cta?cct?ccg?gtt?gtt?gaa?gaa?gcc?acg?cct 11174
Pro?Ser?Ser?Leu?Pro Leu?Pro?Pro?Val?Val?Glu?Glu?Ala?Thr?Pro
930 935 940
gga?gaa?gct?tat?agt?caa?cat?ata?tgt?gtc?caa?gga?tgg?tat?ggt?tgg 11222
Gly?Glu?Ala?Tyr?Ser?Gln?His?Ile?Cys?Val?Gln?Gly?Trp?Tyr?Gly?Trp
945 950 955
gga?tgt?gga?tga?agc?tgt?ggc?agc?tgc?gag?gaa?cgt?agt?cga?cca?tca 11270
Gly?Cys?Gly Ser?Cys?Gly?Ser?Cys?Glu?Glu?Arg?Ser?Arg?Pro?Ser
960 965 970
tat?caa?cca?tcc?aac?act?tga?gat?aaa?tag?aat?gga?aga?aag?tat?ttc 11318
Tyr?Gln?Pro?Ser?Asn?Thr Asp?Lys Asn?Gly?Arg?Lys?Tyr?Phe
975 980 985
tga?cca?aca?tca?aca?g?gcaagatata?tctctcttct?tgagctcgtc?cccaatattg 11374
Pro?Thr?Ser?Thr
990
tactacagag?aaacccagtt?aatttgtaac?accaaccatc?acattctgaa?tcatgtttgc 11434
cttttaattg?tttttatgca?tactctacat?gagagtgcta?tatatatttt?gtttcacttg 11494
cagttgctca?tagtcatttc?acaagataca?tatgcatggt?ggtagactgc?taaaactata 11554
gttcatccta?acatagcatt?attttttgac?tgtttctag?gt ccc?tga?aaa?taa 11607
Gly?Pro Lys
995
ttg?a?gtggcaagct?gttcatttga?aggttaagat?tggcacagtg?atggcacaat 11661
Leu
attaatgaat?ccgagccaga?ctagacggga?agctgtcata?gcattgacta?atctgtaatc 11721
cgatttcaaa?atgcaggata?ttcggatcaa?tttcctcata?tgtcgtctac?tcttttactg 11781
taagtatgac?actttatctt?ttatctcatt?tatgtatctt?tcgtaagtgg?cacagtaata 11841
aggtgacatc?ccattgttat?tcccacacgt?ttgtaaccaa?attctttaat?aattgagtgc 11901
agttctggtg?catgtttgat?tctctgcatt?tggtccaagc?agaggttaca?gcacattatc 11961
aacggagaaa?ataactgaaa?aaaggtgatg?ataaacagaa?tattatgggt?gctgtagcat 12021
aattgactct?gcacctgatc?accctcctca?tgggttggta?agcacaacag?cttgccatct 12081
tattgctgta?caatcttttg?tgaatgatga?taggtgtaat?tggatatgtg?tgctggagaa 12141
aaaatgctca?atccaagtga?caaattgcgc?acttgttttg?atctttaatt?ccctttcaga 12201
gtgatctact?ctaaggacaa?tcttggatca?cccttggatg?aaaaacttaa?ggattggatg 12261
ttggacattt?tacctttgat?aggcctaaaa?acttcatgta?tatattatat?ccaaactaga 12321
ggttataagt?gtgttttggc?tatgtgcatc?atagctatgc?aaatgccggg?agtatgctca 12381
acatagttgt?atcgcctcat?tgtaattgac?tgcgttcaat?aaaatcttcc?attatctaaa 12441
aaatggaaag?cttcaatcca?ccaacagagt?catggccaca?tgattccctc?attggacacc 12501
gaaagattca?aagaaagcag?acttcaggct?gtaaaattgt?ttgtgaaaac?cttcatgttg 12561
catgaatgca?agagtgacct?tgaatatgtt?gcttaatttt?tcagattatg?tgtttcacat 12621
tttctaattt?agtatactgt?gaagtttgcg?ggggaatgta?tagatttagt?tgatgaattc 12681
