Biosynthetic gene of homovanillic acid and application of biosynthetic gene in resisting brown planthopper of rice

Abstract

The invention belongs to the technical field of biology. More specifically, the invention relates to a homovanillic acid biosynthetic gene and an application thereof in resisting brown planthopper. The research finds that the over-expression of the LOCOs03g03720 gene or the LOCOs01g40870 gene in the rice can improve the resistance of the rice to the brown planthopper and can also improve the content of homovanillic acid in the rice, and after the LOCOs03g03720 gene or the LOCOs01g40870 gene is knocked out, the resistance of the rice plant to the brown planthopper is reduced, and the content of the homovanillic acid in the rice is also reduced. According to the invention, the LOCOs03g03720 gene or the LOCOs01g40870 gene is over-expressed in the rice through a gene engineering technology, so that the brown planthopper-resistant rice variety is created, germplasm resources are provided for breeding of the brown planthopper-resistant rice variety, and the application prospect and value are very good.

Description

Technical Field

[0001] The invention belongs to the field of biotechnology, and more specifically relates to a homovanillic acid biosynthesis gene and its application in rice resistance to brown planthoppers. Background Art

[0002] Brown planthopper (Nilaparvata lugens ) is one of the important pests on rice. It mainly affects the normal growth and development of rice by sucking rice juice and spreading diseases. When it occurs seriously, it can cause a large-scale reduction in rice production or even a total crop failure. Chemical pesticides are often used in production to control brown planthoppers, but the long-term and large-scale use of chemical pesticides has caused brown planthoppers to develop medium to high levels of resistance to many commonly used chemical pesticides. Although the use of biological control methods such as parasitic wasps, green muscardine, and Bacillus thuringiensis can control the growth of brown planthopper populations in farmland, the most economical, efficient and green control approach to control rice diseases and insect pests is to use rice varieties with insect-resistant characteristics and give full play to the advantages of insect-resistant genes or insect-resistant metabolites in rice. The research results of the International Rice Research Institute (IRRI) and the rice production practice in Southeast Asia have proved that even rice varieties with only moderate insect resistance can effectively control the population of brown planthoppers below the level that causes harm, so as not to cause actual harm and yield loss to rice. Rice insect-resistant breeding is an important way to control rice brown planthoppers.

[0003] Therefore, in-depth exploration and screening of insect-resistant materials in rice, finding new insect-resistant genes, and developing new rice varieties with highly resistant gene materials are of great significance for the prevention and control of rice pests. Summary of the invention

[0004] The present invention aims to provide a gene resistant to rice brown planthopper, including LOC_Os03g03720 gene and / or LOC_Os01g40870 gene, and improve the resistance of rice to brown planthopper by increasing the expression of the above genes in rice, thereby constructing a rice variety with high resistance to brown planthopper.

[0005] The first object of the present invention is to provide an application of a rice brown planthopper resistance gene in regulating rice resistance to brown planthoppers, wherein the rice brown planthopper resistance gene comprises the LOC_Os03g03720 gene and / or the LOC_Os01g40870 gene.

[0006] The second object of the present invention is to provide the application of the above rice brown planthopper resistance gene.

[0007] The third object of the present invention is to provide an agent and its application that can be used to improve the resistance of rice to brown planthopper.

[0008] A fourth object of the present invention is to provide a method for constructing a rice variety with enhanced resistance to brown planthoppers.

[0009] The fifth object of the present invention is a method for constructing a rice variety rich in high vanillic acid.

[0010] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0011] The present invention has found that overexpression of LOC_Os03g03720 gene or LOC_Os01g40870 gene in rice can improve the resistance of rice to brown planthoppers and can also increase the content of homovanillic acid in rice. Therefore, the present invention claims the following scheme:

[0012] The application of rice brown planthopper resistance genes in regulating rice resistance to brown planthoppers, the rice brown planthopper resistance genes include LOC_Os03g03720 gene and / or LOC_Os01g40870 gene; the nucleotide sequence of LOC_Os03g03720 gene is a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO.3; the nucleotide sequence of LOC_Os01g40870 gene is a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO.6.

[0013] Optionally, the nucleotide sequence of the LOC_Os03g03720 gene is shown in SEQ ID NO.1.

[0014] Optionally, the cDNA sequence of the LOC_Os03g03720 gene is shown as SEQ ID NO.2.

[0015] Optionally, the nucleotide sequence of the LOC_Os01g40870 gene is shown in SEQ ID NO.4.

