Rice transcription factor OsbHLH091 and application of coding gene thereof in disease-resistant breeding

Abstract

The invention discloses an application of a rice transcription factor OsbHLH091 and a coding gene thereof in disease-resistant breeding, and belongs to the field of gene engineering. The amino acid sequence of the rice transcription factor OsbHLH091 is as shown in SEQ ID NO. 2, and the coding gene is as shown in SEQ ID NO. 1. Experiments prove that the coding gene of the rice transcription factor OsbHLH091 is over-expressed in rice, and a transgenic rice plant has higher disease resistance than a receptor rice plant and a rice plant with OsbHLH091 knocked out, which indicates that the OsbHLH091 positively regulates the resistance of the rice to the rice blast. The OsbHLH091 protein and the coding gene thereof found by the invention have important significance on breeding disease-resistant rice varieties, and can be applied to breeding rice with improved disease resistance by molecular design.

Description

Technical Field

[0001] The invention relates to the field of genetic engineering, in particular to the application of rice transcription factor OsbHLH091 and its encoding gene in disease-resistant breeding. Background Art

[0002] Fungal diseases such as rice blast cause large-scale crop yield reductions, so breeding disease-resistant crops is one of the main goals of the planting industry. Many genes related to plant disease resistance have been reported, including effector molecule genes and regulatory genes, which come from crops such as rice, wheat, corn, soybeans, and model plants such as Arabidopsis. Some of them have been used as target genes in crop disease resistance genetic engineering, and disease-resistant rice, wheat, corn, soybeans, etc. have been successfully bred. However, due to differences in genetic background, some genes have been blocked in application, so new related genes are still needed to meet the needs of breeding.

[0003] As one of the most important food crops, improving rice's disease resistance has important theoretical and practical significance. Therefore, seeking new rice disease resistance genes is still the key to current disease resistance breeding. Summary of the invention

[0004] The purpose of the present invention is to provide an application of rice transcription factor OsbHLH091 and its encoding gene in disease resistance breeding to solve the problems existing in the above-mentioned prior art. OsbHLH091 positively regulates the resistance of rice to rice blast.

[0005] To achieve the above object, the present invention provides the following solutions:

[0006] The present invention provides an application of rice transcription factor OsbHLH091 in any of the following:

[0007] (1) Application in controlling rice blast disease;

[0008] (2) Application in the preparation of products for regulating rice blast;

[0009] (3) Application in rice breeding for resistance to rice blast;

[0010] (4) Application in breeding rice resistant to rice blast;

[0011] The amino acid sequence of the rice transcription factor OsbHLH091 is shown in SEQ ID NO. 2. However, the amino acid sequence of the rice transcription factor OsbHLH091 of the present invention is not limited thereto, and may also be a protein derived from the sequence protein and having the same function obtained by replacing and / or deleting and / or adding one or more amino acid residues of the amino acid sequence shown in SEQ ID NO. 2, or a protein having more than 80% identity with the sequence protein and having the same function; or may be a fusion protein obtained by connecting a protein tag to the N-terminus or / and C-terminus of the above protein or derived protein.

[0012] In the above-mentioned proteins or derived proteins, a protein tag refers to a polypeptide or protein fused and expressed together with a target protein using DNA in vitro recombination technology to facilitate the expression, detection, tracing and / or purification of the target protein. The protein tag can be a GFP tag, a Flag tag, a His tag, a MBP tag, a HA tag, a myc tag, a GST tag and / or a SUMO tag, etc.

[0013] In the above-mentioned protein or derived protein, identity refers to the identity of the amino acid sequence. The identity of the amino acid sequence can be determined using a homology search site on the Internet, such as the BLAST page on the NCBI homepage website. For example, in Advanced BLAST2.1, by using blastp as a program, setting the Expect value to 10, setting all Filters to OFF, using BLOSUM62 as a Matrix, setting the Gap existence cost, Per residue gap cost and Lambda ratio to 11, 1 and 0.85 (default values) respectively, and searching for the identity of a pair of amino acid sequences for calculation, the identity value (%) can be obtained.

