Application of capsicum CaVAP27 gene and capsicum CaVAP27 protein in regulation and control of capsaicin content

Abstract

The invention discloses a capsicum CaVAP27 gene and application of a capsicum CaVAP27 protein in regulation and control of capsaicin content. A CDS sequence of the capsicum CaVAP27 gene is shown as SEQ ID NO: 1, and an amino acid sequence of the capsicum CaVAP27 gene is shown as SEQ ID NO: 2. It is found that the expression level of the VAMP related protein gene CaVAP27 is remarkably related to the blister number and the capsaicin content of pepper fruits, the pepper CaVAP27 gene may influence the accumulation level of capsaicin by influencing blister formation, and discovery of the pepper CaVAP27 gene function provides a theoretical basis for creation of pepper varieties for regulating and controlling the capsaicin substance content.

Description

Technical Field

[0001] This invention belongs to the field of crop genetics and breeding technology. Specifically, this invention relates to the application of the CaVAP27 gene and protein in chili peppers in regulating the capsaicin content of chili peppers. Background Technology

[0002] Chili peppers (Capsicum spp.) are the most widely cultivated and highest-value vegetable crop in my country. Capsaicin, a unique active substance in chili peppers, has high economic value. In addition to being used as a food and condiment, it also has significant medicinal, industrial, and military value.

[0003] The biosynthetic pathway and regulatory mechanism of capsaicin are relatively well understood. However, the transport process and molecular regulatory mechanism of capsaicin are still unclear. Therefore, it is very meaningful to explore genes related to capsaicin accumulation. Summary of the Invention

[0004] Based on this, the purpose of this invention is to provide the application of vesicle-associated membrane protein (VAMP) and its encoding gene in regulating the capsaicin content of chili peppers.

[0005] The specific technical solutions for achieving the above-mentioned objectives are as follows.

[0006] In a first aspect, the present invention provides the application of the chili pepper CaVAP27 gene in regulating the capsaicin content of chili peppers or in chili pepper genetic breeding that regulates the capsaicin content, the CDS sequence of which is shown in SEQ ID NO:1.

[0007] In a second aspect, the present invention provides the application of the chili CaVAP27 protein in regulating the capsaicin content of chili peppers or in chili pepper genetic breeding that regulates the capsaicin content, the amino acid sequence of which is shown in SEQ ID NO:2.

[0008] In a third aspect, the present invention provides the application of a silencing vector for the pepper CaVAP27 gene in regulating the capsaicin content of peppers or in pepper genetic breeding for regulating capsaicin content, wherein the CDS sequence of the pepper CaVAP27 gene is shown in SEQ ID NO:1.

[0009] In a fourth aspect, the present invention provides the application of engineered bacteria that transform the silencing vector of the CaVAP27 gene in chili peppers in the genetic breeding of chili peppers that regulate the capsaicin content or regulate the capsaicin content, wherein the CDS sequence of the CaVAP27 gene in chili peppers is shown in SEQ ID NO:1.

[0010] A fifth aspect of the present invention provides a method for reducing the capsaicin content of chili peppers, comprising the following steps: reducing the expression level of the chili pepper CaVAP27 gene in chili pepper plants.

[0011] The inventors of this invention have discovered that the expression level of the chili vesicle-associated protein (VAMP) gene CaVAP27 is significantly correlated with the number of vesicles and capsaicin content in chili fruits. The chili CaVAP27 gene may affect the accumulation level of capsaicin by influencing the formation of vesicles. The discovery of the function of the chili CaVAP27 gene provides a theoretical basis for the creation of chili varieties that regulate the content of capsaicin-like substances. Attached Figure Description

[0012] Figure 1 This diagram illustrates the changes in vesicle tissue on the diaphragm surface of chili peppers with different spiciness levels in Example 1 of this invention. A: Observation of vesicles on the diaphragm surface of chili peppers with different spiciness levels, where Z531 is a sweet pepper; CA59 is a medium-spiciness inbred line; and China Chili 839 is a high-spiciness inbred line. B: Total vesicle area in the ImageJ statistical graph.

