Application, methods, and recombinant vectors of the pepper CaWRKY37 gene or its encoded protein in regulating carotenoid content in peppers.

Abstract

This application relates to the field of crop genetics and breeding technology, and discloses the application, method, and recombinant vector of the pepper CaWRKY37 gene or its encoded protein in regulating the carotenoid content of pepper. The pepper... CaWRKY37 The nucleotide sequence of the gene is shown in SEQ ID NO:1, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO:2. This invention also discloses a method for reducing the carotenoid content of capsaicin by inhibiting or silencing capsaicin. CaWRKY37 Gene expression to reduce capsaicin carotenoid content. The recombinant vector of this invention contains capsaicin... CaWRKY37 Genes or specific fragments thereof. This application found... CaWRKY37 It plays a key regulatory role in the carotenoid metabolic pathway of peppers, which has important theoretical significance and application potential for in-depth analysis of the molecular mechanism of pepper color formation and related molecular breeding.

Description

Technical Field

[0001] This application relates to the field of crop genetics and breeding technology, specifically to a chili pepper. CaWRKY37 Applications, methods, and recombinant vectors of genes or their encoded proteins in regulating capsicum carotenoid content. Background Technology

[0002] chili( Capsicum spp. Capsicum, belonging to the genus Capsicum in the family Solanaceae, is an important vegetable crop. Its fruits are not only rich in active ingredients such as capsaicin and carotenoids, but also exhibit outstanding economic value and nutritional benefits. Fruit color, a key indicator for evaluating chili quality, primarily depends on the composition and accumulation level of carotenoids. Capsanthin and capsicum rubigin account for 30%–80% of this type of pigment and are the main sources of pigments for red fruit formation. Therefore, a thorough understanding of the molecular regulatory mechanisms of carotenoid synthesis is crucial for the genetic improvement of chili fruit color.

[0003] The biosynthesis of carotenoids belongs to the isoprene metabolic pathway, and its transcriptional regulation mechanism is complex, with limited identification of key regulatory factors. Virus-induced gene silencing (VIGS) and other techniques have been widely used for rapid gene function identification. By silencing target genes and combining this with pigment content analysis, their regulatory roles in carotenoid accumulation can be effectively determined. Therefore, using gene silencing techniques to identify and analyze the functions of relevant regulatory genes is of great significance for elucidating the mechanism of pepper fruit color formation. Summary of the Invention

[0004] The present invention aims to solve the problems in the prior art and provide a chili pepper CaWRKY37 Applications, methods, and recombinant vectors of genes or their encoded proteins in regulating capsicum carotenoid content.

[0005] To achieve the above objectives, the first aspect of this application provides a chili pepper. CaWRKY37 The application of genes or their encoded proteins in reducing the carotenoid content of capsicum peppers. CaWRKY37 The nucleotide sequence of the gene is shown in SEQ ID NO:1, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO:2.

[0006] The second aspect of this application provides a chili pepper CaWRKY37 The application of genes or their encoded proteins in genetic breeding for reducing carotenoid content in peppers, wherein the pepper... CaWRKY37 The nucleotide sequence of the gene is shown in SEQ ID NO:1, and the amino acid sequence of the protein it encodes is shown in SEQ ID NO:2.

[0007] A third aspect of this application provides a chili pepper CaWRKY37The application of genes as molecular markers in carotenoid-assisted breeding of peppers, wherein the pepper CaWRKY37 The CDS sequence of the gene is shown in SEQ ID NO:1.

[0008] The fourth aspect of this application provides a method for reducing the carotenoid content of capsaicin, by inhibiting or silencing capsaicin... CaWRKY37 Gene expression is reduced to decrease the carotenoid content in capsicum.

[0009] The above method, preferably, involves using ingredients containing chili peppers. CaWRKY37 Gene recombinant vectors are introduced into pepper tissues to regulate the content of carotenoids in peppers.

[0010] In the above method, preferably, the recombinant vector is a virus-induced gene silencing vector.

[0011] In the above method, preferably, the virus-induced gene silencing vector is pTRV2-CaWRKY37.

[0012] A fifth aspect of this application provides a recombinant vector comprising chili pepper. CaWRKY37 The gene or its specific fragment, the chili pepper CaWRKY37 The CDS sequence of the gene is shown in SEQ ID NO:1; the nucleotide sequence of the specific fragment is shown in SEQ ID NO:5.

