CN1641029A - Wheat disease-resistant gene Lr-L1 and its use - Google Patents

Abstract

The present invention relates to plant gene engineering technology, and is especially one kind of wheat disease resisting gene Lr-L1 and its application. The wheat disease resisting gene Lr-L1 has cDNA of whole length 2326 bp, including 5'-noncoding area of 135 bp, 3'-noncoding area of 249 bp, coding area of 1911 bp, terminator, and polyadenyl acid tail of 28 bp. The 3'-noncoding area includes includes 3 typical tailing signal sequences and can reinforce the tailing process after gene transcription. The wheat disease resisting gene Lr-L1 may be inserted into the downstream of different types of promoter to constitute plant expression vector for plant disease resisting improvement in gene engineering technology, and may be cloned for making wheat gene chip for wheat disease resistance and other research.

Description

Wheat resistance genes Lr-L1 and application thereof

One. technical field: the present invention relates to the plant gene engineering technology field, particularly relate to a kind of wheat resistance genes Lr-L1 and application thereof.

Two. background technology:

Plant transgene research comes from the initial stage eighties, nineteen eighty-three Zambryski has obtained the first routine transfer-gen plant in the world, Horch had created the Ye Panfa in the Agrobacterium-mediated Transformation in 1985, henceforth, set up agrobacterium-mediated transformation respectively, virus-mediated method, the PEG mediated method, electricity swashs perforation method, microinjection, pollen tube passage method, supersonic method, particle bombardment etc., the species of transgenosis success constantly enlarge, relate to more than 50 species totally 110 various plants, wherein, the transgenic plant that agrobacterium-mediated transformation obtains account for more than 85% of transgenic plant sum, the transgenic plant that particle bombardment obtains account for about 10%, dicotyledons is the natural host of Agrobacterium, utilize Agrobacterium Ti-plasmids mediated method to set up the gene transfer system of multiple dicotyledons, and some good foreign genes have been changed over to dicotyledons, bred transformed variety, but utilized agrobacterium-mediated transformation relatively slow the work progress that monocotyledons carries out gene transformation.

Wheat is one of topmost food crop, but its engineered flow of research has lagged behind other crop, and wheat transgenic research starts from the initial stage eighties.Vasil in 1992 etc. utilize the particle gun mediated method to obtain the first routine wheat transgenic plant in the world, and once monarch's happiness, Cheng Zhuomin etc. utilized pollen tube passage method to obtain the wheat transgenic plant respectively in 1994.In afterwards several years, wheat transgenic is studied basically by means of the particle gun mediated method.According to statistics, up to the present obtain in the report of wheat transgenic, particle bombardment accounts for about 90%, and other method only accounts for about in the of 10%, and reason is the method comparative maturity of particle gun mediated method, and agrobacterium-mediated transformation is also relatively more difficult.But, compare with the particle gun mediated method, that agrobacterium-mediated transformation has is simple to operate, cost is low, transformation efficiency is high, good reproducibility, can import advantages such as large fragment DNA, and the gene that imports is generally single copy and integrates, be unlikely to the reticent phenomenon of producer, yet the agriculture bacillus mediated transgenosis of wheat is an a difficult problem in the world always, though Hess, Mooney etc. did trial once, fail to obtain transfer-gen plant.Raising along with gene isolation technology and clone's means, to there be more important gene relevant to be cloned out from now on the wheat improvement, some practical problemss in the while Wheat Production, come out in succession as aphid, head blight, Powdery Mildew, rust, gaeumannomyces graminis disease, banded sclerotial blight, soil-borne disease viral disease, arid, problem such as saline and alkaline, and genetic engineering breeding may be best solution route.

Plant is subjected to the infringement of many pathogens in the process of growth, the phytopathy original of a great variety comprises virus, bacterium, mould and nematode etc.The pathogen invaded plants causes two kinds of results: 1. breeding in host plant of pathogen success causes relevant illness; 2. host plant produces defense response, kills pathogen or stops its growth.Disease is the major obstacle of Wheat Production, and most of diseases of wheat belong to fungal disease, and it is one of approach of control wheat diseases that the external source anti-fungal gene is imported wheat, and agrobacterium-mediated transformation generally commonly used carries out gene transformation to wheat.Number of patent application is 03109435.X, denomination of invention is for disclosing the agriculture bacillus mediated method of wheat being carried out gene transformation of a kind of usefulness in " the agriculture bacillus mediated method of wheat being carried out gene transformation of a kind of usefulness ", it is that WHEAT CALLUS and the Agrobacterium that contains the Ti-plasmids carrier are cultivated altogether, foreign gene on the plasmid vector is transferred in the acceptor gene group, but before conversion, will be carried out resuspended with the nutrient solution that contains ethyl sulfonic acid, Methionin, octopine, glutamine, Syringylethanone, glucose, maltose etc.At present worldwide, fewer comparatively speaking to the research of separation, clone and the disease resistance response of wheat resistance genes.

Three. summary of the invention:

The objective of the invention is: overcome the deficiency that present wheat resistance genes exists, a kind of new wheat resistance genes Lr-L1 and the application of this gene are provided.

Technical scheme of the present invention is:

The full length cDNA sequence of a kind of wheat resistance genes Lr-L1, the nucleotide sequence based composition is:

672a 534c 535g 585t

1 acgcggggaa?ttgaaacact?ctcctcgtcc?tcagaggcct?aactatcaat?agagcatgac

61 tctgactgcg?tacctactcg?ccactccact?tcttcttcct?ctgtgctcac?catcttggta

121 caacagtaca?gaaagatgag?taaattactt?gccatagcac?tgctgctgct?gcatcttatc

181 aaccacaaaa?tcaacatggc?aacggcatgg?gatgatcaag?atttcttcaa?atactgccca

241 ccatcccact?gcagccaaca?tggcccagag?atcaggtatc?ctttctgcct?tgaatccaac

301 aatacatcat?catgtggatg?tagtgggaaa?tcaatcagga?agatagcatg?ctctggccaa

361 gacaccattc?tagtccaccc?agttctttgc?ccatacagtg?tcagcgccat?agattacaag

421 cgttcttcca?tgaagatcac?cccacttgta?gacccctgtt?tggtactcca?gcagaagctc

481 gccatctcca?gaagctcgtc?atctccacag?gttgatgtta?tcaacgatga?gaagccaagt

541 cttgacagat?atttattttg?gagttctaca?atttccctag?tacgctgttc?aagagagttt

601 gcacctggtg?cagccgatgg?gattgctggc?ccagtctcct?gccttagcaa?cataacccac

661 ttcttttatt?tggtgaatgg?ttatgaagac?atgtctattc?ttccgttgga?ctgcaaggtc

721 gtcccaccct?cagatggtgt?cggtggcggt?cagataacca?tgtatatgtt?tgacgaccca

781 atgtcagaca?cgctctcact?gtccttcaag?gaacgcacag?aaagaatcct?cggttttgct

841 gagacgacag?tgtattggta?taattactat?tgcaaaggat?gtgaacgcag?tgggggacgc

901 tgcgcgttca?gctcacaaag?ggatagaaaa?ttctgcatgc?ccggcccaca?tggttcacgt

961 atcaaagtca?ttgcagctac?atcatcagtg?gccgcatttg?tggttctttt?gttaacggtg

1021?gccactgtgc?tttatctttc?actcaagaca?agatataatg?cagagataca?tttgaaggtt

1081?gaaatgttcc?tcaagacata?tggcacatcg?aaacccacaa?ggtacacttt?ctctgaagtt

1141?aagaagatgg?caagacggtt?taaggaaaaa?gtagggcagg?gaggatttgg?gagcgtatac

1201?aaaggtgagc?tgccaaatgg?agtgcccgtg?gcagtcaaga?tgctagagaa?ctctacagga

1261?gagggagaag?cattcatcaa?tgaagttgca?accatcggac?ttatccatca?tgccaatatt

1321?gtccgcctcc?tgggattctg?ctctgaagga?atgaggcggg?ctcttattta?tgaattcatg

1381?cctaatgagt?cactggagaa?atacatattc?tctgacgact?ctaatatttt?tcagaatctt

1441?ctagtaccag?ataaactgct?agatattgct?ttaggcatcg?cccgaggtat?ggagtacttg

1501?catcaagggt?gcaaccagcg?catcctccac?tttgacatca?agcctcacaa?tatcctgctg

1561?gactacaact?tcaatccaaa?gatctcagac?tttggccttg?caaagctgtg?cgcaagggac

1621?caaagcatcg?tgaccttaac?agcagcaaga?ggcacaatgg?gctacattgc?accagagcta

1681?tactcccgga?actttggggg?agtatcgtac?aagtcagacg?tgtacagttt?cgggatgctg

1741?gtgctagaaa?tggtgagcgg?gaggaggaat?tcaggcccaa?gaatcgggag?ccaggacgat

1801?gtttacctcc?ctgagtggat?ctacgagaaa?gtggtcaatg?gggaggactt?ggcgcttact

1861?ttggaaacga?ctgaagaaga?taaagaaaag?gtgaggaagc?tggctatggt?ggcactgtgg

1921?tgtatccagt?ggaacccaag?aaaccggccg?tcgatgacaa?aggtcgtgaa?catgctaact

1981?gggaggcttc?agagtctgca?gatgccacca?aagccttatg?tctcatccga?gaatgaactt

2041?atgccataaa?ttaaagcatg?tcgcacggac?taccatactc?tgtcaggttg?tactttgtcc

2101?acgcgtgcgc?tgcaaataat?agcaaccact?gtgtttcgac?gctttcagtc?agtcaattag

2161?gtctgaagat?gtatgaataa?tcaccggaca?ctagcagcat?gtaatagtgt?cactagttgt

2221?ttgcaagata?ctggtgcttc?catatttatg?tccaaaaact?tgcaatagag?ctgtgtaaaa

2281?taaaataata?atctgtcgaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaa

Wheat resistance genes Lr-L1 encoded protein, its aminoacid sequence is:

1?MSKLLAIALL?LLHLINHKIN?MATAWDDQDF?FKYCPPSHCS?QHGPEIRYPF?CLESNNTSSC

61 GCSGKSIRKI?ACSGQDTILV?HPVLCPYSVS?AIDYKRSSMK?ITPLVDPCLV?LQQKLAISR5

121?SSSPQVDVIN?DEKPSLDRYL?FWSSTISLVR?CSREFAPGAA?DGIAGPVSCL?SNITHFFYLV

181?NGYEDMSILP?LDCKVVPPSD?GVGGGQITMY?MFDDPMSDTL?SLSFKERTER?ILGFAETTVY

241?WYNYYCKGCE?RSGGRCAFSS?QRDRKFCMPG?PHGSRIKVIA?ATSSVAAFVV?LLLTVATVLY

301?LSLKTRYNAE?IHLKVEMFLK?TYGTSKPTRY?TFSEVKKMAR?RFKEKVGQGG?FGSVYKGELP

361?NGVPVAVKML?ENSTGEGEAF?INEVATIGLI?HHANIVRLLG?FCSEGMRRAL?IYEFMPNESL

421?EKYIFSDDSN?IFQNLLVPDK?LLDIALGIAR?GMEYLHQGCN?QRILHFDIKP?HNILLDYNFN

481?PKISDFGLAK?LCARDQSIVT?LTAARGTMGY?IAPELYSRNF?GGVSYKSDVY?SFGMLVLEMV

541?SGRRNSGPRI?GSQDDVYLPE?WIYEKVVNGE?DLALTLETTE?EDKEKVRKLA?MVALWCIQWN

601?PRNRPSMTKV?VNMLTGRLQS?LQMPPKPYVS?SENELMP

Wheat resistance genes Lr-L1 encoded protein, its genes encoding be a receptoroid protein kinase, comprise N-end signal peptide, extracellular domain, middle part membrane spaning domain, highly charged sequence and C-end protein kinase domain.

The application of wheat resistance genes Lr-L1 in the plant disease-resistant improvement.

The application of wheat resistance genes Lr-L1 in the wheat cdna chip manufacturing.

The application of wheat resistance genes Lr-L1 in synthetic new disease-resistant gene.

Positive beneficial effect of the present invention is:

1. transgenosis breeding for disease resistance: wheat resistance genes Lr-L1 can be inserted into downstream of different types of promoter, is built into plant expression vector, carries out the plant disease-resistant improvement by genetic engineering technique.Promotor can be the promotor of constitutive expression, as the promotor of cauliflower mosaic virus 35S, corn ubiquitin promoter Ubi etc.The gene that is connected the composition type expression promoter downstream will be expressed at any tissue and any developmental stage of transgenic plant.Promotor also can be the promotor of pathogenic bacterium inducing expression promoter or tissue specific expression.The gene that is connected under these promotors is only expressed when pathogenic bacteria exists or in particular organization, makes this expression of gene be subjected to more accurate control.

2. the synthetic new disease-resistant gene of design: with wheat resistance genes Lr-L1 and the disease-resistant protein kinase of LRK10, YRK1 more as can be known, these proteic carboxyl terminal protein kinase function territory high conservatives, and having than big-difference at N-terminal extracellular domain.Therefore, similar to the mankind's immunoglobulin (Ig), N-terminal extracellular domain changes the disease-resistant specificity of decision gene.By technology such as rite-directed mutagenesis or gene order reorganization, the gene that can synthetic much has new disease-resistant characteristic is applied in the plant disease-resistant improvement by genetic engineering technique.