[0013] Figure 2 This is a cellular structure analysis of pepper CA59 (C. annuum) at different developmental stages in Example 2 of the present invention. AD are cross-sectional views of pepper CA59 fruits at 16 DPA, 25 DPA, 30 DPA, and 38 DPA, respectively. I, II, and III are magnified views of corresponding parts of the cross-sections (I is the endocarp, II is the septum epidermis, and III is the septum-placental junction). D-IV are magnified views of another septum epidermis in section D.

[0014] Figure 3 This is a cellular structure analysis of Chinese pepper 839 (C. chinense) at different developmental stages in Example 2 of the present invention. AD represents cross-sectional views of Chinese pepper 839 fruit at 16 DPA, 25 DPA, 30 DPA, and 38 DPA, respectively. I, II, and III are magnified views of the corresponding parts of the cross-sections (I is the endocarp, II is the septum epidermis, and III is the septum-placental junction).

[0015] Figure 4 The figure shows the change in capsaicin content before and after removing blisters in Example 3 of the present invention. The data in the figure are expressed as mean ± standard deviation (n=3). The significance analysis was performed using t-test (*, p<0.05; **, p<0.01).

[0016] Figure 5This document presents an evolutionary and conserved domain analysis of Arabidopsis thaliana and pepper VAP family members in Embodiment 4 of the present invention. A: Evolutionary tree of Arabidopsis thaliana and pepper VAP family members. B: Schematic diagram of conserved domains of Arabidopsis thaliana and pepper VAP family members.

[0017] Figure 6 This is a heatmap of the expression of CaVAP family members in Embodiment 4 of the present invention.

[0018] Figure 7 The figures represent the relative expression levels of CaVAP27 in different genotypes and tissues in Example 5 of this invention. Wherein, A: CaVAP27 expression level in different genotypes of pepper; B: CaVAP27 expression level in different tissues of pepper CA59.

[0019] Figure 8 This image shows the in situ expression analysis of the VAMP-related protein gene CaVAP27 and the synthetic gene AT3 in Example 6 of this invention. A: CaVAP27 in situ hybridization signal. B: AT3 in situ hybridization signal. AI / II / III and BI / II / III are magnified views of the corresponding positions in A and B, respectively.

[0020] Figure 9 This document presents the results of silencing CaVAP27 on the capsaicin content in septum in Example 7 of this invention. A: Relative expression levels of CaVAP27 and key genes for capsaicin synthesis in CaVAP27-silenced plants. B: Relative expression levels of MYB31 and CaVAP27 in MYB31-silenced plants. C: Phenotypic comparison between CaVAP27-silenced plants and control plants. D: Capsaicin and dihydrocapsaicin content in CaVAP27-silenced plants. Detailed Implementation

[0021] To facilitate understanding of the present invention, a more complete description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention.

[0022] Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention. The term "and / or" as used in this invention includes any and all combinations of one or more of the associated listed items.

[0023] Unless otherwise specified, all examples were performed under standard experimental conditions, such as those described in Sambrook et al., Molecular Cloning: A Laboratory Manual (Sambrook J & Russel DW, 2013), or as recommended by the manufacturer.

[0024] This invention, based on the analysis of capsaicin spatial distribution in different tissues of chili peppers of different varieties (with varying degrees of spiciness), identified a vesicle-associated membrane protein (VAP) gene, CaVAP27, which is associated with capsaicin accumulation. Its function was then identified as follows: 1. CaVAP27 belongs to the VAP family, possesses a Motile_Sperm domain, and shows high homology with AtVAP27-2, primarily mediating membrane contact. 2. The expression level of the CaVAP27 gene is positively correlated with capsaicin content. 3. In situ hybridization results showed that the CaVAP27 gene is expressed in the septal epidermal cell membrane and around vesicles in chili pepper CA59. 4. Silencing the CaVAP27 gene with VIGS reduced the number of vesicles on the septal surface and decreased capsaicin content. Therefore, the chili pepper CaVAP27 gene may influence capsaicin accumulation levels by affecting vesicle formation. The discovery of the function of the chili pepper CaVAP27 gene provides a theoretical basis for the creation of chili pepper varieties related to capsaicin content.

[0025] In some embodiments of the present invention, the application of the chili CaVAP27 gene in regulating the capsaicin content of chili peppers is disclosed, and its CDS sequence is shown in SEQ ID NO:1.