agtagagcaa?acacaaagta?caggtacaaa?acaataacat?gatgggattc?agatctcaga 12741
aggtgcacag?attcgtaact?gtgaacctac?agaggcacaa?tgcaacaaca?ggtcaccaaa 12801
taatatctca?actgagtaac?aactaaaaat?cattttactt?gaacaacttg?cccaaataga 12861
Resistance gene Pi 36 of rice blast and application sequence table thereof
gtcatcaaaa?ggagaaataa?caacttgccc?aagaataaga?aaatctgatc?caccttaaga 12921
tgaccaccct?agtcatcaaa?atccacatca?cttacaaaac?agatttagga?tcataaccat 12981
caacaacttc?aataacgtaa?ctggtggcct?aggttgtaaa?agaaaacatt?gaaatatgag 13041
tcatggaatt?ccaattcaaa?agacttgagt?gtgcaaatgc?tgaaattaag?atgctcaaca 13101
ttctcagcca?ttttggggat?ggattttgat?ttgagggaga?ggattaataa?tggaaaataa 13161
gatcttcaca?atgatgtgga?ctcttctatc?gccttgtggg?caaatgattt?tgattttcct 13221
atgcttttca?cagttaactg?ctgccctgtt?ccttcgcatg?tgggatctaa?atctgcagta 13281
gcatttgcag?aattggtgtc?gtgatcatag?gcataagctc?catgctggta?aaatgaacaa 13341
ggaaacagtt?aaactcaatt?catgagaaga?aaagaaacag?caatgtggag?agggggagat 13401
ataccgaacc?ttgacaagat?gcagggaaag?tttctctatg?ttaggtgagt?gccgaagcaa 13461
gtacagtagt?ggatagcagc?catcactaaa?aaaccactca?ccaagtgcca?aactcgttag 13521
attgctgaat?atcccacacc?ttggaagatc?acgtagtaat?accttttgga?ccacctgccc 13581
acaagtttta?aactccaaag?atgttaaaga?taggaatata?taaacaacaa?ataagagcaa 13641
ttgtaattta?aagcatttag?cttacatctt?cgtagataga?tgaagacatc?aagtgcaatc 13701
ttgcagcgtt?tgaaagagca?ctgagaatag?tgcagtcaac?atcagaatgt?ccaaaagaga 13761
acctattaag?ggtaaactga?agctgattcc?actgatgata?tgtctaggag?ccgaagattt 13821
ctgtttgcaa?tctcctctac?atgcaggcag?ataaggtttg?gcatatttat?cacaagatgc 13881
tgaaaacctt?ctgggtcatc?ttgagttcta?tctgcagctg?taatggtcag?aatcttcaat 13941
gtagcagagc?aaaacatggt?ggccttgatg?gcacaatgtc?tcaactccag?atgcagtagc 14001
tccgggcaac?cagaaaagag?tttctcgaca?aaacggttat?caatagaaaa?tttgcaaagg 14061
tatacactct?tcaagtgcaa?ggaagtgaag?gggacaggat?atccatctac?agaaaggagc 14121
ttgccatcac?aataattgag?tatgccgaga?gttttaacgt?tgcacctaag?ggcatggcag 14181
acccatggat?ggatgtcaga?ataatcaagg?gaatcatcag?agctgtcata?ggaggtatgg 14241
atacagaaca?tatccaacag?ggcacagctg?cgctgcagga?gcaaattgtc?aacaaacttc 14301
ttaaaatgtt?tcactgtaat?gaagttgcac?gaatcaacat?tgaggtatcc?cacagatgcc 14361
cagacgttct?gccacctccg?agatagcaca?caagtctgca?cagcctcctt?cgctgttagg 14421
aatgtcatga?catggtgcaa?aagttcgtc 14450
<210>2
<211>1056
<212>NBS-LRR
<213〉Oryza paddy rice (Orysa sativa L.)