[0013] Preferably, the recombinant vector described above is a virus-induced gene silencing vector.

[0014] Preferably, the virus-induced gene silencing vector described above is pTRV2-CaWRKY37.

[0015] Compared with the prior art, this application has the following beneficial effects:

[0016] (1) The gene C involved in this invention aWRKY37 Its encoded proteins can regulate the biosynthesis and accumulation of carotenoids in pepper fruits. In the pepper variety *C. annuum* Zhangshugang'S8, this regulation is achieved through silencing... CaWRKY37 Genetic analysis revealed a significant decrease in the total carotenoid content of the fruit during its red-ripe stage, specifically a marked reduction in the content of key components such as lycopene, α-carotene, β-carotene, γ-carotene, and capsanthin. These results indicate that... CaWRKY37 This discovery plays a key regulatory role in the carotenoid metabolic pathway in peppers. It has important theoretical significance and application potential for a deeper understanding of the molecular mechanism of pepper color formation and for carrying out related molecular breeding.

[0017] (2) The invention identifiedCaWRKY37 The genes and their encoded proteins are closely related to the carotenoid content of pepper fruits and can be used as molecular markers for the screening and identification of pepper germplasm resources. Attached Figure Description

[0018] To more clearly illustrate the technical solutions in the embodiments of this application or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of this application. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0019] Figure 1 The control plants and in Example 2 of this invention CaWRKY37 Comparison of fruit color at red ripening stage in gene-silenced plants.

[0020] Figure 2 The figures show the relative expression levels of the CaWRKY37 gene and key genes in the carotenoid biosynthesis pathway in the control plants and silent plants in Example 3 of this invention. The data in the figure are expressed as mean ± standard deviation (n=3). The significance analysis was performed using the t-test (ns, no significant difference). p<0.05; p<0.01; (p<0.001).

[0021] Figure 3 This is a comparison of the content of some carotenoid substances in the control plants and silent plants in Example 3 of the present invention. Detailed Implementation

[0022] To facilitate understanding of this application, the following description will be more comprehensive and detailed in conjunction with the accompanying drawings and preferred embodiments, but the scope of protection of this application is not limited to the following specific embodiments.

[0023] Unless otherwise defined, all technical terms used herein have the same meaning as commonly understood by those skilled in the art. The technical terms used herein are for the purpose of describing particular embodiments only and are not intended to limit the scope of this application.

[0024] Unless otherwise specified, all raw materials, reagents, instruments and equipment used in this invention can be purchased from the market or prepared by existing methods.

[0025] This invention provides an in-depth investigation into the biosynthesis of carotenoids in chili pepper fruits. Carotenoid biosynthesis begins with geraniol pyrophosphate (GGPP), which is catalyzed by enzymes such as phytoene synthase (PSY), phytoene dehydrogenase (PDS), and zeta-carotene dehydrogenase (ZDS) to produce lycopene. Lycopene is a key node in this metabolic pathway, being converted to β-carotene under the action of lycopene β-cyclase (LCYB), or ultimately to capsanthin and capsanthin by capsanthin-capsulopene synthase (CCS). The expression levels of these genes and the activities of these enzymes directly determine the composition and content of carotenoids. Transcription factors play a central regulatory role in carotenoid biosynthesis. They directly activate or inhibit the transcription of these key synthase genes by specifically binding to the promoter regions of target genes (such as PSY and LCYB), thereby precisely controlling the activity and flow of the synthetic pathway. In addition, various transcription factors can cooperate or compete with each other to form a complex regulatory network that integrates endogenous hormones (such as ABA) and external environmental signals (such as light and temperature), ultimately dynamically regulating the composition and accumulation of carotenoids, which directly affects the color, resistance and nutritional value of plants.

[0026] WRKY transcription factors are a key class of transcription regulators in plants, named after the highly conserved heptapeptide sequence (WRKYGQK) in their DNA-binding domain. As one of the most numerous and functionally diverse families of transcription factors in the plant genome, they activate or inhibit downstream gene expression by specifically recognizing and binding to W-box cis-acting elements (with the core sequence TTGACC / T) in the promoter regions of target genes. Structurally, these proteins typically contain a WRKY domain of approximately 60 amino acids, with a conserved WRKYGQK motif at the N-terminus and a zinc finger structure at the C-terminus, all participating in DNA binding and protein-DNA interactions. WRKY transcription factors play a central regulatory role in various plant life processes, participating not only in growth and development, organ senescence, and the synthesis of secondary metabolites, but also extensively mediating plant hormone signal transduction and responses to biotic and abiotic stresses. They are of great significance in enhancing plant environmental adaptability, influencing species distribution patterns, and promoting agricultural and livestock production.

