Method for identifying the number of fruits per plant of pepper and application thereof
Patent Information
- Application Number
- CN202611494464.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-09-23
- Publication Date
- 2026-11-03
AI Technical Summary
In current technologies, the identification of the number of fruits per chili plant mainly relies on traditional field phenotypic identification, which is slow and inefficient, making it difficult to meet the needs of modern high-efficiency molecular breeding, and lacking precise, rapid, and low-cost molecular markers.
Based on genome-wide association analysis, SNP sites that were significantly associated with the number of results per pepper plant were detected. dCAPS molecular markers were developed, and PCR amplification and restriction endonuclease digestion were performed using primer pairs. SNP genotyping was achieved by agarose gel electrophoresis.
This technology enables the rapid and accurate screening of chili pepper materials with high potential for yield per plant during the seedling stage, shortening the breeding cycle, reducing costs, improving selection efficiency, and promoting the development of molecular marker-assisted breeding for high-yield chili peppers.
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Abstract
Description
Technical Field
[0001] This invention relates to the field of molecular marker technology, and in particular to a method for identifying the number of fruits per chili pepper plant and its application. Background Technology
[0002] chili( Capsicum The yield of chili peppers (spp.) is influenced by a combination of traits, including the number of fruits per plant, the weight of a single fruit, the diameter of the fruit, and the thickness of the flesh. Among these, the number of fruits per plant is one of the core factors determining chili pepper yield. Currently, the identification of the number of fruits per plant in chili peppers mainly relies on traditional phenotypic identification in the field, which requires going through the entire growth cycle from sowing and transplanting to growth and maturity. This method is slow, inefficient, and consumes a lot of manpower and resources, making it difficult to meet the industrial demands of modern, efficient molecular breeding.
[0003] With the widespread adoption and application of high-throughput sequencing technology, SNP molecular markers have become mainstream markers for genetic mapping of pepper traits, identification of kinship, and marker-assisted selection breeding due to their wide distribution, high density, genetic stability, and ease of genotyping. Cleaved amplified polymorphic sequences (CAPS) are classic molecular markers developed for SNPs, achieving accurate SNP genotyping by recognizing naturally occurring restriction endonuclease cleavage sites and their variant genotypes. However, in most cases, SNP sites cannot be recognized by conventional commercial restriction endonucleases. Derived cleaved amplified polymorphic sequences (dCAPS) create cleavage sites that can be recognized by restriction endonucleases by artificially introducing mismatched bases into primers, thereby achieving accurate genotyping of target SNP sites. dCAPS molecular markers have advantages such as ease of operation, low cost, and good stability, and possess great application potential.
[0004] Currently, the molecular identification system for the trait of tael count per chili plant is still imperfect, and molecular markers that can accurately, rapidly, and cost-effectively identify the tael count per plant are relatively scarce. The industry still widely relies on traditional field phenotypic identification methods, which suffer from drawbacks such as identification lag, significant environmental influence, high cost, and low efficiency, severely hindering the development of molecular breeding for high-yield chili plants. Therefore, developing a dCAPS molecular marker that is highly specific, stable, easy to operate, and capable of identifying the tael count per plant at the seedling stage is of significant practical importance and application value for shortening the breeding cycle of high-yield chili plants, reducing breeding costs, and promoting the development of molecular marker-assisted breeding technology for high-yield chili plants. Summary of the Invention
[0005] This invention provides a molecular marker, method, and application for identifying the number of fruits per chili pepper plant.
[0006] This invention is based on genome-wide association analysis of 220 natural chili pepper populations. Under a mixed linear model, SNP sites that are significantly associated with the number of chili pepper plants were detected, and the dCAPS molecular marker was developed accordingly.
[0007] Specifically, the present invention provides the following technical solutions.
[0008] In a first aspect, the present invention provides an SNP molecular marker related to the number of fruits per chili pepper plant, the SNP molecular marker containing a nucleotide sequence with a C / G polymorphism at position 204 of the sequence shown in SEQ ID NO.1.