3. gene chip is made: the clone of this wheat resistance genes Lr-L1 can be used for the wheat cdna chip manufacturing, be applied in the research such as disease-resistant wheat.According to Lr-L1 gene cDNA sequence design PCR primer, the full-length cDNA of amplification Lr-L1 gene in containing the clone of Ben Jiyin, or the strong section of the gene specific of amplification coding extracellular domain carry out the making of gene chip.

4.Lr-L1 the expression of gene in the mildew-resistance reaction: the Lr-L1 gene is specific expressed in wheat leaf blade, in young stem and young fringe very micro-expression is only arranged, and does not express at root.When the attack that is subjected to wheat powdery mildew, express significantly enhancing, the enhancing amplitude can reach more than 5 times, and as shown in Figure 1, the express spectra of Lr-L1 changes has proved that it plays an important role in the wheat anti-powdery mildew reaction.

5. the coded proteic characteristics of wheat resistance genes Lr-L1 of the present invention are: a receptoroid protein kinase of this genes encoding (RLK), this receptoroid protein kinase comprises several sections such as N-end signal peptide, extracellular domain, middle part membrane spaning domain and C-end protein kinase domain, and its structure is embroidered disease protein kinase LRK10 with the anti-leaf of the wheat of having cloned, and (Feuillet etc. 1997; The GanBank query ID: Gi|1680686|gb|AAC49629.1|) and the anti-stripe rust protein kinase of wheat YRK1 (the GanBank query ID: Gi|34809101|gb|AAQ82627.1|) highly similar, wheat resistance genes Lr-L1 of the present invention has disease resistance from the molecular structure explanation.

Four. description of drawings:

Fig. 1 is for changing by the expression of quantitative RT-PCR technical Analysis Lr-L1 gene in wheat anti-powdery mildew reaction, and M is a molecular weight standard figure on, 0,1,12,24 etc. digital be that wheat powdery mildew is inoculated induction time; Actin is the wheat actin gene, and rRNA is a ribosome-RNA(rRNA), is the stably express gene, as the contrast of changes in gene expression.The result as seen, the Lr-L1 gene is expressed significantly in white powder germ inoculation back and is strengthened, the white powder germ inoculates back 24 hours expression of gene amounts and reaches when not inoculating about 5 times.

Five. embodiment:

Embodiment one: clone's process of wheat resistance genes Lr-L1:

Wheat Lr-L1 gene is to isolating the very high disease-resistant wheat new lines " 99-2439 " of Powdery Mildew, rust resistance degree.Inducing back 22 hours blade with the white powder germ is the total RNA of material extraction (Yeast Nucleic Acid), further again separating mRNA (messenger RNA(mRNA)), by reverse transcription mRNA being transcribed into cDNA (complementary DNA (cDNA)), is that template is carried out gene clone with this cDNA.According to the conservative VI functional domain of vegetable-protein kinases designed a merger property primer (sequence is: 5 '-(G/A) TC (A/G) AA (G/A) T (G/A) GAG (G/A) ATGCG-3 ' of GTGAGGCTT (G/A) A (T/C)), by rapid amplifying cDNA end technology (5 '-RACE), a lot of products have been obtained, the amplified production of one of them about 1500bp is through reclaiming, clone on pGEM-T Eseay carrier, the order-checking back proves the part that this cDNA fragment is the receptoroid protein kinase gene by sequential analysis.

Use special test kit (SMART ^TMRACE cDNA Amplification Kit) increases.Solution preparation by specification carries out.Amplification program is 94 ℃ of sex change of 5 round-robin 15 seconds, anneals and extends 3 minutes for 72 ℃; 94 ℃ of sex change of 25 round-robin 15 seconds were annealed 30 seconds for 68 ℃, and 72 ℃ were extended 3 minutes; Extended 7 minutes at 72 ℃ at last.Amplified production is electrophoretic separation on 1% sepharose.The sepharose that will contain target DNA fragment cuts down, and reclaims equal-volume chloroform extracting and purifying, the alcohol precipitation of 2 times of volumes with the liquid nitrogen multigelation.The dna fragmentation that reclaims is connected it with the T4-DNA ligase enzyme after dissolving with ultrapure water with pGEM-T Eseay carrier, 4 ℃ of connections are spent the night.Use CaCl ₂The thermal shock method 42 ℃ of thermal shocks 90 seconds, will connect product and be transformed in the intestinal bacteria DH10B bacterial strain.Extract plasmid DNA with plasmid a small amount of extraction method.Precious biotech firm checks order with the ABI377 automatic sequencer in Dalian.Sequencing result with BLASTt and BLASTp computer aided program (Altschul etc. 1997) American National bioinformation center website NCBI ( Http:// www.ncbi.nlm.nih.gov) carry out the homologous sequence inquiry relatively with relevant networking website.Guard the functional domain data analysis in the NCBI website with the method for (2003) such as Marchler-Bauer.On this basis, designed again a gene-specific primer (sequence is: 5 '-ACCTACTCGCCACTCCACTTCTTCTTCCTC-3 '), by rapid amplifying cDNA end technology (3 '-RACE) obtained full length cDNA clone.This gene is inserted on the pGEM-T Esay carrier, and is kept in the intestinal bacteria DH10B bacterial strain.

In GenBank, carry out the similarity inquiry relatively, prove that the full-length cDNA of being cloned is new wheat cdna, called after wheat resistance genes Lr-L1.The preceding 1551bp part of this gene has been deposited among the GenBank as the new gene of wheat and has been logined, and is numbered: AY584533

The structure of wheat resistance genes Lr-L1 is embroidered disease protein kinase LRK10 with the anti-leaf of the wheat of having cloned, and (Feuillet etc. 1997; The GanBank query ID: Gi|1680686|gb|AAC49629.1|) and the anti-stripe rust protein kinase of wheat YRK1 (the GanBank query ID: Gi|34809101|gb|AAQ82627.1|) highly similar, its comparative result is as follows:

(1) comparison of the anti-leaf rust protein kinase of Lr-L1 receptoroid protein kinase and wheat (LRK10):

