Use of asf1 gene in improving plant disease resistance or breeding disease-resistant plants

Abstract

The application discloses a kind of ASF1 The application belongs to the field of genetic engineering. ASF1 The gene includes ASF1A The gene and ASF1B The gene, ASF1A The nucleotide sequence of the gene is shown as SEQ ID NO.1, ASF1B The nucleotide sequence of the gene is shown as SEQ ID NO.2. ASF1A The phenotype of single knockout strain, ASF1B The phenotype of single knockout strain and ASF1A , ASF1B Double knockout strain and the influence on the disease resistance of rice are studied, and the results show that, compared with wild type, the mutant plant is short, wherein the phenotype of double knockout mutant is more obvious than that of single knockout mutant; single knockout strain and double knockout strain can improve the resistance of rice to white leaf rot fungus and rice blast fungus, and the resistance of double knockout strain is stronger, which provides new genetic resources for the breeding research of rice disease resistance.

Description

Technical Field

[0001] This invention belongs to the field of genetic engineering technology, specifically relating to a... ASF1 The application of genes in improving plant disease resistance or cultivating disease-resistant plants. Background Technology

[0002] Plant diseases are one of the major biological stressors restricting global agricultural production, causing huge yield losses and economic damage every year. Traditional disease control mainly relies on chemical pesticides, but long-term use can easily lead to pathogen resistance, environmental pollution, and agricultural product safety issues. Therefore, exploring the plant's own disease-resistant genetic resources and cultivating new crop varieties with broad-spectrum and durable disease resistance through molecular breeding technology has become an important direction for sustainable agricultural development.

[0003] Currently, the main genetic improvement strategies for enhancing plant disease resistance include:

[0004] (1) Introduction and utilization of disease resistance genes: By cloning and transferring specific disease resistance genes, plants can acquire resistance to specific pathogens.

[0005] (2) Regulation of key genes in defense signaling pathways: Overexpression of positive regulators in defense hormone pathways or silencing of negative regulators can enhance the basic resistance of plants.

[0006] (3) Application of pattern recognition receptors: The introduction or overexpression of cell surface receptor kinases can enhance the ability to recognize conserved molecules of pathogens and improve basic resistance.

[0007] (4) Exploration of epigenetic regulatory factors: Recent studies have shown that the regulation of histone modifying enzymes (such as histone acetyltransferase and methyltransferase) can affect plant immune memory, such as systemic acquired resistance.

[0008] However, existing technology has not yet addressed this. ASF1 Reports on the use of genes to improve plant disease resistance. Summary of the Invention

[0009] To address the aforementioned shortcomings of the existing technology, the objective of this invention is to provide a... ASF1 The application of genes in improving plant disease resistance or cultivating disease-resistant plants will broaden the genetic resources for genetic improvement of plant disease resistance.

[0010] The technical solution of the present invention to solve the above-mentioned technical problems is as follows: A method is provided. ASF1 The application of genes in improving plant disease resistance or cultivating disease-resistant plants. ASF1 Genes include ASF1A Genes and ASF1B Gene, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0011] Based on the above technical solution, the present invention can be further improved as follows:

[0012] Furthermore, by knocking out ASF1A Genes and / or ASF1B Genes can be used to enhance plant disease resistance.

[0013] Furthermore, the plant is rice.

[0014] Furthermore, it exhibits resistance to bacterial blight and rice blast.

[0015] One method to improve plant disease resistance includes: knocking out [a specific substance] in the plant. ASF1A Genes and / or ASF1B Genes are used to enhance plant disease resistance; among them, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0016] Furthermore, the plant is rice.

[0017] Furthermore, it exhibits resistance to bacterial blight and rice blast.

[0018] A method for cultivating disease-resistant plants includes: knocking out [a specific disease-resistant component] in the plant. ASF1A Genes and / or ASF1B Genes, by reducing their expression levels, can be used to obtain disease-resistant plants; among them, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2.

[0019] Furthermore, the plant is rice.

[0020] Furthermore, it exhibits resistance to bacterial blight and rice blast.