[0014] In the above proteins or derived proteins, the above 80% identity may be at least 81%, 82%, 85%, 86%, 88%, 90%, 91%, 92%, 95%, 96%, 98%, 99% or 100% identity.

[0015] The present invention also provides the use of the coding gene of rice transcription factor OsbHLH091 in any of the following items:

[0016] (1) Application in controlling rice blast disease;

[0017] (2) Application in the preparation of products for regulating rice blast;

[0018] (3) Application in rice breeding for resistance to rice blast;

[0019] (4) Application in breeding rice resistant to rice blast;

[0020] The nucleotide sequence of the coding gene is shown in SEQ ID NO. 1. The nucleotide sequence of the coding gene is a cDNA or DNA molecule sequence.

[0021] The present invention also provides the use of a recombinant vector containing a gene encoding a rice transcription factor OsbHLH091 in any of the following:

[0022] (1) Application in controlling rice blast disease;

[0023] (2) Application in the preparation of products for regulating rice blast;

[0024] (3) Application in rice breeding for resistance to rice blast;

[0025] (4) Application in breeding rice resistant to rice blast;

[0026] The amino acid sequence of the rice transcription factor OsbHLH091 is shown in SEQ ID NO.2, and the nucleotide sequence of the encoding gene is shown in SEQ ID NO.1.

[0027] The present invention also provides the use of a host bacterium containing a recombinant vector in any of the following:

[0028] (1) Application in controlling rice blast disease;

[0029] (2) Application in the preparation of products for regulating rice blast;

[0030] (3) Application in rice breeding for resistance to rice blast;

[0031] (4) Application in breeding rice resistant to rice blast;

[0032] The recombinant vector is a vector in which the coding gene of the rice transcription factor OsbHLH091 is integrated into the genome, and the nucleotide sequence of the coding gene is shown in SEQ ID NO.1.

[0033] The carrier of the present invention is a plant expression vector, which includes a binary Agrobacterium vector and a vector that can be used for plant microprojectile bombardment, etc. Such as pAHC25, pWMB123, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa or pCAMBIA1391-Xb (CAMBIA company), etc. The plant expression vector can also include the 3' non-translated region of the foreign gene, i.e., include polyadenylic acid signal and any other DNA fragments that participate in mRNA processing or gene expression. The polyadenylic acid signal can guide polyadenylic acid to be added to the 3' end of the mRNA precursor, such as Agrobacterium crown gall induction (Ti) plasmid gene (such as nopaline synthase gene Nos), plant gene (such as rice starch synthase gene) 3' end transcription non-translated region all have similar functions. When using the gene of the present invention to construct a plant expression vector, enhancers, including translation enhancers or transcription enhancers, can also be used. These enhancer regions can be ATG start codons or adjacent region start codons, etc., but must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence.

[0034] The host bacteria of the present invention include yeast, bacteria, algae and fungi.

[0035] Optionally, the method of regulating rice blast disease is that rice transcription factor OsbHLH091 positively regulates rice blast disease.

[0036] The present invention also provides a method for regulating rice blast disease, comprising any of the following methods:

[0037] (1) reducing the resistance of rice to rice blast by knocking out the OsbHLH091 gene in rice;

[0038] (2) improving the resistance of rice to rice blast by overexpressing the OsbHLH091 gene in rice;

[0039] Wherein, the nucleotide sequence of the OsbHLH091 gene is shown as SEQ ID NO.1.

[0040] The present invention also provides a method for cultivating rice blast-resistant rice, comprising the steps of overexpressing the OsbHLH091 gene in rice, increasing the expression level of the OsbHLH091, and obtaining rice blast-resistant rice; wherein the nucleotide sequence of the OsbHLH091 gene is shown in SEQ ID NO.1.