Lr-L1：1 MSKLLAIALLLLHLINHKINMATAWDDQDFFKYCPPSHCSQHGPEIRYPFCLESNNTSS-?59

MSKLL?IALLLL?LINH?I?+ATAWDDQDFFKYCPPS?CSQHGP?IRYP?CLES+NTSS

LRK10：1 MSKLLVIALLLLPLINHGIYLATAWDDQDFFKYCPPSKCSQHGPMIRYPLCLESSNTSSS?60

Lr-L1：60?--CGCSGKSIRKIACSGQDTILVHPVLCPYSVSAIDYKRSSMKITPLVDPCLVLQQKLAI?117

CGC+G+SI?K+ACSGQDTILVHPVL?PYSVSAIDY+RSSMKITPLVDPCLVLQQKL?I

LRK10：61?SSCGCAGRSIWKLACSGQDTILVHPVLGPYSVSAIDYRRSSMKITPLVDPCLVLQQKLII?120

Lr-L1：118?SRSSSSPQVDVINDEKPSLDRYLFWSSTISLVRCSREFAPGAA--DGIAGPVSCLSNITH?175

SRSSSSPQVDVINDEKPS?D F?SS+?++V?CSREF?P?AA D?IAGPVSCLSN?TH

LRK10：121?SRSSSSPQVDVINDEKPSFDENFFESSSATIVHCSREFTPAAAHADSIAGPVSCLSNTTH?180

Lr-L1：176?FFYLVNGYEDMSILPLDCKVVPPSDGVGGGQITMYMFDDPMSDTLSLSFKERTERILGFA?235

FFYLVN EDMSILPLDCKVVP?SD GG +M D?M +F?E ++I FA

LRK10：181?FFYLVNSDEDMSILPLDCKVVPVSDR--GGISLPHMLKDQMF----YNFTETAKKIPSFA?234

r-L1：236?ETTVYWYNYYCKGCERSGGRCAFSSQRDRKFCMPGPHGSRIKVIAATSSVAAFVVLLLTV?295

ET?V?W C+?CE?SG?RCAFSSQRDR+FCMP?PHGS?IKVIAATSSVAAFV?LLLTV

LRK10：235?ETAVSWDEGDCRECELSGRRCAFSSQRDREFCMPDPHGSHIKVIAATSSVAAFVALLLTV?294

Lr-L1：296?ATVLYLSLKTRYNAEIHLKVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKVGQGGFGSVY?355

ATVLYLSLKTRYNAEIH+KVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKVGQGGFGSVY

LRK10：295?ATVLYLSLKTRYNAEIHMKVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKVGQGGFGSVY?354

Lr-L1：356?KGELPNGVPVAVKMLENSTGEGEAFINEVATIGLIHHANIVRLLGFCSEGMRRALIYEFM?415

KGELPNGVPVAVKMLENSTGEGE+FINEVATIGLIHHANIVRLLGFCSEGMRRALIYEFM

LRK10：355?KGELPNGVPVAVKMLENSTGEGESFINEVATIGLIHHANIVRLLGFCSEGMRRALIYEFM?414

Lr-L1：416?PNESLEKYIFSDDSNIFQNLLYPDKLLDIALGIARGMEYLHQGCNQRILHFDIKPHNILL?475

PNESLEKYIFSDDSNIFQNLLVP+KLLDIALGIARGMEYLHQGCNQRILHFDIKPHNILL

LRK10：415?PNESLEKYIFSDDSNIFQNLLVPEKLLDIALGIARGMEYLHQGCNQRILHFDIKPHNILL?474

Lr-L1：476?DYNFNPKISDFGLAKLCARDQSIVTLTAARGTMGYIAPELYSRNFGGVSYKSDVYSFGML?535

DYNFNPKISDFGLAKLCARDQSIVTLTAARGTMGYIAPELYSRNFGGVSYK+DVYSFGML

LRK10：475?DYNFNPKISDFGLAKLCARDQSIVTLTAARGTMGYIAPELYSRNFGGVSYKADVYSFGML?534

Lr-L1：536?VLEMVSGRRNSGPRIGSQDDVYLPEWIYEKVVNGEDLALTLETTEEDKEKVRKLAMVALW?595

VLEMVSGRRNS?PRIGSQDDVYLPEWIYEKV+NGE+LALTLETT+E+K+KVR+LAMVALW

LRK10：535?VLEMVSGRRNSDPRIGSQDDVYLPEWIYEKVINGEELALTLETTQEEKDKVRQLAMVALW?594

Lr-L1：596?CIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKPYVSSENELM?636

CIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKP+VSSENELM

LRK10：595?CIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKPFVSSENELM?635

(2) comparison of Lr-L1 receptoroid protein kinase and wheat stripe rust resisting protein kinase YRK1

Lr-L1：1 MSKLLAIALLLLHLINHKINMATAWDDQDFFKYCPPSHCSQHGPEIRYPFCLESNNTSSC?60

MSKLL?IALLLL?LINH?I?+ATAWDDQDFFKYCPPS+CSQHGPE+R+PFCLES+NTS+

YRK1：1 MSKLLVIALLLLPLINHGIYLATAWDDQDFFKYCPPSNCSGHGPEVRFPFCLESSNTSAV?60

Lr-L1：61 GCSGKS----------IRKIACSGQDTILVHPVLCPYSVSAIDYKRSSMKITPLVDPCLV?110

+ + I?K+AC?GQDTILVHPVL YSVSAIDY+RSS+K+?PLVDPCL+

YRK1：61 AAAAAASCGCSSTDGLIWKLACFGQDTILVHPVLGSYSVSAIDYRRSSVKLIPLVDPCLM?120

Lr-L1：111?LQQKLAISRSSSSPQVDVIND-EKPSLDRYLFWSSTISLVRCSREFAPGA--ADGIAGPV?167

LQQKLAISR SS?QVDVI+ +?P + +LV?CS?EF?P?A AD?IAGPV

YRK1：121 LQQKLAISRRWSSRGVDVIDGMQLPDRQFSILSYKYATLVCCSSEFTPAAVDADSIAGPV?180

Lr-L1：168?SCLSNITHFFYLVNGYEDMSILPLDCKVVPPSDGVGGGQITMYMFDDPMSDT-LSLSFKE?226

SCLSN?T?F?YLV Y+DMSILPLDCKV?P?SDGV?G I?MYM?+DPMS?T SLSFKE

YRK1：181 SCLSNTTLFLYLVAAYKDMSILPLDCKVFPVSDGVSGRLIPMYMLEDPMSGTPFSLSFKE?240

Lr-L1：227?RTERILGFAETTVYWYNYYCKGCERSGGRCAFSSQRDRKFCMPGPHGSRIKVIAATSSVA?286

R?ER+L?FAETTVYW+ C+?CE?SG?RCAFSSQRD FC+P?PHGSRIKVIAATSSVA

YRK1：241 RAERMLSFAETTVYWFGGNCRQCELSGQRCAFSSQRDETFCIPDPHGSRIKVIAATSSVA?300

Lr-L1：287?AFVVLLLTVATVLYLSLKTRYNAEIHLKVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKV?346

AFVVLL+TVATVLY?SLKTRYNAEIHLKVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKV

YRK1：301 AFVVLLVTVATVLYPSLKTRYNAEIHLKVEMFLKTYGTSKPTRYTFSEVKKMARRFKEKV?360

Lr-L1：347?GQGGFGSVYKGELPNGVPVAVKMLENSTGEGEAFINEVATIGLIHHANIVRLLGFCSEGM?406

GQGGFGSVYKGEL?NGVPVAVKMLENSTGEGEAFINEVATIGLI+HANIV+LLGFCSEGM

YRK1：361 GQGGFGSVYKGELQNGVPVAVKMLENSTGEGEAFINEVATIGLIYHANIVQLLGFCSEGM?420

Lr-L1：407?RRALIYEFMPNESLEKYIFSDDSNIFQNLLVPDKLLDIALGIARGMEYLHQGCNQRILHF?466

RRALIYEFMPNESLEKYIFS?DS FQ+LLVPDKLLDIALGIARGMEYLHQGCNQRILHF

YRK1：421 RRALIYEFMPNESLEKYIFSRDSANFQHLLVPDKLLDIALGIARGMEYLHQGCNQRILHF?480

Lr-L1：467?DIKPHNILLDYNFNPKISDFGLAKLCARDQSIVTLTAARGTMGYIAPELYSRNFGGVSYK?526

DIKPHNILLDYNFNPKISDFGLAKLCARDQSIVTLTAARG?MGYIAPELYSRNFGGVSYK

YRK1：481 DIKPHNILLDYNFNPKISDFGLAKLCARDQSIVTLTAARGKMGYIAPELYSRNFGGVSYK?540

Lr-L1：527?SDVYSFGMLVLEMVSGRRNSGPRIGSQDDVYLPEWIYEKVVNGEDLALTLETTEEDKEKV?586

SDVYSFGMLVLEMVSGRRNS?PRIGSQDDVYLPEWIYEKV+NGE+LALTLETT+E+K+KV

YRK1：541 SDVYSFGMLVLEMVSGRRNSDPRIGSQDDVYLPEWIYEKVINGEELALTLETTQEEKDKV?600

Lr-L1：587?RKLAMVALWCIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKPYVSSENELMP?637

+LAMVALWCIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKP+VS?ENE?MP

YRK1：601 SQLAMVALWCIQWNPRNRPSMTKVVNMLTGRLQSLQMPPKPFVSYENEPMP?651

Above comparative result can have disease resistance from molecular structure explanation wheat resistance genes Lr-L1 of the present invention.

Embodiment two: the transgenosis breeding for disease resistance:

Lr-L1 gene enzyme from the pGEM-T easy carrier is scaled off, or increase with high-fidelity Taq-DNA polysaccharase and gene-specific primer., be inserted into again in the pCAMBIA-Bar plant expression vector, thereby be built into pCAMBIA-Ubi-Lr-L1 expression vector plasmid the downstream that Lr-L1 is connected corn ubiquitin efficient promoter Ubi (Ubiquitin-1) with the T4-DNA ligase enzyme.This expression vector is transformed in the intestinal bacteria DH10B bacterial strain breeds, extract the plasmid of this carrier, can carry out the transgenosis breeding for disease resistance.(the high-pressure helium particle gun as Bole company PDS-1000/He) is transformed into high yield, high-quality with pCAMBIA-Ubi-Lr-L1 expression vector plasmid, but in the rataria callus of the wheat breed of disease resistance difference by particle gun.Utilize the selection markers gene on the expression vector---anti-herbicide gene Bar, in being added with the MS solid medium of weedicide PPT, screen, obtain the transgenic calli of anti-PPT.To screen the division culture medium (PPT that contains 3mg/L that the transgenic calli that obtains forwards new induced tissue differentiation to again; The Zeatin of 1mg/L and the IAA of 1mg/L, 1/2 MS solid medium) the middle cultivation, induce to produce new regeneration plant.So just obtained transgenic wheat.Detect by the offspring of round pcr to transfer-gen plant with gene-specific primer, select gene pure, the desirable transgenic strain that disease resistance is good can carry out field planting, popularization.

Embodiment three: the synthetic new disease-resistant gene of design:

Modern DNA synthetic technology can be synthesized full-length gene.Therefore,, the synthetic back of the cDNA of LRK10 genes encoding 1-180 amino-acid residue is connected to the gene cDNA terminal portions with Lr-L1 genes encoding 176 amino acids residues, has so just synthesized brand-new gene according to the difference of Lr-L1 gene and LRK10 gene.Then, whether the encoder block (ORF) by the sequence verification synthetic gene is correct.Gene by checking can further be building up in the plant expression vector, detects by the disease resistance of transgenic technology to synthetic gene.Synthetic gene with good resistance characteristic of disease can be used for the cultivation of transgenosis anti-disease wheat new variety (structure of plant expression vector and genetically modified concrete operations as (1) example as described in).

Embodiment four: gene chip is made:

With the Lr-L1 gene cDNA clone is template, obtains the DNA product with the M13 universal primer on gene-specific primer or the pGEM-T easy carrier by pcr amplification.Product is carried out purifying, after the dissolving, make the wheat cdna chip, be used for the disease-resistant correlative study of Denging according to the gene chip production process.As can put automatically with the BioRobotics of Britain the film instrument with specimens point on 8 * 12 centimetres nylon membrane.2 points of each sample spot, the DNA of each some amount is several nanograms, with house-keeping gene on the time point (as 18sRNA and actin gene) as the stably express genetic contrast.The mRNA extraction back isotropic substance of experimental study material (as ³³P) mark is hybridized with gene chip as probe.Analyze hybridization signal with registering instrument.Analyze Lr-L1 expression of gene variation under the different experimental conditions according to the power of hybridization signal.The relation that research Lr-L1 gene and disease-resistant etc. reacts.

Embodiment five: the expression of Lr-L1 gene in the mildew-resistance reaction:

The concrete grammar of expression analysis: the amplification of (1) stably express crt gene-wheat actin gene (actin): according to sequence (the GenBank query ID: gi:48927617) designed a pair of primer, WAC-F:5 '-GTTCCAATCTATG AGGGATACACGC-3 ' and WAC-R:5 '-GAACCTCCACTGAGAACAACATTACC-3 ' of the actin gene cDNA fragment among the GenBank.Reverse transcription PCR carries out on the Perkin-Elmer9600 gene-amplificative instrament.Reaction solution is 25 μ L systems, comprises the quantitative equivalent cDNA of process of 3 μ L, every kind of gene-specific primer of 0.4 μ mol/L, the dNTP of 200 μ mol/L, 1 times PCR damping fluid, the MgCl of 1.5mmol/L ₂, the Taq-DNA polysaccharase of 1-2 unit.Amplification program is: 94 ℃ of sex change 3 minutes; 94 ℃ of sex change of 18 round-robin 15 seconds were annealed 15 seconds for 55 ℃, and 72 ℃ were extended 1 minute; Extended 7 minutes at 72 ℃ at last.Amplified production is electrophoretic separation in 1% sepharose, the dyeing of yttrium bromide ingot, and ultraviolet lamp is observed, is taken a picture down.The specific amplification products of wheat actin gene is that 422bp is long.(2) amplification of wheat Lr-L1 gene: method is identical with aforesaid method, and just the annealing temperature difference of amplification is 53 ℃.Gene-specific primer is: p1:5 '-TATCAACGATGAGAAGCCAAGT-3 '; P2:5 '-TCCCAAATCCTC CCTGCCCTAC-3 '.The specific amplification products of Lr-L1 gene is that 674bp is long.