[0016] Optionally, the cDNA sequence of the LOC_Os01g40870 gene is shown as SEQ ID NO.5.

[0017] As an optional embodiment, the method for regulating rice brown planthopper resistance includes (1) and (2):

[0018] (1) Improving the resistance of rice to brown planthopper by increasing the expression of the above genes (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice;

[0019] (2) By inhibiting the expression of the above genes (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice, the resistance of rice to brown planthoppers is reduced, which can be used to prepare a rice model with low resistance to brown planthoppers for use in scientific research.

[0020] As an optional embodiment, the method for increasing the expression of the rice brown planthopper resistance gene (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice is: connecting the gene to an overexpression vector and transferring it into Agrobacterium, and using Agrobacterium to infect rice.

[0021] As an optional embodiment, the method for inhibiting the expression of the above-mentioned rice brown planthopper resistance gene (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice includes: inhibiting the expression of LOC_Os03g03720 gene and / or LOC_Os01g40870 gene by RNAi, or knocking out LOC_Os03g03720 gene and / or LOC_Os01g40870 gene in rice by gene editing technology.

[0022] The application of the above rice brown planthopper resistance gene (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) as a target in screening brown planthopper resistance rice varieties.

[0023] Application of the above rice brown planthopper resistance gene in regulating the high vanillic acid content of rice.

[0024] As an optional embodiment, the method for regulating the high vanillic acid content in rice includes (1) and (2):

[0025] (1) increasing the homovanillic acid content in rice by increasing the expression of the above genes (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice;

[0026] (2) By inhibiting the expression of the above genes (LOC_Os03g03720 gene and / or LOC_Os01g40870 gene) in rice, the content of homovanillic acid in rice is reduced.

[0027] Application of the reagent for promoting the expression of the rice brown planthopper resistance gene in improving the resistance of rice to brown planthopper.

[0028] The invention provides a reagent for improving the resistance of rice to brown planthoppers, which contains a reagent for promoting the expression of the rice brown planthopper resistance gene.

[0029] As an alternative embodiment, the reagent includes an overexpression vector containing the above rice brown planthopper resistance gene.

[0030] Application of the rice brown planthopper resistance gene or the reagent in constructing a rice variety with enhanced resistance to brown planthopper.

[0031] Application of the rice brown planthopper resistance gene or the reagent in improving rice resistance to brown planthopper.

[0032] Application of the rice brown planthopper resistance gene or the reagent in preparing a product for improving rice resistance to brown planthoppers.

[0033] Application of the rice brown planthopper resistance gene or the reagent in increasing the high vanillic acid content of rice,

[0034] Application of the rice brown planthopper resistance gene or the reagent in preparing a product for increasing the high vanillic acid content of rice,

[0035] Application of the rice brown planthopper resistance gene or the reagent in constructing homovanillic acid-rich rice.

[0036] The invention provides a method for constructing a rice variety with enhanced resistance to brown planthoppers, and overexpressing the rice brown planthopper resistance gene in a plant.

[0037] The invention provides a method for constructing a rice variety rich in homovanillic acid, and overexpressing the rice brown planthopper resistance gene in a plant.

[0038] As an optional embodiment, the method for overexpressing the rice brown planthopper resistance gene in rice is: constructing an overexpression vector of the rice brown planthopper resistance gene, transferring the overexpression vector into Agrobacterium, and then using Agrobacterium to infect rice.

[0039] The present invention has the following beneficial effects:

[0040] The present invention has found that overexpression of LOC_Os03g03720 gene or LOC_Os01g40870 gene in rice can improve the resistance of rice to brown planthoppers, and can also increase the content of homovanillic acid in rice, while knocking out LOC_Os03g03720 gene or LOC_Os01g40870 gene reduces the resistance of rice plants to brown planthoppers. Using LOC_Os03g03720 gene and / or LOC_Os01g40870 gene as a target can quickly screen rice varieties resistant to brown planthoppers, and can simplify the screening process. In addition, the present invention uses genetic engineering technology to overexpress LOC_Os03g03720 gene or LOC_Os01g40870 gene in rice, creates rice varieties resistant to brown planthoppers, provides germplasm resources for the breeding of rice varieties resistant to brown planthoppers, and has good application prospects and value. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] Figure 1 The mortality rate of 4th instar nymphs and short-winged adults of brown planthopper fed on artificial diets containing different concentrations of homovanillic acid (a shows the mortality rate of 4th instar nymphs of brown planthopper; b shows the mortality rate of short-winged adults of brown planthopper).