<400>2
Met?Glu?Phe?Val?Thr?Gly?Ala?Met?Ser?Ser?Leu?Ile?Pro?Lys?Leu
5 10 15
Gly?Glu?Leu?Leu?Lys?Glu?Lys?Tyr?Asn?Leu?Gln?Lys?Asn?Ile?Arg
20 25 30
Gly?Lys?Ile?Glu?Ser?Leu?Ser?Ala?Glu?Leu?Arg?Arg?Ala?Gln?Ala
35 40 45
Val?Leu?Arg?Met?Val?Gly?Glu?Val?Pro?Gln?Asp?Gln?Gln?Asn?Glu
50 55 60
Leu?Val?Lys?Leu?Trp?Ala?Ser?Asp?Leu?Arg?Glu?Ala?Ser?Tyr?Asp
Resistance gene Pi 36 of rice blast and application sequence table thereof
65 70 75
Met?Glu?Asp?Ile?Ile?Asp?Ala?Phe?Leu?Val?His?Val?Asp?Asp?Asp
80 85 90
Gly?Pro?Lys?Pro?Ala?Asp?Pro?His?Val?Leu?Arg?Arg?Leu?Gly?Lys
95 100 105
Gln?Val?Lys?Lys?Leu?Phe?Lys?Lys?Thr?Lys?His?Arg?Val?Thr?Ile
110 115 120
Ala?Asp?Ala?Ile?Gln?6lu?Met?Glu?Lys?Lys?Leu?Leu?Glu?Ile?Asp
125 130 135
Ala?Arg?His?Gly?Arg?Tyr?Pro?Val?Asp?Asn?Ile?Val?Ile?Pro?Ala
140 145 150
Arg?Pro?Thr?Pro?Ile?Asp?Pro?Arg?Ile?Leu?Asn?Ile?Glu?Lys?Met
155 160 165
Ala?Asn?Asn?Leu?Val?Gly?Ile?Asp?Glu?Pro?Arg?Asp?Glu?Leu?Ile
170 175 180
Lys?Met?Leu?Ser?Gln?His?Glu?Arg?Asp?Asn?Asn?Leu?Asn?Thr?Ser
185 190 195
Asn?Arg?Lys?Thr?Lys?Ile?Val?Tyr?Val?Val?Gly?Met?Gly?Gly?Leu
200 205 210
Gly?Lys?Thr?Thr?Leu?Ala?Thr?Ala?Val?Tyr?Glu?Lys?Ile?Lys?Val
215 220 225
Gly?Phe?Pro?Leu?Asn?Ala?Phe?Val?Pro?Ile?Gly?Gln?Asn?Pro?Asn
230 235 240
Met?Lys?Ala?Ile?Leu?Trp?Asn?Ile?Leu?His?Arg?Leu?Gly?Ser?Glu
245 250 255
Lys?Tyr?Leu?Asn?Cys?Pro?Asn?Ile?Glu?Met?Leu?Thr?Val?Gln?Glu
260 265 270
Leu?Ile?Gly?Glu?Leu?Lys?Gln?Phe?Ile?Lys?Gly?Lys?Arg?Phe?Phe
275 280 285
Ile?Val?Ile?Asp?Asp?Ile?Trp?Asp?Lys?Pro?Ser?Trp?Gln?Ile?Leu
290 295 300
Glu?Ser?Gly?Leu?Gln?Asp?Asn?Asp?Tyr?Gly?Ser?Lys?Ile?Leu?Val
305 310 315
Thr?Thr?Arg?Lys?Ser?Glu?Val?Ala?Thr?Ile?Ile?Ser?Asp?Val?Tyr
320 325 330
Asn?Met?Lys?Pro?Leu?Ser?His?Asp?Asn?Ser?Lys?Glu?Leu?Leu?Tyr
335 340 345
Thr?Arg?Thr?Gly?Ser?Glu?Gly?Lys?Ser?Leu?Asp?Ser?Ser?Ser?Thr
350 355 360
Glu?Ala?Cys?Asp?Lys?Ile?Leu?Lys?Lys?Cys?Ala?Gly?Val?Pro?Leu
365 370 375
Ala?Ile?Ile?Thr?Ile?Ala?Ser?Leu?Leu?Ala?Ser?Arg?Ser?Gly?Leu
380 385 390
Asp?Trp?Ser?Glu?Val?Tyr?Arg?Ala?Ile?Asp?Phe?Gly?Glu?Glu?Asp
395 400 405
Asn?Tyr?Glu?Met?Ala?Asn?Thr?Lys?Arg?Ile?Leu?Ser?Phe?Ser?Tyr
410 415 420
Tyr?Asp?Leu?Pro?Ser?His?Leu?Lys?Asn?Cys?Leu?Leu?Tyr?Leu?Ser
425 430 435
Met?Phe?Pro?Glu?Asp?Tyr?Lys?Ile?Asp?Lys?Asn?His?Leu?Ile?Trp
440 445 450
Met?Trp?Ile?Ala?Glu?Gly?Phe?Val?Pro?Glu?Lys?Gln?Asn?Thr?Asn
455 460 465
Leu?Gly?Leu?Tyr?Glu?Leu?Gly?Glu?Ser?Tyr?Phe?Asn?Glu?Leu?IIe
470 475 480
Asn?Arg?Ser?Met?Ile?Gln?Pro?Ile?Glu?Asn?Glu?Phe?Phe?Gly?Cys
485 490 495
Ile?Glu?Gly?Cys?Arg?Val?His?Asp?Met?Val?Leu?Asp?Leu?Ala?Arg
500 505 510
Ser?Leu?Thr?Ser?Glu?Gln?Asn?Phe?Val?Thr?Val?Leu?Asp?Asn?Asp
515 520 525
Glu?Gln?Arg?Lys?Pro?Glu?Ser?Thr?Asn?Ala?Arg?Arg?Leu?Ala?Leu
530 535 540
His?Arg?Thr?Ser?Ile?Thr?Ser?Tyr?Arg?Phe?Val?Asn?Met?Asp?Met
545 550 555
