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Method for identifying authenticity of pepper varieties and special SSR primer combination thereof

A technology of authenticity and primer set, applied in biochemical equipment and methods, recombinant DNA technology, microbial measurement/inspection, etc., can solve the problems of lack of genomic data support, lack of identification ability and representativeness, etc.

Active Publication Date: 2020-06-12
BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, because the previous SSR molecular identification technology was developed based on a small number of varieties, and there is no genomic data support, its identification ability and representativeness are very insufficient.

Method used

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  • Method for identifying authenticity of pepper varieties and special SSR primer combination thereof
  • Method for identifying authenticity of pepper varieties and special SSR primer combination thereof
  • Method for identifying authenticity of pepper varieties and special SSR primer combination thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0090] Acquisition of SSR Primer Combination for Identifying the Authenticity of Pepper Varieties

[0091] 1. Discovery of 22 SSR loci: These loci were obtained by data mining based on the reference genome Capsicum.annuum.L of the closely related species Capsicum annuum.L and the whole genome resequencing data of 40 germplasm resources.

[0092] 1. Different from other molecular marker identification methods, the method of blindly selecting SSR markers at random, the present invention uses the pepper reference genome, and uses unique, representative, resequencing data of 40 pepper varieties (the resequencing results are not disclosed ), through a specific big data analysis method, the SSR loci of the whole genome are analyzed, and a series of screening conditions such as PIC, MAF, and sequence conservation are screened, and finally a set of SSR loci is obtained. Dimension and other methods were used to select the SSR loci that could fully represent the genome of the crop. Thr...

Embodiment 2

[0111] This example is the validity test of the SSR primer combination developed in Example 1.

[0112] 108 capsicum varieties for testing of the present embodiment are common excellent varieties or part foreign imported varieties, and the specific varieties are as follows:

[0113]

[0114]

[0115]

[0116] 1. Obtaining the genomic DNA of the tested pepper varieties

[0117] The leaves of 108 tested capsicum varieties were extracted by CTAB method (the true leaves of 30 seeds were mixed, that is, the true leaves of 30 seeds for each variety, referring to: the true leaves of 30 different plants for the same variety) leaf mixture) to obtain the genomic DNA of the pepper variety to be tested.

[0118] The operation of the above CTAB method is as follows:

[0119] The leaves of the above-mentioned 108 varieties at the seedling stage were picked respectively, and placed in a freeze dryer (CoolSafe 55-4) to dehydrate; Add 800 μL of CTAB extract (2% CTAB, 1.4mM NaCl, 10...

Embodiment 3

[0136] This embodiment is a method for detecting whether the pepper variety to be tested belongs to the 108 varieties of pepper varieties for testing. The variety of the pepper to be tested is unknown, and it is necessary to obtain whether the variety of the pepper to be tested is the 108 through the detection method of this embodiment. one of the species.

[0137] 1. Obtaining the genomic DNA of the pepper variety to be tested

[0138] The leaves of the pepper varieties to be tested were taken from the experimental base of the Vegetable Research Center of Beijing Academy of Agriculture and Forestry Sciences.

[0139] According to the method of step 1 in the embodiment 2, "leaves of the capsicum species to be tested" were replaced with "leaves of the capsicum species to be tested", and other steps were all unchanged to obtain the genomic DNA of the capsicum species to be tested.

[0140] 2. Configuration of SSR primers and PCR reaction system

[0141] According to the method...

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Abstract

The invention belongs to the field of molecular markers and detecting thereof, and particularly relates to a method for identifying the authenticity of pepper varieties and a special SSR primer combination thereof. SSR loci for identifying the authenticity of the pepper variety are obtained by performing data mining according to a reference genome Capsicum.annum.L of pepper and 40 parts of germplasm resource whole genome re-sequencing data; the SSR primer combination is selected from the group consisting of the first SSR primer pair to the twenty-second SSR primer pair which are used for PCR amplification of the first SSR site to the twenty-second SSR site respectively, and preferably, the first SSR primer pair to the twenty-second SSR primer pair are nucleotide sequences with the homologyof 85%-100% with the nucleotide sequences shown as SEQ ID NO: 1-44 in the sequence table respectively. The method can be used for identifying the authenticity of the pepper variety in the full life cycle from seeds, and technical support is provided for pepper germplasm resource and new variety protection.

Description

technical field [0001] The invention belongs to the field of molecular markers and detection thereof, and in particular relates to a method for identifying the authenticity of pepper varieties and a combination of special SSR primers. Background technique [0002] Capsicum is a plant of Magnolia, Solanaceae Capsicum. It is suitable for raw food, cooking, and as a condiment. It is the most widely planted and edible crop in the world. This species originated from Mexico to Colombia, and then introduced to the world Widely cultivated. [0003] Due to the extensiveness of pepper cultivation and the universality of eating, pepper is a plant that is very popular with human beings. Due to the fact that there are certain differences in the actual demand for peppers in different parts of the world, the breeding direction of peppers is also complicated and endless. At the same time, due to the rich economic benefits it can produce, the current situation is that the pepper varieties o...

Claims

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Application Information

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IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 温常龙杜和山杨静静张建罗江
Owner BEIJING ACADEMY OF AGRICULTURE & FORESTRY SCIENCES
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