[0041] The present invention discloses the following technical effects:

[0042] The inventors of the present invention identified a member of the bHLH transcription factor family in the study of rice immune response and named it OsbHLH091. Experiments have shown that when OsbHLH091 is overexpressed in rice, the transgenic rice exhibits higher disease resistance than the recipient rice; when OsbHLH091 is knocked out in rice, the transgenic rice exhibits lower disease resistance than the recipient rice, indicating that overexpression of OsbHLH091 has the function of improving rice disease resistance. The present invention confirms that OsbHLH091 can positively regulate rice disease resistance, and the OsbHLH091 protein and its encoding gene are of great significance for breeding disease-resistant rice varieties. BRIEF DESCRIPTION OF THE DRAWINGS

[0043] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0044] Figure 1 Schematic diagram of the gene structure of OsbHLH091 and the sgRNA designed at the knockout target site;

[0045] Figure 2 The results of gene sequence identification of OsbHLH091 knockout plants;

[0046] Figure 3 Schematic diagram of the overexpression vector structure of OsbHLH091;

[0047] Figure 4 The results of the detection of OsbHLH091 gene expression in OsbHLH091 overexpressing plants;

[0048] Figure 5Figure 2 is a diagram of the phenotypic changes of wild-type rice Z11 and OsbHLH091 knockout and overexpression plants after inoculation with rice blast; A is a representative picture of lesions on the leaves of Zhonghua11 (ZH11), knockout plants OsbHLH091#KOs, and overexpression plants OsbHLH091#OEs after inoculation with rice blast; B is a representative picture of lesions on the leaves of Zhonghua11 (ZH11), knockout plants OsbHLH091#KOs, and overexpression plants OsbHLH091#OEs after inoculation with rice blast. Statistical results of spot length; C is a representative picture of lesions on the leaves of Zhonghua11 (ZH11), knockout plant OsbHLH091#KOs, and overexpression plant OsbHLH091#OEs after spray inoculation with rice blast fungus; D is the statistical result of the number of lesions on the leaves of Zhonghua11 (ZH11), knockout plant OsbHLH091#KOs, and overexpression plant OsbHLH091#OEs after puncture inoculation with rice blast fungus; Asterisks above the bar graphs indicate significant differences (P<0.05). DETAILED DESCRIPTION

[0049] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0050] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0051] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0052] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0053] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0054] In the study of rice immune response, the applicant identified a member of the bHLH transcription factor family and named it OsbHLH091. bHLH transcription factors play an important role in plant development and biological stress, so previous studies on OsbHLH091 focused on its regulation of plant development. The applicant's research found that compared with the control, the lesions of OsbHLH091 knockout plants became longer, and the lesions of OsbHLH091 overexpressing plants became shorter and fewer, indicating that OsbHLH091 positively regulates the disease resistance of plants and can be used as a target gene for molecular breeding to improve plant disease resistance. The disease resistance function of OsbHLH091 is specifically described in the following examples.

[0055] The experimental methods in the examples of the present invention are conventional methods unless otherwise specified, and are performed according to the techniques or conditions described in the literature in the field or according to the product instructions. The materials, reagents, etc. used in the examples of the present invention are commercially available unless otherwise specified.

[0056] Some experimental materials and reagents involved in the following examples:

[0057] (1) The rice variety Zhonghua 11 (O. Sativa L. spp. japonica, var. zhonghua11, AA genome, ZH11) belongs to the japonica subspecies, as recorded in the following literature: Ni Yuchong, A New Rice Variety Cultivated by Flower Culture—Zhonghua 11, Agricultural Science and Technology Newsletter, 1989, 07, 35; provided by Professor Chen Xuewei, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences.

[0058] (2) The OsbHLH091 overexpression and knockout transgenic rice of the present invention was produced by Boyuan Biotechnology Company.

[0059] (3) Plant binary expression vector pTCRISPR, provided by Associate Professor Tang Yongyan from the State Key Laboratory of Sichuan Agricultural University. Plant binary expression vector pCAMBIA2300, provided by Professor Wu Xianjun from the Rice Research Institute of Sichuan Agricultural University.

[0060] (4) Total RNA extraction kit: TRIzol purchased from Invitrogen, USA, catalog number 15596026.

[0061] Reverse transcription kit: HiScript III RT SuperMix for qPCR (+gDNA wiper) purchased from Vanzyme, China, with the catalog number R323-01.

[0062] Fluorescence quantitative kit: AceQ qPCR SYBR Green MasterMix purchased from Vanzyme, China, with the catalog number Q111-02.

[0063] Homologous recombination kit: ClonExpress II One Step Cloning Kit purchased from Vanzyme, China, with the catalog number C112-01.