[0027] The gene CaWRKY37 is a member of the WRKY family. Its CDS sequence is shown in SEQ ID NO:1, and the amino acid sequence of the encoded protein is shown in SEQ ID NO:2, as follows: SEQ ID NO:1 ATGTTTGTCTATGAATTATTTTCAAGTCTTCAAAAAGTGGCTCATACCAGTTTTTGTGACTTTCACTTACCAACTTCAAATATTTTTCAAGTAAAATGCATATCTAGAAGTTTTAAGTTTCAACTTCAAAATTTATCGGCAAATGGAGCATATATTTGTAGTTTGGTCATTGTTTTTTGTTTCCATACACAGAGATATGTGGAGCATGAAAAGGGGACTTTTTTCTGTTTTTTCTGACAGCTCTGAGTGTCCTGATATTAGGAAGATTGGTCAGAAAGTTGTGTTCAAAGTGCAAATGGAAGGAAAAGCTATAAAGCAGAAAAATGAAGGGCCTCCTTCAGATTGTTGGTCCTGGAGAAAATATGACAGAAACCCATTAAAGGTTCACCTTATCCCAGGGGATACTACAGATGCAGCAGTTCAAAGGGTTGTTCAGCTAAAAAA CAAGTAGAAAGGTGCAGTAAAGATGCATCCTTGTTCATCATCACATACACATCCAGCCATAATCATCCAGGTCCAAACTTGCCTAATAATAAAGACACAGTCACACAAGATTCCACCGCGACTCTGCCGCAGCAAGATCAAGAACCAACAGTGGATAAAGACGTGCCCCTCAAAGATAATGGGGATAGTACTACTACTCCTACTACTGCCACCAGCATCTCC CAAGGCATACCTGAAGAGAATTTGTTCACTGATAGTTTCTTGGGAACAATTTCATATGATGATTTTCTGCCCCTTTCTTACCCTCAACTAATGGAATTCCCAAAATCCGAATTGTCAGAAGAAAATGACTTCTATGATGAACTGGGAGAATTGCAACTACCTCCATCTTCTACGTCCTTCGCAGGCATTTTTGAGGAGGCAATCCTTGTAGATCCCTCTTAG SEQ ID NO:2 MFVYELFSSLQKVAHTSFCDFHLPTSNIFQVKCISRSFKFQLQNLSANGAYICSLVIVFCFHTQRYVSMKRGLFSVFSDSSECPDIRKIGQKVVFKVQMEGKAIKQKNEGPPSDCWSWRKYGQKPIKGSPYPRGYYRCSSSKGCSAK KQVERCSKDASLFIITYTSSHNHPGPNLPNNKDTVTQDSTATLPQQDQEPTVDKDVPLKDNGDSTTTPTTATSISQGIPEENLFTDSFLGTISYDDFLPLSYPQLMEFPKSELSEENDFYDELGELQLPPSSTSFAGIFEEAILVDPS Example 1: Chili Pepper Gene CaWRKY37 Cloning 1. Plant materials and planting environment The tested chili pepper variety was *C. annuum* 'S8' (Zhangshugang), collected from Changsha, Hunan Province, China (28°11′49″N, 112°58′429″E). It was cultivated in an artificial climate chamber under the following specific growing conditions: a photoperiod of 16 hours of light (25°C) and 8 hours of darkness (20°C), and a light intensity of 300 µmol / m² / s. -2-1 The relative humidity was 65%. Fruits at the red-ripe stage were used as material for gene cloning.

[0028] 2. Total RNA extraction and cDNA first-strand synthesis Total RNA was extracted from red-ripe pepper fruits using the Eastep@Super Total RNA Extraction Kit (Promega (Beijing) Biotechnology Co., Ltd., catalog number LS1040). Subsequently, using this RNA as a template, reverse transcription was performed to synthesize the first strand of cDNA according to the operating procedures of Evo Super M-MLV Plus 1stcDNA Synthesis Premix (Aikerui Biotechnology, catalog number AG11623).

