CN104004771A - Magna-porthe grisea avirulence gene AvrPi9, encoded polypeptide and polynucleotide and application thereof - Google Patents
Magna-porthe grisea avirulence gene AvrPi9, encoded polypeptide and polynucleotide and application thereof Download PDFInfo
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- CN104004771A CN104004771A CN201410258019.6A CN201410258019A CN104004771A CN 104004771 A CN104004771 A CN 104004771A CN 201410258019 A CN201410258019 A CN 201410258019A CN 104004771 A CN104004771 A CN 104004771A
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Abstract
The invention discloses a magna-porthe grisea avirulence gene AvrPi9, encoded polypeptide and polynucleotide and application thereof. A nucleotide sequence and an encoded amino acid polypeptide sequence of the magna-porthe grisea new avirulence gene Avr-Pi9 are provided. The gene achieves peculiar inducible expression during plant infection. Based on the gene, the plant with the improved disease resistance can be prepared, or molecular markers tracking magnaporthe oryzae population changes can be prepared.
Description
Technical field
The present invention relates to gene engineering technology field, be specifically related to separating clone and the application of a kind of rice blast fungus nontoxic gene Avr-Pi9.
Background technology
Paddy rice (Oryza sativa L.) is the first food crop of China, is also the most important food crop in the whole world.Rice blast fungus (Magnaporthe oryzae) is the pathogen that causes rice blast, and the whole world is annual because rice blast causes 10%～30% Rice Yield Loss Caused, and even No kernels or seeds are gathered, as in a year of scarcity for serious field.The means of prevention and control rice blast are mainly applying pesticides and plantation resistant variety at present.Applying pesticides exists contaminate environment to increase production cost, affects a series of unfavorable factors such as rice quality, so the selection and popularization of disease-resistant variety is the most economic and effective means of current prevention and control rice blast.Yet, in physical environment, in the selection of rice resistant variety, to depress, rice blast fungus flora is replaced and can be broken through rapidly the new physiological strain of resistant variety direction and form dominant population, finally causes resistant variety resistance " forfeiture ".In production practice, resistant variety is planted after 3-5 continuously, and resistance will obviously fail, disease-resistant cannot persistence.How to monitor physiological races of rice blast fungus and occur dynamically, cultivate the kind of permanent disease-resistant and find more efficiently resistant breeding strategy, become an urgent demand in current production.
Mutual between paddy rice and Pyricularia oryzae meets " gene pairs gene " hypothesis (Flor, 1947 as mechanism; Silue et al., 2000; Bryan et al., 2000; Orbach et al.2000).According to this theory, paddy rice derives from the nontoxic gene (Avr gene) of Pyricularia oryzae by its resistant gene (R gene) specific recognition, thereby excites a series of defense responses in paddy rice body.In the past twenty years, the research of relevant rice blast resistant gene aspect both at home and abroad makes great progress, at least reported 63 blast resisting sites totally 77 major genes (national paddy rice Data centre. rice blast Major resistance gene list [DB/OL] .http: //www.ricedata.cn/gene.).These genes have been accelerated paddy disease-resistant breeding process greatly.Yet the resistance of R gene mediated has strict kind specificity, different R gene and its recognition factors have strict corresponding relation, and the high variability of pathogenic bacteria microspecies virulence easily causes the resistance of R gene mediated to disappear.Address this problem, need to further investigate the essence of interaction between resistant gene and nontoxic gene, new Genetically engineering for plant broad-spectrum disease resistance strategy and method (Wit.1992 is provided then; Bryan et al., 2000; Orbach et al.2000).At present, research with respect to rice blast resistant gene, rice blast fungus nontoxic gene clone aspect relatively lags behind, although nearly 40 nontoxic genes are in the news, but only have 9 nontoxic genes to be cloned, PWL1 (Kang et al. wherein, 1995) and PWL2 (Sweigard et al., 1995) be weeping love grass host specificity, remaining AvrPi-ta (Orbach et al., 2000), ACE1 (Bohnert et al., 2004), Avr1-CO39 (Farman and Leong, 1998), AvrPiz-t (Li et al., 2009), AvrPii, AvrPia and AvrPik/km/kp (Yoshida et al., 2009) these 7 is that control rice blast fungus is pathogenic to paddy rice.Wherein only has the ACE1 non-secretory polyketide synthases gene of having encoded, all the encoded secretory protein of prediction of other 8 nontoxic genes.Between the resistant gene of having cloned at these and nontoxic gene, only have Pi-ta and Avr-Pita, Pia and Avrpia, Piz-t and AvrPiz-t and Pikm and AvrPik|km|kp are paired, and only have the molecular Evidence that has direct interaction between Pi-ta and AvrPi-ta.