[0021] The present invention has the following beneficial effects:

[0022] This invention uses the rice variety Zhonghua 11 (ZH11) as background material and obtains two homozygous rice genetic transformations using CRISPR / Cas9 technology and Agrobacterium-mediated transformation technology. ASF1A Single knockout strain osasf1a-1 , osasf1a-2 2 homozygous ASF1B Single knockout strain osasf1b-1 , osasf1b-2 and 2 homozygous ASF1A , ASF1BDouble knockout strains osasf1ab-1 , osasf1ab-2 Phenotypic analysis revealed that the mutant plants were shorter than the wild type, with the double knockout mutant showing a more pronounced phenotype than the single knockout mutant. Furthermore, the effects of single and double knockout lines on rice resistance to bacterial blight and rice blast were investigated. The results showed that both single and double knockout lines improved rice resistance to bacterial blight and rice blast fungus, with the double knockout lines exhibiting stronger resistance. This provides new genetic resources for rice disease resistance breeding research. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the OsASF1A mutation site in ZH11; where Exon represents an exon and Intron represents an intron.

[0024] Figure 2 for osasf1a-1 A diagram illustrating mutation types.

[0025] Figure 3 for osasf1a-2 A diagram illustrating mutation types.

[0026] Figure 4 This is a schematic diagram of the OsASF1B mutation site in ZH11; where Exon is an exon and Intron is an intron.

[0027] Figure 5 for osasf1b-1 A diagram illustrating mutation types.

[0028] Figure 6 for osasf1b-2 A diagram illustrating mutation types.

[0029] Figure 7 for osasf1ab-1 A diagram illustrating mutation types.

[0030] Figure 8 for osasf1ab-2 A diagram illustrating mutation types.

[0031] Figure 9 Phenotypes of wild-type ZH11 and mutant plants; from left to right, they are ZH11, asf1a-1, asf1b-1, and asf1ab-1.

[0032] Figure 10 for OsASF1A and OsASF1B Results of single and double knockout homozygous materials improving rice resistance to bacterial blight.

[0033] Figure 11 In the case of spray inoculation, OsASF1A and OsASF1B Single and double knockout homozygous materials improved rice resistance to rice blast fungus.

[0034] Figure 12 In the case of punch vaccination, OsASF1A and OsASF1B Single and double knockout homozygous materials improved rice resistance to rice blast fungus. Detailed Implementation

[0035] In this invention ASF1A The nucleotide sequence of a gene ASF1B The nucleotide sequence of a gene ASF1A The amino acid sequence of the gene-encoded protein, ASF1B The amino acid sequences of the protein encoded by the gene are shown below:

[0036] ASF1A The nucleotide sequence of the gene is shown below:

[0037] ATGAGCGTGATCGACATCCTGACGCGGGTGGACGCCATCTGCCAGAAGTACGGCAGGTACGACGCCGAGAAGCTCCACGGCTCCGGCGTCGCCGGCGAGGACCCCTTCGCGCGCCTCTACGCCTCGGTCGACGCTGACCTCAACGAATGCCTCGAGAAAGCGGAGGCGGCGAAGCAGGAGAAGAACCGCGCCACGGTGGTCGCGCTGAACGCGGAGATCCGCGGAACGAAGGCCAAGCTCCTCGAGGAGGATCTGCCCAAGCTGCAGCGGCTCGCCCTGAAGAAGGTTAAAGGGCTTTCAAAAGAAGAACTTGCAATCCGTGGTGATCTGGTTACTGCCTTACCTGATAGGATTCAGTCAATACCAGATGGAAGTGCTACATCATCAAAGAAAACTGGATTATGGGGATCTTCTGGATCTCGTGCAGGAACAGGGATTAAGTTTGACTCAACATATGATCTGGAGTGGAAGCTTATATATGTTGGATCGGCTGAGGATGAAAATTATGATCAGCTGCTTGAGAGTGTGCTTGTTGGCCCTGTGAATGTTGGGACATACCGTTTTGTTCTCCAGGCTGATCCTCCTGACCCGTCAAAGATCCGTAAGGAAGACATAATTGGTGTTATTGTGCTGCTATTAACTTGTTCCTACATGGGTCAAGAGTTCATCAGAGTAGGTTACTATGTGAACAATGACAACGATGATGAGCAGCTTCGGGAGGAGCCTCCTGCGAAGCTGCTGATTGACCGGGTGCAGAGGAACATATTGGCAGACAAGCCCAGCGTCACCAAGTTCCCCATCAATTTCCATCCTGAAACCAGTGCAGGCGCAGGACAAGAGCAGCAGCAGCAACAGCAATCTGGTTCACCTGAAAATCACCCAAACCAAGGTAGCAAGCCCAACCCTGATCAATGA (SEQ ID NO.1).

