EST-SSR labeled primer combination and screening method for vegetable use, grain use and wild soybean genetic diversity analysis and authentication

A technology for genetic diversity and vegetable soybeans, applied in the field of molecular biology, can solve the problems of genetic diversity identification that have not been reported yet

Inactive Publication Date: 2015-09-09
ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, in soybean, although some EST-SSR markers have been developed, most of them are based on grain soybean, and rarely used in vegetable soybean; Soybean molecular marker development and genetic diversity identification have not been reported

Method used

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  • EST-SSR labeled primer combination and screening method for vegetable use, grain use and wild soybean genetic diversity analysis and authentication
  • EST-SSR labeled primer combination and screening method for vegetable use, grain use and wild soybean genetic diversity analysis and authentication
  • EST-SSR labeled primer combination and screening method for vegetable use, grain use and wild soybean genetic diversity analysis and authentication

Examples

Experimental program
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Effect test

Embodiment 1

[0060] The EST-SSR marker primer combination and screening method analyze and identify the genetic diversity of vegetable, grain and wild soybeans. The specific steps are as follows:

[0061] (1) Preparation of germplasm materials.

[0062] As shown in Table 1, the soybean materials utilized in the present invention include 35 parts of vegetable soybeans, 18 parts of grain soybeans and 15 parts of wild soybeans.

[0063] Table 1 Information table of soybean germplasm.

[0064]

[0065] (2) DNA extraction of germplasm samples

[0066] Seedlings of the tested germplasm were grown in the bean breeding base of the Institute of Vegetables, Zhejiang Academy of Agricultural Sciences. Young leaves were collected when the seedlings grew to four leaves, and stored at -70°C after quick-freezing with liquid nitrogen. Total DNA was extracted using a kit (DNeasy Plant Mini Kit, QIAGEN).

[0067] (3) Primer design

[0068] Download soybean EST data from PlantGDB database (http: / / ww...

Embodiment 2

[0075] Below in conjunction with Example 1, the EST-SSR marker primer combination and screening method for vegetable, grain and wild soybean genetic diversity analysis and identification experiment results are analyzed as follows.

[0076] figure 1 For the detection results of EST-SSR labeled fluorescent capillary electrophoresis, through the analysis of 615 pairs of primers, a total of 386 pairs were effectively amplified, and obvious bands could be obtained in the same lane.

[0077] As shown in Table 2, 35 pairs of ETS-SSR markers were screened and used for identification of soybean germplasm resources.

[0078] like figure 2 As shown, 35 servings of vegetable soybeans, 18 servings of grain soybeans and 15 servings of wild soybeans were successfully divided into three categories.

[0079] Table 2 Information table of soybean EST-SSR primers

[0080]

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Abstract

The invention discloses an EST-SSR labeled primer combination which comprises 35 pairs of EST-SSR available primers. A screening method of the EST-SSR labeled primer combination includes the steps of SSR locus screening in a soybean EST sequence, primer design, PCR amplification, product capillary electrophoresis detection and label screening. The EST-SSR labeled primer combination is applied to genetic diversity of vegetable use soybeans, grain use soybeans and wild soybeans; genetic diversity and clustering of the vegetable use soybeans, the grain use soybeans and the wild soybeans are analyzed; germplasm of the vegetable use soybeans, the grain use soybeans and the wild soybeans is quickly and accurately analyzed and distinguished easily, the genetic relationship of the vegetable use soybeans, the grain use soybeans and the wild soybeans is analyzed, and a foundation is laid for directional breeding of the species.

Description

technical field [0001] The present invention belongs to the field of molecular biology, and in particular relates to an EST-SSR marker primer combination and a screening method for analyzing and identifying vegetable, grain and wild soybean genetic diversity. Background technique [0002] Soybean originated in China and has been planted for 5000-6000 years. Since the discovery of wild soybeans, with the improvement and domestication of this species, soybeans have gradually differentiated into two types: grain soybeans and vegetable soybeans. Among them, vegetable soybean not only has a good taste, but also is rich in protein with superior nutrition, a variety of free amino acids and vitamins. It is recognized as a health food today and has a very broad market prospect. So far, China has become the world's largest producer and exporter of vegetable soybeans. However, the identification and breeding of vegetable soybean germplasm resources in my country started relatively la...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12N15/10C12Q1/68
Inventor 徐盛春龚亚明胡齐赞刘娜
Owner ZHEJIANG ACADEMY OF AGRICULTURE SCIENCES
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