[0042] Figure 2 The content of high vanillic acid in the leaves and stems of rice Zhonghua No. 11.

[0043] Figure 3 It is the ratio of the expression levels of related genes in the stems and leaves of rice.

[0044] Figure 4 The mortality rates of 3rd and 5th instar nymphs of brown planthoppers after feeding on leaves of different rice plants (Figure a shows the mortality rate of 3rd instar nymphs of brown planthoppers; Figure b shows the mortality rate of 5th instar nymphs of brown planthoppers; the C04 and F06 gene knockout mutants in the figures represent rice glyceraldehyde phosphate dehydrogenase gene and acetaldehyde dehydrogenase gene knockout mutant plants, respectively).

[0045] Figure 5 The mortality rates of 3rd and 5th instar nymphs of brown planthoppers after feeding on leaves of different rice plants (Figure a shows the mortality rate of 3rd instar nymphs of brown planthoppers; Figure b shows the mortality rate of 5th instar nymphs of brown planthoppers; the C04 and F06 gene overexpression mutants in the figures represent rice overexpression mutant plants of glyceraldehyde phosphate dehydrogenase gene and acetaldehyde dehydrogenase gene, respectively). DETAILED DESCRIPTION

[0046] The present invention is further described below in conjunction with the accompanying drawings and specific examples, but the examples do not limit the present invention in any form. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art.

[0047] Unless otherwise specified, the reagents and materials used in the following examples are commercially available.

[0048] The artificial feed in Example 1 contains amino acids, vitamins, inorganic salts and sucrose. The specific formula is referred to the literature "Fu Q, Zhang Z, Hu C, Lai F, Sun Z, 2001. A chemically defined diet enables continuous rearing of the brown planthopper Nilaparvata lugens (Homoptera: Delphacidae). Appl. Entomol. Zool., 36(1): 111-116.》MMD-1 formula.

[0049] The knockout and overexpression of target genes in Example 2 were commissioned to BioGene Technology Co., Ltd.

[0050] Example 1 Determination of the toxicity of homovanillic acid to brown planthopper

[0051] 1. Experimental Methods

[0052] Take the 4th instar nymphs and short-winged adults of brown planthoppers and put them into insect bottles (350 mL mineral water bottles, remove the bottom of the bottle and stick a 40-mesh gauze), cover the bottle mouth with a double-layer parafilm sealing film, and add artificial feed in the middle of the sealing film for brown planthoppers to eat. Observe the mortality of brown planthoppers every day.

[0053] 2. Experimental Results

[0054] When brown planthoppers were fed with artificial diets containing 0.5-30 ppm homovanillic acid, the mortality rates of 4th instar nymphs and short-winged adults of brown planthoppers on the 6th day increased by 2.5%-45.1% compared with the mortality rates when fed with the control group (with 0 ppm homovanillic acid added).

[0055] When brown planthoppers were fed with artificial diets containing 0, 0.9, and 17 ppm homovanillic acid (i.e., the final concentrations of homovanillic acid in the artificial diets were 0, 0.9, and 17 ppm, respectively), the mortality of 4th instar nymphs and short-winged adults of brown planthoppers was as follows: Figure 1 The results showed that when the 4th instar nymphs of brown planthoppers fed on artificial diets containing the same high vanillic acid content as in rice stems (0.9 ppm), the mortality rate on the 6th day increased by 22.0% compared with the mortality rate of the control group, and when they fed on artificial diets containing the same high vanillic acid content as in rice leaves (17 ppm), the mortality rate on the 6th day increased by 136.4%. This indicates that the secondary metabolite of rice, high vanillic acid, can cause a large number of deaths in brown planthoppers.

[0056] Example 2 Effect of GAPDH or ALDH gene mutation on the content of high vanillic acid in rice plants

[0057] 1. Experimental Methods

[0058] (1) Using the rice variety Zhonghua 11 as the background, its LOC_Os03g03720 gene (glyceraldehyde phosphate dehydrogenase gene) was knocked out and overexpressed.

[0059] The rice variety Zhonghua 11 was used as the background, and its LOC_Os01g40870 gene (acetaldehyde dehydrogenase gene) was knocked out and overexpressed.

