Application of transcription factor OsMYB51 in increasing content of volatile metabolites in rice

Abstract

The invention relates to the technical field of plant genetic engineering, and discloses application of a transcription factor OsMYB51 in increasing the content of volatile metabolites in rice, and the amino acid sequence of the transcription factor OsMYB51 is shown as SEQ ID NO.27. The invention further discloses a preparation method of the transcription factor OsMYB51. According to the invention, through overexpression of the OsMYB51 gene driven by a constitutive promoter in rice, the content of volatile metabolites of rice, especially 2-acetyl-1-pyrroline, is significantly increased. The method has the advantages that OsMYB51 is used as a positive regulatory factor to promote synthesis of rice 2-acetyl-1-pyrroline, and the method can be applied to rice fragrance quality improvement.

Description

Technical Field

[0001] The invention relates to the technical field of plant genetic engineering and discloses the application of transcription factor OsMYB51 in improving the content of volatile metabolites in rice. Background Art

[0002] Plants produce a large number of low molecular weight, lipophilic molecules with high vapor pressure called Volatile Organic Compounds (VOCs). These chemically and biologically rich volatile metabolites contribute to the interaction between plants and their environment by acting as aroma and flavor molecules through interactions with human receptors, and also through interactions with insect receptors. In food: the common vegetables and fruits in daily life are often judged by their flavor and color to determine their maturity and freshness. The volatile substances in fruits are mainly aldehydes, alcohols, esters, etc. In addition to fruits, there are also condiments, such as the flavor source of pepper is terpenes and phellandrene; in cosmetics: the main chemical components of rose essential oil are citronellol, geraniol, etc.; the main substances in daily lotions are linalool, limonene, etc.; creams and lotions often contain lyral I and lyral II; in agriculture: plant volatile substances can repel pests and achieve the purpose of protecting plants. For example, limonene, the main volatile of celery, can be used to repel whiteflies. When tea trees are infected by pests and diseases, they will release acaciaene and ocimene as signal molecules to activate the defense mechanism of nearby plants. Not only that, VOCs have gradually become a key indicator for consumers to measure the quality of crops and their processed products, and their particularity significantly affects the sensory quality of crops. As one of the most important food crops in the world, the aroma of rice can effectively improve the quality and value of rice, and rice with aroma is very popular among consumers. Therefore, the research on aroma genes and their application in genetic breeding have attracted extensive attention from rice geneticists and breeders.

[0003] Studies have shown that 2-acetyl-1-pyrroline (2-AP) is an important characteristic aroma substance of aromatic rice. Due to its low odor threshold, 2-AP can significantly change the aroma of rice even at low concentrations, making it an important component contributing to the unique aroma of rice. So far, the molecular mechanism of rice aroma traits has mainly focused on betaine aldehyde dehydrogenase 2 ( OsBADH2 )superior. OsBADH2Many mutations within the gene can transform non-fragrant rice into fragrant varieties. For example, there is a 2-base pair deletion in exon 1, a 7-base pair deletion in exon 2, an 803-base pair deletion between exon 4 and exon 5, an 8-base pair deletion and 3 single nucleotide polymorphisms (SNPs) in exon 7, a 7-base pair insertion in exon 8, 1 SNP each in exon 10 and exon 13, and a 1-base pair insertion / deletion / SNP in exon 14. All of these mutations result in a reduction or loss of OsBADH2 activity, thereby blocking the conversion of γ-aminobutyraldehyde (GABald) to γ-aminobutyric acid (GABA), which promotes the accumulation of 2-AP.

