CN108130320A - Verticillium dahliae pathogenesis related protein and its encoding gene, application and mutant - Google Patents
Verticillium dahliae pathogenesis related protein and its encoding gene, application and mutant Download PDFInfo
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- C12Y302/01—Glycosidases, i.e. enzymes hydrolysing O- and S-glycosyl compounds (3.2.1)
- C12Y302/01011—Dextranase (3.2.1.11)
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Abstract
The present invention relates to Inducing Toxin From Cotton Verticillium Wilt Pathogen fields, and in particular to verticillium dahliae pathogenesis related protein and its encoding gene, application and mutant.The nucleotide sequence of the gene is as shown in SEQ ID No.2.Present invention screening obtains low pathogenicity mutant T1027, which does not change compared with wild type, but the secretory volume of sporulation quantity and Raw toxin is remarkably decreased, and dextranase is accredited as by the T DNA genes being inserted into.Verticillium dahliae mutant T1027 is the desirable strain of a research verticillium dahliae molecular basis of the pathogenesis, and the expression and modification of gene and protein can be used as target site to have good application potential for designing and screening antifungal medicine.
Description
Technical field
The present invention relates to Inducing Toxin From Cotton Verticillium Wilt Pathogen fields, and in particular to verticillium dahliae pathogenesis related protein
And its encoding gene, application and mutant.
Background technology
Cotton verticillium wilt (Verticillium Wilt) is to influence the major cotton growing area cotton (Gossypium in the world
Hirsutum L.) good quality and high output primary disease.
, there are following several respects in the main reason for cotton verticillium wilt outburst is caused to be caused disaster：Field continuous cropping leads to germ in soil
Amount constantly accumulation；The Variation Pathogenic Bacteria is frequent, and High pathogenicity and defoliation bacterial strain continuously emerge；Antigen-like material is deficient, disease-resistant to educate
Kind lag, varietal resistance are lost fast.In addition, the pathogen pathogenesis is complicated, the Coupling effects of pathogen and host are also unclear
Chu, this is also basis and the difficult point of the area research.Its pathogenesis is disclosed in detail, it is necessary to which separation is caused a disease related to identification
Gene.The research for molecular mechanism of causing a disease both at home and abroad in relation to verticillium dahliae to cotton at present is also seldom, and clearly pathogenic related
Cause is to further investigate its molecular basis of the pathogenesis and pathogen and the element task of cotton plant interaction.
Invention content
Verticillium dahliae low pathogenicity mutant of the present invention for exploitation dimensionally stable, assesses it in key pathogenetic base
Application potential in terms of the functional verification of cause and the research of molecular basis of the pathogenesis completes the present invention.
The object of the present invention is to provide a kind of verticillium dahliae pathogenesis related proteins.
Another object of the present invention is to provide verticillium dahliae pathogenic related gene.
Another object of the present invention is to provide the application of above-mentioned verticillium dahliae pathogenesis related protein.
Another object of the present invention is to provide the application of above-mentioned verticillium dahliae pathogenic related gene.
Another object of the present invention is to provide a kind of method for reducing verticillium dahliae pathogenicity.
Another object of the present invention is to provide the verticillium dahliae mutant that a kind of pathogenicity reduces.
Another object of the present invention is to provide the application of above-mentioned verticillium dahliae mutant.