[0026] In some other embodiments of the present invention, the application of the chili CaVAP27 gene in chili genetic breeding for regulating capsaicin content is disclosed, and its CDS sequence is shown in SEQ ID NO:1.

[0027] In one embodiment, the capsaicinoids include capsaicin and / or dihydrocapsaicin.

[0028] In other embodiments of the present invention, the application of the chili CaVAP27 protein in regulating the capsaicin content of chili peppers is disclosed, and its amino acid sequence is shown in SEQ ID NO:2.

[0029] In other embodiments of the present invention, the application of the chili CaVAP27 protein in chili genetic breeding for regulating capsaicin content is disclosed, and its amino acid sequence is shown in SEQ ID NO:2.

[0030] In one embodiment, the capsaicinoids include capsaicin and / or dihydrocapsaicin.

[0031] In other embodiments of the present invention, the application of a silencing vector for the chili CaVAP27 gene in regulating the capsaicin content of chili peppers is disclosed, wherein the CDS sequence of the chili CaVAP27 gene is shown in SEQ ID NO:1.

[0032] In other embodiments of the present invention, the application of a silencing vector for the chili CaVAP27 gene in chili genetic breeding for regulating capsaicin content is disclosed, wherein the CDS sequence of the chili CaVAP27 gene is shown in SEQ ID NO:1.

[0033] In one embodiment, the capsaicinoids include capsaicin and / or dihydrocapsaicin.

[0034] In other embodiments of the present invention, the application of engineered bacteria transforming a silencing vector of the pepper CaVAP27 gene in regulating the capsaicin content of peppers is disclosed, wherein the CDS sequence of the pepper CaVAP27 gene is shown in SEQ ID NO:1.

[0035] In other embodiments of the present invention, the application of engineered bacteria transformed with a silencing vector of the pepper CaVAP27 gene in pepper genetic breeding for regulating capsaicin content is disclosed, wherein the CDS sequence of the pepper CaVAP27 gene is shown in SEQ ID NO:1.

[0036] In one embodiment, the capsaicinoids include capsaicin and / or dihydrocapsaicin.

[0037] In some other embodiments of the present invention, a method for reducing the capsaicin content of chili peppers is disclosed, comprising the following steps: reducing the expression level of the chili pepper CaVAP27 gene in chili pepper plants.

[0038] In one embodiment, the capsaicinoids include capsaicin and / or dihydrocapsaicin.

[0039] The present invention will be described in detail below with reference to the accompanying drawings and specific embodiments.

[0040] Example 1: The changing patterns of blisters and their correlation with capsaicin accumulation

[0041] Using annual chili peppers Z531 (sweet pepper, germplasm resources collected by the applicant), chili pepper CA59 (medium spiciness, self-pollinated high-generation chili peppers bred by the applicant), and Chinese chili pepper 839 (extremely spicy, germplasm resources collected by the applicant) as materials, the changes in vesicles in their septa at five different developmental stages (16 DPA, 25 DPA, 30 DPA, 35 DPA, and 45 DPA) were observed.

[0042] The results are as follows Figure 1 As shown, in sweet pepper Z531, no vesicular structures were observed throughout the entire developmental period. In contrast, in CA59 and Chinese pepper 839, which contain capsaicin accumulation, vesicles were present, and the number and size of the vesicles increased continuously with the developmental period, stabilizing after 35 DPA. Figure 1 (A) Chinese chili pepper 839 has more large blisters than annual chili pepper CA59. Figure 1 (B in the middle).

[0043] The above results indicate that the presence or absence of blisters is significantly correlated with capsaicin content, and blister development is positively correlated with capsaicin content.

[0044] Example 2: Structural observation of water bubbles

[0045] Paraffin sections of pepper fruits from the CA59 and Chinese pepper 839 at four different developmental stages (16 DPA, 25 DPA, 30 DPA, and 38 DPA) were observed. The results are as follows: Figure 2 and Figure 3 As shown.