[0064] Example 1 Gene OsbHLH091 Induced in Rice Immune Response and Its Cloning

[0065] In the process of studying the immune response of rice, it was found that the expression of OsbHLH091 was induced by rice blast fungus. The expression of OsbHLH091 at different time points after rice blast infection was verified by polymerase chain reaction. The specific method is as follows:

[0066] ZH11 was used as the research material. The experimental group was sprayed with 5×10 5 ml -1 Zhong10-8-14 rice blast fungus spores and 0.1% Tween 20 solution, control group: spray 0.1% Tween 20 solution, and take rice leaves at 12 hours. Sent to Novogene for Ribosme profiling sequencing, and after analysis, it was determined that the translation of the gene was induced by rice blast fungus. After analysis and comparison, the gene encodes a bHLH transcription factor. According to the reference gene sequence of the rice genome, primers were designed:

[0067] OsbHLH091F (SEQ ID NO.3): 5'-ATGGACGGCGGCAGGGTCGG-3';

[0068] OsbHLH091R (SEQ ID NO. 4): 5'-GAACTCATTTTCATGTGGCCT-3'.

[0069] Using ZH11 total cDNA as a template, a DNA band of about 1.2Kb was amplified by the above primers. After sequencing, it was confirmed to be the OsbHLH091 gene belonging to the bHLH family. After alignment of the LOC_Os08g41320 sequence in the rice genome reference sequence and the clone sequence amplified from the ZH11 total cDNA, it was shown that the clone sequence obtained from ZH11 was identical to the LOC_Os08g41320 sequence. The amino acid sequence of the OsbHLH091 protein is shown in SEQ ID NO.2. The CDS sequence of the coding chain of the gene OsbHLH091 encoding the OsbHLH091 protein is shown in SEQ ID NO.1 (see Figure 1).

[0070] Example 2 Construction of OsbHLH091 plant knockout vector

[0071] 1. OsbHLH091 knockout vector pTCRISPR-sgRNA OsbHLH091 Construction

[0072] Using the genome of rice ZH11 as a template, the knockout target sequence was designed as: 5'-AGAATTGTACCCTCTCAGCG-3'. Figure 1 .

[0073] The following primers were synthesized:

[0074] F (SEQ ID NO.5): 5'-TGTG AGAATTGTACCCTCTCAGCG G-3';

[0075] R (SEQ ID NO.6): 5'-AAAAC CGCTGAGAGGGTACAATTCT -3'.

[0076] After denaturation at 95°C, anneal to form a double strand. Digest the pTCRISPR vector with BsaI to recover the linearized vector. Mix the above DNA double strands and the linearized vector according to the following system, perform T4 DNA ligase ligation reaction and incubate at room temperature for 30 minutes.

[0077] Table 1 Connection system

[0078] Reagents Dosage DNA double strand fragment 0.5-4μL Linearized vector 0.5-4μL T4ligase 2μL <![CDATA[ddH2O]]> XμL Total volume 10μL

[0079] The same effect can be achieved within the range shown in Table 1 above. This experiment specifically uses a 10 μL ligation system: 0.5 μL DNA double-stranded fragment, 0.5 μL linearized vector, 2 μL T4 ligase, and ddH2O supplemented to 10 μL. According to the ligation system reaction, pTCRISPR-sgRNA was obtained OsbHLH091 Knockout vector.

[0080] 2. Add the above pTCRISPR-sgRNA OsbHLH091 The knockout vector was transformed into DH5α competent medium. After kanamycin selection, single clones were picked for colony PCR identification. Plasmids were extracted from positive colonies and sequenced to confirm that pTCRISPR-sgRNA was correct. OsbHLH091 Correct, genetic transformation can be carried out.

[0081] 3. OsbHLH091 knockout transgenic rice was completed by Boyuan Biotechnology Company. Genetically stable knockout plants OsbHLH091#KO1 and OsbHLH091#KO2 were sequenced and screened. The results showed that there were insertions or deletions of bases in the target sequence of the knockout plants, see Figure 2 .

