摘要
木根麦冬（Ｏｐｈｉｏｐｏｇｏｎｘｙｌｏｒｒｈｉｚｕｓ）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８０～１６０μｇ。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。
Ophiopogon 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.
出处
《生物多样性》
CAS
CSCD
1996年第2期119-122,共4页
Biodiversity Science
基金
国家自然科学基金
中国科学院系统与进化植物学开放实验室支持项目
关键词
麦冬
木根麦冬
干叶
DNA提取
Ophiopogon xylorrhizus, dried leaves, DNA extraction, RAPD, amplification condition