Lys?Lys?Val?Arg?Ser?Phe?Val?Ala?Thr?Glu?Cys?Asn?Asn?Gly?Asn
560 565 570
Asn?Ser?Val?Ala?Pro?Pro?Arg?Phe?Gln?Val?Leu?Arg?Val?Leu?Ser
575 580 585
Leu?Asp?Lys?Cys?Ser?Gly?Met?Glu?Asp?Tyr?Tyr?Ile?Glu?Ser?Ile
590 595 600
Leu?Gln?Tyr?Ala?Gly?Arg?Leu?Gly?His?Leu?Arg?Cys?Leu?Gln?Leu
605 610 615
Ser?Ser?His?Thr?Glu?Phe?His?Arg?Leu?Pro?Lys?Glu?Leu?Gly?Asp
620 625 630
Resistance gene Pi 36 of rice blast and application sequence table thereof
Leu?Lys?Phe?Leu?Lys?Ile?Leu?Asp?Leu?Gly?Asp?Cys?Gly?Gly?Thr
635 640 645
Ile?Glu?Glu?Leu?Pro?Glu?Glu?Leu?Gly?Leu?Leu?Thr?His?Leu?Leu
650 655 660
Cys?Leu?Arg?Ile?Pro?Arg?Arg?Leu?Glu?Met?Leu?Pro?Ala?Gly?Leu
665 670 675
Ile?Gly?Lys?Leu?Thr?Ser?Leu?Gln?Glu?Leu?Cys?Ile?Ser?Leu?Ile
680 685 690
Glu?Glu?Val?Ala?Val?Arg?His?Phe?Val?Gln?Glu?Leu?Gly?Asn?Leu
695 700 705
Arg?Glu?Leu?Arg?Val?Leu?Tyr?Ala?Lys?Ile?Tyr?Ile?Gly?Leu?Lys
710 715 720
Asp?Glu?Ser?Met?Gln?Arg?Asp?Phe?Leu?Gln?Ser?Leu?Gly?Cys?Leu
725 730 735
His?Lys?Val?His?Thr?Met?Asn?Ile?Tyr?Ser?Ile?Glu?Val?Arg?Glu
740 745 750
Cys?Thr?Arg?Pro?Asp?Ala?Ala?Gly?Ser?Val?Ser?Cys?Pro?Arg?Leu
755 760 765
Trp?Gln?Leu?Ser?Leu?Pro?Cys?Ile?Lys?Phe?Phe?Ser?Leu?Pro?Val
770 775 780
Trp?Ile?Asn?Ser?Ser?Thr?Leu?Gly?Cys?Leu?Ser?His?Leu?Asp?Val
785 790 795
Lys?Val?Gln?Val?Val?Arg?Glu?Gln?Asp?Met?Glu?Thr?Leu?Gly?Arg
800 805 810
Leu?Pro?Met?Leu?Cys?Tyr?Leu?Lys?Leu?Asp?Ser?Gln?Tyr?Thr?Arg
815 820 825
Leu?Ile?Ser?Ile?Lys?Lys?Pro?Ala?Asp?Asp?Gly?Tyr?Phe?Gln?Lys
830 835 840
Leu?Arg?Phe?Phe?Lys?Thr?Pro?His?Ser?Phe?Val?Arg?Phe?Asp?Leu
845 850 855
His?Gly?Cys?Glu?Ser?Ser?Ser?Gly?Ala?Ser?Ser?Phe?Met?Pro?Arg
860 865 870
Leu?Glu?His?Ile?Glu?Phe?Ser?Val?Asp?Val?Arg?Phe?Leu?Lys?Asp
875 880 885
Ala?Ala?Asp?Leu?His?Ala?Gly?Leu?Asn?Asn?Leu?Leu?Ala?Gly?Phe
890 895 900
Asn?Asp?Phe?Gly?Arg?Thr?Ser?Leu?Lys?Ser?Val?Asp?Ala?Arg?Ile
905 910 915
His?Cys?Lys?Asp?Ala?Leu?Ala?Ala?Glu?Val?Gln?Glu?Ala?Glu?Val
920 925 930
Ala?Leu?Ala?Asn?Ala?Ala?His?Gly?His?Pro?Asn?Arg?Pro?Val?Leu
935 940 945
Arg?Thr?Asp?Met?Ala?Tyr?Glu?Arg?Met?Met?Arg?Ser?Pro?Asp?Asp
950 955 960
Asp?Lys?Pro?Thr?Ser?Trp?Ser?Asp?Gln?Lys?Phe?Phe?Glu?Val?His
965 970 975
Val?Tyr?Pro?Ser?Ser?Asn?Asp?His?His?Arg?Tyr?Leu?Ser?Tyr?Leu
980 985 990
Arg?Leu?Leu?Lys?Lys?Pro?Arg?Leu?Glu?Lys?Leu?Ile?Val?Asn?Ile
995 1000 1005
Tyr?Val?Ser?Lys?Asp?Gly?Met?Val?Gly?Asp?Val?Asp?Glu?Ala?Val
1010 1015 1020
Ala?Ala?Ala?Arg?Asn?Val?Val?Asp?His?His?Ile?Asn?His?Pro?Thr
1025 1030 1035
Leu?Glu?Ile?Asn?Arg?Met?Glu?Glu?Ser?Ile?Ser?Asp?Gln?His?Gln
1040 1045 1050
Gln?Val?Pro?Glu?Asn?Asn
1055
Claims (9)
1, a kind of resistance gene Pi 36 of rice blast encoded protein matter, it is characterized in that its aminoacid sequence shown in SEQ ID NO:2, or this sequence is replaced, disappearance or add one or several amino-acid residue and the amino acid polypeptide with identical function that forms.
