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Molecular identification method of cytoplasm male sterile restoring line

A technique for male sterility and restorer lines, which is applied in the fields of primers and molecular identification for amplifying them, can solve the problems of high consumption, poor accuracy, speed up the breeding and improvement process of cotton restorer lines, etc. clear effect

Inactive Publication Date: 2015-02-11
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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Problems solved by technology

[0005] The purpose of the present invention is to solve the problems of long time, high consumption and poor accuracy of the field identification and detection method for the selection of cotton cytoplasmic male sterile restorer lines in the conventional breeding process, and to provide a fast molecular method for cotton cytoplasmic male sterile restorer lines Identification methods, which can speed up the process of cotton restorer line breeding and improvement, while ensuring the purity of restorer line seeds

Method used

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  • Molecular identification method of cytoplasm male sterile restoring line
  • Molecular identification method of cytoplasm male sterile restoring line
  • Molecular identification method of cytoplasm male sterile restoring line

Examples

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Embodiment 1

[0030] Example 1 Establishment of Molecular Identification Method for Cotton Cytoplasmic Male Sterility Restorer Line

[0031] 1. Cotton seed DNA extraction

[0032] The restorer lines Zhonghui 46 and Zhonghui 80, the three-line hybrid Zhongmiansuo 83, and the conventional cotton varieties Zhongmiansuo 45 and Zhongmiansuo 69 were dehulled, and each seed was crushed separately and transferred 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, adjust the pH value to 8.0, autoclave; add 1%β before use -Mercaptoethanol), after vortex mixing, 65 ℃ water bath for 30 minutes, shake gently at intervals of about 10 minutes; after the water bath, add 800 μl phenol:chloroform:isoamyl alcohol (25:24:1), mix up and down until no separation layer (gentle action for sufficient time), centrifuge at 12000rpm for 10min; absorb the supernatant (about 800μl) and transfer it to another 2ml centrifuge tube, add 2μl RN...

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Abstract

The invention belongs to the technical filed of biology, and relates to a molecular identification method of a cytoplasm male sterile restoring line, and concretely provides a CAPS-labeled primer used for identifying the cytoplasm male sterile restoring line, and a nucleotide sequence is shown as SEQ ID NO. 2 and SEQ ID NO.3. The method employs the CAPS label for a polymorphism detection of molecule level of a cytoplasm male sterile restoring line material, The method has the advantages of fast identification speed, high accuracy and simple operation, and is very suitable for indoor identification of purity of the cytoplasm male sterile restoring line material and molecular marker-assisted selection of a new restoring line material.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a molecular marker for identifying a cotton cytoplasmic male sterile restorer line, a primer for amplifying it and a molecular identification method. Background technique [0002] The research and utilization of heterosis has played a significant role in improving cotton yield, quality and resistance. At present, hybrid cotton has been widely planted in seedling and transplanting areas such as the Yangtze River Basin in my country; hybrid cotton is being vigorously developed in cotton areas in the Yellow River Basin, and even direct cotton areas such as Xinjiang. However, artificial emasculation and assisted pollination hybrid seed production method is the main production method of hybrid cotton seeds (accounting for more than 90%). With the needs of the country's urbanization development, the labor costs in the main cotton producing areas and their surrounding areas have increased ye...

Claims

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Application Information

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IPC IPC(8): C12N15/11C12Q1/68
Inventor 吴建勇邢朝柱郭立平戚廷香
Owner INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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