[0004] The biosynthesis of VOCs depends on the content and availability of carbon, nitrogen and sulfur and the energy provided by primary metabolism. The biosynthesis of various VOCs can only be separated from a few major metabolic pathways. According to their biosynthetic sources, all VOCs are divided into four major categories, including terpenoids, phenylpropanoids / benzenes, fatty acid derivatives and amino acid derivatives. Terpenoids are derived from two common five-carbon precursors, namely isopentanyl diphosphate and its allyl isomers and dimethylallyl diphosphate. Terpenoids are widely present in nature. For example, the highest content in plants is terpenoids, coumarins, sterols, fatty acids and sterol esters. In plants, terpenoid synthesis involves two independent pathways, namely the mevalonic acid (MVA) pathway and the methylerythrose phosphate (MEP) pathway. The MVA pathway is mainly responsible for the production of volatile sesquiterpenes (C15), while the MEP pathway is responsible for providing precursors for volatile hemiterpenes (C5), monoterpenes (C10) and diterpenes (C20). Phenylpropanoids are a variety of volatile organic compounds synthesized by plants from phenylalanine and tyrosine. They can be divided into many categories, including phenylpropanol, coumarin and lignin. Phenylpropanoids / benzenes are widely present in the plant kingdom and can protect against ultraviolet rays, invasion by herbivores and pathogens. Fatty acid derivatives are derived from fatty acids. Fatty acids originally referred to even-carbon saturated straight-chain carboxylic acids that existed in the form of triglycerides in ordinary oils and fats, and later expanded to unsaturated fatty acids such as oleic acid, linoleic acid and linolenic acid. Common ones in plants include 1-hexanal, nonanal and methyl jasmonate. Amino acid metabolism can be divided into branched-chain amino acid metabolism and aromatic amino acid metabolism according to the different metabolic pathways of the substrate. Leucine, isoleucine and valine belong to branched-chain amino acids, while alanine and tyrosine belong to aromatic amino acids.

[0005] MYB transcription factors are the largest transcription factor family in plants and are widely present in eukaryotes. MYB affects taste and flavor by regulating aroma, astringency and pungency. MsMYB in spearmint inhibits the activity of geranyl diphosphate synthase (GPPS) by binding to the promoter region of GPPS, thereby limiting the accumulation of monoterpenes. SlMYB75 in tomatoes can promote anthocyanin accumulation and enhance the production of volatile aromas in tomato fruits. Overexpression of SlMYB75 can activate the promoters of LOXC, AADC2 and TPS genes, thereby increasing the synthesis of various aromatic volatiles such as aldehydes, phenylpropanoid derivatives and terpene volatiles. The HcMYB gene in ginger flower can directly bind to the promoters of the underlying structural volatile synthesis genes (HcTPS1, HcTPS3, HcTPS10 and HcBSMT2), thereby participating in the regulatory mechanism of terpenoid and benzene biosynthesis in ginger flower. AtMYB21 and AtMYB24 in Arabidopsis are involved in the production of the sesquiterpene volatile (E)-β-caryophyllene. OfMYB1R114 and OfMYB1R70 in the floral organs of Osmanthus fragrans are involved in accelerating the formation of β-ionone, while OfMYB1R201 can reduce the synthesis of β-ionone. The volatile metabolites of rice are closely related to the aroma quality, but the specific molecular mechanism of its regulation is not clear, and the regulatory network formed has always been one of the hot and difficult research points at home and abroad. Summary of the invention

[0006] To solve the above problems, one of the objects of the present invention is to provide an application of transcription factor OsMYB51 in increasing the content of volatile metabolites in rice; a second object of the present invention is to provide a method for increasing the content of volatile metabolites in rice.

[0007] The technical solution adopted by the present invention to achieve the technical purpose is: The invention discloses an application of transcription factor OsMYB51 in increasing the content of volatile metabolites in rice. The amino acid sequence of the transcription factor OsMYB51 is shown in SEQ ID NO.27.

[0008] Preferably, the nucleotide sequence of the transcription factor OsMYB51 is shown as SEQ ID NO.26.

[0009] Preferably, the volatile metabolites include ethanol, vinyl acetate, 2,3-butylene oxide, isovaleraldehyde, 2-methylbutanal, 2-ethylfuran, 2-methyl-2-butenal, cis-2-methyl-2-butanal, tetrahydropyrrole, 1,3-octadiene, 2-acetyl-1-pyrroline, heptanol, 1-octen-3-ol, 2-n-pentylfuran, undecane, phenylethyl alcohol, L-menthol, nonanol, trans-isopiperenol, decanal, D-carvone, 2-undecenal, vanillin, hexadecanal, and n-octadecane.

[0010] More preferably, the volatile metabolite is 2-acetyl-1-pyrroline.

[0011] Preferably, the transcription factor OsMYB51 inhibits the rice 2-acetyl-1-pyrroline pathway OsBADH2 Gene expression promotes the synthesis of volatile metabolites.

[0012] A method for increasing the content of volatile metabolites in rice comprises: overexpressing a transcription factor OsMYB51 in rice, wherein the amino acid sequence of the transcription factor OsMYB51 is shown in SEQ ID NO.27.

