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Cowpea chloroplast SSR molecule labeled polymorphic primers and screening method thereof, and method for identifying genetic relationship of cowpeas

A technology of polymorphic primers and molecular markers, applied in the determination/testing of microorganisms, biochemical equipment and methods, DNA preparation, etc., can solve the problem of incomplete germplasm resources of cowpea, and achieve a comprehensive method for identifying the genetic relationship of cowpea , Small molecular weight, not easy to recombine

Active Publication Date: 2014-07-09
JIANGHAN UNIVERSITY
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  • Summary
  • 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 SSR molecule labeled polymorphic primers and screening method thereof, and method for identifying genetic relationship of cowpeas
  • Cowpea chloroplast SSR molecule labeled polymorphic primers and screening method thereof, and method for identifying genetic relationship of cowpeas
  • Cowpea chloroplast SSR molecule labeled polymorphic primers and screening method thereof, and method for identifying genetic relationship of cowpeas

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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.); Base 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.), The search conditions are set as follows: the minimum number of repeats for a si...

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 index (PIC) calculation reference formula (Botstein D, White RL, Skolnick M, Davis RW. Con...

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Abstract

The invention discloses cowpea chloroplast SSR molecule labeled polymorphic primers and a screening method thereof and a method for identifying genetic relationship of cowpeas, belonging to the technical field of molecular biology of crops. The cowpea chloroplast SSR molecule labeled polymorphic primers are described in the specification. The method for identifying polymorphism of the cowpea chloroplast SSR molecule labeled primers comprises the following steps: extracting genomic DNAs of cowpeas of different varieties; mixing designed primers and the genomic DNAs and carrying out PCR amplification and electrophoretic separation; and acquiring the cowpea chloroplast SSR molecule labeled polymorphic primers. The method for identifying the genetic relationship of cowpeas comprises the following steps: extracting genomic DNA of a cowpea; carrying out PCR amplification and electrophoretic separation by using the cowpea chloroplast SSR molecule labeled polymorphic primers and recording statistical results; and carrying out genetic relationship analysis on the results. The polymorphic primers can reflect genetic information of plasmon of cowpeas and can be used for research on germplasm resource classification, genetic relationship identification, genetic diversity analysis and the like of cowpeas.

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 basic units consisting of 1-6 nucleotides repeated many times in the genome, which is widely distributed in different positions of the genome, and the length is general...

Claims

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Application Information

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