[0009] The SNP sites of the SNP molecular markers provided by this invention correspond to the pepper genome ( C. annuum The SNP at position 213,021,209 on chromosome 12 (cv. CM334v1.6) exhibits a C / G polymorphism. When the genotype at this locus is GG, it corresponds to the trait of high single-plant yield.
[0010] Preferably, the nucleotide sequence of the SNP molecular marker is as shown in SEQ ID NO.1, and the polymorphic site is located at position 204 of the sequence shown in SEQ ID NO.1, with a polymorphism of C / G.
[0011] Preferably, the genotype of the polymorphic site of the SNP molecular marker is GG, which corresponds to a high number of single-plant results.
[0012] Secondly, the present invention provides primer pairs for amplifying the SNP molecular markers described in the first aspect above.
[0013] Preferably, the nucleotide sequences of the primer pair are shown in SEQ ID NO.2-3.
[0014] The primer pairs described above are dCAPS primer pairs. By introducing mismatched bases, the amplification products can be processed by restriction endonucleases. Bgl II (AGATCT) specific recognition, thereby achieving SNP genotyping based on conventional gel electrophoresis.
[0015] Thirdly, the present invention provides the application of the SNP molecular markers described in the first aspect or the primer pairs described in the second aspect in the preparation of detection reagents, detection kits or gene chips for identifying peppers with high yield per plant.
[0016] Preferably, the reagent, kit, or gene chip contains the primer pair described in the second aspect.
[0017] Fourthly, the present invention provides any of the following applications of the SNP molecular markers described in the first aspect or the primer pairs described in the second aspect: (1) Identify or screen chili pepper materials with high potential for high yield per plant; (2) Molecular marker-assisted breeding of the number of fruits per chili plant; (3) Variety improvement of the number of fruits per chili plant.
[0018] Preferably, the application includes: detecting the genotype at position 204 of the nucleotide sequence shown in SEQ ID NO.1 in the genome of the pepper to be tested; if the genotype is GG, it is determined to be material with high potential for high yield per plant.
[0019] Fifthly, the present invention provides any one of the following applications of SNP molecular markers or detection primers or reagents for the detection of said SNP molecular markers: (1) Identify or screen chili pepper materials with high potential for high yield per plant; (2) Molecular marker-assisted breeding of the number of fruits per chili plant; (3) Variety improvement of the number of fruits per chili plant; The polymorphic site of the SNP molecular marker is located at position 213,021,209 on chromosome 12 of the pepper genome, with a polymorphism of C / G; the version number of the pepper genome is... C . annuum cv. CM334 v1.6.
[0020] The nucleotide sequences of the detection primers for the SNP molecular marker are shown in SEQ ID NO.2-3.
[0021] In a sixth aspect, the present invention provides a method for identifying peppers with a high number of fruits per plant, the method comprising: detecting the genotype in the genome of the pepper to be identified corresponding to position 213,021,209 on chromosome 12 of the reference genome; if the genotype is GG, then it is identified as a pepper material with a high number of fruits per plant; The reference genome is version number [version number missing]. C . annuum The chili pepper genome cv. CM334 v1.6.
[0022] Preferably, the detection of the genotype corresponding to position 213,021,209 on chromosome 12 of the reference genome in the chili pepper genome to be identified is performed by using the chili pepper genome to be identified as a template, performing PCR amplification using primer pairs as shown in SEQ ID NO.2-3, analyzing the PCR amplification products, and determining the genotype.
[0023] Preferably, the method includes: (1) Extract genomic DNA from the peppers to be tested; (2) Using the genomic DNA of the pepper to be tested as a template, PCR amplification was performed using the dCAPS primer pair shown in SEQ ID NO.2-3; (3) Analyze the PCR amplification products to determine the genotype of the SNP molecular markers mentioned above, and identify pepper materials with high single-plant yield based on the genotype of the SNP molecular markers.
