Indel Marker of Cotton Cytoplasmic Male Sterile Restorer Line and Its Identification Method

A male sterility, cytoplasmic technology, applied in biochemical equipment and methods, recombinant DNA technology, microbial determination/inspection, etc., can solve the problems of long time, poor accuracy, large consumption, etc. , easy to detect effects

Active Publication Date: 2019-02-01
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The purpose of the present invention is to provide a method for identifying cotton cytoplasmic male sterile restorer line InDel markers and its application for the problems of long time, large consumption, and poor accuracy of field identification and detection methods for cotton restorer line selection in the conventional breeding process.

Method used

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  • Indel Marker of Cotton Cytoplasmic Male Sterile Restorer Line and Its Identification Method
  • Indel Marker of Cotton Cytoplasmic Male Sterile Restorer Line and Its Identification Method
  • Indel Marker of Cotton Cytoplasmic Male Sterile Restorer Line and Its Identification Method

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Embodiment 1

[0039] Example 1 Molecular Identification Method of Cotton Cytoplasmic Male Sterile Restorer Line

[0040] 1. DNA extraction

[0041] Referring to the mature cotton CTAB DNA extraction technology, restorer lines Zhonghui 46 (CGMCC5166, disclosed in patent application No. 201210092777.6) and Zhonghui 80 (CGMCC5167, disclosed in patent application No. Peel off the shells of 5 seeds of Cotton Institute 83 and conventional cotton varieties Zhongmiansuo 45 and Zhongmiansuo 69, grind each seed separately and transfer to a 2ml centrifuge tube; add 1000 μl DNA extraction solution (0.05M Tris-Cl , 0.01MEDTA, 2% SDS, 1% PVP, 0.5% sorbitol, 0.705M NaCl, PH = 8.0, autoclaved; add 1% β-mercaptoethanol before use), vortex and mix well, 65 ℃ water bath for 30min, Shake the centrifuge tube gently at intervals of 10 minutes; after the water bath, add 800 μl phenol: chloroform: isoamyl alcohol (25:24:1), mix up and down until there is no layering (gentle action, enough time), centrifuge at 120...

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Abstract

The invention belongs to the field of biotechnologies, and relates to an InDel marker for cotton cytoplasm male sterility restoring lines.The length of the InDel marker is 102bp, and nucleotide sequences are specific insert fragments shown as SEQ ID No.1.The invention further provides a method for identifying molecules by the aid of the InDel marker.The method includes steps of carrying out PCR (polymerase chain reaction) by the aid of specific primer pairs, amplifying DNA (deoxyribonucleic acid) of to-be-tested cotton materials and judging the cotton materials according to amplification results on the basis of standards that each cotton material with a single amplified 267bp band does not contain restoring gene loci; carrying out homozygosis on restoring gene loci of each cotton material with a single amplified 369bp band; carrying out heterozygosis on restoring gene loci of each cotton material with double simultaneously amplified 267bp and 369bp bands.The InDel marker and the method have the advantages that molecular-level polymorphism detection is carried out on the cotton cytoplasm male sterility restoring line materials by the aid of the InDel marker, identification result bands are clear, stable and reliable and are easy to detect, and accordingly the InDel marker and the method are applicable to cotton cytoplasm male sterility restoring line material purity indoor identification and molecular-marker-assisted selective breeding and improvement.

Description

technical field [0001] The invention belongs to the field of biotechnology, and specifically relates to a method and application for identification using the InDel marker of a cotton cytoplasmic male sterility restorer line. Background technique [0002] The utilization of heterosis plays an important role in increasing cotton yield, improving quality, improving resistance, etc., and will continue to play an important role in promoting cotton production. At present, the main production method of hybrid cotton seeds is still artificial detasseling assisted pollination hybrid seed production method (accounting for more than 90%). With the needs of the country's urbanization development, labor costs in the main cotton producing areas and surrounding areas are increasing year by year, resulting in higher and higher hybrid production costs, and many related seed companies engaged in hybrid cotton seed production are unsustainable, thus limiting Further large-scale popularization...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6895C12N15/11
CPCC12Q1/6895C12Q2600/13C12Q2600/156
Inventor 吴建勇邢朝柱张梦郭立平戚廷香
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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