Rice bHLH transcription factor OsbHLH024 and application of coding gene of rice bHLH transcription factor OsbHLH024 in disease-resistant breeding

Abstract

The invention discloses an application of a rice bHLH transcription factor OsbHLH024 and a coding gene thereof in disease-resistant breeding, and belongs to the technical field of gene engineering. The amino acid sequence of the rice bHLH transcription factor OsbHLH024 is as shown in SEQ ID NO. 2, and the nucleotide sequence of the gene OsbHLH024 is as shown in SEQ ID NO. 1. Experiments prove that through overexpression of the OsbHLH024 gene in the Zhonghua 11, the resistance of rice to rice blast, banded sclerotial blight and leaf blight can be remarkably improved; the OsbHLH024 gene is knocked out, and the resistance of rice to rice blast, banded sclerotial blight and leaf blight can be remarkably reduced. Therefore, the OsbHLH024 gene can positively regulate the disease resistance of the plant, which is of great significance to the cultivation of disease-resistant rice varieties.

Description

Technical Field

[0001] This invention relates to the field of genetic engineering technology, and in particular to the application of rice bHLH transcription factor OsbHLH024 and its encoding gene in disease-resistant breeding. Background Technology

[0002] Rice blast and other fungal diseases pose significant threats to crop production, easily leading to large-scale yield reductions, quality declines, and even regional food production crises. Therefore, cultivating crop varieties with superior disease resistance has always been one of the core breeding goals in the global crop industry. Based on existing disease-resistant gene resources, researchers have applied some high-quality disease-resistant genes as target genes in crop disease-resistant genetic engineering breeding practices, successfully breeding new varieties of rice, wheat, corn, and soybeans with significantly improved disease resistance, effectively mitigating the production damage caused by some fungal diseases. However, due to the continuous mutation and differentiation of pathogenic fungi, some disease-resistant genes have lost their resistance, making it difficult to fully meet current disease-resistant breeding needs. Therefore, continuously exploring new disease-resistant genes and enriching the disease-resistant gene resource library remains a crucial task that urgently needs to be promoted in the field of crop disease-resistant breeding.

[0003] Rice yield and quality are directly related to overall food security. Based on practical production and scientific research needs, in-depth exploration of rice disease resistance-related genes will help breed new rice varieties with broad-spectrum disease resistance, high yield and high quality, effectively reducing production losses caused by fungal diseases. This has extremely strong theoretical research value and significant practical production significance. Summary of the Invention

[0004] The purpose of this invention is to provide the application of rice bHLH transcription factor OsbHLH024 and its encoding gene in disease resistance breeding, in order to solve the problems existing in the prior art, and to overexpress... OsbHLH024 Genes can enhance rice's resistance to rice blast, providing a theoretical basis and scientific evidence for breeding rice with high resistance to rice blast.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides the application of rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; The amino acid sequence of the rice bHLH transcription factor OsbHLH024 is shown in SEQ ID NO.2; the disease resistance includes the resistance of rice to rice blast, rice sheath blight and leaf blight.

[0006] The amino acid sequence of the rice bHLH transcription factor OsbHLH024 of the present invention is not limited to the above sequence. It can also be a derivative protein with the same function obtained by substituting 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 with more than 80% identity and the same function as the protein shown in SEQ ID NO.2 or the derivative protein. It can also be a fusion protein obtained by attaching a protein tag to the N-terminus and / or C-terminus of all the above-mentioned proteins.

[0007] In the aforementioned proteins, the protein tag refers to a polypeptide or protein fused with the target protein using in vitro DNA recombination technology for expression, detection, tracing, and / or purification of the target protein. The protein tag may be a GFP tag, Flag tag, His tag, MBP tag, HA tag, myc tag, GST tag, and / or SUMO tag, etc.

[0008] In the above-mentioned proteins, identity refers to the identity of the amino acid sequences. The identity of amino acid sequences can be determined using homology search sites on the Internet, such as the BLAST page on the NCBI homepage. For example, in Advanced BLAST 2.1, using blastp as the program, setting the Expect value to 10, setting all filters to OFF, using BLOSUM62 as the matrix, setting the Gap existence cost, Per residue gap cost, and Lambda ratio to 11, 1, and 0.85 (default values) respectively, and performing an identity search on a pair of amino acid sequences to calculate the identity value (%), then the identity value can be obtained.

[0009] In the aforementioned proteins, the 80% or more identity can be at least 81%, 82%, 85%, 86%, 88%, 90%, 91%, 92%, 95%, 96%, 98%, 99%, or 100% identity.