[0024] Preferably, the PCR amplification reaction program is as follows: 95℃ pre-denaturation for 8-12 min; 95℃ denaturation for 18-22 s, 54℃-60℃ annealing for 18-22 s, 72℃ extension for 30 s-60 s, for 36-40 cycles.
[0025] More preferably, the PCR amplification reaction program is as follows: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 20 s, 56℃ annealing for 20 s, 72℃ extension for 40 s, 39 cycles.
[0026] Preferably, the 15 μL reaction system for PCR amplification is: 7.5 μL of 2×Taq Master Mix, 0.3 μL each of the primers shown in SEQ ID NO. 2-3, 0.6 μL of DNA template (100 ng / ul), and 6.6 μL of ddH2O.
[0027] Preferably, the analysis of the PCR amplification products is performed using restriction endonucleases. Bgl After digesting the PCR amplification product with enzyme II, the digested product is detected by electrophoresis, and the genotype is determined based on the banding pattern.
[0028] Preferably, a restriction endonuclease is used. Bgl The reaction system (10 μL) for the II enzyme digestion PCR amplification product includes: 4.0 μL ddH2O, 0.7 μL 10 × Buffer O, Bgl II 0.3 μL, PCR amplification product 5 μL; enzyme digestion reaction conditions: 37℃ for 8 h.
[0029] Preferably, if the PCR amplification product has a 207 bp characteristic band after enzyme digestion, then the genotype is GG, and it is identified as a pepper material with a high single-plant yield.
[0030] The beneficial effects of the present invention include at least the following: (1) Based on genome-wide association analysis, this invention detected SNP sites that were significantly associated with the number of single plant results in natural pepper populations. The developed molecular markers have the advantages of strong universality and high accuracy.
[0031] (2) The dCAPS molecular marker and its primer pair provided by the present invention only require conventional PCR amplification and restriction endonuclease digestion. SNP typing can be achieved by agarose gel electrophoresis. The operation is simple, the cost is low, and the results are reliable. No sequencing or fluorescently labeled probes are required, making it suitable for screening large-scale pepper breeding populations.
[0032] (3) The SNP molecular markers of the present invention can be used to identify the genotype of chili pepper materials during the seedling stage. Without waiting for the plant to complete the entire growth period, materials with high potential for single plant yield can be screened in advance, which greatly shortens the breeding cycle, improves selection efficiency, and reduces field costs.
[0033] (4) The SNP molecular marker and identification method provided by this invention provides a directly applicable technical tool for molecular marker-assisted breeding of the number of fruits per chili plant, which has important industrial application value for accelerating the breeding process of high-yield chili varieties. Attached Figure Description
[0034] To more clearly illustrate the technical solutions in this invention 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 invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.
[0035] Figure 1 This is a box plot and bee colony plot showing the best linear unbiased estimator (BLUE) for the number of individual plants at the SNP12.209 locus with genotypes CC and GG, determined based on GBS (genotyping-by-sequencing) in Example 1 of this invention. Each point in the bee colony plot represents the BLUE value for the number of individual plants of a pepper germplasm. The red diamonds inside the box plot represent the mean of that genotype. This indicates the Wilcoxon rank-sum test (Mann-Whitney U test) under... p <0.0001.