Verticillium dahliae pathogenesis related protein according to the present invention, amino acid sequence is as shown in SEQ ID No.1
MKTAILSGLSLLAVAMAQQVGTEEPEVHPKITWKRCTGTNGSNCATVNGEIVIDA
NWRWAHNVGGYENCFDGNDWTGLCTGAEDCAKNCAVEGANYGATYGVSTSGD
ALTLKFVTQHNFGTNIGSRTYLMENESKYQMFTLNNNELAFDVDLSTIPCGMNSA
LYLVPMKPDGGLSDEPNNKAGAKYGTGYCDAQCARDLKFINGKGNIVGWEPSPT
DDNAGVGAMGSCCAEIDIWESNRESFAFTPHACKNNSYHVCTDTTCGGTYSEDR
YGGGCDANGCDYNPYRLGNTEFYGQGKTVDTRSKFTVISRFRDNVSEQVFIQNG
NVIIPPKPTLAGLTQFNAAITPEFCTAYPTIFGDRNRHDEIGGHTQLNAAYRLPMVL
VLSVWADHFANMLWLDSIYPPERRGEPGVARGRCPETGRTPADVIRNHPNASVTW
SNIRFGPIGSTHNTSGAVSPPVTPPSSTTRPVTPPTTTTRAATTTTRVAPPTTTTASTG
GPTAPKWGQCGGQGWTGPTVCAAGSTCQAQNQWYSQCL
Verticillium dahliae pathogenic related gene according to the present invention encodes above-mentioned verticillium dahliae and causes a disease related egg
In vain.
Verticillium dahliae pathogenic related gene according to the present invention, the nucleotide sequence such as SEQ ID of the gene
Shown in No.2
ATGAAGACTGCGATTCTCAGCGGCCTCTCCCTGCTGGCCGTGGCCATGGCCCA
GCAGGTCGGCACCGAGGAGCCCGAGGTTCACCCCAAGATCACCTGGAAGCGC
TGCACCGGCACCAACGGCAGCAACTGTGCTACCGTCAACGGTGAGATCGTCAT
CGACGCCAACTGGCGCTGGGCCCACAACGTCGGCGGCTACGAGAACTGCTTCG
ACGGCAACGACTGGACCGGTCTCTGCACCGGCGCCGAGGACTGCGCCAAGAA
CTGCGCCGTCGAGGGTGCCAACTACGGCGCCACCTACGGTGTCTCGACCAGCG
GCGATGCCCTCACCCTGAAGTTCGTCACCCAGCACAACTTTGGCACCAACATT
GGCTCGCGCACCTACCTCATGGAGAACGAGTCCAAGTACCAGATGTTCACCCT
GAACAACAACGAGCTGGCCTTCGACGTCGACCTGTCGACCATCCCCTGCGGCA
TGAACTCGGCCCTCTATCTCGTCCCCATGAAGCCCGATGGTGGTCTCTCTGACG
AGCCCAACAACAAGGCTGGTGCCAAGTACGGTACCGGTTACTGTGACGCCCAG
TGCGCCCGTGACCTGAAGTTCATCAACGGCAAGGGCAACATTGTCGGCTGGGA
GCCCTCGCCCACCGACGACAACGCCGGTGTCGGTGCCATGGGATCTTGCTGCG
CCGAGATCGACATCTGGGAGTCCAACCGCGAGTCGTTCGCCTTCACCCCCCAC
GCCTGCAAGAACAACAGCTACCACGTCTGCACCGACACCACCTGCGGCGGTAC
CTACTCCGAGGACCGCTACGGCGGTGGCTGCGACGCCAACGGCTGCGACTACA
ACCCTTACCGCCTCGGCAACACCGAGTTCTACGGCCAGGGCAAGACGGTTGAC
ACACGCTCCAAGTTCACCGTCATCTCCCGCTTCCGTGACAACGTCTCCGAGCA
GGTCTTCATCCAGAACGGCAACGTCATCATTCCCCCCAAGCCCACTCTCGCCGG
CCTGACCCAGTTCAACGCCGCCATCACCCCCGAGTTCTGCACGGCCTACCCCA
CCATCTTCGGTGACCGCAACCGCCACGACGAGATTGGCGGACACACTCAGCTC
AACGCTGCCTACCGCCTGCCCATGGTCCTCGTCCTGTCTGTCTGGGCCGACCAC
TTCGCCAACATGCTCTGGCTCGACTCCATCTACCCCCCCGAGCGCCGTGGCGAG
CCCGGTGTCGCCCGTGGCCGCTGCCCCGAGACCGGCCGCACCCCCGCCGATGT
CATCCGCAACCACCCCAACGCCTCCGTCACCTGGTCCAACATCCGCTTCGGCCC
CATCGGCTCCACCCACAACACCTCCGGCGCCGTCAGCCCCCCGGTCACCCCTC
CCAGCTCGACCACTCGCCCCGTCACGCCCCCTACCACCACCACCCGCGCCGCC
ACCACCACCACCCGCGTTGCTCCTCCCACCACCACCACCGCCTCCACCGGCGG
CCCTACCGCTCCCAAGTGGGGCCAGTGCGGTGGCCAGGGCTGGACCGGCCCTA
CCGTCTGCGCCGCCGGCTCTACCTGCCAGGCCCAGAACCAGTGGTACTCTCAG TGCCTGTAA
The present invention also provides the carriers for including above-mentioned verticillium dahliae pathogenic related gene.