[0038] ASF1B The nucleotide sequence of the gene is shown below:

[0039] ATGAGCGCGGTGAACATAACCAACGTGGCGGTGCTGGACAACCCCACCGCCTTCCTCAATCCCTTCCAGTTCGAGATCTCCTACGAGTGCCTCATCCCCCTCGACGACGATCTGGAGTGGAAGCTTATATATGTTGGATCTGCTGAAGATGAAAATTATGACCAACAATTAGAGAGTGTGCTTGTTGGCCCCGTCAATGTTGGGACCTACCGTTTTGTACTCCAGGCTGACCCACCGGATCCCTCAAAGATCCGTGAAGAAGACATAATCGGCGTCACTGTGCTGCTATTGACATGTTCCTACATGGGACAGGAGTTCATGAGAGTAGGTTACTATGTGAACAATGATTATGACGATGAGCAACTGAGGGAGGAACCCCCGGCAAAGCTTCTAATAGACAGGGTGCAGAGGAACATTCTGGCTGACAAGCCCCGAGTCACCAAGTTCCCAATCAACTTCCATCCTGAACCCAGTACGAGCGCAGGGCAGCAGCAGCAGGAGCCACAGACAGCTTCGCCGGAAAACCACACAGGCGGTGAAGGAAGTAAGCCCGCTGCTGATCAATGA (SEQ ID NO.2).

[0040] ASF1A The amino acid sequence of the protein encoded by the gene is as follows:

[0041] MSVIDILTRVDAICQKYGRYDAEKLHGSGVAGEDPFARLYASVDADLNECLEKAEAAKQEKNRATVVALNAEIRGTKAKLLEEDLPKLQRLALKKVKGLSKEELAIRGDLVTALPDRIQSIPDGSATSSKKTGLWGSSGSRAGTGIKFDSTYDLEWKLIYVGSAEDENYDQLLESVLVGPVNVGTYRFVLQADPPDPSKIRKEDIIGVIVLLLTCSYMGQEFIRVGYYVNNDNDDEQLREEPPAKLLIDRVQRNILADKPSVTKFPINFHPETSAGAGQEQQQQQQSGSPENHPNQGSKPNPDQ (SEQ ID NO.3).

[0042] ASF1B The amino acid sequence of the protein encoded by the gene is shown below:

[0043] MSAVNITNVAVLDNPTAFLNPFQFEISYECLIPLDDDLEWKLIYVGSAEDENYDQQLESVLVGPVNVGTYRFVLQADPPDPSKIREEDIIGVTVLLLTCSYMGQEFMRVGYYVNNDYDDEQLREEPPAKLLIDRVQRNILADKPRVTKFPINFHPEPSTSAGQQQQEPQTASPENHTGGEGSKPAADQ (SEQ ID NO.4).

[0044] The examples given below are for illustrative purposes only and are not intended to limit the scope of the invention. Unless otherwise specified, conditions in the examples are performed under standard conditions or as recommended by the manufacturer. Reagents or instruments whose manufacturers are not specified are all commercially available products.

[0045] Example 1:

[0046] This embodiment mainly provides ASF1A , ASF1B The specific process for preparing single-knockout and double-knockout mutant materials is as follows:

[0047] I. Obtaining Rice ASF1 Gene sequence was obtained, plant knockout expression primer sequences were designed, knockout expression vectors were constructed, and genetic transformation was performed to obtain positive plants. The specific process is as follows:

[0048] Each ASF1A , ASF1BThe gene's accession number was entered into the CRISPR-GE website (http: / / skl.scau.edu.cn / home / ) for CRISPR / Cas9 knockout target prediction. Simultaneously, suitable target sequences were selected from the early coding region and conserved protein kinase regions as gRNA targets. These target sequences were then entered into the CRISPR RGEN Tools website (http: / / www.rgenome.net / cas-offinder / ) for off-target analysis, and the targets were introduced via PCR amplification. Finally, a pYLCRISPR / Cas9-mediated CRISPR / Cas9 knockout expression vector for this gene was constructed using a one-step cloning method. After successful sequencing, the recombinant plasmid was transformed into Agrobacterium tumefaciens (EHA105) competent cells using a freeze-thaw method. Transgenic material was obtained through Agrobacterium-mediated rice genetic transformation using the rice variety Zhonghua 11 (ZH11) as background material. DNA was extracted from the obtained T0 generation transgenic plants, and the CRISPR / Cas9 vector-specific primers were used to detect whether they were positive plants (the constructed knockout expression vector was sent to Boyuan Biotechnology Co., Ltd. for genetic transformation, and positive plants were obtained).

[0049] The gRNAs and primer sequences used in constructing the knockout expression vector are shown below:

[0050] ASF1A-sgRNA1: CGCGGAGATCCGCGGAACGAAGG (SEQ ID NO.5);

[0051] ASF1A-sgRNA2:AAAGAAGAACTTGCAATCCGTGG (SEQ ID NO.6);

[0052] ASF1B-sgRNA1:TCTTCTTCACGGATCTTTGAGGG (SEQ ID NO.7);

[0053] ASF1B-sgRNA2: CGGCAAAGCTTCTAATAGACAGG (SEQ ID NO. 8).

[0054] Primer sequences used to amplify ASF1AU6c and U3gRNA:

[0055] ASF1AU6c-F: 5'-tcagCGCGGAGATCCGCGGAACGA-3' (SEQ ID NO.9);

[0056] ASF1AU6c-R: 5'-aaacTCGTTCCGCGGATCTCCGCG-3' (SEQ ID NO. 10);

[0057] ASF1AU3-F: 5'-ggcaAAGAAGAACTTGCAATCCG-3' (SEQ ID NO. 11);

[0058] ASF1AU3-R: 5'-aaacCGGATTGCAAGTTCTTCTT-3' (SEQ ID NO. 12).

[0059] Primer sequences used to amplify ASF1BU6a and U3gRNA:

[0060] ASF1BU6a-F: 5'-gccgTCTTCTTCACGGATCTTTGA-3' (SEQ ID NO. 13);

[0061] ASF1BU6a-R: 5'-aaacTCAAAGATCCGTGAAGAAGA-3' (SEQ ID NO. 14);

[0062] ASF1BU6b-F: 5'-gttgCGGCAAAGCTTCTAATAGAC-3' (SEQ ID NO. 15);

[0063] ASF1BU6b-R: 5'-aaacGTCTATTAGAAGCTTTGCCG-3' (SEQ ID NO. 16).

[0064] The specific primer sequences for detecting positive plants are shown below:

[0065] ASF1A-cas1-F: 5'-TGATTGCCTCCCTTCCTCTC-3' (SEQ ID NO. 17);

[0066] ASF1A-cas1-R: 5'-ATGCCATATGCTAAATTTAG-3' (SEQ ID NO. 18);

[0067] ASF1A-cas2-F: 5'-ACTTTCCTCGGCATAACAA-3' (SEQ ID NO. 19);

[0068] ASF1A-cas2-R: 5'-TCAAACTTAATCCCTGTTC-3' (SEQ ID NO. 20);

[0069] ASF1B-cas1-F: 5'-ACTCAGATTACAGAACTAT-3' (SEQ ID NO. 21);

[0070] ASF1B-cas1-R: 5'-ACTGGGTTCAGGATGGAAG-3' (SEQ ID NO. 22);

[0071] ASF1B-cas2-F: 5'-GTTGGGACCTACCGTTTTG-3' (SEQ ID NO. 23);

[0072] ASF1B-cas2-R: 5'-GCATCAAAATGTTAGCATC-3' (SEQ ID NO. 24).

