Molecular identification method for cotton restoring line containing sterile cytoplasm of gossypium harknessii

A restorer line and cotton technology, applied in the field of molecular identification of cotton restorer line purity identification and molecular marker-assisted breeding, can solve the problems of poor accuracy, high consumption, long time, etc., and achieve the effect of clear and fast result bands

Inactive Publication Date: 2012-08-01
INST OF COTTON RES CHINESE ACAD OF AGRI SCI
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] The purpose of the present invention is to solve the problems in the prior art that the field identification and detection method of the cotton restorer line selection containing Harknessy cotton cytoplasmic male sterile cytoplasm takes a long time, consume

Method used

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  • Molecular identification method for cotton restoring line containing sterile cytoplasm of gossypium harknessii
  • Molecular identification method for cotton restoring line containing sterile cytoplasm of gossypium harknessii
  • Molecular identification method for cotton restoring line containing sterile cytoplasm of gossypium harknessii

Examples

Experimental program
Comparison scheme
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Example Embodiment

[0027] Example 1 The molecular identification method of the sterile cytoplasmic cotton restoring line of Hacknesi cotton

[0028] 2.1 DNA extraction

[0029] Restorer lines Zhonghui 46 (preservation number: CGMCC5166) and Zhonghui 80 (preservation number: CGMCC5167) containing the sterile cytoplasm of Hackneysian cotton, China Cotton Institute 12, China Cotton Institute 41, China Cotton Institute 45 A total of 6 materials from China Cotton Institute 69 were dehulled, and each seed was individually crushed and transferred to a 2ml centrifuge tube; 1000μl DNA extraction solution (0.05M Tris-Cl, 0.01 MEDTA, 2% SDS, 1% PVP, 0.5 % Sorbitol, 0.705M NaCl, adjust the pH to 8.0, autoclave; add 1% β-mercaptoethanol before use, the% in the system represents the volume ratio), vortex and mix, and then bath at 65°C for 30 minutes with an interval of about 10 minutes Shake gently; after the end of the water bath, add 800μl of phenol: chloroform: isoamyl alcohol (volume ratio 25:24:1), mix upsid...

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Abstract

The invention belongs to the field of biological technique, relates to a molecular identification method for the purity of a cotton restoring line, and particularly provides a primer of a SSR (Simple Sequence Repeat) marker for identifying the cotton restoring line containing the sterile cytoplasm of gossypium harknessii. The primer is as shown in nucleotide sequences such as SEQ ID No.1 and 2 and/or SEQ ID No. 3 and 4. The SSR marker is adopted for carrying out polymorphism detection of molecular level on the cotton restoring line material containing the sterile cytoplasm of the gossypium harknessii. The molecular identification method is fast in identification speed, high in accuracy and simple in operation and is very suitable for the indoor identification of the cotton restoring line and the assisted-selective breeding of molecular markers.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a molecular identification method for cotton restorer line purity identification and molecular marker-assisted breeding. Background technique [0002] The research and utilization of heterosis has a significant effect on increasing the yield, quality and resistance of cotton. At present, most cotton farmers in my country's seedling and transplanting areas have planted hybrid cotton; and the cotton areas in the Yellow River Basin, and even direct cotton areas such as Xinjiang are also vigorously developing hybrid cotton. However, the production mode of cotton hybrid seeds is mainly artificial detasseling assisted pollination hybrid seed production method (accounting for more than 90%). With the increase of labor costs in major cotton production areas and their surrounding areas year by year, the cost of seed production has become higher and higher, which greatly limits th...

Claims

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Application Information

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