The present invention also provides mutation said genes to reduce the application of Pathogenicity of Verticillium dahliae of Cotton.
The present invention also provides the verticillium dahliaes that said gene is mutated.
The method according to the present invention for reducing verticillium dahliae pathogenicity includes the above-mentioned verticillium dahliae of mutation and causes a disease
The step of GAP-associated protein GAP or above-mentioned verticillium dahliae pathogenic related gene.
Specific embodiment according to the present invention starts with from verticillium dahliae T-DNA insertional mutagenesis libraries, passes through life
Object measuring shape and Pathogenic Tests are obtained compared with wild type, and sporulation quantity and Raw toxin significantly reduce, and to cotton
The T-DNA insertion mutation body T1027 that pathogenicity significantly weakens, deposit number are：CCTCC M 2017655.Experiment confirms should
Mutant strain is the mono- insertion mutation bodies of T-DNA.
Specific embodiment according to the present invention by TAIL-PCR technologies and the genome database of VdLs.17, obtains
The flanking sequence of T-DNA insertions was obtained, comparison result is shown as dextranase, and the successful clone from wild-type strain Vd080
Pathogenic related gene GLU is obtained.
The present invention is flat using the T-DNA insertional mutagenesis libraries of verticillium dahliae High pathogenicity sclerotium type bacterial strain Vd080
Platform quantitatively dips in bacterium solution method using bottomless paper pot and determines mutant pathogenicity, and analyze its biological characteristics, and screening obtains low
Pathogenicity mutant T1027, which does not change compared with wild type, but the secretion of sporulation quantity and Raw toxin
Amount is remarkably decreased, and dextranase is accredited as by the T-DNA genes being inserted into.Verticillium dahliae mutant T1027 is one and grinds
Study carefully the desirable strain of verticillium dahliae molecular basis of the pathogenesis, the expression and modification of gene and protein can be used as target site
For designing and screening antifungal medicine, has good application potential.
Description of the drawings
Fig. 1 detects hygromycin gene electrophoretogram, wherein M in Positive mutants body for PCR:DL2000DNA maker；Swimming lane
1-25:Positive mutants body；Swimming lane 26:Vd080.
Fig. 2 shows that Southern hybridization checks T-DNA is inserted into copy number, and wherein swimming lane 6 is T1027.
Fig. 3 shows the colonial morphology of wild type Vd080 and mutant T1027 in PDA culture medium.
Fig. 4 wild types Vd080 and mutant T1027 are to cotton Pathogenic Tests.