[0013] Preferably, the nucleotide sequence of the transcription factor OsMYB51 is shown as SEQ ID NO.26.

[0014] Preferably, the method for overexpressing the transcription factor OsMYB51 in rice comprises: inserting the transcription factor OsMYB51 into a plant transformation plasmid containing the overexpression gene function to obtain a plant overexpression vector of the transcription factor OsMYB51.

[0015] More preferably, the plant overexpression vector is constructed using the Gateway system, and the desired gene fragment is amplified using rice cDNA as a template, and the primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6.

[0016] More preferably, pH2GW7 is used as the starting vector of the plant overexpression vector.

[0017] Compared with the prior art, the present invention can achieve the following beneficial effects: The present invention discloses the application of transcription factor OsMYB51 in increasing the content of volatile metabolites in rice. Studies have found that OsMYB51 inhibits the rice 2-AP pathway OsBADH2 The expression of the gene promotes the synthesis of volatile metabolites. Overexpression of the gene can significantly increase the content of volatile metabolites, indicating that OsMYB51 transcription factor, as a positive regulatory factor, has important guiding significance and broad market prospects for improving rice aroma quality. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 The real-time fluorescence quantitative PCR provided in the embodiment of the present invention detects the expression of OsMYB51 in rice leaves of overexpressed and knocked-out mutant strains OsBADH2Gene expression; OsMYB51-OE13, OsMYB51-OE23: different transgenic rice lines with overexpression OE-MYB51; wild type: wild type-ZH11 (A: OsMYB51 gene expression in rice leaves of overexpression lines; B: OsMYB51 gene expression in rice leaves of overexpression and knockout mutant lines OsBADH2 gene expression level).

[0019] Figure 2 A bar graph of the 2-AP content in the grains of OsMYB51 overexpression and mutant strains and a line graph of the 2-AP standard curve determined by gas chromatography-mass spectrometry (GC-MS) provided in an embodiment of the present invention; OsMYB51-OE13, OsMYB51-OE23: different transgenic rice lines OE-MYB51 representing overexpression; osmyb51-14, osmyb51-15: different transgenic rice lines CR-OsMYB51 representing knockout mutants; wild type: wild type-ZH11 (A: bar graph of the 2-acetyl-1-pyrroline content in leaves of OsMYB51 overexpression and mutant strains; B: standard curve of 2-acetyl-1-pyrroline).

[0020] Figure 3 A heat map of the content of volatile metabolites in the grains of OsMYB51 overexpressing strains measured by gas chromatography-mass spectrometry (GC-MS) provided in an embodiment of the present invention; OsMYB51-OE13-1, OsMYB51-OE13-2, OsMYB51-OE23-1, and OsMYB51-OE23-2 represent different transgenic rice lines OE-OsMYB51 overexpressing; wild type: wild type-ZH11.

[0021] Figure 4 The present invention provides an embodiment and verifies the binding of OsMYB51 by dual luciferase reporter assay (Dual-LUC) proOsBADH2 , thereby transcriptionally inhibiting the expression of OsBADH2; proOsBADH2 :express OsBADH2 Promoter OsBADH2 promoter (A: vector schematic; B: OsMYB51 OsBADH2 promoter-mediated dual-luciferase reporter gene activity).

[0022] Figure 5 The yeast one-hybrid experiment (Y1H) provided in the present invention verifies OsMYB51 and OsBADH2 ProOsBADH2: indicates OsBADH2 The promoter OsBADH2 promoter (A: OsBADH2Motif analysis of the promoter region of the gene; B: Results of yeast one-hybrid experiment of OsMYB51). DETAILED DESCRIPTION

[0023] Hereinafter, embodiments of the present invention will be described with reference to the accompanying drawings. In the following description, the same modules are represented by the same reference numerals. In the case of the same reference numerals, their names and functions are also the same. Therefore, the detailed description thereof will not be repeated.

[0024] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the accompanying drawings and specific embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and do not constitute a limitation of the present invention.

[0025] Example 1 Rice OsMYB51 gene and OsBADH2 Promoter cloning 1. Extraction of Total RNA from Rice Leaves Take two pieces of fresh rice leaves with a length of 3-5 cm, freeze them quickly in liquid nitrogen, and grind them into powder. Add 1 ml of Trizol to the sample, and extract total RNA according to the kit method of Quanshijin Company. Use agarose gel electrophoresis to detect RNA quality, and use NanoDrop 2000 spectrophotometer to detect RNA concentration.