2, protein according to claim 1 is characterized in that aminoacid sequence is shown in SEQ ID NO:2.
3, the nucleotide sequence of the described proteinic resistance gene Pi 36 of rice blast of coding claim 1.
4, resistance gene Pi 36 of rice blast according to claim 3, its nucleotide sequence is shown in SEQID NO:1.
5, a kind of carrier that contains claim 3 or 4 described genes.
6, the described carrier transgenic plant transformed of claim 5.
7, the molecule marker of claim 3 or 4 described genes generations.
8, claim 1 or the 2 described protein application in preparation resisting rice blast bacteria medicine.
9, the described molecule marker of claim 7 has application in the paddy rice of disease resistance in seed selection to rice blast.
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN101050233B (en) * | 2007-03-16 | 2010-11-10 | 华南农业大学 | Nontoxic gene of rice blast bacterium AvrP139, and application |
CN102041262B (en) * | 2009-10-22 | 2012-12-19 | 华南农业大学 | Rice blast resistance gene Pik-p and application thereof |
CN105524154A (en) * | 2016-01-14 | 2016-04-27 | 华南农业大学 | Application of rice blast resistance related gene OsCOL9 |
CN106480051A (en) * | 2015-08-24 | 2017-03-08 | 中国种子集团有限公司 | Recombinant nucleic acid fragment RecCR010113 and its detection method |
CN109468400A (en) * | 2018-12-07 | 2019-03-15 | 袁隆平农业高科技股份有限公司 | Rice blast resistant gene Pi36 codominant marker and application |
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2006
- 2006-03-06 CN CNB200610034048XA patent/CN100556916C/en not_active Expired - Fee Related
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN101050233B (en) * | 2007-03-16 | 2010-11-10 | 华南农业大学 | Nontoxic gene of rice blast bacterium AvrP139, and application |
CN102041262B (en) * | 2009-10-22 | 2012-12-19 | 华南农业大学 | Rice blast resistance gene Pik-p and application thereof |
CN106480051A (en) * | 2015-08-24 | 2017-03-08 | 中国种子集团有限公司 | Recombinant nucleic acid fragment RecCR010113 and its detection method |
CN106480051B (en) * | 2015-08-24 | 2020-06-19 | 中国种子集团有限公司 | Recombinant nucleic acid fragment RecCR010113 and detection method thereof |
CN105524154A (en) * | 2016-01-14 | 2016-04-27 | 华南农业大学 | Application of rice blast resistance related gene OsCOL9 |
CN105524154B (en) * | 2016-01-14 | 2019-07-09 | 华南农业大学 | The application of rice blast resistance related gene OsCOL9 |
CN109468400A (en) * | 2018-12-07 | 2019-03-15 | 袁隆平农业高科技股份有限公司 | Rice blast resistant gene Pi36 codominant marker and application |
CN109468400B (en) * | 2018-12-07 | 2022-03-22 | 袁隆平农业高科技股份有限公司 | Rice blast resistance gene Pi36 codominant molecular marker and application thereof |
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