[0036] Figure 2This is the agarose gel electrophoresis result of the identification of pepper germplasm genotypes using SNP molecular marker detection primers for PNPP12.209 in Examples 1 and 2 of this invention; where M is the DNA Marker, and the figure shows the 250 bp band of the DNA Marker; 1-80 are the pepper germplasms to be tested; 81-82 are pepper germplasms with the known genotype GG, with a 207 bp characteristic band; 83-84 are pepper germplasms with the known genotype CC, with a 249 bp characteristic band. bp feature bands; 1, A010; 2, A011; 3, A012; 4, A018; 5, A026; 6, A031; 7, A039; 8, A047; 9, A063; 10, A089; 11, A093; 12, A103; 13, A108; 14, A119; 15, A150; 16, A154; 17, A156; 18, A161; 19, A164; 20, A168; 21, A171; 22, A174; 23, A177; 24, A185; 25, A252; 26, A264; 27, B010; 28, B015; 29, B018; 30, B021; 31, B032; 32, B036; 33, B112; 34, B118; 35, B119; 36, B134; 37, B166; 38, B308; 39, B309; 40, B316; 41, B324; 42, B346; 43, B3 71; 44, B425; 45, B438; 46, B442; 47, BJ10; 48, C004; 49, C005; 50, C024; 51, C031; 52, C032; 53, C040; 54 , C064; 55, C099; 56, C109; 57, C165; 58, CM334; 59, WY003; 60, WY086; 61, WY120; 62, WY266; 63, WY267; 6 4, WY287; 65, ZHE10; 66, ZJ30; 67, WY089; 68, A120; 69, A131; 70, A133; 71, A128; 72, A068; 73, A197; 74, A200; 75, A208; 76, B007; 77, B008; 78, B025; 79, B027; 80, B037; 81, B135; 82, B137; 83, A008; 84, A013.
[0037] Figure 3 The BLUE values represent the number of individual chili pepper plants with genotypes CC and GG at the SNP12.209 locus identified based on the molecular marker PNPP12.209 in Example 2 of this invention; each point in the bee colony diagram represents the BLUE value of the number of individual chili pepper plants; the red diamonds inside the box plot represent the mean of the genotype. This indicates the Wilcoxon rank-sum test (Mann-Whitney U test) under... p <0.001. Detailed Implementation
[0038] To make the objectives, technical solutions, and advantages of this invention clearer, the technical solutions of this invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this invention. All other embodiments obtained by those skilled in the art based on the embodiments of this invention without creative effort are within the scope of protection of this invention.
[0039] Unless otherwise specified, all experimental materials, reagents and instruments used in the following examples are commercially available, and the experimental procedures were performed in accordance with conventional molecular biology methods (such as Sambrook et al., Molecular Cloning Laboratory Manual) or relevant product instructions.
[0040] Example 1: Development of molecular markers for the number of fruits per chili pepper plant 1. Genome-wide association analysis of the number of fruits per chili pepper plant This invention uses 220 chili germplasm samples ( C . annuum The association analysis population (220 chili pepper germplasm accessions) includes breeding lines, local varieties, and transitional germplasm, primarily covering germplasm resources from 28 provinces in China, with approximately 10% sourced from 9 other countries: the United States, the United Kingdom, Hungary, Indonesia, Italy, Japan, the Netherlands, South Korea, and Thailand. For detailed information on these 220 chili pepper germplasm accessions, please refer to the literature "Xinjie Yuan". Xuejun Chen, Kunhua Zhou, Gang Lei, Yueqin Huang, Gege Li, YuFang, Jubin Wang and Rong Fang Genetic, hormonal, and transcriptomic analyses highlight the crucial role of phytohormones in first branch angle regulation in pepper. Table S1 of BMC Plant Biology, 2025, (2025) 25:1492. Genotyping of the above populations was performed using GBS (genotyping-by-sequencing) technology: restriction endonucleases... Eco RI and Nla Genomic DNA was digested with enzyme III, ligated with tagged sequencing adapters, and then a small fragment library was constructed for PE125 paired-end sequencing. After quality control, a total of 955,772 high-quality variant sites were obtained, including 919,743 SNPs and 36,029 Indels, which were used for subsequent genome-wide association studies.
[0041] The number of fruits per chili pepper plant was counted during the peak fruiting period in three different environments: E1 (Yichun, Jiangxi, China, 115.13°E, 28.24°N, spring 2021), E2 (Sanya, Hainan, China, 109.08°E, 18.37°N, winter 2022), and E3 (Yichun, Jiangxi, China, 115.13°E, 28.24°N, winter 2023). To reduce environmental variation, improve the accuracy of trait estimation, and enhance the ability to detect true genotype-trait associations, the best linear unbiased estimator (BLUE) for each of the three environments was calculated using the R package lme4 (version 1.1-26) for subsequent analysis. Association analysis was performed on the phenotypic and genotypic data of 220 pepper accessions using GEMMA software (v0.98.1). Significantly associated SNPs were detected under a mixed linear model, including the SNP sites described in this invention, corresponding to the pepper genome (…). C . annuum The SNP at position 213,021,209 on chromosome 12 (cv. CM334 v1.6) with a C / G polymorphism ( p = 4.78×10 -10 The site was named SNP12.209, and the phenotypic variation explained by this site was 10.09%. The sequence shown in SEQ ID NO.1 is the nucleotide sequence containing the SNP site SNP12.209, which is located at position 204 of the nucleotide sequence shown in SEQ ID NO.1.