Summary of the invention
The cloning process and the molecule marker that the object of this invention is to provide avirulence gene of rice blast.
In a first aspect of the present invention, a kind of polypeptide of avirulence gene of rice blast AvrPi9 coding is provided, this polypeptide is selected from lower group:
(a) there is the polypeptide of SEQ ID NO:3 aminoacid sequence;
(b) SEQ ID NO:3 aminoacid sequence process is one or more (as 1-50, preferably 1-30, more preferably 1-10, more preferably 1-5,1-3 more preferably) replacement, disappearance or the interpolation of amino-acid residue form, and have can with the protein of rice blast resistant gene Pi9 coding do mutually function by (a) derivative polypeptide.
In another aspect of this invention, provide avirulence gene of rice blast AvrPi9, it is characterized in that, it comprises a nucleotide sequence, and this nucleotide sequence is selected from lower group:
The polynucleotide of coding said polypeptide; Polynucleotide with above-mentioned polynucleotide complementation.
In another preference, described polynucleotide are:
The polynucleotide with nucleotide sequence shown in SEQ ID NO:1; The polynucleotide with nucleotide sequence shown in SEQ ID NO:2;
In another aspect of this invention, provide a kind of carrier, it contains described polynucleotide.
In another aspect of this invention, provide a kind of genetically engineered host cell, it contains described carrier; Or the polynucleotide described in being integrated with in its genome.
In another aspect of this invention, utilize described aminoacid sequence or nucleotide sequence or carrier or genetically engineered host cell, for cultivating disease-resistant plants kind.
In another aspect of this invention, utilize described aminoacid sequence or nucleotide sequence, for the identification of the infection ability of Pyricularia oryzae, or for the preparation of the marker that detects plant Pyricularia oryzae toxicity population distribution.These markers include but not limited to SNP (mononucleotide polymorphic), SSR (simple sequence repetition polypeptide), RFLP (restriction enzyme length polypeptide), CAP (cutting amplified fragments polypeptide).
In another aspect of this invention, the purposes of the molecular marked compound described in providing, for the planting structure distribution of rice varieties.
The invention has the beneficial effects as follows:
The present invention obtains a kind of new avirulence gene of rice blast AvrPi9, and the structure and function result of study based on described gene can be widely used in novel agrochemical exploitation, physiological races of rice blast fungus monitoring and the theoretical investigation of rice blast Molecular interaction; Sequence based on described gene, can set up the Markers for Detection system of the pathogenic variation of monitoring Pyricularia oryzae natural population; Understand the variation of described gene in the natural population of field dynamic, thereby instruct rice varieties planting structure distribution and rotation, effectively to control the generation of disease; Utilize described gene and corresponding disease-resistant gene and suitable promotor to assemble, build the disease-resistant engineering carrier of broad spectrum durable, and and then Introduced into Rice or other plant cell, cultivate the disease-resistant rice varieties of broad spectrum durable.
Accompanying drawing explanation
Fig. 1 is the sudden change F+strain that the present invention passes through R01-1 in 26 hybrid bacterial strains of 192 specific molecular markers for identification of R01-1, and according to the comparison of pcr amplification band and result statistics, the systematic evolution tree that adopts adjacent method (Neighbor-joining) to build;
Fig. 2 is natural bacterial strain scanning result and the gene structure of AvrPi9 candidate gene, and avirulent strains R88-002 1434 exists with No. 1435 genes and consistent with in Guy11; In virulent strain R01-1, two genes all exist still No. 1434 gene inside to have transposon Mg-SINE to insert;
Fig. 3 is that the present invention identifies the complementary function of AvrPi9 candidate gene, candidate gene 1434 and 1435 is imported to respectively in virulent strain R01-1 and 75-5, and the conversion bacterial strain that each transformation event obtains utilizes the Tp309 single-gene system (Pi9-Tp309) of Tp309 and importing disease-resistant gene Pi9 to carry out pathogenic evaluation.Fig. 3 A is R01-1 transformant inoculation the result, and Fig. 3 B is 75-5 transformant inoculation the result.Result shows, R1434 is exactly AvrPi9 gene.