[0010] The present invention also provides the use of the gene encoding the rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; The CDS sequence of the gene is shown in SEQ ID NO.1; the disease resistance includes the resistance of rice to rice blast, rice sheath blight and leaf blight.

[0011] The present invention also provides the use of a recombinant vector containing a gene encoding the rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; The CDS sequence of the gene is shown in SEQ ID NO.1; the disease resistance includes the resistance of rice to rice blast, rice sheath blight and leaf blight.

[0012] The present invention also provides the use of a host bacterium containing a gene encoding the rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; The host bacterium's genome integrates a gene encoding the rice bHLH transcription factor OsbHLH024, the CDS sequence of which is shown in SEQ ID NO.1; the disease resistance includes the rice's resistance to rice blast, sheath blight, and leaf blight.

[0013] Preferably, the regulation is as follows: overexpressing the gene encoding the rice bHLH transcription factor OsbHLH024 to improve rice disease resistance; knocking out the gene encoding the rice bHLH transcription factor OsbHLH024 to reduce rice disease resistance.

[0014] The recombinant expression vectors involved in this invention include binary Agrobacterium vectors and vectors that can be used for plant microbombardment. Examples include pAHC25, pWMB123, pBin438, pCAMBIA1302, pCAMBIA2301, pCAMBIA2300, pCAMBIA1301, pCAMBIA1300, pBI121, pCAMBIA1391-Xa, or pCAMBIA1391-Xb (CAMBIA). The plant expression vectors may also contain the 3' untranslated region of a foreign gene, i.e., containing a polyadenylated signal and any other DNA fragment involved in mRNA processing or gene expression. The polyadenylated signal can guide the addition of polyadenylated acid to the 3' end of the mRNA precursor; similar functions exist in the untranslated regions transcribed at the 3' end of Agrobacterium crown gall-inducing (Ti) plasmid genes (such as the carmine synthase gene Nos) and plant genes (such as the rice starch synthase gene). When constructing plant expression vectors using the genes of this invention, enhancers, including translational enhancers or transcriptional enhancers, may also be used. These enhancer regions may be ATG start codons or adjacent region start codons, etc., but they must be the same as the reading frame of the coding sequence to ensure the correct translation of the entire sequence.

[0015] In addition to Escherichia coli, the host bacteria involved in this invention can also be other bacteria, yeast, algae or fungi.

[0016] This invention also provides a method for regulating disease resistance in rice, including overexpression or knockout in rice. OsbHLH024 Genes regulate the resistance of rice to diseases; The OsbHLH024 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the diseases include rice blast, rice sheath blight, and rice leaf blight.

[0017] Preferably, the overexpression of the OsbHLH024 Genes enhance the resistance of the rice to diseases; knocking out the gene... OsbHLH024 The gene reduces the rice's resistance to disease.

[0018] The present invention also provides a method for cultivating highly disease-resistant rice, comprising overexpressing in rice OsbHLH024 Genes, to improve the OsbHLH024 The steps for obtaining highly disease-resistant rice by controlling gene expression levels; wherein, the... OsbHLH024 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the disease resistance is resistance to rice blast, sheath blight and leaf blight.

[0019] In addition to rice, the recipient plant of this invention can also be other monocotyledonous or dicotyledonous plants.

[0020] The present invention discloses the following technical effects: This invention identified a bHLH transcription factor, named OsbHLH024. Experiments demonstrated that the OsbHLH024 protein enhances disease resistance in rice. This was achieved through overexpression in Zhonghua 11 (ZH11). OsbHLH024 Gene overexpression in transgenic plants resulted in higher disease resistance than the recipient. (Explanation) OsbHLH024 Genes positively regulate plant disease resistance, and the OsbHLH024 protein and its encoding gene are of great significance for breeding disease-resistant rice varieties. Attached Figure Description

