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SSR molecular marker primer set for identifying purity of variety Shandong cotton research number 34 and application thereof

A molecular marker and variety purity technology, applied in the field of biotechnology, can solve the problems affecting the development of the market sales reputation of fine varieties, the decline of the heterosis of hybrid varieties, the influence of the purity of fine varieties, etc., so as to improve the efficiency of electrophoresis detection and the detection speed. , economical and simple effect

Inactive Publication Date: 2016-06-22
SHANDONG COTTON RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the process of cotton hybrid seed production, artificial detasselling is not complete or omission of emasculation often occurs, false hybrids often appear, leading to the decline of heterosis in hybrid varieties, causing huge economic losses to production. At the same time, some criminals or seeds in the sales process In order to pursue high profits, business units substitute inferior products for superior ones, artificial adulteration, etc., which seriously affect the purity of fine varieties, which in turn affects the market reputation and industrialization development of fine varieties

Method used

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  • SSR molecular marker primer set for identifying purity of variety Shandong cotton research number 34 and application thereof
  • SSR molecular marker primer set for identifying purity of variety Shandong cotton research number 34 and application thereof
  • SSR molecular marker primer set for identifying purity of variety Shandong cotton research number 34 and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] DNA was extracted, PCR amplification was performed using SSR molecular markers, the amplified products were subjected to non-denaturing polyacrylamide gel electrophoresis, the results were counted and characteristic primers were screened.

[0043] 1. Cotton genomic DNA extraction

[0044] The experimental materials in this example are Lumianyan No. 34 variety and Lumianyan No. 34 parent parent (purchased from Shandong Luyi Cotton Technology Co., Ltd.).

[0045]Soak dry cotton seeds at room temperature for about 20 hours, peel off the seed coat, put the germinated seed kernels into a 2ml centrifuge tube, add 700 μl of lysis buffer (composition: 2wt% CTAB, 0.02mol / LEDTA, 1.4mol / LNaCl, 0.1mol / LTris-HCl, 2wt% PVP, 1wt% β-mercaptoethanol), and crush the seeds with a tissue grinder. Water bath at 65°C for 20 minutes, and shake once every 10 minutes or so. Centrifuge at 12,000 r / min for 8 min (15°C), take 500 μl of supernatant, add 0.7 times the volume of pre-cooled isoprop...

Embodiment 2

[0056] Use the 5 SSR molecular marker characteristic primers screened out in Example 1 to carry out the purity identification of Lumianyan No. 34 commodity seeds, the steps are as follows:

[0057] 1. DNA extraction

[0058] The cotton genome DNA extraction method is the same as that in Example 1.

[0059] 2. SSR-PCR amplification

[0060] Select the SSR molecular marker primer set in Example 1 to carry out PCR amplification respectively, reaction system:

[0061] 10×PCRBuffer1μL, 10mMdNTPMix0.2μL, specific SSR forward and reverse primers (10μmol / L) 0.6μL each, 5UTaq DNA polymerase 0.1μL, test sample DNA (20~200ng / μL) 2μL, ddH 2 O5.6 μL.

[0062] Reaction procedure:

[0063] Pre-denaturation at 94°C for 5min; denaturation at 94°C for 40s, annealing at 55°C for 45s, extension at 72°C for 50s, cycle 32 times; extension at 72°C for 5min; storage at 4°C.

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Abstract

The invention relates to an SSR molecular marker primer set for identifying the purity of Lumianyan No. 34 variety and its application. The primer set consisted of 5 pairs of primers, namely: primers for molecular marker NAU3995, primers for molecular marker MUCS101, primers for molecular marker DPL0917, primers for molecular marker DPL0528, and primers for molecular marker NAU4926. The present invention screens and studies the hybrid Lumianyan 34 and its parental genome DNA characteristic SSR primers, and identifies that the bands with specific markers of the father and mother can be produced simultaneously in the first generation of hybrids, and the band type is clear and repeatable. 5 pairs of characteristic SSR primers of Lumianyan No. 34 with strong reliability constitute the SSR molecular marker primer set for identifying the purity of Lumianyan No. 34 variety. This primer set can be used to quickly detect the seed purity of Lumianyan No. 34 variety.

Description

technical field [0001] The invention relates to a SSR molecular marker primer set for identifying the purity of Lumianyan No. 34 variety and its application. The characteristic primer set of Lumianyan No. 34 is used to identify the variety purity of Lumianyan No. 34 samples, which belongs to biotechnology technology field. Background technique [0002] Cotton is an important economic crop in our country and plays an important role in the development of our national economy and society. Compared with conventional cotton varieties, cotton hybrid varieties have star advantages in terms of vitality, growth potential, stress resistance and high yield. Therefore, they are favored by the majority of cotton farmers. Their promotional planting area basically covers the cotton area in the Yangtze River Basin, which is helpful for increasing cotton production. It plays an important role in stabilizing cotton production and increasing the income of cotton farmers. The purity of hybrid...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68C12N15/11
CPCC12Q1/6895C12Q2600/156
Inventor 刘国栋王芙蓉张军张传云张景霞陈煜周娟
Owner SHANDONG COTTON RES CENT
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