生物多样性 ›› 1996, Vol. 04 ›› Issue (2): 119-122.  DOI: 10.17520/biods.1996020

• 论文 • 上一篇    下一篇

木根麦冬(Ophiopogon xylorrhizus)干叶提取DNA用于RAPD分析

张大明,陈新露,邓峥嵘   

  1. 1) (中国科学院植物研究所系统与进化植物学开放研究实验室,  北京 100093)
    2) (北京农学院园林系,  北京 102206)
  • 收稿日期:1995-03-30 修回日期:1995-07-17 出版日期:1996-05-20 发布日期:1996-05-20

DNA isolated from dried leaves of Ophiopogon xylorrhizus for Random Amplified Polymorphic DNA(RAPD) analysis

Zhang Daming, Chen Xinlu, Deng Zhengrong   

  1. 1) Laboratory of Systematic and Evolutionary Botany , the Chinese Academy of Sciences ,Beijing 100093
    2) Department of Landscape Architecture ,Beijing Agricultural College ,Beijing 102206
  • Received:1995-03-30 Revised:1995-07-17 Online:1996-05-20 Published:1996-05-20

摘要: 木根麦冬(Ophiopogon xylorrhizus)是我国珍稀濒危植物,分布仅限于云南西双版纳雨林,在植物系统学和保护生物学研究中具有独特的意义。随机扩增多态DNA(RAPD)方法是揭示群体遗传多样性的高效、简便方法,但一般均以新鲜材料提取总DNA,对一些分布边远地区物种难以采用此法。本文研究从木根麦冬干叶片中提取总DNA,进行RAPD分析。样品取自4个居群、49个个体。选取生长旺盛的叶片,在野外用硅胶快速干燥保存样品。采用高盐低pH值法提取总DNA,每克鲜重所得的干叶可得80~160 μg。通过对模板DNA的各种处理和PCR扩增程序的调整,解决了扩增片段边缘弥散、界线模糊、产率低等问题,获得了理想的扩增带型。这一成果对其它从野外直接采样的干叶提取DNA进行RAPD研究具有指导意义。

AbstractOphiopogon xylorrhizus is an endangered species,restrictedly distributed in tropical rain forest of Xishuangbanna,Yunnan,China,whihc is especially significant to studies on systematics and conservation biology of plants.RAPD is a rapid and simple method to reveal genetic diversity of populations.DNA as templets was generally isolated from fresh materials,which has difficulties in dealing with materials from remote districts.Total DNAs of 49 individuals among 4 populations were extracted using High salt low pH method from Ophiopogon xylorrhizus leaves dried through silica gel in the field.Quantity of the DNA ranged from 80 to 160 μg/dried leaves of 1 gram of fresh leaves.Compared with DNA from fresh materials,the DNA from dried leaves was not even in molecular weight and partly took place reduction,which affected PCR amplification.Good results and high production of amplification were achieved through different treatments to templet DNAs and adjustment of PCR programs.This research lays the foundation for extracting DNA from other tax using materials dried in the field for RAPD analyses.