[0021] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0022] Figure 1 for OsbHLH024 A schematic diagram of the gene structure and sgRNA location; Figure 2 for OsbHLH024 Knockout (KO) plant gene sequence identification results; Figure 3 for OsbHLH024 Detection of expression levels in overexpression (OE) plants; asterisks above the bars indicate significant differences (P<0.05). Figure 4 For comparison, OsbHLH024 Phenotypic effects of puncture inoculation on gene-overexpressing and knockout plants; A represents three-week-old Zhonghua 11 (ZH11). OsbHLH024 -KO1、 OsbHLH024 -KO2、 OsbHLH024 -OE1 and OsbHLH024 - Representative lesion images and statistical results of OE2 plant leaves 5 days after spray inoculation with rice blast fungus (physiological race zhong10-8-14); B represents three-week-old Zhonghua 11 (ZH11). OsbHLH024 -KO1、 OsbHLH024 -KO2、 OsbHLH024 -OE1 and OsbHLH024 - Representative lesion images and statistical results of OE2 plant leaves 5 days after inoculation with rice blast fungus (physiological race zhong10-8-14); C represents 10-week-old Zhonghua 11 (ZH11). OsbHLH024 -KO1、 OsbHLH024 -KO2、 OsbHLH024 -OE1 and OsbHLH024 -Representative lesion images and statistical results of leaves of OE2 plants after spray inoculation with Rhizoctonia solani (physiological race AG-1-IA); D represents 10-week-old Zhonghua 11 (ZH11). OsbHLH024 -KO1、 OsbHLH024 -KO2、 OsbHLH024 -OE1 and OsbHLH024 -Representative lesion images and statistical results of leaves of OE2 plants after being punctured and inoculated with Bacillus subtilis (physiological race POX99); the asterisks above the bars indicate significant differences (P<0.05). Detailed Implementation

[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0024] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0025] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0026] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0027] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0028] The rice variety Zhonghua 11 (O.Sativa L.spp.japonica, var. zhonghua11, AA genome, ZH11) belongs to the japonica subspecies and is described in the following literature: Ni Yuchong, New rice variety cultivated by flower cultivar—Zhonghua 11, Agricultural Science and Technology Newsletter, 1989.07.35; provided by Professor Chen Xuewei of Sichuan Agricultural University.

[0029] OsbHLH024 The knockout and overexpression of transgenic rice was accomplished by Boyuan Biotechnology Co., Ltd.

[0030] The plant binary expression vector pTCRISPR was provided by Associate Professor Tang Yongyan of the State Key Laboratory of Sichuan Agricultural University.

[0031] The plant binary expression vector pCambia2300 was provided by Professor Chen Xuewei of the State Key Laboratory of Sichuan Agricultural University.

[0032] Total RNA Extraction Kit: Purchased from Invitrogen TRIzol, catalog number 15596026.

[0033] Reverse transcription kit: HiScript III RT SuperMix for qPCR (+gDNA wiper) purchased from Vanzyme (China), catalog number R323-01.

[0034] Homologous recombination kit: ClonExpress II One StepCloning Kit, catalog number C112-01, purchased from Vanzyme China.

[0035] Example 1: Rice bHLH transcription factor OsbHLH024 and its clone A bHLH transcription factor, OsbHLH024, was screened in rice. Primers were designed based on the reference gene sequence of the rice ZH11 genome. OsbHLH024F: 5'-ATGGATGAGGTGTGGTGCAG-3' (SEQ ID NO. 3); OsbHLH024R: 5'-CTGTGCTGCCGTGATCGCGA-3' (SEQ ID NO. 4).

[0036] Using ZH11 total cDNA as a template, a DNA band of approximately 1365 bp was amplified using the aforementioned primers. Sequencing confirmed that it belongs to the bHLH transcription factor family. OsbHLH024 Gene. Alignment of the LOC_Os01g39330 (OsbHLH024) sequence from the rice genome reference sequence with the clone sequence obtained from total cDNA amplification of ZH11 showed that the clone sequence obtained from ZH11 was identical to the LOC_Os01g39330 (OsbHLH024) sequence. The amino acid sequence of the OsbHLH024 protein is shown in SEQ ID NO.2. The gene encoding the OsbHLH024 protein. OsbHLH024 (See Figure 1 The CDS sequence of the DNA molecule is shown in SEQ ID NO.1.

[0037] OsbHLH024 The CDS sequence (SEQ ID NO.1) of the gene is as follows:

[0038] The amino acid sequence (SEQ ID NO.2) of the OsbHLH024 protein is as follows: MDEVWCSLDSMPADIAAGIHQPPPPDLHDPSFWPAFADCAASFIAGGGGDNACFDELMAGGSSGDTRMVAMDDGDDGSGFLVGDAEAEHLMLSSSSPSSSLSSGRSLSIDSAGSMSSFSLDAAAALAMSTLAVPHPYPPPVAHGMFASGDGGGGGGGAVDDHEDAIMRAMMAVISSASASPSSSGGSASSPTPFSRDSGAHHQPAGQPAMAAPQQPRGGNGGHVVVKS SSSSGGLAVPMDQKPGGGGRGRQQEEAAAASATNSSQLYHMMSERKRREKLNDSFHTLRSLLPPCSKKDKTTVLINAAKYLKSLETEITELEGTNTKLEKHIAGGGGAADAAM RARRAQQRAKVQISKAADSQSQQLVSLTVMVMVECDVVELVLHILECLRWMKEISVLSVYADTYSPQLLLKAIANIKLQIVGGDWNEASFHEAMTKAANDATISCAPLAITAAQ .