[0026] 2. Cloning of Rice Genes The extracted total RNA was used as a template to synthesize cDNA using a one-step reverse transcription kit from Beijing Quanshijin Company; PCR amplification primers were designed based on the OsMYB51 gene sequence, and the primer sequences are shown in SEQ ID NO.1 and SEQ ID NO.2, see Table 1.

[0027] Table 1 Cloning primers for rice OsMYB51

[0028] The obtained cDNA was used as a template for PCR amplification, and the procedure was: 94℃ pre-denaturation for 2min, 94℃ denaturation for 15s, 60℃ annealing for 20s, 72℃ extension for 1min, repeated 35 cycles; 72℃ final extension for 5min; stored at 25℃. PCR amplification was performed with the OsMYB51 gene primers in Table 1, and the products were excised and recovered and sequenced to obtain the rice OsMYB51 sequence. The sequencing results showed that the obtained OsMYB51 gene was 930bp in length, and its nucleotide sequence was shown in SEQ ID NO.26; the obtained OsMYB51 gene encodes 310 amino acids, and its amino acid sequence is shown in SEQ ID NO.27.

[0029] 3. Cloning of Rice Promoter Refer to the CTAB extraction method of plant genome to extract rice leaf DNA. proOsBADH2 PCR amplification primers were designed based on the 1646 bp sequence of the promoter. The primer sequences are shown in SEQ ID NO.3 and SEQ ID NO.4, see Table 2.

[0030] Table 2 Rice proOsBADH2 Amplification primers

[0031] The obtained DNA was used as a template for PCR amplification, and the procedure was 94℃ pre-denaturation for 5min, 94℃ denaturation for 15s, 58℃ annealing for 20s, 72℃ extension for 2min, repeated 35 cycles; 72℃ final extension for 5min; stored at 25℃. The PCR amplification product was recovered by gel excision and sequencing to obtain rice proOsBADH2 The sequencing results showed that the obtained proOsBADH2 The total length of the promoter sequence is 1646 bp, and its sequence is shown in SEQ ID NO.28.

[0032] Example 2 Construction of plant overexpression vector The construction of plant overexpression vectors was carried out using the Gateway system. First, the desired gene fragment was amplified using rice cDNA as a template. The primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.6, as shown in Table 3. The gel recovery product with the pDONR207 recombination linker was obtained by PCR, and BP (LR) enzyme, pH2GW7+pUbi (Smedley MA, Harwood WA. Gateway-compatible plant transformation vectors. Methods Mol Biol. 2015;1223:3-16.doi: 10.1007 / 978-1-4939-1695-5_1. PMID: 25300827.), and water were added to form a 2µL system (see Table 4), and the mixture was connected in a 25℃ dry bath for 4 hours, and transformed into DH5α. The final vector pH2GW7_OE was obtained after sequencing verification.

[0033] Table 3 Amplification primers for rice overexpression vector

[0034] Table 4 BP, LR reaction system

[0035] Example 3 Construction of plant gene knockout vector To construct a plant gene knockout vector, the gene editing target site was first designed using the online software CRISPR-P2.0 (http: / / crispr.hzau.edu.cn / CRISPR2), and a three-target site strategy was adopted. A 50 μL PCR amplification system was used, and transformation and colony PCR identification were performed after restriction digestion and ligation. The target site sequences of OsMYB51 are shown in SEQ ID NO.7-9 (see Table 5). The OsMYB51 editing sequence was amplified with SEQ ID NO.10 and SEQ ID NO.11, and then connected to pH-Ubi-cas9-7 (purchased from Addgene; catalog number: 106331) to obtain a plant transformation plasmid containing the OsMYB51 gene editing function of CRISPR / Cas9. The recombinant plasmid was the knockout vector (the vector was constructed by Boyuan Biotechnology Co., Ltd.).