Fig. 4 is that the present invention loses PCR detected result and the Function Identification result figure of transformant to AvrPi9 gene function.Fig. 4 A loses the PCR detected result of transformant to AvrPi9 gene function, be respectively the transformant RJ3-53 that knocks out the transformant RJ1-43 of R1434, the transformant RJ2-7 that knocks out R1434 and RJ1435 and RJ2-15, knocks out RJ1435.Fig. 4 B is to losing transformant Function Identification result, and experimental result has further confirmed that the R1434 in R88-002 is exactly AvrPi9 gene.
Embodiment
Below in conjunction with the drawings and specific embodiments, further illustrate content of the present invention, but should not be construed as limitation of the present invention.If do not specialize, the conventional means that the technique means adopting in embodiment is well known to those skilled in the art.
Embodiment 1: carry determining of nontoxic gene AvrPi9 wild-type rice blast fungus bacterial strain R88-002.
R01-1 is the avrPi9 mutant (Kim BR, et al., 2004) that the method by the vaccinization of 26 AvrPi9 strain mixtures screens.Therefore, can very reasonably infer that R01-1 should derive from 26 hybrid bacterial strains.The present invention identifies the sudden change F+strain of R01-1 in 26 hybrid bacterial strains by 192 specific molecule markers of R01-1 of exploitation (table 1), result is as shown in table 2, compare with R01-1, bacterial strain R88-002 does not observe polymorphic, and bacterial strain 85-242 polymorphism is the abundantest.According to the comparison of pcr amplification band and result statistics, adopt adjacent method (Neighbor-joining) constructing system evolutionary tree (accompanying drawing 1), finally determine that R88-002 is the wild-type of R01-1 bacterial strain.
Table 1 is for the identification of the PCR primer of R01-1 and 26 bacterial strain sibships
Table 2.192 pair primer is polymorphic situation .NA in 26 bacterial strains: without amplified production; AD: product varies in size.
Embodiment 2:R88-002 and R01-1 genome sequence determination and comparison.
The present invention adopts the order-checking of Illumina platform to carry out gene order-checking to R88-002 and R01-1, and for reaching the assembling genome of equal quality, the present invention carries out 100 times of coverage order-checkings to them, and order-checking amount reaches about 4.5Gb.On the whole, the genome of R88-002 and R01-1 assembling is at Genome Size, and the aspects such as predicted gene number and secretory protein number and other rice blast fungus genomes that checked order are without significant difference.In order to facilitate comparison and analysis, the present invention also introduces two other AvrPi9 avirulent strains 70-15 and Ina168 of having announced sequence.R88-002 is AvrPi9 avirulent strains and R01-1 is AvrPi9 virulent strain.According to the rice blast fungus nontoxic gene experience of having cloned, courageously suppose AvrPi9 coding secretory protein, and the mechanism that nontoxic gene function is lost is the afunction that has large Indel to cause at gene elmination, gene or gene regulating region (upstream 1kb).AvrPi9 candidate gene should be the secreted protein gene of R88-002 like this, and upstream 1kb regulation and control region is consistent with 70-15 and Ina168 gene, and has larger difference with R01-1.
The present invention uses Repeatmask3.3 (http://www.repeatmasker.org to rice blast fungus genomic data, species=" magnaporthe grisea ") filter, the tumor-necrosis factor glycoproteins storehouse of use is from RepBasehttp: //www.girinst.org/).Predictive genes has been used the fgenesh module (species: magnaporthe of Molquest; Sequence length is greater than 20aa).R88-002 has 12344 predicted genes, secretory protein prediction is used SignalP4.1 detection to contain 1764 of signal peptide genes, then use the Protcomp-AN module predicted protein cellular localization of Molquest, contain secretion signal gene 1015, finally with TMHMM2.0, filter out the albumen of two above membrane spaning domains, finally remaining 1008 genes are prediction minute albumen.Transfer these 1008 secreted protein genes and upstream 1Kb regional sequence as inquiry storehouse, blastn comparison R01-1/70-15 and In168 genome, transfer consistent with 70-15/In168 gene and exist 23 secreted protein genes of larger difference as AvrPi9 candidate gene with R01-1, detailed results is in Table 3.For these 23 candidate genes, we get rid of sequence assembly and compare the false positive gene causing through careful human interpretation, and design specific aim primer is got rid of the gene that has variation in avirulent strains.Nature bacterial strain scanning (seeing Fig. 3 A and B) result avirulent strains R88-002 1434 exists with No. 1435 genes and consistent with in Guy11; In virulent strain R01-1, two genes all exist still No. 1434 gene inside to have transposon Mg-SINE to insert; In virulent strain 75-5, two genes do not exist.Above result tentatively determines that 1434 and No. 1435 genes (gene structure is shown in Fig. 3 B) of R88-002 are AvrPi9 emphasis candidate genes, then by following its function of functional complementation experimental verification.
