CN100549029C - One plant disease resistance-related SGT1 and encoding gene and application - Google Patents

Abstract

The invention discloses a plant disease resistance-related and encoding gene thereof and application.This plant disease resistance-related protein, its aminoacid sequence is shown in SEQ ID NO:2.Gene of the present invention can change the disease resistance that improves plant in unifacial leaf or the dicotyledons over to, for the good operation basis is laid in the plant broad spectrum antidisease breeding.

Description

One plant disease resistance-related SGT1 and encoding gene and application

Technical field

The present invention relates to a plant disease resistance-related SGT1 and encoding gene and application, plant disease resistance-related protein SGT1 and encoding gene and the application of couchgrass in the middle of particularly a kind of the deriving from.

Background technology

Host's disease resistance response is the result that a series of signal genetic expression, startup host defense response were discerned, activated to direct or indirect product of host's disease-resistant gene and the direct or indirect product of cause of disease nontoxic gene mutually.The modal mode of disease resistance response is: by the mutual recognition reaction of plant disease resistance genes and cause of disease, the stimulating activity oxidative burst, produce reactive oxygen intermediate (reactive oxygen intermediates, ROI) and anaphylaxis (hypersensitivereaction, HR), cause infecting position peripheral cell programmed cell death, excite a series of defence expression of gene, produce tissue or complete stool disease resistance, this resistance of inducing generation be called systemic acquired resistance (systemicacquired resistance, SAR).

In dicotyledonous and monocotyledons, need many signal genes to participate in when disease-resistant (R) gene mediated, activation defensive raction.In recent years in Arabidopis thaliana, barley, tobacco, tomato, identify an important disease-resistant signal gene SGT1, between R gene and active oxygen excite.The albumen of SGT1 genes encoding has following structural domain: 1 repeating structure territory (tetratricopeptide repeat domain, TPR), 1 CS primitive (CHORD and SGT1) and 1 SGS (SGT1-specific motif) and 2 variable regions (VR1 and VR2).SGT1 is one of composition of SCF ubiquitin lytic enzyme complex body, in the plant disease-resistant defensive raction, can when pathogen infection, regulate host's programmed cell death, excite a series of defence expression of gene, produce SAR, start comprehensive defense response of plant, make plant performance disease resistance (Austin M J et al.Science.2002,295:2077, Azevedo C et al.Science.2002,295:2073, Holt BF, Belkhadir Y, Dangle Jl.Antagonistic control of disease resistance protein stability in plantimmune system.Science, 2005,309:929-932).It has participated in the disease resistance response of barley CC-NBS-LRR class disease-resistant gene Mla6 mediation.In Arabidopis thaliana, the resistance that SGT1 gives the disease-resistant albumen of a plurality of TIR-NBS-LRR is essential.People such as Peart in 2002 have further studied the function of SGT1 in the tobacco disease resistance reaction, find that the disease resistance response that SGT1 gives for NBS-LRR class and a plurality of other resistance protein all is essential.Research shows that also SGT1 also plays an important role in non-host-resistance.Compare with some important disease-resistant signaling molecules of previous evaluation, SGT1 may be an important widely disease-resistant signaling molecule in plant disease-resistant.

Present various fungi, virus, Micobial Disease regular meeting cause the reduction of important crop yield and quality, are the important factors that the important farm crop of restriction produce.Gas borne fungus diseases such as silborne fungal diseases such as wheat hypochnus, head blight, wheat powdery mildew, stripe rust and virus disease are serious day by day in causing harm of China wheat main producing region, gently then cause wheat yield 20-30%, heavy then underproduction 40-50%, even total crop failure.Cultivating disease-resistant variety is the first-elected approach of the above-mentioned disease of control.Because the variation of pathogenic bacteria physiological strain often makes host's disease-resistant gene forfeiture resistance.Therefore, the important disease-resistant signal conduction gene SGT1 of clone is applied to the broad spectrum antidisease gene engineering from the plant of immunity or high anti-multiple diseases, and inducing plant produces SAR, makes plant produce resistance to multiple cause of disease, will address the above problem.This strategy theoretically should be more effective than importing single disease-resistant gene.