Main reference document of the present invention:

1.Feuillet?C，Schachermayr?G，Keller?B.1997.Molecular?cloning?of?a?newreceptor-like?kinase?gene?encoded?at?the?Lr10?disease?resistance?locus?of?wheat.Plant?J，11：45-52

2.Altschul?S?F，Madden?T?L，Schffer?A?A，Zhang?J，Zhang?Z，Miller?W，LipmanD?J.1997.Gapped?BLAST?and?PSI-BLAST：a?new?generation?of?protein?databasesearch?program.Nucleic?Acids?Res，25：3389-3402.

3.Marchler-Bauer?A，Anderson?J?B，DeWeese-Scott?C，Fedorova?N?D，Geer?L?Y，He?S，Hurwitz?D?I，Jackson?J?D，Jacobs?A?R，Lanczycki?C?J，Liebert?C?A，Liu?C，Madej?T，Marchler?G?H，Mazumder?R，Nikolskaya?A?N，Panchenko?A?R，Rao?B?S，Shoemaker?B?A，Simonyan?V，Song?J?S，Thiessen?P?A，Vasudevan?S，Wang?Y，Yamashita?R?A，Yin?J?J，Bryant?S?H.2003.“CDD：a?curated?Entrez?database?ofconserved?domain?alignments”.Nucleic?Acids?Res.31：383-387

Sequence table

＜110〉Agricultural University Of He'nan

＜120〉wheat resistance genes Lr-L1 and application thereof

<160>2

<210>1

<211>2326

<212>DNA

＜213〉common wheat (Triticum aestivum L.)

<400>1

acgcggggaa?ttgaaacact?ctcctcgtcc?tcagaggcct?aactatcaat?agagcatgac 60

tctgactgcg?tacctactcg?ccactccact?tcttcttcct?ctgtgctcac?catcttggta 120

caacagtaca?gaaagatgag?taaattactt?gccatagcac?tgctgctgct?gcatcttatc 180

aaccacaaaa?tcaacatggc?aacggcatgg?gatgatcaag?atttcttcaa?atactgccca 240

ccatcccact?gcagccaaca?tggcccagag?atcaggtatc?ctttctgcct?tgaatccaac 300

aatacatcat?catgtggatg?tagtgggaaa?tcaatcagga?agatagcatg?ctctggccaa 360

gacaccattc?tagtccaccc?agttctttgc?ccatacagtg?tcagcgccat?agattacaag 420

cgttcttcca?tgaagatcac?cccacttgta?gacccctgtt?tggtactcca?gcagaagctc 480

gccatctcca?gaagctcgtc?atctccacag?gttgatgtta?tcaacgatga?gaagccaagt 540

cttgacagat?atttattttg?gagttctaca?atttccctag?tacgctgttc?aagagagttt 600

gcacctggtg?cagccgatgg?gattgctggc?ccagtctcct?gccttagcaa?cataacccac 660

ttcttttatt?tggtgaatgg?ttatgaagac?atgtctattc?ttccgttgga?ctgcaaggtc 720

gtcccaccct?cagatggtgt?cggtggcggt?cagataacca?tgtatatgtt?tgacgaccca 780

atgtcagaca?cgctctcact?gtccttcaag?gaacgcacag?aaagaatcct?cggttttgct 840

gagacgacag?tgtattggta?taattactat?tgcaaaggat?gtgaacgcag?tgggggacgc 900

tgcgcgttca?gctcacaaag?ggatagaaaa?ttctgcatgc?ccggcccaca?tggttcacgt 960

atcaaagtca?ttgcagctac?atcatcagtg?gccgcatttg?tggttctttt?gttaacggtg 1020

gccactgtgc?tttatctttc?actcaagaca?agatataatg?cagagataca?tttgaaggtt 1080

gaaatgttcc?tcaagacata?tggcacatcg?aaacccacaa?ggtacacttt?ctctgaagtt 1140

aagaagatgg?caagacggtt?taaggaaaaa?gtagggcagg?gaggatttgg?gagcgtatac 1200

aaaggtgagc?tgccaaatgg?agtgcccgtg?gcagtcaaga?tgctagagaa?ctctacagga 1260

gagggagaag?cattcatcaa?tgaagttgca?accatcggac?ttatccatca?tgccaatatt 1320

gtccgcctcc?tgggattctg?ctctgaagga?atgaggcggg?ctcttattta?tgaattcatg 1380

cctaatgagt?cactggagaa?atacatattc?tctgacgact?ctaatatttt?tcagaatctt 1440

ctagtaccag?ataaactgct?agatattgct?ttaggcatcg?cccgaggtat?ggagtacttg 1500

catcaagggt?gcaaccagcg?catcctccac?tttgacatca?agcctcacaa?tatcctgctg 1560

gactacaact?tcaatccaaa?gatctcagac?tttggccttg?caaagctgtg?cgcaagggac 1620

caaagcatcg?tgaccttaac?agcagcaaga?ggcacaatgg?gctacattgc?accagagcta 1680

tactcccgga?actttggggg?agtatcgtac?aagtcagacg?tgtacagttt?cgggatgctg 1740

gtgctagaaa?tggtgagcgg?gaggaggaat?tcaggcccaa?gaatcgggag?ccaggacgat 1800

gtttacctcc?ctgagtggat?ctacgagaaa?gtggtcaatg?gggaggactt?ggcgcttact 1860

ttggaaacga?ctgaagaaga?taaagaaaag?gtgaggaagc?tggctatggt?ggcactgtgg 1920

tgtatccagt?ggaacccaag?aaaccggccg?tcgatgacaa?aggtcgtgaa?catgctaact 1980

gggaggcttc?agagtctgca?gatgccacca?aagccttatg?tctcatccga?gaatgaactt 2040

atgccataaa?ttaaagcatg?tcgcacggac?taccatactc?tgtcaggttg?tactttgtcc 2100

acgcgtgcgc?tgcaaataat?agcaaccact?gtgtttcgac?gctttcagtc?agtcaattag 2160

gtctgaagat?gtatgaataa?tcaccggaca?ctagcagcat?gtaatagtgt?cactagttgt 2220

ttgcaagata?ctggtgcttc?catatttatg?tccaaaaact?tgcaatagag?ctgtgtaaaa 2280

taaaataata?atctgtcgaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaa 2326

<210>2

<211>637

<212>PRT

＜213〉common wheat (Triticum aestivum L.)

