Method for identifying hybrid soybeans seeds by molecular markers

A technology for soybean seeds and molecular markers, which is applied in the directions of biochemical equipment and methods, microbial determination/inspection, DNA/RNA fragments, etc., to achieve the effects of convenient operation, simple method and simple operation method.

Active Publication Date: 2012-01-25
JILIN ACAD OF AGRI SCI
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Problems solved by technology

[0004] The invention provides a method for identifying hybrid soybean seeds with molecular markers, which solves many defects in the existing identification methods for seed purity

Method used

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  • Method for identifying hybrid soybeans seeds by molecular markers
  • Method for identifying hybrid soybeans seeds by molecular markers
  • Method for identifying hybrid soybeans seeds by molecular markers

Examples

Experimental program
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Embodiment 1

[0043] The experimental materials were hybrid soybean seeds and their respective parent material seeds.

[0044] The experimental method is to extract the genomic DNA of the above-mentioned experimental materials, use SSR molecular markers to perform PCR amplification, conduct gel electrophoresis on the amplified products, count the results and screen the characteristic primers, and use the characteristic primers to identify the purity of the hybrid species to be detected.

[0045] Soybean Seed Genomic DNA Extraction

[0046] (1) Take a hybrid soybean seed and place it in a mortar, add liquid nitrogen and grind it into a powder, take 0.2g of the powder and pour it into a 1.5 ml centrifuge tube;

[0047] (2) Add 600ml 65°C preheated 1×CTAB buffer (50 mM Tris-Cl pH8.0, 0.7 M NaCl, 10 mM EDTA pH8.0, 1% CTAB) and 40ml mercaptoethanol to the centrifuge tube; Stir the sample powder and the buffer solution quickly so that it does not form a lump, bathe in 65°C water for 30 minutes...

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Abstract

The invention discloses a method for identifying the purity of hybrid soybeans seeds by molecular markers, comprising the following steps of: using identified hybrid soybean varieties improved by RN cytoplasmic male sterility triple system and parental gene groups DNA thereof as a template, by SSR (simple sequence repeat) analysis, establishing SSR specificity fingerprint of the hybrid soybean and respective parents, successfully obtaining codominant an SSR marker which can distinguish the parents and the hybrid seeds definitely, and carrying out seed purity detection of the identified hybridsoybean variety improved by the RN cytoplasmic male sterility triple system according to the specificity markers. The result and the field identification are consistent by random group identificationof the hybrid soybean varieties. The method is simple and convenient to operate, the seed purity of the hybrid soybean varieties improved by the RN cytoplasmic male sterility triple system can be fast, accurately and reliably identified by experiment means through the screened specific molecular markers, so that the risk causing possible production reduction due to insufficient purity of the seeds in the sales process of the hybrid soybean seeds can be reduced.

Description

technical field [0001] The invention discloses a method for detecting hybrid soybean seeds by molecular markers, which is suitable for identifying the purity of hybrid soybean seeds bred by RN type cytoplasmic male sterility "three lines", and belongs to the technical field of plant molecular marker detection. Background technique [0002] Utilizing heterosis is one of the most effective measures to greatly increase soybean yield. my country is in the leading position in the field of soybean heterosis research. The Jilin Academy of Agricultural Sciences used the "three lines" of cytoplasmic male sterility to breed and approve the world's first soybean hybrid "Hybrid Dou No. 1" in 2002, which has increased yield compared with the control 20.9%. Afterwards, Jilin Academy of Agricultural Sciences bred and approved Hybrid Beans No. 2, No. 3, No. 4 and No. 5, Anhui Academy of Agricultural Sciences bred and approved Zayoudou No. 1 and No. 2, and Fuyang Institute of Agricultural Sc...

Claims

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Application Information

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IPC IPC(8): C12Q1/68C12N15/11
Inventor 张春宝赵丽梅王玉民彭宝张伟龙张伟张井勇赵晓明
Owner JILIN ACAD OF AGRI SCI
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