Couchgrass in the middle of the wheat kindred plant has manyly to the useful important gene of wheat improvement, as has characteristics such as anti-wheat rust, Powdery Mildew, head blight, virus disease, banded sclerotial blight.

Summary of the invention

The purpose of this invention is to provide a plant disease resistance-related and encoding gene thereof and application.

Plant disease resistance-related protein provided by the present invention, name is called TiSGT1, and couchgrass (Thinopyrum intermedium) in the middle of deriving from is the protein with one of following aminoacid sequence:

1) the SEQ ID № in the sequence table: 2 amino acid residue sequence;

2) with SEQ ID № in the sequence table: 2 amino acid residue sequence is through replacement and/or disappearance and/or the interpolation and the protein relevant with plant disease-resistant of one or several amino-acid residue.

Wherein, sequence 2 is made up of 373 amino-acid residues in the sequence table.From the 1st of the aminoterminal of sequence 2 to 373 amino acids residues is the TiSGT1 gene, has the proteic functional domain of plant SGT1: 1 repeating structure territory (tetratricopeptide repeat domain, TPR) be positioned at the 21st to the 98th amino acids district, 1 CS primitive (CHORD and SGT1) is positioned at 173-260 amino acids district and 1 SGS (SGT1-specificmotif) structural domain is positioned at 290-371 amino acids district.

The encoding gene of above-mentioned plant disease resistance-related protein (TiSGT1) also belongs to protection scope of the present invention.

The cDNA gene of above-mentioned plant disease resistance-related protein can have one of following nucleotide sequence:

1) SEQ ID № in the sequence table: 1 dna sequence dna;

2) SEQ ID № in the code sequence tabulation: the polynucleotide of 2 protein sequences;

3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 1 dna sequence dna hybridization that limits.

The rigorous condition of above-mentioned height can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.

Wherein sequence 1 is made up of 1449 deoxynucleotides in the sequence table, from 5 ' and the 16th to the 1137th deoxynucleotide of end is encoding sequence (ORF).

Contain expression carrier of the present invention, clone and host bacterium and all belong to protection scope of the present invention.

TiSGT1 gene of the present invention can be building up in the existing plant expression vector with existing method, can add any promotor that comprises constitutive promoter, strengthens promotor, inducible promoter, tissue-specific promoter, etap specificity promoter before it transcribes super beginning Nucleotide.For the ease of identifying and screen to changeing TiSGT1 gene plant cell or plant, can process employed carrier, as the antibiotic marker thing (gentamicin, kantlex etc.) that adds the alternative mark (Bar gene, gus gene, luciferase genes etc.) of plant or have resistance.By the plant transformed host both can be monocotyledons, also can be dicotyledons, as: paddy rice, wheat, corn, cucumber, tomato, willow, turfgrass or lucerne place etc.Carry that TiSGT1 expression carrier of the present invention can Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity be led, conventional biological method transformed plant cells or tissue such as agriculture bacillus mediated by using, and plant transformed become plant through tissue cultivating, obtain the plant that disease resistance improves.

Experimental result shows, induces the expression enhancing of TiSGT1 down white powder germ and barly yellow dwarf virus.The expressed intact sequence (ORF) of TiSGT1 gene is replaced gus gene sequence among the unifacial leaf constructive expression carrier pAHC25, made up the transgene carrier pASGT1 that monocotyledons efficiently expresses, this gene can be expressed under the Ubi of carrier promoters driven; PASGT1 imports wheat with TiSGT1 by transgene carrier, detects the disease resistance of transgenic wheat, shows that TiSGT1 can improve the resistance of wheat to the white powder germ.Gene of the present invention can also change the resistance that improves in other unifacial leaves or the dicotyledons the other plant disease over to, for the good operation basis is laid in the plant broad spectrum antidisease breeding.