[0042] 2. Development of the molecular marker PNPP12.209 for the number of fruits per chili pepper plant The genotype of SNP12.209 was not identified in 80 out of 220 pepper germplasms. Based on GBS identification, the genotypes of SNP12.209 at this locus were CC and GG, with 52.88 ± 38.19 and 218.93 ± 133.65 per plant, respectively, showing a highly significant difference between the two genotypes. p <0.0001, Figure 1Among them, GBS identified 13 pepper germplasms with the genotype GG at SNP12.209. These germplasms included A139, A250, B005, B135, B137, B138, B306, B432, BJ30, BJ31, C153, WY035, and WY081, with BLUE values for the number of fruits per plant of 309.42, 109.00, 68.11, 127.33, and 196, respectively. The scores for 67, 160.92, 223.00, 327.92, 107.33, 532.42, 104.00, 371.07, and 208.92, except for B005 which had a single-plant result count of 68.11, all other 12 GG genotype germplasms had single-plant result counts higher than 80 in the three environmental comprehensive assessments (BLUE values), indicating that this SNP locus can identify pepper germplasms with high single-plant result counts with high accuracy.
[0043] Based on the aforementioned SNP 12.209 site, this invention further designed dCAPS primer pairs 12.209-F and 12.209-BgR, whose nucleotide sequences are shown in SEQ ID NO.2 and SEQ ID NO.3, respectively. PCR amplification using these primer pairs yielded products containing restriction endonucleases. Bgl The recognition site of II can be obtained through Bgl II enzyme digestion and electrophoresis were used to genotype the C / G genotype of SNP12.209, thereby constructing the dCAPS molecular marker PNPP12.209.
[0044] To verify the effectiveness of the above-mentioned dCAPS primer pairs, two chili germplasms with genotypes GG and CC at the SNP12.209 locus were randomly selected from the 220 chili germplasms mentioned above. Using the genomic DNA of these germplasms as templates, PCR amplification was performed using primer pairs 12.209-F / 12.209-BgR (SEQ ID NO. 2-3). The PCR reaction system consisted of 15 μL: 7.5 μL of 2×TaqMaster Mix, 0.3 μL each of 10 μM forward and reverse primers (12.209-F and 12.209-BgR), 0.6 μL of DNA template (100 ng / μL), and ddH2O to a final volume of 15 μL. The reaction program was: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 20 s, 56℃ annealing for 20 s, and 72℃ extension for 40 s, for a total of 39 cycles. Of these, 2×Taq Master Mix was purchased from Nearshore Protein Technology Co., Ltd. (item number E005-02B).
[0045] The PCR products were subjected to enzyme digestion for identification. The enzyme digestion system (10 μL) consisted of: 4.0 μL ddH2O, 0.7 μL 10×Buffer O, and restriction endonuclease. Bgl II 0.3 μL, PCR product 5 μL; enzyme digestion reaction conditions: incubation at 37℃ for 8 h. Bgl II was purchased from Thermo Scientific (catalog number ER0081). 3 μL of the enzyme digestion product was electrophoresed on a 3.5% agarose gel. Results showed that both GG genotype germplasms exhibited a characteristic 207 bp band (a 42 bp band also existed, but it was weak and did not affect genotype determination, so it was not shown in the figure), while both CC genotype germplasms showed a single 249 bp band. Figure 2 Lanes 81-82 represent the GG genotype, and lanes 83-84 represent the CC genotype. These results indicate that the dCAPS primer pair can accurately distinguish between the C and G genotypes of SNP12.209, and it is named PNPP12.209. In practical applications, the 249 bp and 207 bp bands can be used as the primary interpretation criteria.