[0060] After knocking out and overexpressing the glyceraldehyde phosphate dehydrogenase gene and acetaldehyde dehydrogenase gene in rice, respectively, plants with glyceraldehyde phosphate dehydrogenase gene knockout (LOC_Os03g03720), plants with glyceraldehyde phosphate dehydrogenase gene overexpression, plants with acetaldehyde dehydrogenase gene knockout (LOC_Os01g40870) and plants with acetaldehyde dehydrogenase gene overexpression were obtained.

[0061] The nucleotide sequence of the LOC_Os03g03720 gene (glyceraldehyde phosphate dehydrogenase gene) is shown in SEQ ID NO.1, SEQ ID NO.1:

[0062]

[0063] The cDNA sequence of LOC_Os03g03720 gene (glyceraldehyde phosphate dehydrogenase gene) is shown in SEQ ID NO.2, SEQ ID NO.2:

[0064]

[0065] The amino acid sequence of the LOC_Os03g03720 gene (glyceraldehyde phosphate dehydrogenase gene) is shown in SEQ ID NO.3, SEQ ID NO.3:

[0066] MATHAALAASRIPATARLHSKAASKQRVDFADFSGLRPGSCSISHAAREASFSDVLGSQLVARATGENAVRAPAEAKLKVAINGFGRIGRNFLRCWHERENSPLEVVVVND SGGVRNASHLLKYDSMLGTFKADVKIVDDQTISVDGKLIKVVSNRDPLKLPWAELGIDIVIEGTGVFVDGPGAGKHIQAGAKKVIITAPAKGADIPTYVLGVNEGDYSHEVA NIISNASCTTNCLAPFVKILDEEFGIVKGTMTTTHSYTGDQRLLDASHRDLRRARAAALNIVPTSTGAAKAVALVLPQLKGKLNGIALRVPTPNVSVVDLVINTVKTGITA DDVNAAFRKAAAGPLSGILDVCDVPLVSVDFRCSDVSSTIDASLTMVMGDDMVKVVAWYDNEWGYSQRVVDLAHLVASKWPGAAVQGSGDPLEDFCKDNPETDECKVYEN*.

[0067] The nucleotide sequence of the LOC_Os01g40870 (acetaldehyde dehydrogenase gene) gene is shown in SEQ ID NO.4, SEQ ID NO.4:

[0068]

[0069] The cDNA sequence of the LOC_Os01g40870 (acetaldehyde dehydrogenase gene) gene is shown in SEQ ID NO.5, SEQ ID NO.5:

[0070]

[0071] The amino acid sequence of the LOC_Os01g40870 (acetaldehyde dehydrogenase gene) gene is shown in SEQ ID NO.6, SEQ ID NO.6:

[0072] MGSTGDCNGKAAAGGGGLVVPEIKFTKLFINGEFVDAASGKTFKTRDPRTGDVLAHIAEADKADVDLAVKAAREAFEHGKWPRMSGYERSRVMNKLADLVEQHADELAALDGADAGKLLTLGKIID MPAAAQMMRYYAGAADKIHGESLRVAGKYQGYTLREPIGVVGVIIPWNNFPTMMFFLKVSPALAAGCTIVVKPAEQTPLSALYYAHLAKLAGVPDGVINVVPGFGPTAGAALSSHMDVDSVAFTGSAE IGRAIMESAARSNLKNVSLELGGKSPMIVFDDADVDMAVSLSSLAVFFNKGEICVAGSRVYVQEGIYDEFVKKAVEAAKNWKVGDPFDAATNMGPQVDKVQFERVLKYIEIGKNEGATLLTGGKPTG DKGYYIEPTIFVDVKEEMTIAQEEIFGPVMSLMKFKTVEEAIEKANCTKYGLAAGIVTKNLNIANMVSRSVRAGTVWVNCYFAFDPDAPFGGYKMSGFGRDQGMVAMDKYLQVKTVITAVPDSPWY*.

[0073] (2) Determination of homovanillic acid content and target gene expression level in rice

[0074] HPLC was used to detect the high vanillic acid content in the wild-type rice Zhonghua No. 11, as well as the high vanillic acid content in plants with glyceraldehyde phosphate dehydrogenase gene knockout, glyceraldehyde phosphate dehydrogenase gene overexpression, acetaldehyde dehydrogenase gene knockout and acetaldehyde dehydrogenase gene overexpression based on this background.