[0029] 3. CaWRKY37 PCR amplification and cloning of genes Based on the predicted sequence of the CaWRKY37 gene from the Zhangshugang chili genome database, specific primers were designed using PrimerPremier 5 software to amplify its full-length coding region. The primer sequences are as follows: Upstream primer CaWRKY37-F: 5'- ATGTTTGTCTATGAATTATTTTCAAGTCTTCAAAAAGTGGC-3' (as shown in SEQ ID NO:3); Downstream primer CaWRKY37-R: 5'- CTAAGAGGGATCTACAAGGATTGC-3' (as shown in SEQ ID NO:4).

[0030] Using the cDNA obtained in step 2 as a template, PCR amplification was performed using 2 × Phanta Max Master Mix (Dye Plus) (Nanjing Novizan Biotechnology Co., Ltd., catalog number P525). The specific PCR reaction system and procedure were followed according to the reagent instructions. After separation of the PCR products by 1% agarose gel electrophoresis, the target fragment was recovered using the SteadyPureDNA Gel Recovery Kit (Aikerui Biotechnology, catalog number AG21006).

[0031] Based on the TOPO cloning principle, a recombinant cloning vector containing a full-length CDS sequence was constructed using the 5-min TA / Blunt-Zero Cloning Kit (Nanjing Novizan Biotechnology Co., Ltd., catalog number C601). The recombinant product was transformed into E. coli DH5α competent cells, and the cells were plated on LB agar plates containing 50 mg / L kanamycin using the heat shock method and cultured overnight at 37°C until positive single clones grew. Positive clones were then picked and sequenced.

[0032] Sequencing results showed that the obtained fragment was 1000 bp in length and contained the complete CaWRKY37 coding region sequence, the nucleotide sequence of which is shown in SEQ ID NO:1 and the amino acid sequence of the encoded protein is shown in SEQ ID NO:2.

[0033] Example 2: Silent Genes CaWRKY37 Construction and genetic transformation of VIGS vector 1. Construction of VIGS carrier The CaWRKY37-specific fragment for VIGS technology was identified using the gene CDS sequence on the Solanaceae gene website https: / / vigs.solgenomics.net / . The specific fragment is nucleotides 400 to 699 of the sequence shown in SEQ ID NO:1, with a length of 300 bp. The nucleotide sequence is shown in SEQ ID NO:5, as follows: SEQ ID NO:5 GGATACTACAGATGCAGCAGTTCAAAGGGTTGTTCAGCTAAAAAACAAGTAGAAAGGTGCAGTAAAGATGCATCCTTGTTCATCACATACACATCCAGCCATAATCATCCAGGTCCAAACTTGCCTAATAATAAAGACACAGTCACA CAAGATTCCACCGCGACTCTGCCGCAGCAAGATCAAGAACCAACAGTGGATAAAGACGTGCCCCTCAAAGATAATGGGGATAGTACTACTACTCCTACTACTGCCACCAGCATCTCCCAAGGCATACCTGAAGAGAATTTGTTCACTGAT Using the recombinant cloning plasmid (containing the full-length CDS shown in SEQ ID NO:1) that was correctly sequenced in Example 1 as a template, specific primers were designed using Primer Premier 5 software to amplify the silencing target fragment of the CaWRKY37 gene.

[0034] The amplification primer sequences are as follows: Upstream primer (VIGS-CaWRKY37-F): 5′-tgaggagaagagcccGGATACTACAGATGCAGCAG -3′ (as shown in SEQ ID NO:6); Downstream primer (VIGS-CaWRKY37-R): 5′- gtcgacgacaagacccATCAGTGAACAAATTCTCTTCAGG -3′ (as shown in SEQ ID NO:7); PCR amplification, gel electrophoresis, and gel recovery were performed as described in Example 1.

[0035] The recovered PCR product was inserted into the pTRV2 vector using homologous recombination. Specific steps: The pTRV2 vector was linearized using the Small1 restriction endonuclease, and the linearized vector was recovered via gel electrophoresis. The linearized vector and the recovered PCR product were mixed at a 1:3 molar ratio, and homologous recombinase (Jinsha Biotechnology, catalog number SC612) was added. The mixture was incubated at 50°C for 15 min.