[0046] Example 2: Application of molecular markers for the number of fruits per chili plant To verify the accuracy and effectiveness of the molecular marker PNPP12.209, 80 of the 220 pepper germplasm GBS accessions from Example 1 in which the genotype of the SNP12.209 locus was not identified ( Figure 2 Germplasm samples numbered 1-80 (the genotype of SNP 12.209 at this site is unknown) were used for identification. The number of individual plants identified in the three environmental conditions described in Example 1 was used, and the BLUE value was used for subsequent analysis. DNA was extracted from the germplasm samples using the CTAB method, and amplified using primer pairs 12.209-F and 12.209-BgR. The PCR system consisted of: 7.5 μL of 2×Taq Master Mix, 0.3 μL each of 10 μM primers 12.209-F and 12.209-BgR, 0.6 μL of DNA template, and 6.6 μL of ddH2O. The PCR reaction program was: 95℃ pre-denaturation for 10 min; 95℃ denaturation for 20 s, 56℃ annealing for 20 s, 72℃ extension for 40 s, for 39 cycles. The PCR product digestion system was: 4.0 μL ddH₂O, 0.7 μL 10 × Buffer O. Bgl II 0.3 μL, PCR product 5 μL. After digestion at 37℃ for 8 h, 3 μL of the digestion product was electrophoresed on a 3.5% agarose gel for detection. Figure 2(lanes 1-80). The results showed that A011, A093, A128, A197, B037, B324, B371, CM334, WY086, WY089, and ZHE10 were of the GG genotype, with a single 207 bp characteristic band (of which 42 bp band was weak and not shown), while the rest were of the CC genotype, with a 249 bp characteristic band. It is worth noting that CM334 had only 15.42 fruits per plant in this invention, and field observations showed that it had a large number of flowers but a low fruit set rate. The candidate gene at SNP12.209 may be the gene that determines the yield of reproductive organs in plants. Even if the genotype of CM334 at SNP12.209 is the high-yielding genotype (GG), the combined effect of other genetic factors (such as functional defects or acceptance barriers in the female reproductive system such as stigma, style, or ovules) may have led to its failure to exhibit the phenotypic characteristic of a high number of fruits per plant. Of course, it cannot be ruled out that the SNP12.209 locus is linked to nearby functional genes, with a certain distance, leading to minor biases in its identification. However, regardless of the genetic mechanism, the dCAPS molecular marker developed based on this SNP locus in this invention has a significant identification effect on the high single-plant yield trait at the population level: the single-plant yield BLUE values of the other genotypes GG (A011, A093, A128, A197, B037, B324, B371, WY086, WY089, and ZHE10) are all greater than 80. The single-plant yield of the GG genotype germplasm at SNP12.209 (mean BLUE value 139.35 ± 75.68) is significantly higher than that of the CC genotype (mean BLUE value 52.52 ± 36.78). p <0.001, Figure 3 ).
[0047] In conclusion, using the SNP12.209 genotype as the basis for judgment, both sequencing and molecular marker PNPP12.209 can identify chili germplasm with a high number of single plant results (more than 80) with high accuracy (overall accuracy rate of 91.67%). This can significantly improve the efficiency and accuracy of molecular marker-assisted breeding for chili yield and has important application value.
[0048] It should be noted that the "high number of fruits per plant" described in this invention is a relative criterion derived from phenotypic evaluation of 220 pepper germplasms under three different environments. In this study population, the BLUE values of materials with the GG genotype were generally higher than 80, and statistically significantly higher than those of materials with the CC genotype. Therefore, this threshold was used as a reference benchmark for evaluating the potential for high number of fruits per plant corresponding to this locus. As a typical quantitative trait, the absolute value of the number of fruits per plant is affected by factors such as cultivation environment and year. In practice, the absolute value may fluctuate under different environments, but the relative advantage of the GG genotype reflected by this locus remains stable in multiple environments.