[0075] Preparation of standard curve: Weigh an appropriate amount of homovanillic acid sample and add it to a solvent (a mixture of methanol and potassium dihydrogen phosphate, V:V = 25:75) to prepare homovanillic acid solutions with concentrations of 0.5, 1.5, 4.5, 13.5, 40.5, and 121.5 ppm. First, take 20 μL of the solvent for sample loading to obtain the peak area of ​​the solvent, and then take 20 μL of the prepared solution for sample loading to obtain the peak area of ​​the sample solution. The peak area of ​​the homovanillic acid sample is the peak area of ​​the sample solution minus the peak area of ​​the solvent. The standard curve of the standard sample is obtained by performing linear regression analysis between the concentration of homovanillic acid and its peak area.

[0076] HPLC detection conditions: DAD detector. C18 chromatographic column: 250*4.6mm; 5μm. Column temperature 25℃, flow rate: 1ml / min. Injection volume: 20μL. Wavelength: 280nm. Mobile phase: methanol: potassium dihydrogen phosphate (0.01mol / L) = 25:75 elution.

[0077] The leaves and stems of rice were ground into powder with liquid nitrogen respectively, a certain amount (3 g) of the sample was weighed into a centrifuge tube, an appropriate amount of methanol (30 mL) was added, and the sample was placed in an oscillator for thorough shaking and mixing for 1-2 min, and then centrifuged at 4000 r / min for 10 min, and the supernatant was transferred to a new centrifuge tube. The above operation was repeated several times, the supernatants were combined and mixed, dried with nitrogen at room temperature, and the supernatant was taken after dissolving an appropriate amount of methanol and filtered through a 0.22 μm filter membrane, and 20 μL of the sample was taken for liquid chromatography detection.

[0078] At the same time, fluorescence quantitative PCR was used to detect the expression levels of target genes in the leaves of wild-type, knockout mutant and overexpression mutant plants of Zhonghua No. 11.

[0079] 2. Experimental Results

[0080] HPLC detection of homovanillic acid content in leaves and stems of wild-type rice Zhonghua No. 11 showed that the content of homovanillic acid in wild-type rice Zhonghua No. 11 was as follows: Figure 2 As shown, the results showed that the content of homovanillic acid in rice leaves and stems was 16.3-19.6 ppm and 0.57-1.2 ppm, respectively, and the content of homovanillic acid in the stems was only 3%-7% of that in the leaves.

[0081] Fluorescence quantitative PCR was used to detect the expression levels of glyceraldehyde phosphate dehydrogenase gene (LOC_Os03g03720 gene) and acetaldehyde dehydrogenase gene (LOC_Os01g40870 gene) in the leaves and stems of wild-type rice Zhonghua No. 11. The test results are as follows Figure 3As shown, the results showed that the expression levels of glyceraldehyde phosphate dehydrogenase gene and acetaldehyde dehydrogenase gene in rice stems were only 0.59 times and 0.01 times of the expression levels in leaves, which indicates that the content of high vanillic acid in rice leaves is high, and the expression level of glyceraldehyde phosphate dehydrogenase gene (LOC_Os03g03720 gene) or acetaldehyde dehydrogenase gene (LOC_Os01g40870 gene) related to its biosynthesis is also high.

[0082] Therefore, detecting the expression levels of glyceraldehyde phosphate dehydrogenase gene (LOC_Os03g03720 gene) and acetaldehyde dehydrogenase gene (LOC_Os01g40870 gene) can reflect the high vanillic acid content in rice, and the expression levels of glyceraldehyde phosphate dehydrogenase gene (LOC_Os03g03720 gene) and acetaldehyde dehydrogenase gene (LOC_Os01g40870 gene) can be used as detection indicators of rice resistance to brown planthoppers.

[0083] HPLC and fluorescence quantitative PCR were performed on the leaves and stem tissues of the knockout or overexpression plants of glyceraldehyde phosphate dehydrogenase gene (LOC_Os03g03720 gene) or aldehyde dehydrogenase gene (LOC_Os01g40870 gene). The results are shown in Table 1. The results showed that compared with the wild-type plants, the homovanillic acid content in rice leaves decreased by 84% after knockout of glyceraldehyde phosphate dehydrogenase or aldehyde dehydrogenase gene, and the homovanillic acid content in rice stems increased by 27% after overexpression of glyceraldehyde phosphate dehydrogenase or aldehyde oxidase gene. This indicates that regulating the expression of glyceraldehyde phosphate dehydrogenase or aldehyde dehydrogenase gene in rice can achieve the regulation of the biosynthesis of homovanillic acid, a secondary metabolite of rice.