Embodiment 3: the complementation of candidate gene is identified.
Candidate gene of the present invention is complementary identifies that primer and the carrier background of vector construction are as shown in table 3.Use KOD high-fidelity enzyme, utilize 12655CF/12655CR and 12654CF/12654CR respectively R1434 and R1435 to be increased out from R88-002, and be cloned into the T-easy carrier of buying from Quan Shi King Company.Recycling Sac I and Asc I recombinate gene fragment to pCSN43 carrier, respectively called after pR55 and pR54.Use the method for PEG protoplast transformation that carrier construction is imported to (Sweigard et al.1995) in virulent strain R01-1 and 75-5.Wherein, in R01-1, R1434 gene inside has Mg-SINE to insert, and R1435 is the same with R88-002, therefore, only carries out the complementation of pR55.And two genes do not exist in 75-5, therefore, carry out respectively the complementation of pR55 and pR54.
The Tp309 single-gene system (Pi9-Tp309) that the conversion bacterial strain that the present invention obtains each transformation event utilizes Tp309 and imports disease-resistant gene Pi9 carries out pathogenic evaluation, with R01-1 and R88-002 in contrast.Result shows, R01-1 has pathogenic to Pi9-Tp309, but after importing the R1434 gene of R88-002, transformant (Fig. 3 A, pR55-1/pR55-5) shows the nontoxicity identical with F+strain R88-002.Virulent strain 75-5, after importing the R1434 gene (Fig. 3 B, 127-5-1-15/127-5-1-22) of R88-002, obtains the nontoxicity to Pi9 equally.But the R1435 gene (Fig. 3 B, 127-5-3-9) that imports R88-002 can not complementary Pi9 nontoxicity.Equally, transform the nontoxicity that the contrast transformant (Fig. 3 B, 127-5-4-5) of pCSN43 empty carrier can not complementary Pi9, got rid of the impact of transformation experiment on complementary result itself.These results show, R1434 is exactly AvrPi9 gene.
Embodiment 4: knock out the afunction experiment of AvrPi9 gene.
In order further to confirm the function of AvrPi9, applicant, carried out the gene knockout methodological function property the lost experiment based on homologous recombination gene substitution.Recombinant dna fragment utilizes primer as shown in table 3 to carry out respectively pcr amplification, recycling Overlapping PCR is by RJ1LB/HPH/RJ1RB, RJ1LB/HPH/RJ2RB, RJ3LB/HPH/RJ2RB tri-fragments connect, obtain RJ1, RJ2, RJ3PCR product, the PCR product purification test kit that Yong Zi Omega company buys afterwards carries out purifying.Utilize the method for PEG protoplast transformation, import in avirulent strains R88-002, subsequently the transformant of gene substitution restructuring is carried out the Molecular of double PCR technology, result shows, RJ1-43, RJ2-7/RJ2-15, RJ3-53 is the transformant (Fig. 4 A) of homologous recombination.
Utilize equally the Tp309 single-gene system (Pi9-Tp309) of Tp309 and importing disease-resistant gene Pi9 to carry out pathogenic evaluation, above-mentioned knock-out bacterial strain is carried out to artificial inoculation.Result shows, knock out the transformant RJ1-43 of R1434 and knock out R1434 and the nontoxic function of transformant RJ2-7/RJ2-15 of RJ1435 completely loses, and the bacterial strain RJ3-53 nontoxicity that only knocks out RJ1435 do not change (Fig. 4 B).Experimental result has further confirmed that the R1434 in R88-002 is exactly AvrPi9 gene.
The carrier that table 3. Function Identification of the present invention is used or PCR fragment and primer thereof
Sequence and the structure of embodiment 5:AvrPi9 gene.