[0073] 2. DNA was extracted from positive plants using the CTAB method. Primers (SEQ ID NO. 13-20) were designed 100-200 bp before and after the PAM sequence of the genome for PCR amplification, followed by third-generation sequencing. The mutation type was analyzed based on the sequencing results. The specific process is as follows:

[0074] Positive plants were planted in the experimental field. T1 generation seedlings were harvested individually, and DNA was extracted from leaves. [The text then abruptly shifts to a different topic:] ...and then... ASF1A , ASF1B Primers were designed near two and four target sites in single and double knockout mutants to detect the knockout status. Two homozygous primers were obtained for each knockout mutant. ASF1A The single knockout line OsASF1A is denoted as osasf1a-1 , osasf1a-2 (See details) Figure 1-3 ), 2 homozygous ASF1B The single knockout line OsASF1B is denoted as osasf1b-1 , osasf1b-2 (See details) Figure 4-6 ), 2 homozygous ASF1A , ASF1B The double knockout strains OsASF1A / OsASF1B are denoted as osasf1ab-1 , osasf1ab-2 (See details) Figure 7-8 ).

[0075] Figure 2 middle osasf1a-1 The mutation is based on Figure 1 Based on the PAM sequence 1, 99 bases are deleted after its 17th base; Figure 3 middle osasf1a-2 The mutation is based on Figure 1 Based on the PAM sequence 1, an A base is inserted after its 17th base.

[0076] Figure 5 middle osasf1b-1 The mutation is based on Figure 4 Based on the PAM sequence 1, replace the 16th base G with A; Figure 6 middle osasf1b-2 The mutation is based on Figure 4 Based on the PAM sequence 2, the 18th-20th bases are deleted.

[0077] Figure 7 middle osasf1ab-1 The mutation is to delete 99 bases after the 17th base in the PAM sequence 1 of OsASF1A, and to replace the 16th base G in the PAM sequence 1 of OsASF1B with A. Figure 8 middle osasf1ab-2 The mutation involves inserting an A base after the 17th base in the PAM sequence 1 of OsASF1A, and deleting the 18th-20th bases in the PAM sequence 2 of OsASF1B.

[0078] III. The knockout strains were grown in the field, and the plant phenotypes were observed and photographed. The results are as follows: Figure 9 As shown. By Figure 9 It can be seen that, compared with the wild type, the mutant plants are shorter, and the phenotype of the double knockout mutant is more obvious than that of the single knockout mutant.

[0079] Example 2:

[0080] This embodiment mainly studies OsASF1A , OsASF1B The negative regulation of rice resistance to bacterial blight by genes occurs in the following process:

[0081] Bacterium oxysporum Xoo. PX099A (physiological race PX099A) was cultured on YEB solid medium at 28°C for 2-3 days, the bacterial cells were washed off with sterile water, and the OD of the *Rhizoctonia solani* suspension was adjusted. 600 To achieve a pH of 0.8-1.0, use sterile scissors to pick up the Bacillus subtilis suspension and then apply it to wild-type rice (WT). osasf1a-1 , osasf1a-2 , osasf1b-1 , osasf1b-2 , osasf1ab-1 and osasf1ab-2 Cut the material from the same location on a fully unfolded leaf and inoculate it with bacterial blight pathogen. Survey and statistics were conducted 14 days later, and the results were as follows: Figure 10 As shown.

[0082] Depend on Figure 10 It can be seen that, ASF1A and ASF1B Single gene knockout ( osasf1a-1 , osasf1a-2 , osasf1b-1 , osasf1b-2 ) and double knockout homozygous materials ( osasf1ab-1 , osasf1ab-2 This improved the resistance of rice to bacterial blight pathogen, and the double knockout... osasf1ab-1 , osasf1ab-2 It has stronger resistance to bacterial blight.

[0083] The composition of the YEB medium is as follows: 5g sucrose, 10g tryptone, 5g yeast extract, 0.5g MgSO4·7H2O, 2% agar, pH 7.2-7.4, and sterile water to a final volume of 1L.

[0084] Example 3:

[0085] This embodiment mainly studies OsASF1A , OsASF1B The negative regulation of rice resistance to rice blast fungus by genes occurs in the following process:

[0086] Inoculate the filter paper containing the rice blast fungus strain "Guy11" into the center of a starch yeast culture medium plate and incubate it upside down in a 26℃ biochemical incubator for 5-6 days. Pick freshly grown mycelial blocks and inoculate them onto rice bran culture medium plates, incubate them at a constant temperature of 26℃ for 7-8 days. After the mycelium has covered the culture dish, scrape off the aerial mycelium with a sterile glass slide, and then place the plate in a 25℃ constant temperature and light incubator for 2-3 days of conidial production. Then, collect the conidia and prepare a spore suspension for subsequent inoculation.