Description of drawings

Fig. 1 is the gel electrophoresis figure of amplification TiSGT1 gene 5 ' end fragment from the middle couchgrass of white powder germ inductive

The TiSGT1 proteose intention that Fig. 2 obtains for the BLASTP by GenBank

Fig. 3 is couchgrass TiSGT1 expression analysis result in the middle of the barley yellow dwarf inductive

Fig. 4 is couchgrass TiSGT1 expression analysis result in the middle of the white powder germ inductive

Fig. 5 is a pASGT1 vector construction synoptic diagram

The PCR of screening-gene Bar detects gel electrophoresis figure in Fig. 6 part transfer-gen plant

Fig. 7 is the wheat plant of commentaries on classics pASGT1 and the incidence photo of adjoining tree inoculation Powdery Mildew

Embodiment

Experimental technique among the following embodiment if no special instructions, is ordinary method.

The acquisition of embodiment 1, TiSGT1 full-length cDNA

With the blade inoculation white powder germ of the middle couchgrass around growing, after 24 hours, use total RNA of the method extraction blade of TRIZOL, synthesize the 1st chain of cDNA according to the method that the precious biotech firm in Dalian provides, and carry out pcr amplification as template with it.According to barley, paddy rice, Arabidopis thaliana, conservative section of tobacco SGT1 gene and wheat EST design a pair of primer SGT1-F:5 '-CACCTCGACGCAGACATG-3 ' and SGT1-R:5 '-CTC (A/C) AGCTTATCCCAGTC-3 ', PCR reaction cumulative volume is 25 μ L, comprises 2 μ L cDNA (50ng), 100 μ mol/LdNTP, 0.4 every primer of μ mol/L, 1U Hotstar Taq enzyme and 1 * Ex PCR damping fluid (all available from the precious biotech firm in Dalian).The PCR response procedures is: 95 ℃ of pre-sex change 5min of elder generation; 95 ℃ of 30s then, 56 ℃ of 1min, 72 ℃ of 1min, 35 circulations; Last 72 ℃, 10min mends flat terminal.Amplified production separates through agarose gel electrophoresis, and the result shows to amplify an about 1.0kb band from the middle couchgrass cDNA of inoculation white powder germ as shown in Figure 1. Swimming lane 1 and 2 bands among Fig. 1 for amplification; M is a dna molecular amount standard (available from Huamei Bio-Engrg Co.); The stripe size that arrow refers among the figure is 1.0kb.This band is reclaimed, be cloned on the pMD18-T carrier (the precious biotech firm in Dalian), carry out sequencing analysis.Sequential analysis shows that this sequence is SGT1 gene 5 ' end fragment.According to the Auele Specific Primer that obtains SGT1 gene fragment order design 3 '-RACE, the RACE program is carried out according to the specification sheets of 3 '-RACE test kit that introgen company provides.3 '-RACE Auele Specific Primer RP ₁:

5 '-AGCAAGCCCAAATACAGG-3 ', RP ₂: 5 '-GCGTGGTTGTTGACTTTG-3 '; Use RP earlier ₁AUAP primer (downstream primer in upstream primer and the test kit, sequence: 5 '-GGCCACGCGTCGACTAGTAC-3 '), with 3 '-the AP primer of RACE test kit carries out the first round from middle the couchgrass blade RNA reverse transcription synthetic first chain cDNA of powder germ as template from inoculation and increases, and then make template with 50 times first round PCR product of dilution, with RP ₂(upstream primer) and AUAP downstream primer carry out second and take turns pcr amplification, reclaim purified pcr product, connect and are cloned on the pMD18-T carrier, by bacterium colony PCR, select 3 positive colony order-checkings, obtain 3 ' terminal sequence of TiSGT1.