AvrPi9 full length cDNA sequence (SEQ ID NO:1) and genome sequence (SEQ ID NO:2) are as follows.Gene order analysis to AvrPi9 shows, AvrPi9 comprises 2 exons and 1 intron.
AvrPi9 full length cDNA sequence (SEQ ID NO:1):
atgcagttctctcagatcctcaccgtcttgttccttggcgtctccgtcagcgcccttcccgccggcggtctgcccggcagccctggcagcgctgtccagaggtgccactgcccccctcgtggctcccacgcccacggctccctcgccgctcgggaggaagcgcccgaggccgaaggtgacgccaagatttccgcccgctacacctgccccaactgccacaagacgggcaaaggctgcgatgatggctggtgccaagtcgaaaagacgcactgg
AvrPi9 genome sequence (SEQ ID NO:2, comprises intron and exon):
Aaaaaaaaaaaaaaagacaaacacgggctgacagtgacaaattctcccagtggtaccggagtcggtttggttcacgagtcgcagcccgccggcgagattgtcagagaagtccgatcagaagctctggctgttattgacaagcttaaagcgcagctcacagtgccgtgagttttccatgtcctgaaacacgttgttggtggtgggctgcggctttagctacgttcagagtcatgacatcaacaccgcccccctcgcacataagcacatggtgaggtgggctcggttaacgcttaaagaccgcatcccaaaatcccatgtgtgcttatcgcgtgatttctttttgttcttgtttttttttccttttttttgttttctcctgtgctgccgtctattccaagttacttccctcggcatggtcaagctatgaataggtctaggtagtcgccgcggcggttgtgggagtgcctcgctaccaccgaagatatccactccccacaactcgaagctgcgcggcgatcctaataaccgcatgactggtctgggtggtattgttgcacgaaaagggtaaaacccaaaccatgcacttgggagcagaacaacataggtccactgctccatcttgtttggccttctttgtggcctactgctgatcccccctttccagaatcgctacatttgttcttggacaggacaaaaatatgacctgtgtaagcataaagtaacattattgcatgatcataccagatctgatatagcagcccatgagtcctgtctgctttgcttaccccctcccaactccactatccagtcctatactcgacgtgaccaaaaggcgcttcggggcgtggactatagtctgatgagtgcggttacgagtcggcaggtgggtttatcatcagtcacgcctcatgacggtctggccgattggttcgttgggggctggagttggaacgcacttgttttgagtgtcgccatgaacatcattctcctacactggggctctcctacactggggctgtcctacactgagaaaagatattgggcacctgttcgacgagtataaaggccgcctttcgctgccgaagaaatcacttcctctgctccttcctatccccatccacaggattccagctattcgacaaccccttctctcttttctttatctcttggctttttttccttactgtcactctctttctctctctctctgtcttccttctagtcattcctttggcaaagttctttttttccttccagcccaac (promoter and 5 - UTR sequence)
Atgcagttctctcagatcctcaccgtcttgttccttggcgtctccgtcagcgccct tcccgccggcggtctgcccggcagccctggcagcgctgtccagaggtgccactgcc cccctcgtggctcccacgcccacggctccctcgccgctcgggaggaagcgcccgag gccgaaggtgacgccaagatttccgcccgctacacctgccccaactgccacaagac gggcaaaggctgcgatgatgg (exons 1)
Gtaaataagcaaccccccatccccttgaaatcctttttgcggtcatgttgctgacg tctttgatag (introne 1)
Ctggtgccaagtcgaaaagacgcactggtag (exon 2)
Aggaaaagtcaatcgagtaattatacaggttagtgcttgtactagtgctctttctaataacaatccatagtatattatactaagtagtaagcacattcgcctttaattaattctggtatcagtaatgcgcagaagggtgttgatgctggggcttcaaaagtcgggccactcgcattatcgcataattgcgaccaagttaaaagctggtcaattttccaacaatagtaagctaagatgttggttaatactagtatggccttggtaataaataactcttctgcaagcagagaacaactatgtcacaaagtttcaacaaatatatagagtcgctgtctgctttggaaattataccccgtctcttctcccccttaccatcgccgcaaccattcgatattcgccttcttcttcttttgtcttttttctttcttcttttct (3 - UTR sequence and termination)
Sequence and the structure of embodiment 6:AvrPi9 nontoxic protein.
