Cowpea Chloroplast Microsatellite Molecular Marker Polymorphism Primer, Screening Method, and Genetic Relationship Identification Method

A polymorphic primer and molecular marker technology, which can be used in the determination/inspection of microorganisms, biochemical equipment and methods, DNA preparation, etc., can solve the problem of incomplete cowpea germplasm resources, and achieve a comprehensive method for identifying cowpea kinship. , not easy to reorganize, the effect of comprehensive response

Active Publication Date: 2017-01-25
JIANGHAN UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] In order to solve the problem of incomplete identification of cowpea germplasm resources in the prior art, the embodiment of the present invention provides a polymorphic primer of cowpea chloroplast microsatellite molecular markers, a screening method thereof, and a method for identifying genetic relationship

Method used

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  • Cowpea Chloroplast Microsatellite Molecular Marker Polymorphism Primer, Screening Method, and Genetic Relationship Identification Method
  • Cowpea Chloroplast Microsatellite Molecular Marker Polymorphism Primer, Screening Method, and Genetic Relationship Identification Method
  • Cowpea Chloroplast Microsatellite Molecular Marker Polymorphism Primer, Screening Method, and Genetic Relationship Identification Method

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Embodiment 1

[0049] The embodiment of the present invention provides a polymorphic primer for microsatellite molecular markers of cowpea chloroplast.

[0050] 1. Polymorphic primer screening of chloroplast microsatellite molecular markers

[0051]Download the full sequence of the cowpea (V.unguiculata) chloroplast genome (NC_018051) from the GenBank public database (http: / / www.ncbi.nlm.nih.gov / ), analyze the full sequence of the cowpea chloroplast genome, and screen chloroplast microsatellite molecules sequence. Single-base microsatellites were searched using the software Tandem Repeats Finder4.04 (Benson G.Tandem repeats finder: a program to analyze DNA sequences[J].Nucleic Acids Research,1999,27(2):573-580.); Microsatellite molecules were analyzed using SSRhunter1.3 software (Li Qiang, Wan Jianmin. SSRHunter, the development of a localized SSR site search software [J]. Genetics, 2005, 27(5):808-810.), search The conditions are set as follows: the minimum number of repeats for a single-...

Embodiment 2

[0072] The embodiment of the present invention provides a kind of method that utilizes the polymorphism primer of the cowpea chloroplast microsatellite molecular marker that embodiment one provides to identify cowpea genetic relationship, specifically as follows:

[0073] Sample: 13 parts of samples provided for testing in Example 1 of the present invention, including 10 parts of cowpea materials and 3 parts of kidney bean materials (3 parts of kidney beans are respectively codes J9, J13, B20 in Table 2 of Embodiment 1), and extract Genomic DNA of 13 samples to detect the generality of polymorphic primers of microsatellite molecular markers in cowpea chloroplasts in kidney bean.

[0074] First, the polymorphic primers of cowpea chloroplast microsatellite molecular markers provided in Example 1 of electrophoresis detection are mixed with the genomic DNA of the sample to amplify to obtain an amplified product, and the amplified product is passed through a denaturing polyacrylamid...

Embodiment 3

[0080] The embodiment of the present invention provides a method for analyzing the genetic diversity of cowpea by using the polymorphic primers of 5 pairs of cowpea chloroplast microsatellite molecular markers provided in Example 1 to reveal the current status of genetic diversity of the tested cowpea samples, based on 5 pairs of cowpea The polymorphic primers of chloroplast microsatellite molecular markers and the diversity information detected by 10 cowpea samples were analyzed by Popgene1.32 software, wherein:

[0081] Calculate the main genetic diversity parameters observed heterozygosity (observed heterozygosity, Ho);

[0082] Expected heterozygosity (expected heterozygosity, He);

[0083] number of alleles (Na);

[0084] Effective number of alleles (=Effective number of alleles, Ne);

[0085] Shannon coefficient (Shannon's Information index, I).

[0086] Polymorphism information content (polymorphism information index, PIC) calculation reference formula (Botstein D, W...

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Abstract

The invention discloses a polymorphic primer of a cowpea chloroplast microsatellite molecular marker, a screening method thereof, and a method for identifying genetic relationship, and belongs to the technical field of crop molecular biology. The polymorphic primers of the cowpea chloroplast microsatellite molecular markers. The method for identifying the polymorphism of cowpea chloroplast microsatellite molecular marker primers includes: extracting genomic DNA of different varieties of cowpea; mixing the designed primers with genomic DNA and performing PCR amplification and electrophoresis separation; obtaining the chloroplast microsatellite molecular marker of cowpea Polymorphic primers. The method for identifying cowpea kinship includes: extracting cowpea genomic DNA; using polymorphic primers of cowpea chloroplast microsatellite molecular markers to perform PCR amplification, electrophoresis separation, and statistical results; and analyzing the results for kinship. The polymorphic primers can reflect the genetic information of the cowpea cytoplasmic genome, and can be used in researches such as the classification of cowpea germplasm resources, the identification of genetic relationship, and the analysis of genetic diversity.

Description

technical field [0001] The invention relates to the technical field of crop molecular biology, in particular to a polymorphism primer of a cowpea chloroplast microsatellite molecular marker, a screening method thereof, and a method for identifying genetic relationship. Background technique [0002] Germplasm resources are also called genetic resources. Accurate identification and identification of germplasm resources is the basis for breeding and genetic improvement. [0003] In the identification process of cowpea germplasm resources, the SSR (Simple Sequence Repeat, simple repeat sequence) molecular marker method of cowpea nuclear DNA is used to identify the differences at the molecular level between different varieties. SSR is also called microsatellite DNA, which refers to It is a piece of DNA composed of 1-6 nucleotides repeated multiple times in the genome, widely distributed in different positions of the genome, and the length is generally below 200bp. Studies have s...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 潘磊陈禅友李依
Owner JIANGHAN UNIVERSITY
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