[0039] Example 2: Construction of OsbHLH024 plant knockout vector 1. Construction OsbHLH024 Knockout vector pTCRISPR- sgRNA OsbHLH024 The knockout target sequence is designed as follows: TGGCCGCCGTTCCCTCCACG (SEQ ID NO. 5).

[0040] And synthesize the following primers: F1: 5'-TGTG TGGCCGCCGTTCCCTCCACG G-3' (SEQ ID NO.6); R1: 5'-AAAAC CGTGGAGGGAACGGCGGCCA -3' (SEQ ID NO.7).

[0041] After denaturation at 95 °C, annealing was performed to form double strands. The pTCRISPR vector was digested with BsaI and the linearized vector was recovered. The above double-stranded DNA and the linearized vector were mixed according to the following system, and the T4 DNA ligase ligation reaction was performed and incubated at room temperature for 30 minutes. The ligation system is shown in Table 1.

[0042] Table 1 Connection System Note: The same effect can be achieved within the range shown in the table above.

[0043] After linking using the above reaction system, pTCRISPR- sgRNA OsbHLH024-Target Knock out the carrier.

[0044] 2. pTCRISPR- sgRNA OsbHLH024-Target The knockout vector was transformed into DH5α competent cells. After kanamycin selection, single colonies were picked for colony PCR identification. Plasmids were extracted from positive colonies and sequenced to confirm pTCRISPR- sgRNA OsbHLH024-Target Yes, genetic transformation can be performed.

[0045] 3. OsbHLH024 The knockout of transgenic rice was accomplished by Boyuan Biotechnology Co., Ltd. Genetically stable knockout lines were sequenced and screened. OsbHLH024 -KO1 and OsbHLH024 -KO2, the results showed that one or more bases were missing at the target sequence in the knockout lines, see Figure 2 .

[0046] Example 3: Construction of OsbHLH024 plant expression vector 1. OsbHLH024 Construction of the overexpression vector pCambia2300-35S:OsbHLH024 Using cDNA obtained by reverse transcription of total RNA from rice ZH11 as a template, the cloning primers were: F: 5'- TATCCAGATCCAGTGGGATCC ATGGATGAGGTGTGGTGCAG-3' (SEQ ID NO. 8); R: 5'- CGCACTAGTAAGCTTGGTACC CTGTGCTGCCGTGATCGCGA-3' (SEQ ID NO. 9).

[0047] PCR amplification OsbHLH024Full-length cDNA was obtained by double digestion of the pCambia2300-35S:eGFP-nosT vector with BamHI and KpnI, followed by linearization. The recovered PCR products were then processed according to the system shown in Table 2. OsbHLH024 Full-length cDNA and linearized vector were mixed and homologous recombination was performed.

[0048] Table 2 Note: The same effect can be achieved within the range shown in the table above.

[0049] After recombination according to the above homologous recombination reaction system, the pCambia2300-35S:OsbHLH024 recombinant vector was obtained.

[0050] 2. The above-mentioned pCambia2300-35S:OsbHLH024 recombinant vector was transformed into DH5α competent cells. After kanamycin screening, single clones were selected for colony PCR identification. After plasmid extraction and sequencing of positive colonies, pCambia2300-35S:OsbHLH024 was confirmed to be correct and could be genetically transformed.

[0051] 3. OsbHLH024 The overexpression of transgenic rice was completed by Boyuan Biotechnology Co., Ltd. The expression level of the target gene was detected, and genetically stable overexpression lines were screened. OsbHLH024 -OE1 and OsbHLH024 -OE2, the results showed that the expression of the target gene in overexpressing plants was significantly increased compared with that in wild-type plants, see [link to relevant documentation]. Figure 3 .

[0052] Example 4: OsbHLH024 participates in regulating plant disease resistance In plants, regarding OsbHLH024 The research primarily focuses on plant tolerance to heat stress. The inventor discovered... OsbHLH024 It plays an important role in regulating plant disease resistance responses. Therefore, it was tested. OsbHLH024 The impact on disease resistance in rice.

