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几种濒危植物及其近缘类群总DNA的提取与鉴定
ISOLATION AND CHARACTERIZATION OF TOTAL DNA FROM SEVERAL ENDANGERED SPECIES AND THEIR ALLIES
【摘要】 用低pH 介质,高盐沉淀蛋白质方法成功地从银杉(Cathaya argyrophylla Chun etKuang)、矮牡丹(Paeonia suffruticosa var. spontanea)、南川升麻(Cim icifuga nanchuanensisHsiao)、裂叶沙参(Adenophora lobophylla)的同属种泡沙参(A. potaninii)等植物中提取和部分纯化了细胞总DNA,并对其产率、质量和纯度作了鉴定。此方法的关键是用了一个低pH提取介质,它能有效防止组织破碎及沉淀大量材料时的电离化作用及酚化合物的进一步氧化。所得DNA 不需经氯化铯梯度离心或柱层析,直接可用于限制性片断长度多态性(RFLP)及随机扩增的DNA多态性(RAPD)等分子水平的遗传标记。为检测濒危植物的遗传多样性提供了一套迅速、简便和可靠的技术方案
【Abstract】 A low pH extraction medium with high salts, which avoids ionization and subsequent oxidation of phenolic compounds during tissue grinding and precipitation of large amounts of materials, were successfully used to obtain total DNA from Cathaya argyrophylla Chun et Kuang, Paeonia suffruticosa var. spontanea, Cimicifuga nanchuanensis Hsiao, Adenophora potaninii (Congeneric species with A. lobophylla) etc. The DNA yields, quality and purity were characterized. These isolated DNA could be used directly for RFLP and RAPD analysis which are useful as molecular genetic markers without sedimentation in cesium chloride gradient or column chromatography. A fast, inexpensive and reliable procedure has been developed for detecting the genetic diversity of endangered plants.
【Key words】 Isolation of total DNA; Low pH extraction medium; Digestion with restriction enzyme; RAPD;
- 【文献出处】 植物学报 ,Acta Botanica Sinica , 编辑部邮箱 ,1994年07期
- 【分类号】Q946.2
- 【被引频次】376
- 【下载频次】929