[0046] In the CA59 septal tissue of chili peppers, the epidermal cells are slender and palisade-like, tightly packed, with deeply stained nuclei (obvious hematoxylin staining); the cytoplasm of the septal epidermal cells is deep red (significant eosin staining), indicating a possible accumulation of large amounts of alkaline substances. A vesicular structure forms on the outer side of the epidermal cells (…). Figure 2 The B-II and C-II molecules gradually expand and merge with each other during development, forming sheet-like fluid storage spaces. Figure 2 The D-IV vesicles lack cell nuclei (hematoxylin staining is absent), indicating they are not independent cellular tissues. At the junction of the placenta and septum, the cell morphology of this layer shows significant changes. The placental epidermal cells are flattened and tightly packed, lacking vesicular structures on the outer side, forming a stark contrast with the septal epidermis. Figure 2 (A-III, B-III, C-III, D-III). The inner epidermal cells of the fruit are more loosely arranged than the septate epidermal cells, with an elliptical cell shape and no vesicles on the outer surface. Larger thin-walled cells are present in the lower layer of the epidermal cells, gradually increasing in size as the fruit develops. Figure 2 In the AI, BI, CI), by 38 DPA, large thin-walled cells shrink and collapse. Figure 2The arrow at DI indicates that it may be involved in water storage and turgor pressure maintenance during fruit enlargement.

[0047] The cellular structure of the septum and placenta in Chinese chili pepper 839 is similar to that of CA59: the placental epidermal cells are flat and tightly arranged. Figure 3 In the A-III, B-III, C-III, and D-III subtypes, the septal epidermal cells are slender, palisade-like, and tightly packed, with deeply stained nuclei (obvious hematoxylin staining), forming continuous vesicular structures on the outer side. Unlike CA59, the septal epidermal cells of Chinese pepper 839 show vesicular structures at 16 DPA, and by 25 DPA, the vesicles have begun to gradually enlarge and fuse, forming larger cavities. Figure 3 The A-II, B-II, C-II, and D-II in the Chinese pepper 839 are shown. At 16 DPA, the epidermal cells of the pulp are oval in shape, loosely arranged, and do not form vesicular structures on the outer side. Figure 3 In the AI ​​(Alternative Acidity) of the pulp, at 25 DPA, the inner epidermal cells of the pulp have a similar cell structure to the septum, exhibiting a slender and tightly packed arrangement, while larger vesicles appear on the outer side. Figure 3 (BI, CI, DI in the text). The absence of large thin-walled cells in the lower layer of the inner epidermis of the flesh of Chinese pepper 839 indicates that its flesh has a lower water storage capacity compared to CA59, which is consistent with the thin and wrinkled phenotype of Chinese pepper 839 flesh.

[0048] Example 3: Confirmation of vesicles as the main storage tissue for capsaicin

[0049] The septa of the CA59 inbred line of one-year-old chili peppers, the septa of the Chinese chili pepper 839, and the vesicles in the pulp were treated. Specifically, the vesicles were punctured, and the tissue fluid was absorbed using filter paper. The capsaicin content before and after vesicle removal was then measured. The results are as follows: Figure 4 As shown in the results, the capsaicin content at different developmental stages decreased significantly after the blisters were removed. This phenomenon suggests that blisters play an important role in the storage of capsaicin, and that blisters may serve as a storage structure for capsaicin.

[0050] Further analysis revealed that the capsaicin content decreased more significantly in the later stages of development than in the early stages. This may be because the vesicles are larger in the later stages of development, thus storing more capsaicin. Therefore, the loss of capsaicin is more significant when the vesicles are removed. This result further supports the hypothesis that the vesicles serve as a capsaicin storage structure.

[0051] The results of this embodiment demonstrate that vesicles are indeed the main storage tissue for capsaicin. This discovery is of great significance for understanding the accumulation mechanism of capsaicin in chili pepper fruits.

[0052] Example 4: Discovery of genes related to capsaicin accumulation in vesicle-associated membrane protein-associated protein (VAP)

[0053] Transcriptome sequencing analysis was performed on five tissue sites (Sep_T, Sep_B, Pla_S, Pla_I, and Pulp) of the CA59 pepper fruit at three distinct periods: high expression of capsaicin synthesis genes (16 DPA), rapid capsaicin accumulation (25 DPA), and peak capsaicin content (45 DPA). WGCNA analysis combined with differential expression analysis was used to screen for candidate genes related to capsaicin transport and regulation. Weighted co-expression network analysis (WGCNA) was performed on the expression patterns of all genes. At power=12, R²=0.86 and the average connectivity was less than 100, indicating a scale-free network distribution suitable for hierarchical clustering tree construction. Based on gene expression patterns, all genes were divided into 24 modules. The MEturquoise and MEred modules showed high correlation with capsaicin content. The genes in the MEred module are mainly highly expressed in the septum. In this module, several capsaicin accumulation-related genes were found, including the vesicle-associated membrane protein-associated protein (VAP) gene Capann_ca59V2g03217.1 (named CaVAP27), whose 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.