First round PCR reaction conditions: 94 ℃ of 1min of elder generation; 94 ℃ of 30s then, 58 ℃ of 30S, 72 ℃ of 1min 30S totally 10 circulations; 94 ℃ of 30s again, 56 ℃ of 30S, 72 ℃ of 1min30S totally 25 circulations; Last 72 ℃ of 5min.Second takes turns the PCR reaction conditions: 94 ℃ of pre-sex change 2min of elder generation; 94 ℃ of 30S then, 63 ℃ of 30S, 72 ℃ of 2min totally 10 circulations; 94 ℃ of 30S again, 61 ℃ of 30S, 72 ℃ of 2min totally 25 circulations; Last 72 ℃ of 5min.

5 ' terminal sequence of TiSGT1 and 3 ' terminal sequence after external splicing, are shown through order-checking, obtain the cDNA sequence of 1449bp, nucleotide sequence with sequence 1 in the sequence table.

For verifying that the cDNA that this splicing obtains is the TiSGT1 full-length cDNA, design 1 couple of Auele Specific Primer TiF:5 '-CACCTCGACGCAGACATG-3 ' again, TiR:5 '-TTAATACTCCCACTTCTTGAG-3 ', cDNA with the middle couchgrass of inoculation white powder germ is a template, carry out pcr amplification, to obtain the total length ORF of this gene.Wherein the reaction system of PCR is: 2 μ L cDNA (50ng), 100 μ mol/L dNTP, every primer of 0.4 μ mol/L, 1U Hotstar Taq enzyme and 1 * Ex PCR damping fluid (all available from the precious biotech firm in Dalian).The PCR response procedures is: 95 ℃ of pre-sex change 5min of elder generation; 95 ℃ of 30s then, 56 ℃ of 1min, 72 ℃ of 1min, 35 circulations; Last 72 ℃, 10min mends flat terminal.Amplified production carries out agarose electrophoresis to be separated, and reclaims and is cloned on the pGEM-T carrier, carries out sequencing analysis, and sequencing result shows that this pcr amplification product has the nucleotide sequence of the total length ORF of sequence 1 in the sequence table, called after TiSGT1.TiSGT1 open reading frame (encoding sequence) is the 16th to the 1137th nucleotide sequence of 5 ' end of sequence 1 in sequence table, the TiSGT1 of 373 amino acid residue sequences (sequence 2 in the sequence table) of encoding, and molecular weight is 41KD, pI=4.87.

The TiSGT1 full length gene ORF sequence that obtains is carried out the BLASTP of GenBank, compare, the result as shown in Figure 2, Fig. 2 shows that TiSGT1 has 1 repeating structure territory (tetratricopeptide repeatdomain, TPR) be the amino acid residue sequence that is positioned at from the aminoterminal 21-98 position of sequence 2, it is the amino acid residue sequence that is positioned at from the aminoterminal 173-260 position of sequence 2 that 1 CS primitive (CHORD and SGT1) is arranged, also constitute 1 p23 structural domain from the aminoterminal 175-297 of sequence 2 amino acids residue sequence, and 1 SGS (SGT1-specific motif) structural domain is to be positioned at from the aminoterminal 290-371 of sequence 2 amino acids residue sequence.Wherein TPR participates in multiple functions such as albumen interphase interaction, folding of p23 energy binding molecule companion heat shock protein hsp90, participation multiple protein, the effect of CS domain is still unintelligible, may interact with the CHORD tool, and SGS domain is the peculiar structural domain of SGT1-like proteins.

The expression characterization of embodiment 2, TiSGT1 detects

With the blade of the middle couchgrass around the growth, inoculate white powder germ, band barley yellow dwarf aphid 0,6,12,24 respectively, after 48 hours, with total RNA of Trizol method extraction blade, with DNaseI enzymolysis, purifying RNA, reverse transcription becomes cDNA.Earlier, make the initial concentration unanimity (being homogenization) of amplification template mRNA/cDNA with constructive expression's Actin gene primer each sample RNA that increases, then with the primer SG-F1 of TiSGT1:

5 '-CACCTCGACGCAGACATG-3 ', SG-R2:5 '-CTCCCACTTCTTGAGCTC-3 ' carries out sxemiquantitative RT-PCR amplification and gel electrophoresis analysis.Analytical results as shown in Figure 3, Figure 4, the result shows that the expression of TiSGT1 gene transcription is subjected to the back of inducing of barly yellow dwarf virus, white powder germ to express enhancing, under normal operation, TiSGT1 has trace expression, white powder germ, band barley yellow dwarf attacked by aphids are after 6 hours, TiSGT1 expression of gene amount begins to increase, and reaches maximum value in 12 hours infecting.Explanation is induced the expression enhancing of TiSGT1 down at barly yellow dwarf virus and white powder germ.

The disease resistance analysis of embodiment 3, commentaries on classics TiSGT1 dna triticum plant

CDNA with the TiSGT1 gene is a template, uses the primer that has the SmaI/SacI enzyme recognition site: upstream primer SGL-SF:5 ' TT CCCGGGCACCTCGACGCAGACATG-3 ' (sequence that underscore marks is represented the SmaI enzyme recognition site), downstream primer SGL-SR:5 '-CC

TCATTAATACTCCCACTTC-3 ' ( The sequence that marks is represented the SacI enzyme recognition site) (the PCR response procedures is: 95 ℃ of pre-sex change 5min of elder generation to carry out pcr amplification; 95 ℃ of 30s then, 56 ℃ of 1min, 72 ℃ of 1min, 35 circulations; Last 72 ℃, 10min mends flat terminal), acquisition adds the TiSGT1 full length gene ORF in SmaI and SacI site, reclaim amplified production, be connected on the pMD18-T carrier, show that through order-checking this recombinant vectors is the pMD18-T that contains the TiSGT1 gene, its called after pTSGT1.PTSGT1 cuts with SmaI and SacI enzyme, reclaim the TiSGT1 gene fragment, the TiSGT1 gene is inserted between the SmaI and SacI recognition site of pAHC25, replace gus gene among the monocotyledons expression vector pAHC25, (transform by pUC8 and to form to monocotyledons expression vector pAHC25 TiSGT1 is gene constructed, contain 2 expression cassettes, the 1st expression cassette has corn UBIQUITIN promotor, Exon, Intron, GUS, the Nos terminator, the GUS two ends have SmaI and SacI restriction enzyme site, the 2nd expression cassette has corn UBIQUITIN promotor, Exon, Intron, Bar, the Nos terminator) between SmaI and the SacI recognition site, form recombinant vectors, identifying through order-checking, the total length ORF that contains the TiSGT1 gene, the carrier called after pASGT1 (building process as shown in Figure 5) that direction of insertion is correct.This pASGT1 carrier is controlled by the UBI promotor.This carrier also has 1 Bar expression casette that is subjected to the control of Ubi promotor in the downstream, can be to utilize weedicide bialaphos (Bialaphos) screening transformation tissue culture plant that resistance is provided in the follow-up work.

With particle gun pASGT1 is bombarded 3200 of the callus of wheat breed poplar wheat 12, the callus after the bombardment is aftertreatment 16h on the osmotic pressure substratum.Callus is transferred to SD2 (add VB in the inorganic salt composition of MS substratum ₁1mg/L, asparagus fern door acid amides 150mg/L, 2,4-D2mg/L), 26 ℃, the dark cultivation recovers to cultivate for 2 weeks.Callus after recover cultivating transferred to (the 1/2MS substratum is additional: NAA1mg/L+KT1mg/L in the differentiation screening culture medium, Bialaphos2-5mg/L), 24-26 illumination cultivation 14d, callus broken up transfer to behind the seedling in the growth screening culture medium (the 1/2MS substratum is additional: Bialaphos 2-3mg/L), and the 24-26 illumination cultivation.(the 1/2MS substratum is additional: 0.5mg/L NAA), flowerpot is arrived in the conversion transplantation of seedlings of height of seedling 7-8cm and well developed root system, grow in the greenhouse to transfer to the strong seedling culture base through the conversion seedling of 2-3 Bialaphos screening.Survive plant 168 strains.In tri-leaf period, every strain is got 1 blade and is extracted the PCR evaluation that genomic dna carries out transgene, and inoculate the white powder germ subsequently, bacterial classification is that the Beijing area wheat powdery mildew (Erysiphe graminisf.sp.tritici) that the greenhouse is gathered mixes microspecies, adopts the live body frictional engagement to shake off the method inoculation of spore.Begin to carry out seedling stage and the disease-resistant evaluation that becomes the strain phase after inoculating for 2 weeks.Be decided to be high anti-(0 grade) with no sorus on the blade and necrotic plaque person seedling stage, and the person is decided to be susceptible to have the sorus on the blade; Become the disease-resistant classification of strain phase to be divided into 9 grades according to international wheat and corn improvement center and national standard (State Standard of the People's Republic of China's pesticide field efficacy medicine test standard () GB/T1798.22-200, bactericidal agent for preventing and treating cereal class Powdery Mildew).

