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大麻性别的RAPD和SCAR分子标记

陈其军, 韩玉珍, 傅永福, 赵德刚, 国凤利, 孟繁静, 刘卫平

中国农业大学生物学院,农业部植物生理生化开放实验室.北京 100094

摘要

利用随机扩增多态性DNA(random amplified polymorphic DNA,RAPD)技术获得与大麻性别连锁的分子标记。将10株雄性大麻或10株雌性大麻的单个DNA样品等量混合分别组成雄性或雌性DNA池(DNA pool),以提供具有相同遗传背景的雌、雄性DNA样品。每个随机引物分别用三个不同的循环程序进行PCR扩增。在30个随机引物中,用引物S401扩增得到一条约2.5kb雄性多态性片段。对该片段进行了克隆和序列分析,并根据序列分析结果将上述RAPD分子标记转化为重复性和特异性更好的SCAR(sequence characterized amplified regions)分子标记。

关键词： 性别; 分子标记; RAPD; SCAR; 大麻

RAPD and SCAR molecular markers of sexuality in the dioecious species Cannabis sativa L.

Chen Qijun, Han Yuzhen, Fu Yongfu, Zhao Degang, Guo Fengli, Meng Fanjing, Liu Weiping

College of Biology,China Agricultural University.Beijing 100094

Abstract

In this study the random amplified polymorphic DNA (RAPD) technque was employed with the objective of finding markers linked to sex determination in hemp. Ten male individual DNA samples or ten female individual DNA samples or ten female individual DNA sampes were prepared and contributed to two DNA pools: one was male DNA pool and another female DNA pool, with the intention of providing a common genetic background for each pool and leaving the sex-linked materials as the primary difference between the two pools. A total of 30 10-mer primers was tested with at least three different PCR cycling procedures. A male-associated fragment with a length of about 2.5 kb was generated with S401 primer. The fragment was cloned and sequenced. In order to convert the RAPD marker into SCAR (sequence characterized amplified regions) marker, one 20-mer and another 22-mer specific primers were constructed and used for PCR amplifying. The male-linked dominant SCAR marker was obtained, which would favor the screening of all-gynoecious hemp lines.

Key words: Sexuality;Molecular markers;RAPD;SCAR;Cannabis sativa L.

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引用本文：

陈其军, 韩玉珍, 傅永福, 赵德刚, 国凤利, 孟繁静, 刘卫平. 大麻性别的RAPD和SCAR分子标记[J]. 生理学报 2001; 53 (2): .

Chen Qijun, Han Yuzhen, Fu Yongfu, Zhao Degang, Guo Fengli, Meng Fanjing, Liu Weiping. RAPD and SCAR molecular markers of sexuality in the dioecious species Cannabis sativa L.. Acta Physiol Sin 2001; 53 (2): (in Chinese with English abstract).