Verticillium dahliae T1027 (Verticillium dahliae) was preserved in Chinese allusion quotation on November 03rd, 2017
Type culture collection (CCTCC) (address：Wuhan, China Wuhan University postcode 430072), preserving number is CCTCC No:M
2017655。
Specific embodiment
The structure of 1 verticillium dahliae T-DNA insertional mutagenesis libraries of embodiment
Using the verticillium dahliae Vd080 for being isolated from Hebei Xinji grave illness field as starting strain, will carry
The Agrobacterium of pCTHyg binary vectors, using the method for agrobacterium mediation converted (ATMT), structure capacity is 3000
A T-DNA insertional mutagenesis libraries for carrying hygromycin (HPH) resistance screening label.It is sun whether to have hygromycin label
The property important screening foundation of transformant, with hygromycin special primer Hyg-F/Hyg-R (Hyg-F： 5′-
GCCAAGCTTGCATGCCTGCAGGTC-3′；Hyg-R：5′-GCGCTCGAG
TCCTCTAGAAAGAA GGATTAC-3 ') amplify obtain 895bp specific band be positive transformant (figure
1).Detected using Southern hybridization techniques, in mutant T-DNA be inserted into copy number number (Fig. 2).
The acquisition of 2 low pathogenicity mutant T1027 bacterial strains of embodiment
It is differential host with upland cotton susceptible variety Ji cotton 11, bacterium solution method is quantitatively connect using vermiculite sandy soil bottomless bowl of paper, surveys
The pathogenicity of verticillium dahliae mutant is determined.It is screened after primary dcreening operation and secondary screening two-wheeled, it is extremely significantly reduced to obtain pathogenicity
Mutant T1027, for referring to 50 compared with wild type relative disease, the relative disease of T1027 refers to only 8.7, and pathogenicity greatly reduces, table
Fig. 3 is shown in type comparison.
3 mutant T1027 biology shape analyses of embodiment
The observation of 3.1 colonial morphologies and the measure of growth rate
Using wild-type strain Vd080 as control, the prominent of a concentration of 1 × 107/mL of 5 μ L is dripped in the center of PDA culture medium
Variant spore suspension, each 5 repetitions of bacterial strain, 25 DEG C of constant temperature quiescent cultures 10 days record the growthform of bacterium colony.And point
Colony growth diameter is not measured using crossing method in 5d, 7d, 9d and 11d, calculates growth rate.
The results show that being compared with wild type Vd080, mutant T1027 sclerotium yield drastically declines, B16 cell amount
It is significantly reduced.But both growth rates reach unanimity, and are 4.5mm/d.
3.2 sporulation quantities and Raw toxin assay
Using wild-type strain Vd080 as control, looked into formula culture medium in 40mL and drip 5 μ L mutant spore suspensions, 25
DEG C, after 150rpm shaken cultivations 6d, its sporulation quantity is measured by blood counting chamber.
After continuing shaken cultivation 19d later, bacterium solution 13000rpm is taken, centrifuges 30min.Supernatant is taken, according to Coomassie brilliant blue
G-250 methods make standard curve by standard protein of bovine serum albumin, with microplate reader (Synergy HT, BioTek companies of the U.S.)
The protein concentration of Amber box policy is measured as Raw toxin content.
The results show that due to the insertion of T-DNA, the sporulation quantity of T1027 is by the 4.2 × 10 of wild type7A/ml is reduced to
1.3×107A/ml, the range of decrease are up to 67%.The secretory volume of Raw toxin is also greatly reduced, wild type 58.7ml/L, and T1027
Only 39%, 22.5ml/L of wild type.
Identification and the clone of gene are inserted into 4 mutant T1027 of embodiment by T-DNA
According to DNA sequences on the inside of the T-DNA left arms (LB) of binary vector pCTHyg (used in building ATMT libraries) and right arm (RB)
Row, using the Primer Blast online tools in NCBI separately design 3 ' and 5 ' step shifting primer R-SP1, R-SP2, R-SP3 and
L-SP1, L-SP2 and L-SP3 (table 1), degenerate primer AP1, AP2, AP3 and AP4 that Genome Walking kits provide,
Step moves primer and is used as template amplification T-DNA left and right arms flanking sequences in pairs with random primer.The primer sequence is referring to table 1, institute
It is carried out with response procedures and reaction system according to the method for Genome Walking kits.After reaction, above-mentioned is taken
One takes turns, the reaction product of the second wheel, third round is detected on 1% agarose gel electrophoresis.3 ' ends and 5 ' ends the are separately recovered
Three-wheel TAIL-PCR specific products, cloning and sequencing.Sequencing result is first compared two-by-two with T-DNA, if there is repetitive sequence,
Just and the genome database (http of verticillium dahliae VdLs.17 that shares online of BROAD research institutes of the U.S.://www.
broadinstitute.org/annotation/genome/verticillium_dahliae/
MultiDownloads.html it) is compared, and extracts the essential information of pathogenic related gene.