Bar gene on the conversion carrier in the commentaries on classics pASGT1 plant is carried out PCR to be detected.

The Bar gene PCR detects the primer:

Bar-R：5’-CTTCAGCAGGTGGGTGTAGAGCGTG-3’

Bar-F：5’-CCTGCCTTCATACGCTATTTATTTGCC-3’

The amplification system of cumulative volume 25 μ l: 10 * PCR buffer, 2.5 μ l, MgCl ₂2mmol/L, dNTPs0.2mmol/L each, each 0.4 μ mol/L of primer Bar-R/Bar-F, Taq archaeal dna polymerase (sky is a Time Inc.) 1U, the about 400ng of template DNA, moisturizing to 25 μ l.

Amplification program: 94 ℃ of sex change 5min; 94 ℃ of 45sec, 62 ℃ of 45sec, 72 ℃ of 1min, 45 circulations; 72 ℃ of 10min.Amplified production 1% agarose gel electrophoresis, EB dyeing, ultraviolet is taken pictures.Bar gene PCR test section result as shown in Figure 6, amplification obtains the positive transgenic plant of plant of Bar gene band of 450bp.M is 100bp molecular weight standard (a Fa Tejie company product) among Fig. 6; Swimming lane 1 is a Bar gene fragment among the plasmid pASGT1; Swimming lane 2-9 is for changeing the plant of test positive in the pASGT1 plant; Swimming lane 10 is to detect after the transgenosis to be non-male plant; Swimming lane 11 is ddH ₂O; Arrow indication clip size is 450bp.

The result shows, have 32 strains and change the pASGT1 poplar wheat 12 plant PCR detection positive, wherein there are 20 strains to change the pASGT1 gene, PCR detects male T0 for the high mildew-resistance of plant (0 grade), and not genetically modified poplar wheat 12 high sense Powdery Mildews (7-9 level) (Fig. 7), part is changeed the TiSGT1 gene, PCR detects male T0 plant inoculation barly yellow dwarf virus (Zhang Zengyan, Xin Zhiyong, old filial piety, Wang Xiaoping, Liu Jingfang, Du Lipu. come from the specific PCR mark of wheat resistance to yellow dwarf gene of L1 and the research of assistant breeding. Acta Agronomica Sinica, 2000,28 (4): 486-491), identifying 2 strains from commentaries on classics pASGT1 poplar wheat 12 plant of 20 plant height mildew-resistances changes also resistance to yellow dwarf of pASGT1 plant, illustrates that TiSGT1 gene overexpression in wheat can give wheat to Powdery Mildew hybrid bacterial strain height resistance with to yellow dwarf resistance.