[0036] For E. coli transformation, referencing Example 1, PCR identification of the selected positive single colonies was performed using 2 × Rapid Taq Master Mix (Nanjing Novizan Biotechnology Co., Ltd., catalog number P222). The recombinant plasmid that was identified and verified by sequencing was named pTRV2-CaWRKY37 and used for subsequent experiments.

[0037] 2. Agrobacterium-mediated VIGS injection The pTRV2-CaWRKY37 plasmid and the empty vectors pTRV1 and pTRV2 were transformed into Agrobacterium GV3101 competent cells using a freeze-thaw method. The transformed cells were plated on LB agar plates containing 50 mg / L kanamycin and 50 mg / L rifampin and incubated at 28°C for 48 h. Single colonies were picked and inoculated into liquid medium. Positive monoclonal Agrobacterium cultures, confirmed by PCR, were mixed with an equal volume of sterilized 50% glycerol (1:1) and stored at -80°C.

[0038] Bacterial suspensions carrying pTRV1, pTRV2 empty vectors, and the pTRV2-CaWRKY37 recombinant plasmid were inoculated into 30 mL of LB broth. The broth was supplemented with 50 mg / L kanamycin, 50 mg / L rifampin, 10 mM MES, and 20 μM AS, and incubated overnight at 28 °C and 180 rpm. After centrifugation to collect the bacterial cells, they were resuspended in injection suspension (containing 10 mM MgCl2, 10 mM MES, and 200 μM AS), and the OD600 was adjusted to 0.8. The suspensions were then incubated at 28 °C in the dark for 3 hours. The bacterial suspensions containing pTRV1 were then thoroughly mixed with bacterial suspensions containing either the pTRV2 empty vector (as a control) or pTRV2-CaWRKY37 at a 1:1 ratio.

[0039] Select 'S8' pepper seedlings with uniform growth and fully expanded cotyledons. Using a needleless syringe, inject the mixed bacterial solution into the cotyledons from the underside of the leaves until obvious infiltration spots appear. Injected plants were cultured in the dark and under humid conditions for 48 hours, then transferred to normal light conditions (16 hours light / 8 hours dark, 25°C) for continued growth. At least 16 seedlings were injected for each treatment.

[0040] 3. Silence efficiency test and phenotypic observation Approximately 30 days after injection (during the flowering and fruiting stage of the plant), RNA was extracted from newly emerging leaves, and the expression level of the CaWRKY37 gene was detected by qRT-PCR to confirm the silencing efficiency. When the fruits of the silenced plants reached the red-ripe stage, RNA was extracted from the red-ripe fruits, and the expression level of the CaWRKY37 gene was detected by qRT-PCR to confirm the silencing efficiency. Simultaneously, fruit color changes were observed and recorded, and images were acquired for analysis.

[0041] The results are as follows Figure 1 As shown, compared with the control plants injected with the pTRV2 empty vector, the CaWRKY37 silent plants showed a significant reduction in red pigment accumulation in the ripe fruit, and the fruit peel turned yellow.

[0042] Example 3: CaWRKY37 Molecular identification and carotenoid content analysis of silent plants 1. Fruit sample collection and RNA extraction Fruits at the red-ripe stage were collected from control plants injected with the empty pTRV2 vector and silent plants injected with pTRV2-CaWRKY37 in Example 2 (3 plants per treatment, 3 fruits per plant, pooled as one biological replicate, for a total of 3 biological replicates). After flash freezing in liquid nitrogen, the fruits were stored at -80°C. Total RNA extraction was performed using the same method as in Example 1.

[0043] 2. Real-time quantitative PCR (qRT-PCR) analysis use Evo M-MLV reverse transcription reagent premix (for qPCR) (Aikerui Biotechnology, catalog number AG11706) reverse transcribes the above RNA into cDNA.

[0044] Specific quantitative primers for the CaWRKY37 gene and key genes in the carotenoid synthesis pathway (PSY, PDS, ZDS, LCYB, LCYE, CHYB, ZEP, CYP97A, CYP97C) were designed using Primer Premier 5 software. These primers were then used as internal reference genes for chili peppers. Caz04g20840 The expression levels of [a specific substance] were normalized. The primer sequences are as follows: Table 1. Specific quantitative primer sequences

[0045] qRT-PCR reactions were performed on a QuantStudio 5 real-time quantitative PCR instrument using ChamQ UniversalSYBR qPCR Master Mix (Nanjing Novizan Biotechnology Co., Ltd., catalog number Q711). Three technical replicates were set up for each sample.