<400>2

Met?Ser?Lys?Leu?Leu?Ala?Ile?Ala?Leu?Leu?Leu?Leu?His?Leu?Ile

1 5 10 15

Asn?His?Lys?Ile?Asn?Met?Ala?Thr?Ala?Trp?Asp?Asp?Gln?Asp?Phe

20 25 30

Phe?Lys?Tyr?Cys?Pro?Pro?Ser?His?Cys?Ser?Gln?His?Gly?Pro?Glu

35 40 45

Ile?Arg?Tyr?Pro?Phe?Cys?Leu?Glu?Ser?Asn?Asn?Thr?Ser?Ser?Cys

50 55 60

Gly?Cys?Ser?Gly?Lys?Ser?Ile?Arg?Lys?Ile?Ala?Cys?Ser?Gly?Gln

60 70 75

Asp?Thr?Ile?Leu?Val?His?Pro?Val?Leu?Cys?Pro?Tyr?Ser?Val?Ser

80 85 90

Ala?Ile?Asp?Tyr?Lys?Arg?Ser?Ser?Met?Lys?Ile?Thr?Pro?Leu?Val

95 100 105

Asp?Pro?Cys?Leu?Val?Leu?Gln?Gln?Lys?Leu?Ala?Ile?Ser?Arg?Ser

110 115 120

Ser?Ser?Ser?Pro?Gln?Val?Asp?Val?Ile?Asn?Asp?Glu?Lys?Pro?Ser

125 130 135

Leu?Asp?Arg?Tyr?Leu?Phe?Trp?Ser?Ser?Thr?Ile?Ser?Leu?Val?Arg

140 145 150

Cys?Ser?Arg?Glu?Phe?Ala?Pro?Gly?Ala?Ala?Asp?Gly?Ile?Ala?Gly

155 160 165

Pro?Val?Ser?Cys?Leu?Ser?Asn?Ile?Thr?His?Phe?Phe?Tyr?Leu?Val

170 175 180

Asn?Gly?Tyr?Glu?Asp?Met?Ser?Ile?Leu?Pro?Leu?Asp?Cys?Lys?Val

185 190 195

Val?Pro?Pro?Ser?Asp?Gly?Val?Gly?Gly?Gly?Gln?Ile?Thr?Met?Tyr

200 205 210

Met?Phe?Asp?Asp?Pro?Met?Ser?Asp?Thr?Leu?Ser?Leu?Ser?Phe?Lys

215 220 225

Glu?Arg?Thr?Glu?Arg?Ile?Leu?Gly?Phe?Ala?Glu?Thr?Thr?Val?Tyr

230 235 240

Trp?Tyr?Asn?Tyr?Tyr?Cys?Lys?Gly?Cys?Glu?Arg?Ser?Gly?Gly?Arg

245 250 255

Cys?Ala?Phe?Ser?Ser?Gln?Arg?Asp?Arg?Lys?Phe?Cys?Met?Pro?Gly

260 265 270

Pro?His?Gly?Ser?Arg?Ile?Lys?Val?Ile?Ala?Ala?Thr?Ser?Ser?Val

275 280 285

Ala?Ala?Phe?Val?Val?Leu?Leu?Leu?Thr?Val?Ala?Thr?Val?Leu?Tyr

290 295 300

Leu?Ser?Leu?Lys?Thr?Arg?Tyr?Asn?Ala?Glu?Ile?His?Leu?Lys?Val

305 310 315

Glu?Met?Phe?Leu?Lys?Thr?Tyr?Gly?Thr?Ser?Lys?Pro?Thr?Arg?Tyr

320 325 330

Thr?Phe?Ser?Glu?Val?Lys?Lys?Met?Ala?Arg?Arg?Phe?Lys?Glu?Lys

335 340 345

Val?Gly?Gln?Gly?Gly?Phe?Gly?Ser?Val?Tyr?Lys?Gly?Glu?Leu?Pro

350 355 360

Asn?Gly?Val?Pro?Val?Ala?Val?Lys?Met?Leu?Glu?Asn?Ser?Thr?Gly

365 370 375

Glu?Gly?Glu?Ala?Phe?Ile?Asn?Glu?Val?Ala?Thr?Ile?Gly?Leu?Ile

380 385 390

His?His?Ala?Asn?Ile?Val?Arg?Leu?Leu?Gly?Phe?Cys?Ser?Glu?Gly

395 400 405

Met?Arg?Arg?Ala?Leu?Ile?Tyr?Glu?Phe?Met?Pro?Asn?Glu?Ser?Leu

410 415 420

Glu?Lys?Tyr?Ile?Phe?Ser?Asp?Asp?Ser?Asn?Ile?Phe?Gln?Asn?Leu

425 430 435

Leu?Val?Pro?Asp?Lys?Leu?Leu?Asp?Ile?Ala?Leu?Gly?Ile?Ala?Arg

440 445 450

Gly?Met?Glu?Tyr?Leu?His?Gln?Gly?Cys?Asn?Gln?Arg?Ile?Leu?His

455 460 465

Phe?Asp?Ile?Lys?Pro?His?Asn?Ile?Leu?Leu?Asp?Tyr?Asn?Phe?Asn

470 475 480

Pro?Lys?Ile?Ser?Asp?Phe?Gly?Leu?Ala?Lys?Leu?Cys?Ala?Arg?Asp

485 490 495

Gln?Ser?Ile?Val?Thr?Leu?Thr?Ala?Ala?Arg?Gly?Thr?Met?Gly?Tyr

500 505 510

Ile?Ala?Pro?Glu?Leu?Tyr?Ser?Arg?Asn?Phe?Gly?Gly?Val?Ser?Tyr

515 520 525

Lys?Ser?Asp?Val?Tyr?Ser?Phe?Gly?Met?Leu?Val?Leu?Glu?Met?Val

530 535 540

Ser?Gly?Arg?Arg?Asn?Ser?Gly?Pro?Arg?Ile?Gly?Ser?Gln?Asp?Asp

545 550 555

Val?Tyr?Leu?Pro?Glu?Trp?Ile?Tyr?Glu?Lys?Val?Val?Asn?Gly?Glu

560 565 570

Asp?Leu?Ala?Leu?Thr?Leu?Glu?Thr?Thr?Glu?Glu?Asp?Lys?Glu?Lys

575 580 585

Val?Arg?Lys?Leu?Ala?Met?Val?Ala?Leu?Trp?Cys?Ile?Gln?Trp?Asn

590 595 600

Pro?Arg?Asn?Arg?Pro?Ser?Met?Thr?Lys?Val?Val?Asn?Met?Leu?Thr

605 610 615

Gly?Arg?Leu?Gln?Ser?Leu?Gln?Met?Pro?Pro?Lys?Pro?Tyr?Val?Ser

620 625 630

Ser?Glu?Asn?Glu?Leu?Met?Pro

635 637

Claims (6)

1. wheat resistance genes Lr-L1, it is characterized in that: the full length cDNA sequence of wheat resistance genes Lr-L1, the nucleotide sequence based composition is: 672a 534c 535g 585t