[0087] The first step is spray inoculation, specifically: when the rice seedlings reach the three-leaf stage, use an air compressor connected to a spray nozzle for spray inoculation. Prepare a suspension of rice blast fungus conidia using a 0.02% Tween 20 solution, with a final conidia concentration of 1.5 × 10⁻⁶. 5 The inoculation rate was [number] cells / mL. After inoculation, the plants were placed in a constant temperature and humidity greenhouse at 22-23℃ in the dark for 24 hours, then removed and placed in a constant temperature greenhouse with humidity above 80% for further cultivation. Five to seven days after inoculation, the disease incidence in rice was investigated using a visual inspection method (referencing the standards of the International Rice Research Institute (IRRI)). The type of lesion reaction on each rice leaf was investigated and recorded. If different lesion reaction types appeared on the same plant's leaves, the highest reaction type was used as the standard. The results are as follows: Figure 11 As shown. By Figure 11 It can be seen that single knockout ( osasf1a-1 , osasf1a-2 , osasf1b-1 , osasf1b-2 ) and double knockout homozygous materials ( osasf1ab-1 , osasf1ab-2 Both can improve resistance to rice blast fungus, and double knockout... osasf1ab-1 , osasf1ab-2 It has stronger resistance to rice blast fungus.

[0088] The second method is inoculation by punching holes. Specifically, when the rice seedlings have grown to a leaf width of 1-2 cm, use a 2mm punch head on a Deli belt puncher to punch holes in the leaves, avoiding the veins. Use a 10μL pipette to take 6μL of a Tween 20 solution with a final concentration of 0.02% to prepare a suspension of rice blast fungus conidia. The final spore concentration is 2×10⁻⁶. 4 The number of spores / mL was determined by securing the spore leaves to the wound with transparent adhesive. After inoculation, the plants were placed in a constant temperature and humidity greenhouse at 22-23℃ for 24 hours in darkness, then removed and placed in a constant temperature greenhouse with humidity above 80% for further cultivation. Ten days after inoculation, the disease incidence in rice was investigated using a visual inspection method (referencing the standards of the International Rice Research Institute (IRRI)). The lesion reaction type on each rice leaf was investigated and recorded. If different lesion reaction types appeared on the same plant's leaves, the highest reaction type was used as the standard. The results are as follows: Figure 12 As shown. By Figure 12 It can be seen that single knockout ( osasf1a-1 , osasf1a-2 , osasf1b-1 , osasf1b-2 ) and double knockout homozygous materials ( osasf1ab-1 , osasf1ab-2 Both can improve resistance to rice blast fungus, and double knockout... osasf1ab-1 , osasf1ab-2 It has stronger resistance to rice blast fungus.

[0089] The starch-yeast culture medium used above consists of: 10g soluble starch, 2g yeast extract, 18g agar, pH 6.0-6.5, and sterile water to a final volume of 1L.

[0090] Composition of rice bran culture medium: 20g rice bran, 2.5g yeast extract, 18g agar, pH value of 6.0-6.5, and sterile water to a final volume of 1L.

[0091] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. ASF1 The application of genes in improving plant disease resistance or cultivating disease-resistant plants, the aforementioned ASF1 Genes are ASF1A Genes and ASF1B Gene, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.

1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2; the application is by knocking out... ASF1A Genes and / or ASF1B Genes are used to enhance the disease resistance of plants, the plant being rice, and the disease resistance being resistance to bacterial blight and rice blast.

2. A method for improving plant disease resistance, characterized in that, By knocking out the plant ASF1A Genes and / or ASF1B Genes are used to enhance plant disease resistance; among them, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.

1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2; the plant is rice, and the disease resistance is resistance to bacterial blight and rice blast.

3. A method for cultivating disease-resistant plants, characterized in that, By knocking out the plant ASF1A Genes and / or ASF1B Genes, by reducing their expression levels, can be used to obtain disease-resistant plants; among them, ASF1A The nucleotide sequence of the gene is shown in SEQ ID NO.

1. ASF1B The nucleotide sequence of the gene is shown in SEQ ID NO.2; the plant is rice, and the disease resistance is resistance to bacterial blight and rice blast.

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