The nontoxic protein aminoacid sequence of AvrPi9 genes encoding is as shown in SEQ ID NO:3.Nontoxic gene AvrPi9 1 polypeptide being formed by 92 amino-acid residues of encoding.Use SignalP4.1 to carry out structure elucidation to the albumen of this genes encoding, result shows that this albumen has the feature of secretory protein, and 1-18 amino acid region of its N-terminal is signal peptide region.
AvrPi9 aminoacid sequence (SEQ ID NO:3):
mQFSQILTVLFLGVSVSA(signal peptide sequence)
LPAGGLPGSPGSAVQRCHCPPRGSHAHGSLAAREEAPEAEGDAKISARYTCPNCHKTGKGCDDGWCQVEKTHW
Claims (7)
1. an avirulence gene of rice blast AvrPi9, is characterized in that, it is: the polynucleotide with nucleotide sequence shown in SEQ ID NO:1; The polynucleotide with nucleotide sequence shown in SEQ ID NO:2.
2. the polypeptide that described in claim 1, avirulence gene of rice blast AvrPi9 encodes, is characterized in that: described polypeptide is selected from lower group:
(a) there is the polypeptide of SEQ ID NO:3 aminoacid sequence;
(b) SEQ ID NO:3 aminoacid sequence is formed through replacement, disappearance or the interpolation of one or more amino-acid residues, and have can with the protein of rice blast resistant gene Pi9 coding do mutually function by (a) derivative polypeptide.
3. with the polynucleotide that described in claim 1, avirulence gene of rice blast AvrPi9 is equal to, it is characterized in that: the polynucleotide of polypeptide described in coding claim 2; Polynucleotide with above-mentioned polynucleotide complementation.
4. an application of avirulence gene of rice blast AvrPi9 described in claim 1, is characterized in that being applied to building the carrier that contains the polynucleotide sequence described in claim 1 or 3.
5. according to any one application in cultivating disease-resistant plants kind of claim 1-3.
6. the application of carrier according to claim 4 in cultivating disease-resistant plants kind.
7. utilize any one application in detecting plant Pyricularia oryzae toxicity population distribution and rice varieties planting structure distribution of claim 1-3.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104498612A (en) * | 2014-12-24 | 2015-04-08 | 安徽省农业科学院水稻研究所 | Molecular marker closely linked with rice blast Pi9 gene and application thereof |
| CN112662804A (en) * | 2021-01-25 | 2021-04-16 | 中国水稻研究所 | Primer group, kit and method for detecting pathogenic variation of avirulence gene AvrPi9 of magnaporthe grisea |
| CN114410731A (en) * | 2021-12-02 | 2022-04-29 | 吉林省农业科学院 | Grate screen for screening rice blast resistant materials and application thereof |
| CN115852028A (en) * | 2022-11-01 | 2023-03-28 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Triple PCR primer group, kit and detection method for detecting avirulence gene of rice blast germs |
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| CN110904124B (en) * | 2019-10-25 | 2020-12-11 | 华南农业大学 | Magnaporthe grisea avirulence gene AvrPit and application thereof |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498612A (en) * | 2014-12-24 | 2015-04-08 | 安徽省农业科学院水稻研究所 | Molecular marker closely linked with rice blast Pi9 gene and application thereof |
| CN112662804A (en) * | 2021-01-25 | 2021-04-16 | 中国水稻研究所 | Primer group, kit and method for detecting pathogenic variation of avirulence gene AvrPi9 of magnaporthe grisea |
| CN112662804B (en) * | 2021-01-25 | 2022-05-10 | 中国水稻研究所 | Primer group, kit and method for detecting pathogenic variation of avirulence gene AvrPi9 of rice blast |
| CN114410731A (en) * | 2021-12-02 | 2022-04-29 | 吉林省农业科学院 | Grate screen for screening rice blast resistant materials and application thereof |
| CN114410731B (en) * | 2021-12-02 | 2024-02-27 | 吉林省农业科学院 | Grate screen for screening rice blast resistance material and application thereof |
| CN115852028A (en) * | 2022-11-01 | 2023-03-28 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Triple PCR primer group, kit and detection method for detecting avirulence gene of rice blast germs |
| CN115852028B (en) * | 2022-11-01 | 2024-02-09 | 安徽省农业科学院植物保护与农产品质量安全研究所 | Triple PCR primer group, kit and detection method for detecting rice blast fungus non-toxic genes |
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