It is compared by TAIL-PCR technologies, and with the genome database of VdLs.17, learns mutant T1027 T-
DNA is inserted into flank sequence gene and is encoded to VDAG_00752, is dextranase positioned at verticillium dahliae functional annotation.Design is special
Different primer T1027 (ATGAAGACTGCGATTCTCAG/TTACAGGCACTGAGA GTACC) is cloned from wild type Vd080 should
Gene learns that the gene includes 1602 pairs of bases, intronless, and the nucleotide sequence of gene is compiled as shown in SEQ ID No.2
533 amino acid of code, amino acid sequence is as shown in SEQ ID No.1.
1 TAIL-PCR special primer lists of table
Sequence table
<110>The Chinese Academy of Agriculture Science and Technologys Cotton Research Institute
<120>Verticillium dahliae pathogenesis related protein and its encoding gene, application and mutant
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 533
<212> PRT
<213>Verticillium dahliae T1027 (Verticillium dahliae T1027)
<400> 1
Met Lys Thr Ala Ile Leu Ser Gly Leu Ser Leu Leu Ala Val Ala Met
1 5 10 15
Ala Gln Gln Val Gly Thr Glu Glu Pro Glu Val His Pro Lys Ile Thr
20 25 30
Trp Lys Arg Cys Thr Gly Thr Asn Gly Ser Asn Cys Ala Thr Val Asn
35 40 45
Gly Glu Ile Val Ile Asp Ala Asn Trp Arg Trp Ala His Asn Val Gly
50 55 60
Gly Tyr Glu Asn Cys Phe Asp Gly Asn Asp Trp Thr Gly Leu Cys Thr
65 70 75 80
Gly Ala Glu Asp Cys Ala Lys Asn Cys Ala Val Glu Gly Ala Asn Tyr
85 90 95
Gly Ala Thr Tyr Gly Val Ser Thr Ser Gly Asp Ala Leu Thr Leu Lys
100 105 110
Phe Val Thr Gln His Asn Phe Gly Thr Asn Ile Gly Ser Arg Thr Tyr
115 120 125
Leu Met Glu Asn Glu Ser Lys Tyr Gln Met Phe Thr Leu Asn Asn Asn
130 135 140
Glu Leu Ala Phe Asp Val Asp Leu Ser Thr Ile Pro Cys Gly Met Asn
145 150 155 160
Ser Ala Leu Tyr Leu Val Pro Met Lys Pro Asp Gly Gly Leu Ser Asp
165 170 175
Glu Pro Asn Asn Lys Ala Gly Ala Lys Tyr Gly Thr Gly Tyr Cys Asp
180 185 190
Ala Gln Cys Ala Arg Asp Leu Lys Phe Ile Asn Gly Lys Gly Asn Ile
195 200 205
Val Gly Trp Glu Pro Ser Pro Thr Asp Asp Asn Ala Gly Val Gly Ala
210 215 220
Met Gly Ser Cys Cys Ala Glu Ile Asp Ile Trp Glu Ser Asn Arg Glu
225 230 235 240
Ser Phe Ala Phe Thr Pro His Ala Cys Lys Asn Asn Ser Tyr His Val
245 250 255
Cys Thr Asp Thr Thr Cys Gly Gly Thr Tyr Ser Glu Asp Arg Tyr Gly
260 265 270
Gly Gly Cys Asp Ala Asn Gly Cys Asp Tyr Asn Pro Tyr Arg Leu Gly
275 280 285
Asn Thr Glu Phe Tyr Gly Gln Gly Lys Thr Val Asp Thr Arg Ser Lys
290 295 300
Phe Thr Val Ile Ser Arg Phe Arg Asp Asn Val Ser Glu Gln Val Phe
305 310 315 320
Ile Gln Asn Gly Asn Val Ile Ile Pro Pro Lys Pro Thr Leu Ala Gly
325 330 335
Leu Thr Gln Phe Asn Ala Ala Ile Thr Pro Glu Phe Cys Thr Ala Tyr
340 345 350
Pro Thr Ile Phe Gly Asp Arg Asn Arg His Asp Glu Ile Gly Gly His