Sequence table

<160>2

<210>1

<211>1449

<212>DNA

＜213〉couchgrass (Thinopyrum intermedium) in the middle of

<400>1

cacctcgacg?cagacatggc?cgccgccgcc?gcgtcggatc?tggagagcaa?ggccaaggag 60

gccttcgtcg?acgacgactt?cgagctggcc?gccgagctct?acacccaggc?catcgaggct 120

ggcccagcca?ccgccgaact?ctacgccgac?cgagcccagg?cgcacatcaa?gctgggcagt 180

tacactgagg?ctgtagctga?tgccaacaaa?gcaattgaac?ttgatccttc?gatgcataaa 240

gcataccttc?ggaagggctc?tgcttgcatc?aagctggagg?aataccaaac?tgcaaaggct 300

gctcttgaag?tgggttcttc?ttatgcatct?ggcgactcga?ggtttactcg?tctgatgaag 360

gagtgtgatg?atcgtattgc?tgaggaggct?agccaggttc?cagtaaagaa?tgccgctgca 420

gctgttgctc?catctacatc?ttccggggcc?acaactgtgg?ctactgaagc?tgaggaccag 480

gatggtgcaa?atatggagaa?tgcacaacca?acggtagaag?tgccaagcaa?gcccaaatac 540

aggcatgact?actacaatac?tcctacagaa?gtggtactga?ctatatttgc?taagggtgtt 600

ccagctgaca?gcgtggttgt?tgactttggt?gaacagatgc?tgagtgtctc?aattgaactt 660

cctggtgagg?aaccatacca?ttttcagcct?cgtctgtttt?caaagataat?cccagataag 720

tgcaagtata?ctgtgttgtc?tacaaaggtc?gaaatgcgcc?ttgcaaaagc?tgagccagta 780

acttggacat?cattggatta?tactggtaaa?ccaaaggctc?ctcagaagat?caatgtacca 840

gctgaatcag?cccagaggcc?atcttatcca?tcattaaaat?ccaaaaagga?ctgggataag 900

cttgaagctg?aagtgaaaaa?acaggagaag?gacgaaaaac?ttgatggtga?tgctgcattg 960

aacaaatttt?tccgtgaaat?ttacagtgat?gctgatgaag?atatgcgtag?agcaatgatg?1020

aagtcttttg?tggagtctaa?tggaaccgtt?ctctcaacca?actggaaaga?tgtcgggaaa?1080

aagactgttg?aaggcagccc?tcctgatgga?atggagctca?agaagtggga?gtattaatga?1140

gattcagatc?atccatgttg?taaatcctgg?tgatgagtgt?taacttggct?gcagacttgc?1200

aaatcagtgt?cggtcggatt?atgtgtgttg?ctcttcatta?tgtttccgta?tttgtaagct?1260

gtttgcgtgg?caggttagct?tttaagtact?tatcctagtt?ttgcgagcta?cagctggtaa?1320

tagagtaagc?catacctgtg?catctgtggt?atggaagtgc?aaaacgtccg?ttgtaactgg 1380

tgccaaaact?atcatctgtg?aaacggtgct?attttttgtc?tgaaaaaaaa?aaaaaaaaaa 1440

aaaaaaaaa 1449

<210>2

<211>373

<212>PRT

＜213〉couchgrass (Thinopyrum intermedium) in the middle of

<400>2

Met?Ala?Ala?Ala?Ala?Ala?Ser?Asp?Leu?Glu?Ser?Lys?Ala?Lys?Glu?Ala

1 5 10 15

Phe?Val?Asp?Asp?Asp?Phe?Glu?Leu?Ala?Ala?Glu?Leu?Tyr?Thr?Gln?Ala

20 25 30

Ile?Glu?Ala?Gly?Pro?Ala?Thr?Ala?Glu?Leu?Tyr?Ala?Asp?Arg?Ala?Gln

35 40 45

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

50 55 60

Lys?Ala?Ile?Glu?Leu?Asp?Pro?Ser?Met?His?Lys?Ala?Tyr?Leu?Arg?Lys

65 70 75 80

Gly?Ser?Ala?Cys?Ile?Lys?Leu?Glu?Glu?Tyr?Gln?Thr?Ala?Lys?Ala?Ala

85 90 95

Leu?Glu?Val?Gly?Ser?Ser?Tyr?Ala?Ser?Gly?Asp?Ser?Arg?Phe?Thr?Arg

100 105 110