[0084] Table 1 Expression levels of target genes and homovanillic acid content in wild-type, knockout and overexpression plants

[0085]

[0086] Note: The relative expression of the target gene in the above table is the ratio of the expression level in the mutant to that in the wild type, "+" indicates upregulation or increase, and "-" indicates downregulation or decrease. The change in the content of homovanillic acid is the rate of change of the content of homovanillic acid, and the specific calculation formula is:

[0087]

[0088] Example 3 Determination of the toxicity of plants with mutations in glyceraldehyde phosphate dehydrogenase or acetaldehyde dehydrogenase genes related to homovanillic acid biosynthesis to brown planthoppers

[0089] 1. Experimental Methods

[0090] The rice used in this experiment was the wild type Zhonghua No. 11 and mutant plants with glyceraldehyde phosphate dehydrogenase or acetaldehyde dehydrogenase gene knocked out or overexpressed. The 3rd and 5th instar nymphs of brown planthoppers were placed in small insect bags, which were covered on the leaves or stems of rice so that the pests could only feed on the leaves or stems of rice in the insect bags. The death of brown planthoppers in different insect bags was observed every day.

[0091] 2. Experimental Results

[0092] Mortality of brown planthoppers after feeding on rice leaves of target gene knockout mutants Figure 4 As shown, the results show that:

[0093] Compared with feeding only on wild-type rice leaves, the mortality rate of third-instar nymphs of brown planthoppers that feed only on rice leaves with glyceraldehyde phosphate dehydrogenase gene knockout mutant can be reduced by 75%, and the mortality rate of fifth-instar nymphs can be reduced by 70%; the mortality rate of third-instar nymphs of brown planthoppers that feed only on rice leaves with acetaldehyde dehydrogenase gene knockout mutant can be reduced by 67.5%; the mortality rate of fifth-instar nymphs can be reduced by 67%.

[0094] Mortality of brown planthoppers after feeding on rice leaves of target gene overexpression mutants Figure 5 The results showed that if only the rice stems of the plants overexpressing glyceraldehyde phosphate dehydrogenase gene were fed, the mortality of the 3rd instar nymphs of the brown planthopper could increase by 40%, and the mortality of the 5th instar nymphs could increase by 50%; if only the rice stems of the plants overexpressing acetaldehyde oxidase gene were fed, the mortality of the 3rd instar nymphs of the brown planthopper could increase by 70%, and the mortality of the 5th instar nymphs could increase by 78%.

[0095] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. Application of a rice brown planthopper resistance gene in regulating rice resistance to brown planthoppers, characterized in that: The rice brown planthopper resistance gene includes LOC_Os03g03720 gene and / or LOC_Os01g40870 gene; the nucleotide sequence of LOC_Os03g03720 gene is a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO.3; the nucleotide sequence of LOC_Os01g40870 gene is a nucleotide sequence encoding the amino acid sequence shown in SEQ ID NO.

6.

2. Use of the rice brown planthopper resistance gene as claimed in claim 1 as a target in screening brown planthopper resistance rice varieties.

3. Use of the rice brown planthopper resistance gene of claim 1 in regulating the high vanillic acid content of rice.

4. Use of the agent for promoting the expression of the rice brown planthopper resistance gene according to claim 1 in improving the resistance of rice to brown planthoppers.

5. An agent for improving the resistance of rice to brown planthoppers, characterized in that: Contains an agent for promoting the expression of the rice brown planthopper resistance gene as claimed in claim 1.

6. Use of the rice brown planthopper resistance gene of claim 1 or the reagent of claim 5 in constructing a rice variety with enhanced resistance to brown planthoppers, or in improving the resistance of rice to brown planthoppers.

7. Use of the rice brown planthopper resistance gene of claim 1 or the reagent of claim 5 in the preparation of a product for improving rice resistance to brown planthoppers.

8. Use of the rice brown planthopper resistance gene of claim 1 or the reagent of claim 5 in increasing the high vanillic acid content of rice, or in preparing a product with increased high vanillic acid content of rice, or in constructing rice rich in high vanillic acid.

9. A method for constructing a rice variety with enhanced resistance to brown planthoppers, characterized in that: Overexpressing the rice brown planthopper resistance gene of claim 1 in a plant.

10. A method for constructing a rice variety rich in high vanillic acid, characterized in that: Overexpressing the rice brown planthopper resistance gene of claim 1 in a plant.

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