[0054] SEQ ID NO:1

[0055]

[0056] SEQ ID NO:2

[0057] MTNTQLVEIQPRELKFTFEVKKQSSCAVHVANITDQYVAFKVKTTSPKKYCVRPNIGVVKPKSTYDFTVTMQAQRTAPSDMQCKDKFLIQCTIVPFGTSEEEIPPSMFNKDNGKYIEEECKLKVVLVSPPQSPVLQPANGISKQGASIETLMQQEKFPSGVENLPPIQTVVKNN KDYKYEEEEEPSLGKIADVKNLNFAKNTESNIDEVAENNKGADFDAEEEDPRLTNTVEDVKLNSPNNRESNTNEVVESRNLDMEIKLLLSKDVEELNFKINSLDSQLIEAECIIAKLKEEERSTIKDIEILKQELALLRTKGVERRAQVGFPPLFLCMVGLICLTIGYLVRA*

[0058] Members of the VAP family were identified, and their basic characteristics were analyzed. Sequences of Arabidopsis VAP family members were obtained using the Arabidopsis genome website, and the pepper CA59 genome was subjected to BLAST, resulting in 13 pepper CaVAP family members. Consistent with the characteristics of Arabidopsis VAP members, pepper VAP members all contain only one Motile_Sperm domain, mainly involved in membrane contact, intracellular transport, and signal transduction. Figure 5 ).

[0059] The expression pattern of the CaVAP27 gene (Capann_ca59V2g03217.1) was analyzed using transcriptome data. The results showed that CaVAP27 is specifically expressed on the septum, and highly expressed at 16 DPA and 25 DPA. Figure 6 This aligns with the pattern of capsaicin accumulation.

[0060] Example 5: Exploring the correlation between the CaVAP27 gene in chili peppers and capsaicin content

[0061] The relative expression level of CaVAP27 was detected in the pulp, septum, and placental tissues of nine different capsaicin levels of chili peppers, as well as in the roots, stems, leaves, flowers, and seeds of the annual chili pepper CA59.

[0062] The results are as follows Figure 7As shown, the results indicate that the expression pattern of CaVAP27 is related to capsaicin levels, with high expression mainly in the septum and also in the pulp of Chinese chili inbred lines with high capsaicin content, while expression in sweet peppers is extremely low. Figure 7 (A) Compared to septal tissue, CaVAP27 expression levels are lower in root, stem, leaf, flower, and seed tissues, indicating that CaVAP27 is specifically expressed in the septum, consistent with the distribution pattern of capsaicin. Figure 7 (B in the middle).

[0063] The results of this embodiment suggest that CaVAP27 may be involved in the capsaicin accumulation process.

[0064] Example 6: Exploration of the spatiotemporal expression pattern of the CaVAP27 gene in pepper fruit

[0065] Pepper CA59 fruits with a DPA of 25 were quickly placed in FAA fixative for paraffin sectioning. The 5′ fragments with the highest specificity for the target genes (CaVAP27, AT3) were selected for probe preparation, and the probe sequences are shown in Table 1.

[0066] Table 1

[0067]

[0068] The probes were labeled using the DIG RNA Labeling Kit according to the instructions. Plant paraffin sections were prepared through fixation, dehydration, clearing, paraffin embedding, and sectioning. After dewaxing in xylene, hybridization and staining were performed. The hybridized sections were then examined and photographed under an optical microscope (Olympus).