355 360 365
Thr Gln Leu Asn Ala Ala Tyr Arg Leu Pro Met Val Leu Val Leu Ser
370 375 380
Val Trp Ala Asp His Phe Ala Asn Met Leu Trp Leu Asp Ser Ile Tyr
385 390 395 400
Pro Pro Glu Arg Arg Gly Glu Pro Gly Val Ala Arg Gly Arg Cys Pro
405 410 415
Glu Thr Gly Arg Thr Pro Ala Asp Val Ile Arg Asn His Pro Asn Ala
420 425 430
Ser Val Thr Trp Ser Asn Ile Arg Phe Gly Pro Ile Gly Ser Thr His
435 440 445
Asn Thr Ser Gly Ala Val Ser Pro Pro Val Thr Pro Pro Ser Ser Thr
450 455 460
Thr Arg Pro Val Thr Pro Pro Thr Thr Thr Thr Arg Ala Ala Thr Thr
465 470 475 480
Thr Thr Arg Val Ala Pro Pro Thr Thr Thr Thr Ala Ser Thr Gly Gly
485 490 495
Pro Thr Ala Pro Lys Trp Gly Gln Cys Gly Gly Gln Gly Trp Thr Gly
500 505 510
Pro Thr Val Cys Ala Ala Gly Ser Thr Cys Gln Ala Gln Asn Gln Trp
515 520 525
Tyr Ser Gln Cys Leu
530
<210> 2
<211> 1602
<212> DNA
<213>Verticillium dahliae T1027 (Verticillium dahliae T1027)
<400> 2
atgaagactg cgattctcag cggcctctcc ctgctggccg tggccatggc ccagcaggtc 60
ggcaccgagg agcccgaggt tcaccccaag atcacctgga agcgctgcac cggcaccaac 120
ggcagcaact gtgctaccgt caacggtgag atcgtcatcg acgccaactg gcgctgggcc 180
cacaacgtcg gcggctacga gaactgcttc gacggcaacg actggaccgg tctctgcacc 240
ggcgccgagg actgcgccaa gaactgcgcc gtcgagggtg ccaactacgg cgccacctac 300
ggtgtctcga ccagcggcga tgccctcacc ctgaagttcg tcacccagca caactttggc 360
accaacattg gctcgcgcac ctacctcatg gagaacgagt ccaagtacca gatgttcacc 420
ctgaacaaca acgagctggc cttcgacgtc gacctgtcga ccatcccctg cggcatgaac 480
tcggccctct atctcgtccc catgaagccc gatggtggtc tctctgacga gcccaacaac 540
aaggctggtg ccaagtacgg taccggttac tgtgacgccc agtgcgcccg tgacctgaag 600
ttcatcaacg gcaagggcaa cattgtcggc tgggagccct cgcccaccga cgacaacgcc 660
ggtgtcggtg ccatgggatc ttgctgcgcc gagatcgaca tctgggagtc caaccgcgag 720
tcgttcgcct tcacccccca cgcctgcaag aacaacagct accacgtctg caccgacacc 780
acctgcggcg gtacctactc cgaggaccgc tacggcggtg gctgcgacgc caacggctgc 840
gactacaacc cttaccgcct cggcaacacc gagttctacg gccagggcaa gacggttgac 900
acacgctcca agttcaccgt catctcccgc ttccgtgaca acgtctccga gcaggtcttc 960
atccagaacg gcaacgtcat cattcccccc aagcccactc tcgccggcct gacccagttc 1020
aacgccgcca tcacccccga gttctgcacg gcctacccca ccatcttcgg tgaccgcaac 1080
cgccacgacg agattggcgg acacactcag ctcaacgctg cctaccgcct gcccatggtc 1140
ctcgtcctgt ctgtctgggc cgaccacttc gccaacatgc tctggctcga ctccatctac 1200
ccccccgagc gccgtggcga gcccggtgtc gcccgtggcc gctgccccga gaccggccgc 1260
acccccgccg atgtcatccg caaccacccc aacgcctccg tcacctggtc caacatccgc 1320
ttcggcccca tcggctccac ccacaacacc tccggcgccg tcagcccccc ggtcacccct 1380
cccagctcga ccactcgccc cgtcacgccc cctaccacca ccacccgcgc cgccaccacc 1440
accacccgcg ttgctcctcc caccaccacc accgcctcca ccggcggccc taccgctccc 1500
