RAPD Markers Developed for Identifying Male Sterility in Tagetes erecta

文献类型： 外文期刊

第一作者： Wang, Zhigang

作者： Wang, Zhigang;Yin, Dongsheng;Wang, Ping;Zhao, Jingyun;Pan, Baitao;Zhang, Huihua

作者机构：

关键词： Tagetes erecta;male sterility;RAPD;molecular marker

期刊名称：XXIII INTERNATIONAL EUCARPIA SYMPOSIUM, SECTION ORNAMENTALS, COLOURFUL BREEDING AND GENETICS - PT II

ISSN： 0567-7572

年卷期： 2010 年 855 卷

页码：

收录情况： SCI

摘要： Various experimental protocols for RAPD analysis of genes related to fertility in Tagetes erecta were studied and an optimum procedure was developed for setting up a stable system. The CTAB and SDS method were compared to extract DNA from different tissues of Tagetes erecta. The quality of DNA extracted was evaluated by spectrophotometry and PCR. Results showed that total DNA extracted from young leaves by CTAB was suitable for the analysis of RAPD. The concentration of template DNA, MgCl2, buffer and dNTPs were examined. Amplifications were then carried out in a 20 mu l volume containing 5 mu l diluted template DNA, 2 mu l 10x PCR buffer, 2.5 mmol MgCl2, 0.3 mmol dNTP, 0.3 mu mol primer and 1.50 U Taq DNA polymerase. PCR were operated under the following program: 5 min at 94 degrees C before amplification, 40 cycles of 94 degrees C for 30 s, 37 degrees C 1 min, 72 degrees C 1 min, followed by one cycle of 72 degrees C for 1 min. By using this system, 100 sterile and fertile plants of W205, a two type male sterile line in Tagetes erecta were tested. Two hundred eighty eight bands were obtained using 64 RAPD primers, among which one band of 980 bp derived from the primer G-02(5'-GGCACTGAGG-3') was cosegregating with fertile plants. This RAPD marker named as G-02-980 could be used to identify male and fertile seedlings.

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