[0069] The results are as follows Figure 8 As shown, the results indicate that in pepper fruits, the AT3 in situ hybridization signal mainly appears in the septal epidermal cells (…). Figure 8 B-II in the placenta and pulp epidermal cells showed no signal ( Figure 8 The presence of BI and B-III indicates that capsaicin is synthesized within the septal epidermal cells; while the transcriptional signal of CaVAP27 is mainly concentrated around the vesicles and diffusely expressed in the intercellular spaces. Figure 8 The signal intensity of the outer membrane of the A-II-1 septum epidermal cells was significantly higher than that of the placenta and pulp epidermal cells, suggesting that it may be related to vesicle formation.

[0070] Example 7: Effect of silencing the CaVAP27 gene in chili peppers on capsaicin content

[0071] Agrobacterium bacterial suspensions carrying pTRV2 (stored in the applicant's laboratory, commercially available), pTRV2-CaVAP27 (or capsaicin synthesis-related genes), and pTRV1 (stored in the applicant's laboratory, commercially available) vectors were mixed at a 1:1 volume ratio and shaken well. CA59 pepper seedlings of about 30 days old with basically uniform growth were selected, and the mixed bacterial suspension was injected into the back of the leaves with a syringe (needle discarded) until the leaves appeared water-soaked. After incubation in the dark for 48 hours, the seedlings were transferred to normal culture. Samples were taken at 20 and 45 days after flowering to detect the expression level and capsaicin content.

[0072] The results are as follows Figure 9 As shown, the results are as follows:

[0073] In CaVAP27-silenced plants, CaVAP27 expression was significantly reduced, and the expression levels of capsaicin synthesis-related genes were also significantly decreased. Figure 9 In CaMYB31 silenced plants, the expression level of the pepper CaVAP27 gene was also significantly reduced (A). Figure 9 In CaVAP27 silent plants, capsaicin and dihydrocapsaicin levels decreased, and vesicle volume and number decreased. Figure 9 The results (C, D) indicate that CaVAP27 may regulate capsaicin accumulation by influencing capsaicin storage structure.

[0074] The technical features of the above embodiments can be combined in any way. For the sake of brevity, not all possible combinations of the technical features in the above embodiments are described. However, as long as there is no contradiction in the combination of these technical features, they should be considered to be within the scope of this specification.

[0075] The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.

Claims

1. Use of a Capsicum annuum CaVAP27 protein in regulating the content of capsaicinoids in Capsicum annuum, the amino acid sequence of which is shown as SEQ ID NO:

2.

2. Use of a Capsicum annuum CaVAP27 protein in regulating the content of capsaicinoids in Capsicum annuum genetic breeding, the amino acid sequence of which is shown as SEQ ID NO:

2.

3. Use of a Capsicum annuum CaVAP27 gene in regulating the content of capsaicinoids in Capsicum annuum, the CDS sequence of which is shown as SEQ ID NO:

1.

4. Use of a Capsicum annuum CaVAP27 gene in regulating the content of capsaicinoids in Capsicum annuum genetic breeding, the CDS sequence of which is shown as SEQ ID NO:

1.

5. Use of a silencing vector of a Capsicum annuum CaVAP27 gene in reducing the content of capsaicinoids in Capsicum annuum, the CDS sequence of the Capsicum annuum CaVAP27 gene being shown as SEQ ID NO:

1.

6. Use of a silencing vector of a Capsicum annuum CaVAP27 gene in regulating the content of capsaicinoids in Capsicum annuum genetic breeding, the CDS sequence of the Capsicum annuum CaVAP27 gene being shown as SEQ ID NO:

1.

7. Use of an engineered bacterium transformed with a silencing vector of a Capsicum annuum CaVAP27 gene in reducing the content of capsaicinoids in Capsicum annuum, the CDS sequence of the Capsicum annuum CaVAP27 gene being shown as SEQ ID NO:

1.

8. Use of an engineered bacterium transformed with a silencing vector of a Capsicum annuum CaVAP27 gene in regulating the content of capsaicinoids in Capsicum annuum genetic breeding, the CDS sequence of the Capsicum annuum CaVAP27 gene being shown as SEQ ID NO:

1.

9. The method of claim 1, 3, 5, or 7, wherein, The capsaicinoids include capsaicin and / or dihydrocapsaicin.

10. A method of reducing the capsaicinoid content of a pepper, characterized in that, The method comprises the following steps: Reducing the expression amount of the Capsicum annuum CaVAP27 gene in the Capsicum annuum plant.

Images