Leu?Met?Lys?Glu?Cys?Asp?Asp?Arg?Ile?Ala?Glu?Glu?Ala?Ser?Gln?Val

115 120 125

Pro?Val?Lys?Asn?Ala?Ala?Ala?Ala?Val?Ala?Pro?Ser?Thr?Ser?Ser?Gly

130 135 140

Ala?Thr?Thr?Val?Ala?Thr?Glu?Ala?Glu?Asp?Gln?Asp?Gly?Ala?Asn?Met

145 150 155 160

Glu?Asn?Ala?Gln?Pro?Thr?Val?Glu?Val?Pro?Ser?Lys?Pro?Lys?Tyr?Arg

165 170 175

His?Asp?Tyr?Tyr?Asn?Thr?Pro?Thr?Glu?Val?Val?Leu?Thr?Ile?Phe?Ala

180 185 190

Lys?Gly?Val?Pro?Ala?Asp?Ser?Val?Val?Val?Asp?Phe?Gly?Glu?Gln?Met

195 200 205

Leu?Ser?Val?Ser?Ile?Glu?Leu?Pro?Gly?Glu?Glu?Pro?Tyr?His?Phe?Gln

210 215 220

Pro?Arg?Leu?Phe?Ser?Lys?Ile?Ile?Pro?Asp?Lys?Cys?Lys?Tyr?Thr?Val

225 230 235 240

Leu?Ser?Thr?Lys?Val?Glu?Met?Arg?Leu?Ala?Lys?Ala?Glu?Pro?Val?Thr

245 250 255

Trp?Thr?Ser?Leu?Asp?Tyr?Thr?Gly?Lys?Pro?Lys?Ala?Pro?Gln?Lys?Ile

260 265 270

Asn?Val?Pro?Ala?Glu?Ser?Ala?Gln?Arg?Pro?Ser?Tyr?Pro?Ser?Leu?Lys

275 280 285

Ser?Lys?Lys?Asp?Trp?Asp?Lys?Leu?Glu?Ala?Glu?Val?Lys?Lys?Gln?Glu

290 295 300

Lys?Asp?Glu?Lys?Leu?Asp?Gly?Asp?Ala?Ala?Leu?Asn?Lys?Phe?Phe?Arg

305 310 315 320

Glu?Ile?Tyr?Ser?Asp?Ala?Asp?Glu?Asp?Met?Arg?Arg?Ala?Met?Met?Lys

325 330 335

Ser?Phe?Val?Glu?Ser?Asn?Gly?Thr?Val?Leu?Ser?Thr?Asn?Trp?Lys?Asp

340 345 350

Val?Gly?Lys?Lys?Thr?Val?Glu?Gly?Ser?Pro?Pro?Asp?Gly?Met?Glu?Leu

355 360 365

Lys?Lys?Trp?Glu?Tyr

370

Claims (9)

1, a plant disease resistance-related, its aminoacid sequence is shown in SEQ ID NO:2.

2, the encoding gene of the described plant disease resistance-related protein of claim 1.

3, encoding gene according to claim 2 is characterized in that: the nucleotide sequence of the cDNA gene of described plant disease resistance-related protein is shown in SEQ ID NO:1.

4, contain claim 2 or 3 described expression carrier.

5, the transgenic cell line that contains claim 2 or 3 described genes.

6, the host bacterium that contains claim 2 or 3 described genes.

7, described plant disease resistance-related protein of claim 1 and encoding gene thereof the application in improving disease resistance of plant; Described disease resistance is powder mildew resistance and yellow dwarf resistance.

8, application according to claim 7 is characterized in that: described raising disease resistance of plant is for improving the powder mildew resistance of wheat.

9, application according to claim 7 is characterized in that: described raising disease resistance of plant is for improving the yellow dwarf resistance of wheat.

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