五味子的DNA提取及RAPD鉴定研究.pdfVIP

中国农学通报 2011,27(02):395-399 Chinese Agricu ltural Science Bulletin 五味子的DNA 提取及 RAPD 鉴定研究 张庆 田，许培磊 ，李 昌禹，艾 军 （中国农业科学院特产研究所，吉林 132109） 摘 要：采用 CTAB 、SD S、改 良CTAB 法对五味子总DNA 提取进行研究，并对总DNA 进行电泳分析和含 量测定，结果表 明以改 良CTAB 法提取五味子叶片DNA 纯度最好 ，以此 DNA 为模板可获得条带清晰、 重复性好 的RAPD 扩增结果。从 14 条重复性好 的引物 中共扩增 出67条带，平均每个 引物 4.8 个片段，多 态性位点数为 58，比例为 86.6% 。UPGMA 聚类分析 的结果与五味子形态学分类结果相似，10 份五味子 优系在遗传相似系数 0.69 处可划分为 3 个类群 ，该RAPD 技术体系可以很好地用于 10份五味子种质资 源的鉴定 。 关键词：五味子；改 良CTAB 法 ；RAPD ；鉴定 中图分类号：S567.23+9 文献标志码 ：A 论文编号：2 010-203 1 DNA Ext ra ct ion a nd RAPD Ident ification of Sch isan dr a ch in en sis (Turc .) Baill. Zhang Qingtian, Xu Peilei, Li Changyu, Ai Jun (In stitute of Sp ecial Wild Econ omic Animal and Plant Scien ce, CAAS. Jilin 132 109) Ab st ra ct : Gen omic DNA of Sch isan dra chin en sis (Turc .) Baill. was extracted by means of three different DNA extraction methods, CTAB, SDS and improved CTAB methods. Th e purity an d quality of DNA obtained by these methods were compared , and th e quality was evalu ated by agarose gel electrophoresis. Th e results showed that DNA samp le obtained by improved CTAB method h ad the h igh est purity an d qu ality. Good repeatability of RAPD amplification cou ld be obtained with th is DNA template. Fourteen random primers of good repeatability gen erated 67 bands, with an average of 4.8 fragments per primer , and 86.6% of them were polymorph ic. UPGMA an alysis sh owed similar results with the morphological classification. The accessions could b e divided into three groups at th e coefficient 0.69. The RAPD technology system can be used to identify 10 Schisandra chinensis (Turc .) Baill. accessions. Key wor d s