[0036] Table 5 Target site design information

[0037] Table 6 Amplification primers

[0038] Example 4 Stable genetic transformation of rice mediated by Agrobacterium tumefaciens The plant overexpression vector and plant gene knockout vector of OsMYB51 were introduced into Agrobacterium tumefaciens EHA105 for stable genetic transformation of rice. The specific steps are as follows: Agrobacterium positive single clone identification: Take 1 μL of plasmid and add it to 50 μL EHA105 Agrobacterium tumefaciens competent cells, mix well, add 1 mL YEB liquid medium after electroporation, shake and culture for 30 min at 30°C and 180 rpm on a shaker, inoculate it into YEB solid medium, and culture it in the dark at 28°C for 48 h. After PCR identification, positive Agrobacterium single clones were obtained, and the identification primer sequences are shown in SEQ ID NO.5, SEQ ID NO.6, SEQ ID NO.10, SEQ ID NO.11, SEQ ID NO.12 and SEQ ID NO.13, see Table 3, Table 6 and Table 7.

[0039] Rice genetic transformation: Select complete, plump and uniform rice grains, disinfect with 70% alcohol for 2 minutes, wash with sterile water, disinfect with 30% sodium hypochlorite for 10 minutes, wash with sterile water, place in 1 / 2MS medium, and culture under light for 7-8 days. The grown callus is infecting with positive Agrobacterium bacteria solution for 25 minutes, placed on sterile filter paper to dry in the shade, spread on co-culture medium, and co-cultured in the dark at 25℃ for 2.5-3 days. The callus is evenly placed on the screening medium, screened and cultured in the dark at 25℃ for 2-3 weeks, transferred to the pre-differentiation medium, cultured at room temperature with 14h / 10h light and dark alternation for two weeks, and transferred to the differentiation medium. The rice callus that has grown to 2 leaves or 2 leaves and one heart is transferred to the rooting medium. When the rice seedlings grow to about 10cm, they are placed in clean water for 3-4 days of seedling hardening and moved into soil pots (the transgenic material is created by Boyuan Biotechnology Co., Ltd.).

[0040] The T0 generation positive plants were identified by PCR, and the identification primer sequences were shown in SEQ ID NO.12, SEQ ID NO.13, SEQ ID NO.14, SEQ ID NO.15, SEQ ID NO.16 and SEQ ID NO.17 as shown in Table 7. The T1 generation positive plants were obtained by self-pollination for subsequent experiments and analysis of the content of related metabolites.

[0041] Table 7 PCR primers for positive plant identification

[0042] Quantitative PCR was used to determine the expression of OsMYB51 and OsBADH2 The gene expression status and the detection primer sequences are shown in SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20 and SEQ ID NO.21, see Table 8.

[0043] Table 8 qPCR identification primers

[0044] Test results such as Figure 1 As shown in Figure 2, the expression level of OsMYB51 gene in rice plants of the OE-OsMYB51 overexpression line was significantly increased, while OsBADH2 The expression of the gene was significantly reduced, indicating that OsMYB51 could inhibit OsBADH2 Gene expression.

[0045] Example 5 Determination of the content of volatile metabolites in rice leaves Gas chromatography-mass spectrometry (GC-MS) can meet the requirements for accurate qualitative and quantitative determination of volatile metabolites. Its operation process is briefly as follows: Sample preparation: The harvested mature kernels were shelled and 1 g of sample was extracted by adding 300 μL ddH2O for GC-MS analysis.

[0046] Volatile metabolites were detected by gas chromatography system (7890AGC, Agilent Technologies) and triple quadrupole mass spectrometry system (Agilent 7000D triple quadrupole mass detector). HP-5 mass spectrometry capillary column (30 mm* 0.25 mm id, 0.25 mm film thickness; Agilent Technologies) was used to separate volatile metabolites. The initial column temperature was 50 °C, maintained for 2 min, the temperature was increased by 5 °C / min to 200 °C, maintained for 2 min, and then the temperature was increased by 20 °C / min to the final temperature of 250 °C, maintained for 2 min. The injection temperature was 230 °C, the splitless mode was used, and the He flow rate was 1.0 ml / min (99.999%). In full scan mode, the scan range was 35-500 m / z.

[0047] The results are as follows Figure 2 and 3 As shown in Figure 2, the 2-AP content in the OsMYB51 overexpression strain was 7-8 μg•kg -1 In the mutant strain of OsMYB51 and ZH11, the 2-AP content was 0 μg•kg -1 .