[0082] Example 3 Construction of OsbHLH091 plant expression vector

[0083] 1. Construction of OsbHLH091 overexpression vector pCAMBIA2300-OsbHLH091

[0084] The cDNA obtained by reverse transcription of total RNA of rice ZH11 was used as template, and the cloning primers were:

[0085] F (SEQ ID NO.7): 5'- GGACAGGGTACCCGGGGATCC ATGGACGGCGGCAGGGTCG G-3′;

[0086] R (SEQ ID NO.8): 5'- GGTACTAGTGTCGACTCTAGA GAACTCCATTTTTCATGTGGC CT-3'.

[0087] The full-length cDNA of OsbHLH091 was obtained by PCR amplification, and the pCAMBIA2300-35S-eGFP-OCS vector was double-digested with BamHI and XbaI to recover the linearized vector. The full-length cDNA of OsbHLH091 recovered by PCR and the linearized vector were mixed according to the following system for homologous recombination.

[0088] Table 2 Homologous recombination reaction system

[0089] Reagents Dosage DNA double strand fragment 0.5-5μL Linearized vector 0.5-5μL 5×CEIIBuffer 4μL ExnaseII 2μL <![CDATA[ddH2O]]> XμL Total volume 20μL

[0090] The same effect can be achieved within the range shown in Table 2. This experiment specifically uses a 20 μL homologous recombination reaction system: 1 μL DNA double-stranded fragment, 1 μL linearized vector, 4 μL 5×CE II Buffer, 2 μL Exnase II, and ddH2O is added to 20 μL. Recombination was performed according to the homologous recombination reaction system to obtain the pCAMBIA2300-OsbHLH091 recombinant vector, see Figure 3 .

[0091] 2. The above-mentioned pCAMBIA2300-OsbHLH091 recombinant vector was transformed into DH5α competent cells. After kanamycin screening, single clones were picked for colony PCR identification. After the plasmids were extracted from the positive colonies and sequenced correctly, it was confirmed that pCAMBIA2300-OsbHLH091 was correct and genetic transformation could be carried out.

[0092] 3. OsbHLH091 overexpression transgenic rice was completed by Boyuan Biotechnology Company. The expression of the target gene was detected and genetically stable overexpression plants OsbHLH091#OE1 and OsbHLH091#OE2 were screened. The results showed that the expression of the target gene in the overexpression plants was significantly increased compared with the expression of the wild-type target gene, see Figure 4 .

[0093] Example 4 OsbHLH091 is involved in regulating plant disease resistance

[0094] In plants, research on OsbHLH091 mainly focuses on development and abiotic stress regulation. The applicant first discovered that the bHLHs protein OsbHLH091 is involved in regulating jasmonic acid synthesis in rice and is a regulatory factor in the jasmonic acid pathway. Jasmonic acid is known as a defense hormone and plays an important role in regulating plant disease resistance. Therefore, the effect of OsbHLH091 on rice disease resistance was tested.

[0095] Treatment of seedlings during inoculation:

[0096] (1) Test materials

[0097] The three-week-old Zhonghua 11 (ZH11), the genetically stable OsbHLH091 knockout lines OsbHLH091#KO1 and OsbHLH091#KO2 with ZH11 background, and the genetically stable OsbHLH091 overexpression lines OsbHLH091#OE1 and OsbHLH091#OE2 with ZH11 background.

[0098] (2) Experimental methods

[0099] Select seeds with full grains, place them in a conical bottle filled with tap water, and place them in a dark incubator at 37°C for germination, changing the water every day. After 2 days, select white seeds and place them in a 96-well seedling plate, place the 96-well seedling plate on a float, and grow them in Hoagland nutrient solution. After 21 days, select the second-to-last rice leaf with the same growth and size, pierce it and inoculate 5 μL of 5×10 5 ml -1 The spores of Zhong10-8-14 rice blast fungus were sprayed on the leaves of rice leaves with a concentration of 5×10 5 ml -1The 10-8-14 rice blast fungus spores were used and the number of lesions was counted after 5 days.