[0049] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.
Claims
1. SNP molecular markers associated with the number of fruits per chili pepper plant, characterized in that, The SNP molecular marker contains a nucleotide sequence with a C / G polymorphism at position 204, as shown in SEQ ID NO.
1.
2. The SNP molecular marker according to claim 1, characterized in that, The nucleotide sequence of the SNP molecular marker is shown in SEQ ID NO.1, and the polymorphic site is located at position 204 of the sequence shown in SEQ ID NO.1, with a polymorphism of C / G.
3. The SNP molecular marker according to claim 1 or 2, characterized in that, The genotype of the polymorphic site of the SNP molecular marker is GG, which corresponds to a high number of single-plant results.
4. Primer pairs for amplifying the SNP molecular markers according to any one of claims 1 to 3; Preferably, the nucleotide sequences of the primer pair are shown in SEQ ID NO.2-3.
5. The use of the SNP molecular marker according to any one of claims 1 to 3 or the primer pair according to claim 4 in the preparation of detection reagents, detection kits or gene chips for identifying peppers with high single-plant yield; Preferably, the reagent, kit, or gene chip contains the primer pair as described in claim 4.
6. Any of the following applications of the SNP molecular marker according to any one of claims 1 to 3 or the primer pair according to claim 4: (1) Identify or screen chili pepper materials with high potential for high yield per plant; (2) Molecular marker-assisted breeding of the number of fruits per chili pepper plant; (3) Variety improvement of the number of fruits per chili plant.
7. The application according to claim 6, characterized in that, The application includes: detecting the genotype at position 204 of the nucleotide sequence shown in SEQ ID NO.1 in the genome of the pepper to be tested; if the genotype is GG, it is determined to be material with high potential for high yield per plant.
8. Any of the following applications of SNP molecular markers or detection primers or reagents for the SNP molecular markers: (1) Identify or screen chili pepper materials with high potential for high yield per plant; (2) Molecular marker-assisted breeding of the number of fruits per chili pepper plant; (3) Variety improvement of the number of fruits per chili plant; in, The polymorphic site of the SNP molecular marker is located at position 213,021,209 on chromosome 12 of the pepper genome, with a polymorphism of C / G; the version number of the pepper genome is [missing information]. C. annuum cv. CM334 v1.6; Preferably, the nucleotide sequence of the detection primer for the SNP molecular marker is shown in SEQ ID NO.2-3.
9. A method for identifying peppers with a high number of fruits per plant, characterized in that, The method includes: detecting the genotype in the genome of the pepper to be identified corresponding to position 213,021,209 on chromosome 12 of the reference genome; if the genotype is GG, it is identified as a pepper material with a high number of fruits per plant; The reference genome is version number [version number missing]. C. annuum The chili pepper genome cv. CM334 v1.
6.
10. The method for identifying high-yield pepper plants according to claim 9, characterized in that, The method for detecting the genotype corresponding to positions 213,021,209 on chromosome 12 of the reference genome in the chili pepper genome to be identified is as follows: using the chili pepper genome to be identified as a template, PCR amplification is performed using primer pairs shown in SEQ ID NO.2-3, and the PCR amplification products are analyzed to determine the genotype; Preferably, the PCR amplification reaction program includes: 95℃ pre-denaturation for 8-12 min; 95℃ denaturation for 18-22 s, 54℃-60℃ annealing for 18-22 s, 72℃ extension for 30 s-60 s, 36-40 cycles; Preferably, the analysis of the PCR amplification products is performed using restriction endonucleases. Bgl After digesting the PCR amplification product with enzyme II, the digestion product is detected by electrophoresis, and the genotype is determined based on the banding pattern. Preferably, if the PCR amplification product has a 207 bp characteristic band after enzyme digestion, then the genotype is GG, and it is identified as a pepper material with a high single-plant yield.