aagtggggcc agtgcggtgg ccagggctgg accggcccta ccgtctgcgc cgccggctct 1560
acctgccagg cccagaacca gtggtactct cagtgcctgt aa 1602
Claims (9)
1. verticillium dahliae pathogenesis related protein, which is characterized in that its amino acid sequence is as shown in SEQ ID No.1.
2. verticillium dahliae pathogenic related gene, which is characterized in that encode verticillium dahliae described in claim 1 and cause a disease
GAP-associated protein GAP.
3. verticillium dahliae pathogenic related gene according to claim 2, which is characterized in that the nucleotide of the gene
Sequence is as shown in SEQ ID No.2.
4. include the carrier of the verticillium dahliae pathogenic related gene described in claim 2.
5. gene described in claim 2 is mutated to reduce the application of Pathogenicity of Verticillium dahliae of Cotton.
6. the verticillium dahliae that gene described in claim 2 is mutated.
A kind of 7. method for reducing verticillium dahliae pathogenicity, which is characterized in that the method includes mutation claim 1 institutes
The step of stating verticillium dahliae pathogenic related gene described in verticillium dahliae pathogenesis related protein or claim 2.
8. verticillium dahliae T-DNA insertion mutation body bacterial strains, which is characterized in that its deposit number is：CCTCC M
2017655。
9. verticillium dahliae T-DNA insertion mutation body bacterial strains are in Pathogenicity of Verticillium dahliae of Cotton research side described in claim 8
The application in face.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106011269A (en) * | 2016-07-06 | 2016-10-12 | 中国农业科学院棉花研究所 | Method for rapid identification of cotton verticillium wilt resistance by means of real-time quantitative PCR |
| CN110923250A (en) * | 2019-11-13 | 2020-03-27 | 中国农业科学院棉花研究所 | Application of cotton verticillium wilt resistance related gene GhSDH1-1 |
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| CN104805095A (en) * | 2014-12-18 | 2015-07-29 | 中国农业科学院棉花研究所 | Application of cotton verticillium wilt pathogenicity-related gene CYC8 |
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| CN106011269B (en) * | 2016-07-06 | 2019-12-10 | 中国农业科学院棉花研究所 | Method for rapidly identifying cotton verticillium wilt resistance by using real-time quantitative PCR |
| CN110923250A (en) * | 2019-11-13 | 2020-03-27 | 中国农业科学院棉花研究所 | Application of cotton verticillium wilt resistance related gene GhSDH1-1 |
| CN110923250B (en) * | 2019-11-13 | 2021-12-24 | 中国农业科学院棉花研究所 | Application of cotton verticillium wilt resistance related gene GhSDH1-1 |
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Application publication date: 20180608 |