1 acgcggggaa?ttgaaacact?ctcctcgtcc?tcagaggcct?aactatcaat?agagcatgac

61 tctgactgcg?tacctactcg?ccactccact?tcttcttcct?ctgtgctcac?catcttggta

121 caacagtaca?gaaagatgag?taaattactt?gccatagcac?tgctgctgct?gcatcttatc

181 aaccacaaaa?tcaacatggc?aacggcatgg?gatgatcaag?atttcttcaa?atactgccca

241 ccatcccact?gcagccaaca?tggcccagag?atcaggtatc?ctttctgcct?tgaatccaac

301 aatacatcat?catgtggatg?tagtgggaaa?tcaatcagga?agatagcatg?ctctggccaa

361 gacaccattc?tagtccaccc?agttctttgc?ccatacagtg?tcagcgccat?agattacaag

421 cgttcttcca?tgaagatcac?cccacttgta?gacccctgtt?tggtactcca?gcagaagctc

481 gccatctcca?gaagctcgtc?atctccacag?gttgatgtta?tcaacgatga?gaagccaagt

541 cttgacagat?atttattttg?gagttctaca?atttccctag?tacgctgttc?aagagagttt

601 gcacctggtg?cagccgatgg?gattgctggc?ccagtctcct?gccttagcaa?cataacccac

661 ttcttttatt?tggtgaatgg?ttatgaagac?atgtctattc?ttccgttgga?ctgcaaggtc

721 gtcccaccct?cagatggtgt?cggtggcggt?cagataacca?tgtatatgtt?tgacgaccca

781 atgtcagaca?cgctctcact?gtccttcaag?gaacgcacag?aaagaatcct?cggttttgct

841 gagacgacag?tgtattggta?taattactat?tgcaaaggat?gtgaacgcag?tgggggacgc

901 tgcgcgttca?gctcacaaag?ggatagaaaa?ttctgcatgc?ccggcccaca?tggttcacgt

961 atcaaagtca?ttgcagctac?atcatcagtg?gccgcatttg?tggttctttt?gttaacggtg

1021?gccactgtgc?tttatctttc?actcaagaca?agatataatg?cagagataca?tttgaaggtt

1081?gaaatgttcc?tcaagacata?tggcacatcg?aaacccacaa?ggtacacttt?ctctgaagtt

1141?aagaagatgg?caagacggtt?taaggaaaaa?gtagggcagg?gaggatttgg?gagcgtatac

1201?aaaggtgagc?tgccaaatgg?agtgcccgtg?gcagtcaaga?tgctagagaa?ctctacagga

1261?gagggagaag?cattcatcaa?tgaagttgca?accatcggac?ttatccatca?tgccaatatt

1321?gtccgcctcc?tgggattctg?ctctgaagga?atgaggcggg?ctcttattta?tgaattcatg

1381?cctaatgagt?cactggagaa?atacatattc?tctgacgact?ctaatatttt?tcagaatctt

1441?ctagtaccag?ataaactgct?agatattgct?ttaggcatcg?cccgaggtat?ggagtacttg

1501?catcaagggt?gcaaccagcg?catcctccac?tttgacatca?agcctcacaa?tatcctgctg

1561?gactacaact?tcaatccaaa?gatctcagac?tttggccttg?caaagctgtg?cgcaagggac

1621?caaagcatcg?tgaccttaac?agcagcaaga?ggcacaatgg?gctacattgc?accagagcta

1681?tactcccgga?actttggggg?agtatcgtac?aagtcagacg?tgtacagttt?cgggatgctg

1741?gtgctagaaa?tggtgagcgg?gaggaggaat?tcaggcccaa?gaatcgggag?ccaggacgat

1801?gtttacctcc?ctgagtggat?ctacgagaaa?gtggtcaatg?gggaggactt?ggcgcttact

1861?ttggaaacga?ctgaagaaga?taaagaaaag?gtgaggaagc?tggctatggt?ggcactgtgg

1921?tgtatccagt?ggaacccaag?aaaccggccg?tcgatgacaa?aggtcgtgaa?catgctaact

1981?gggaggcttc?agagtctgca?gatgccacca?aagccttatg?tctcatccga?gaatgaactt

2041?atgccataaa?ttaaagcatg?tcgcacggac?taccatactc?tgtcaggttg?tactttgtcc

2101?acgcgtgcgc?tgcaaataat?agcaaccact?gtgtttcgac?gctttcagtc?agtcaattag

2161?gtctgaagat?gtatgaataa?tcaccggaca?ctagcagcat?gtaatagtgt?cactagttgt

2221?ttgcaagata?ctggtgcttc?catatttatg?tccaaaaact?tgcaatagag?ctgtgtaaaa

2281?taaaataata?atctgtcgaa?aaaaaaaaaa?aaaaaaaaaa?aaaaaa

2. wheat resistance genes Lr-L1 according to claim 1 is characterized in that: wheat resistance genes

The Lr-L1 encoded protein, its aminoacid sequence is:

1 MSKLLAIALL?LLHLINHKIN?MATAWDDQDF?FKYCPPSHCS?QHGPEIRYPF?CLESNNTSSC

61 GCSGKSIRKI?ACSGQDTILV?HPVLCPYSVS?AIDYKRSSMK?ITPLVDPCLV?LQQKLAISRS

121?SSSPQVDVIN?DEKPSLDRYL?FWSSTISLVR?CSREFAPGAA?DGIAGPVSCL?SNITHFFYLV

181?NGYEDMSILP?LDCKVVPPSD?GVGGGQITMY?MFDDPMSDTL?SLSFKERTER?ILGFAETTVY

241?WYNYYCKGCE?RSGGRCAFSS?QRDRKFCMPG?PHGSRIKVIA?ATSSVAAFVV?LLLTVATVLY

301?LSLKTRYNAE?IHLKVEMFLK?TYGTSKPTRY?TFSEVKKMAR?RFKEKVGQGG?FGSVYKGELP

361?NGVPVAVKML?ENSTGEGEAF?INEVATIGLI?HHANIVRLLG?FCSEGMRRAL?IYEFMPNESL

421?EKYIFSDDSN?IFQNLLVPDK?LLDIALGIAR?GMEYLHQGCN?QRILHFDIKP?HNILLDYNFN

481?PKISDFGLAK?LCARDQSIVT?LTAARGTMGY?IAPELYSRNF?GGVSYKSDVY?SFGMLVLEMV

541?SGRRNSGPRI?GSQDDVYLPE?WIYEKVVNGE?DLALTLETTE?EDKEKVRKLA?MVALWCIQWN

601?PRNRPSMTKV?VNMLTGRLQS?LQMPPKPYVS?SENELMP

3. wheat resistance genes Lr-L1 according to claim 2, it is characterized in that: wheat resistance genes Lr-L1 encoded protein, its genes encoding be a receptoroid protein kinase, comprise N-end signal peptide, extracellular domain, middle part membrane spaning domain, highly charged sequence and C-end protein kinase domain.

4. the application of wheat resistance genes Lr-L1 in the plant disease-resistant improvement.

5. the application of wheat resistance genes Lr-L1 in the wheat cdna chip manufacturing.

6. the application of wheat resistance genes Lr-L1 in synthetic new disease-resistant gene.