[0048] Example 6 Interaction between OsMYB51 and proOsBADH2 and verification of the action site The OsMYB51 cloned in Example 1 was connected to the vector pGADT7 (Cat. No.: HG-VJC0483, Changsha Abiwei Biotechnology Co., Ltd.) and pB7WG2.0 (Dual-LUC) (Cat. No.: VT13019, Qingdao Cresbo Biotechnology Co., Ltd.) to obtain the recombinant vectors OsMYB51-pGADT7 (yeast single hybrid) and OsMYB51-pB7WG2.0 (Dual-LUC). The upstream 1600 bp proOsBADH2 sequence was constructed into the vector pHIS2 (Cat. No.: HG-VJC0481, Changsha Abiwei Biotechnology Co., Ltd.) and pGEXT4-1 pGEXT4-1 (Cat. No.: HG-VYA0221, Changsha Abiwei Biotechnology Co., Ltd.) to obtain proOsBADH2-pHIS2 and proOsBADH2-pGEXT4-1. The construction of the vectors was based on the Gateway system in Example 2. The interaction was verified using the mature Y1H and Dual-LUC experimental systems. The Y1H system uses the Y187 yeast competent system. The pGADT7 and pHIS2 containing the target gene can grow plaques normally on the three-deficient plates with 3-AT, which can determine the interaction between DNA and protein. Dual-LUC was verified in tobacco leaves, and the binding of DNA and protein was determined by fluorescent signals. The vector and primer sequences are shown in SEQ ID NO.22, SEQ ID NO.23, SEQ ID NO.24 and SEQ ID NO.25, see Table 9.

[0049] Table 9 Vector and primer information

[0050] The results are as follows Figure 4 and Figure 5 As shown, OsMYB51 can bind to OsBADH2 The promoter region of the gene significantly inhibits its expression.

[0051] It should be understood that the various forms of processes shown above can be used to reorder, add or delete steps. For example, the steps described in the disclosure of the present invention can be performed in parallel, sequentially or in different orders, as long as the desired results of the technical solution disclosed in the present invention can be achieved, and this document does not limit this.

[0052] The above specific implementations do not constitute a limitation on the protection scope of the present invention. It should be understood by those skilled in the art that various modifications, combinations, sub-combinations and substitutions can be made according to design requirements and other factors. Any modification, equivalent substitution and improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. Application of transcription factor OsMYB51 in increasing the content of volatile metabolites in rice, characterized in that: The amino acid sequence of the transcription factor OsMYB51 is shown in SEQ ID NO.

27.

2. The use according to claim 1, characterized in that: The nucleotide sequence of the transcription factor OsMYB51 is shown as SEQ ID NO.

26.

3. The use according to claim 1 or 2, characterized in that: The volatile metabolites include ethanol, vinyl acetate, 2,3-butylene oxide, isovaleraldehyde, 2-methylbutanal, 2-ethylfuran, 2-methyl-2-butenal, cis-2-methyl-2-butanal, tetrahydropyrrole, 1,3-octadiene, 2-acetyl-1-pyrroline, heptanol, 1-octen-3-ol, 2-n-pentylfuran, undecane, phenylethyl alcohol, L-menthol, nonanol, trans-isopiperenol, decanal, D-carvone, 2-undecenal, vanillin, hexadecanal, and n-octadecane.

4. The use according to claim 1 or 2, characterized in that: The volatile metabolite is 2-acetyl-1-pyrroline.

5. The use according to claim 1 or 2, characterized in that: The transcription factor OsMYB51 inhibits the rice 2-acetyl-1-pyrroline pathway OsBADH2 Gene expression promotes the synthesis of volatile metabolites.

6. A method for increasing the content of volatile metabolites in rice, characterized in that: include: The transcription factor OsMYB51 is overexpressed in rice, and the amino acid sequence of the transcription factor OsMYB51 is shown in SEQ ID NO.

27.

7. The method according to claim 6, characterized in that The nucleotide sequence of the transcription factor OsMYB51 is shown as SEQ ID NO.

26.

8. The method according to claim 6 or 7, characterized in that: The method for overexpressing the transcription factor OsMYB51 in rice comprises: inserting the transcription factor OsMYB51 into a plant transformation plasmid containing an overexpression gene function to obtain a plant overexpression vector of the transcription factor OsMYB51.

9. The method according to claim 8, characterized in that The plant overexpression vector was constructed using the Gateway system, and the desired gene fragment was amplified using rice cDNA as a template. The primer sequences are shown in SEQ ID NO.5 and SEQ ID NO.

6.

10. The method according to claim 9, characterized in that pH2GW7 was used as the starting vector for plant overexpression vector.

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