[0046] Use 2 -ΔΔCt The relative expression levels of each gene were calculated using the method with the expression level of the control plant as the baseline (set as 1). Data are expressed as mean ± standard deviation (n=3), and significance was analyzed by t-test (p<0.05; p<0.01; p<0.001).

[0047] The results are as follows Figure 2 As shown, compared with the control plants, the expression level of the CaWRKY37 gene in the fruits of CaWRKY37-silenced plants was significantly reduced. (p<0.001), and the expression levels of key carotenoid synthesis genes PSY, PDS, LCYB, CHYB, and CYP97C were also significantly decreased. This indicates that CaWRKY37 may affect carotenoid synthesis by regulating the expression of these structural genes.

[0048] 3. Determination of carotenoid content To determine the effect of gene silencing on carotenoid accumulation, fruit samples were collected from control plants and the CaWRKY37 silencing group at the red-ripe stage. These samples were then sent to Wuhan Maiwei Metabolic Biotechnology Co., Ltd. for qualitative and quantitative analysis of carotenoid components using ultra-high performance liquid chromatography (UPLC) (ExionLC™ AD, https: / / sciex.com.cn / ) and tandem mass spectrometry (MS / MS) (QTRAP® 6500+, https: / / sciex.com.cn / ). Three biological replicates were performed for each sample. Results are expressed as mean ± standard deviation, and t-tests were used to analyze the significance of differences between the two groups. Results are as follows: Figure 3 As shown, compared with the control plants, the total carotenoid content in the red-ripe fruit of CaWRKY37-silenced plants was significantly reduced (p<0.01), with the contents of major components such as lycopene, α-carotene, β-carotene, γ-carotene, and capsanthin all significantly decreased (p<0.01). Combined with gene expression data and color phenotype, this indicates that silencing the CaWRKY37 gene inhibits the expression of key genes in the carotenoid synthesis pathway, leading to reduced carotenoid accumulation and ultimately resulting in a yellow fruit color.

[0049] The above are merely preferred embodiments of this application. It should be noted that this application is not limited to the above embodiments. For those skilled in the art, several improvements and modifications can be made without departing from the principles of this application. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should also be considered within the scope of protection of this application.

Claims

1. Chili peppers CaWRKY37 The application of a gene or its encoded protein in reducing the carotenoid content of capsaicin, characterized in that, The pepper CaWRKY37 The nucleotide sequence of the gene is shown in SEQ ID NO: 1, and the amino acid sequence of the encoded protein is shown in SEQ ID NO:

2.

2. Chili peppers CaWRKY37 The application of a gene or its encoded protein in genetic breeding for reducing carotenoid content in peppers, characterized by: The pepper CaWRKY37 The nucleotide sequence of the gene is shown in SEQ ID NO: 1, and the amino acid sequence of the encoded protein is shown in SEQ ID NO:

2.

3. Chili peppers CaWRKY37 The application of genes as molecular markers in carotenoid-assisted breeding of peppers is characterized by, The chili pepper CaWRKY37 The CDS sequence of the gene is shown in SEQ ID NO:

1.

4. A method for reducing the carotenoid content of capsaicin, characterized in that, By inhibiting or silencing chili peppers CaWRKY37 Gene expression is reduced to decrease the carotenoid content in capsicum.

5. The method as described in claim 4, characterized in that, By including chili peppers CaWRKY37 Gene recombinant vectors are introduced into pepper tissues to regulate the content of carotenoids in peppers.

6. The method as described in claim 5, characterized in that, The recombinant vector is a virus-induced gene silencing vector.

7. The method as described in claim 6, characterized in that, The virus-induced gene silencing vector is pTRV2-CaWRKY37.

8. A recombinant vector, characterized in that, The recombinant vector contains chili peppers. CaWRKY37 The gene or its specific fragment, the chili pepper CaWRKY37 The CDS sequence of the gene is shown in SEQ ID NO:1; the nucleotide sequence of the specific fragment is shown in SEQ ID NO:

5.

9. The recombinant vector as described in claim 8, characterized in that, The recombinant vector is a virus-induced gene silencing vector.

10. The recombinant vector as described in claim 9, characterized in that, The virus-induced gene silencing vector is pTRV2-CaWRKY37.

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