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中国沙漠 ›› 2006, Vol. 26 ›› Issue (5): 826-831.

• 生物与生态 • 上一篇    下一篇

耐旱苔藓植物DNA提取及优化RAPD、 ISSR反应体系的建立

张道远1, 张元明1, 曹 同2   

  1. 1.中国科学院新疆生态与地理研究所 吐鲁番沙漠植物园, 新疆 乌鲁木齐 830011; 2.上海师范大学 生命与环境科学学院, 上海 200234
  • 收稿日期:2004-12-16 修回日期:2005-07-07 出版日期:2006-09-20 发布日期:2006-09-20

Extraction of Xeric Bryophyte Genomic DNA and Establishment of Optimal RAPD/ ISSR Programmes

ZHANG Dao-yuan1, ZHANG Yuan-ming1, CAO Tong2   

  1. 1.Turpan Eremophytes Botanical Garden, Xinjiang Ecology and Geography Research Institute, Chinese Academy of Sciences, Urumqi 830011, China; 2.Shanghai Normal University, ShangHai 200234, China
  • Received:2004-12-16 Revised:2005-07-07 Online:2006-09-20 Published:2006-09-20

摘要: 耐旱苔藓植物常常单个个体矮小、生物量低,如何从细小的单个个体中有效提取总DNA是进一步开展居群遗传多样性研究的关键。本研究以古尔班通古特沙漠广泛分布的刺叶墙藓(Tortula desertorum)为对象,使用快速提取法、2×CTAB法及DNeasy plant mini kit试剂盒提取法等3种方法对刺叶墙藓单个个体的总DNA进行提取。结果表明,2×CTAB法提取的DNA纯度高,凝胶电泳显示无明显降解现象,适宜作为PCR扩增的模板。利用所提取的单个个体DNA为模板,建立了优化的RAPD、ISSR反应体系。

关键词: 苔藓植物, 刺叶墙藓, DNA提取, RAPD, ISSR反应体系

Abstract: Extracting high quality genomic DNA from single shoots of xeric bryophytes has been the bottleneck for further study on the genetic diversity. In the paper, three DNA extraction methods, simple protocol, 2 CTAB method and DNeasy plant mini kit, were tested, with Tortula desertorum as an example. Among which, 2 CTAB method could get pure DNA and was ready to be used as templates for PCR amplification. In addition, optimal PCR amplification programmes and suitable primers were firstly established when using RAPD and ISSR as DNA markers.

Key words: xeric bryophyte, Tortula desertorum, DNA extraction methods, RAPD/ISSR marker

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