[0100] The results showed that there was no significant difference in the growth of ZH11, OsbHLH091#KO and OsbHLH091#OE under hydroponic conditions. The experiment was repeated three times. Each time, 20 plants were planted. Statistics showed that after puncture inoculation, the length and number of lesions of OsbHLH091#KO plants under the ZH11 background were significantly increased compared with ZH11, and they were more sensitive to pathogens (see Figure 5 ), while overexpression of OsbHLH091 in ZH11 significantly reduced the length and number of lesions, and significantly increased the disease resistance of the plants (see Figure 5 ). This indicates that OsbHLH091 positively regulates the disease resistance of plants, and its encoding gene OsbHLH091 can be used as a target gene for molecular breeding to improve plant disease resistance.

[0101] SEQ ID NO.1 is as follows:

[0102]

[0103] SEQ ID NO.2 is as follows:

[0104] MDGGRVGGDYISSLLSSSSPRLDFGVPVLDAIVAPGGGGGGGDCGLDKLCGDPGFAERAARLSSFNNGGGGVGQRYGGAGAGLFGMPPPAPGDFAGGGSREASVSDPASSAMKDAAAANAKKRKSTAAAAAAAKGKGKEPPVGEEKESDGKRCKTGNGEKESSVKPKAEQAGSDSSVEDGGGGGQKQGKGKNAKPVEPPKDYVH VRARRGQATDSHSLAERVRRERISQRMKVLQDLVPGCNKVIGKALMLDEIINYVQSLQRQVEFLSMKLATVNPLDFSNLPTLLQKDMFQACGPSASSVFSL ESSNSAFRFAEQGDVFQQFAQNSMESQCTLNQLDLALSQATNAAQYAFQDGTAGANLQQRNFWEDDLQSVFHIENGQSQENGVSAPNFHGQQQAGHMKMEF.

[0105] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. Application of rice transcription factor OsbHLH091 in any of the following: (1) Application in controlling rice blast disease; (2) Application in the preparation of products for regulating rice blast; (3) Application in rice breeding for resistance to rice blast; (4) Application in breeding rice resistant to rice blast; in, The amino acid sequence of the rice transcription factor OsbHLH091 is shown in SEQ ID NO.

2.

2. Use of the gene encoding rice transcription factor OsbHLH091 in any of the following: (1) Application in controlling rice blast disease; (2) Application in the preparation of products for regulating rice blast; (3) Application in rice breeding for resistance to rice blast; (4) Application in breeding rice resistant to rice blast; in, The nucleotide sequence of the coding gene is shown in SEQ ID NO.

1.

3. Use of a recombinant vector containing a gene encoding rice transcription factor OsbHLH091 in any of the following: (1) Application in controlling rice blast disease; (2) Application in the preparation of products for regulating rice blast; (3) Application in rice breeding for resistance to rice blast; (4) Application in breeding rice resistant to rice blast; in, The amino acid sequence of the rice transcription factor OsbHLH091 is shown in SEQ ID NO.2, and the nucleotide sequence of the encoding gene is shown in SEQ ID NO.

1.

4. Use of host bacteria containing recombinant vectors in any of the following: (1) Application in controlling rice blast disease; (2) Application in the preparation of products for regulating rice blast; (3) Application in rice breeding for resistance to rice blast; (4) Application in breeding rice resistant to rice blast; in, The recombinant vector is a vector in which the coding gene of the rice transcription factor OsbHLH091 is integrated into the genome, and the nucleotide sequence of the coding gene is shown in SEQ ID NO.

1.

5. The use according to any one of claims 1 to 4, characterized in that: The method for regulating rice blast disease is that rice transcription factor OsbHLH091 positively regulates rice blast disease.

6. A method for regulating rice blast disease, characterized in that: This includes any of the following methods: (1) reducing the resistance of rice to rice blast by knocking out the OsbHLH091 gene in rice; (2) improving the resistance of rice to rice blast by overexpressing the OsbHLH091 gene in rice; Wherein, the nucleotide sequence of the OsbHLH091 gene is shown as SEQ ID NO.

1.

7. A method for cultivating rice resistant to rice blast, characterized in that: The method comprises the steps of overexpressing the OsbHLH091 gene in rice, increasing the expression level of the OsbHLH091, and obtaining rice resistant to rice blast; wherein the nucleotide sequence of the OsbHLH091 gene is shown in SEQ ID NO.1.

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