[0053] Seedling puncture inoculation treatment: The test material was Zhonghua 11 (ZH11), with ZH11 as the background and genetically stable. ZH11 Knockout strains OsbHLH024- KO1 and OsbHLH024 -KO2 ( Figure 2 ), with ZH11 as the background and genetically stable OsbHLH024 Overexpression lines OsbHLH024- OE1 and OsbHLH024 -OE2 ( Figure 3Select plump seeds and place them in an Erlenmeyer flask filled with tap water. Germinate the seeds in a 37°C dark incubator, changing the water daily. After two days, select seeds that have sprouted and sow them in seedbeds. After 21 days, apply a 2.5×10⁻⁶ solution. 5 pcs·mL -1 The spores of the rice blast fungus isolate Zhong10-8-14 were used for spray inoculation. Simultaneously, seeds showing signs of sprouting were selected and placed in 96-well seedling trays, which were then placed on floats and grown in Hoagland nutrient solution. After 21 days, the second-to-last rice leaf of uniform growth and size was selected, punctured, and inoculated with 5 μL of a 5×10⁻⁶ concentration. 5 pcs·mL -1 The spores of *Blastomyces oryzae* (Zhong10-8-14) were examined, and the length of lesions was counted after 5 days. Statistical analysis showed that, compared to ZH11, lesions treated with puncture and spray inoculation had significantly shorter lesions against the ZH11 background. OsbHLH024- KO1 and OsbHLH024-KO2 plants had more and longer lesions, indicating weakened disease resistance. Figure 4 (AB). Compared to ZH11, in the ZH11 background OsbHLH024- OE1 and OsbHLH024 -OE2 plants have fewer lesions and significantly shorter lesion lengths, indicating a marked increase in disease resistance. Figure 4 (AB).

[0054] At the same time, 10-week-old rice planted in the field was treated with Rhizoctonia solani (Bacillus subtilis). R. solani isolate AG-1-IA) and Bacillus subtilis ( Xanthomonas oryzae pv. Oryzae Field inoculation was performed using POX99, and statistical analysis showed that, compared to ZH11, the bacterial count under the ZH11 background was significantly lower. OsbHLH024- The lesions on KO1 and OsbHLH024-KO2 plants were longer, and their disease resistance was weakened. Figure 4 (CD). Compared to ZH11, in the ZH11 background. OsbHLH024- OE1 and OsbHLH024 -OE2 plants showed a significant reduction in lesion length and a marked increase in disease resistance. Figure 4 Medium CD).

[0055] The above results indicate that OsbHLH024 positively regulates plant disease resistance, and its encoding gene... OsbHLH024 It can be used as a target gene in molecular breeding to improve plant disease resistance.

[0056] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Application of rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; wherein The amino acid sequence of the rice bHLH transcription factor OsbHLH024 is shown in SEQ ID NO.2; the disease resistance includes the rice's resistance to rice blast, rice sheath blight, and leaf blight.

2. The use of the gene encoding the rice bHLH transcription factor OsbHLH024 as described in claim 1 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; wherein The CDS sequence of the gene is shown in SEQ ID NO.1; the disease resistance includes the resistance of rice to rice blast, rice sheath blight and leaf blight.

3. The application of a recombinant vector containing the gene encoding the rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; wherein, The CDS sequence of the gene is shown in SEQ ID NO.1; the disease resistance includes the resistance of rice to rice blast, rice sheath blight and leaf blight.

4. The use of host bacteria containing the gene encoding the rice bHLH transcription factor OsbHLH024 in any of the following: (1) Application in regulating disease resistance in rice; (2) Application in the breeding of disease-resistant rice varieties; wherein The host bacterium's genome integrates a gene encoding the rice bHLH transcription factor OsbHLH024, the CDS sequence of which is shown in SEQ ID NO.1; the disease resistance includes the rice's resistance to rice blast, sheath blight, and leaf blight.

5. Use according to any one of claims 1 to 4, wherein the compound is of formula (I) ###00001### (I) or a pharmaceutically acceptable salt thereof. The regulation is as follows: overexpressing the gene encoding the rice bHLH transcription factor OsbHLH024 to improve rice disease resistance; knocking out the gene encoding the rice bHLH transcription factor OsbHLH024 to reduce rice disease resistance.

6. A method for regulating disease resistance in rice, characterized in that, This includes overexpression or knockout in rice. OsbHLH024 Genes regulate the resistance of rice to diseases; The OsbHLH024 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the diseases include rice blast, rice sheath blight, and rice leaf blight.

7. The method as described in claim 6, characterized in that, Overexpression OsbHLH024 Genes enhance the resistance of the rice to diseases; knocking out the gene... OsbHLH024 The gene reduces the rice's resistance to disease.

8. A method for cultivating highly disease-resistant rice, characterized in that, Including overexpression in rice OsbHLH024 Genes, to improve the OsbHLH024 The steps for obtaining highly disease-resistant rice by controlling gene expression levels; wherein, the... OsbHLH024 The nucleotide sequence of the gene is shown in SEQ ID NO.1; the disease resistance is resistance to rice blast, sheath blight and leaf blight.

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