耐旱苔藓植物DNA提取及优化RAPD、ISSR反应体系的建立

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

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RAPDI、SSR反应体系。

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.