含笑属RAPD反应条件优化及遗传多态性分析  

Genetic Diversity Analysis and Optimization of RAPD Reaction System of Michelia

作　　者：唐历波[1] 周鹤峰[1] 陈东[1] 

机构地区：[1]遵义医学院珠海校区生物工程系,广东珠海519043

出　　处：《安徽农业科学》2009年第1期46-48,64,共4页Journal of Anhui Agricultural Sciences

基　　金：遵义医学院校区启动基金(KY200603)

摘　　要：[目的]确立含笑属植物RAPD反应的最优条件,确定7种含笑属植物的亲缘关系。[方法]采用改进的CTAB法提取木兰科含笑属7种植物叶总DNA,进行RAPD分析,分别研究了模板DNA、引物、dNTP、Mg2+和TaqDNA聚合酶用量对RAPD反应的影响。[结果]RAPD最优反应体系为:反应体积20μl,内含125 ng模板DNA,0.3μmol/L引物,0.5 UTaq聚合酶,1.5 mmol/LMgCl2,0.1 mmol/L dNTP,2μl10×buffer。应用Nei距离法计算各种间的相似系数,采用UPGMA法进行聚类分析,7种含笑种可分为3大类。[结论]筛选的最佳RAPD反应条件可为含笑属植物的分类和遗传多样性分析提供理论依据。[ Objective ] The optimization of RAPD reaction condition were established to determine the relationship of seven species of Michelia. [Method] The improved CrAB method was applied to extracted genomic DNA in the leaves of 7 species of Michelia. Using the method of single factor in the experiment, the effects of different concentration of MgCl2, dNTP,template DNA, random primers and Taq DNA polylnerase on RAPD reaction were studied. [Result] The optimization of RAPD reaction system with total volume of 20 μl including 125 ng template DNA, 0.3 μmol/L primer, 0.5 U the Taq polymerase, 1.5 mmol/L MgC12, 0.1 mmol/L dNTP and 2 μl 10x buffer were established. The Nei＇s genetic similarity coefficient was calculated by DPS software, 7 species of Michelia were divided into three groups with UPGMA method. [ Conclusion] The result provided the theory foundation for Michelia Umn classification and genetic diversity analysis.

关 键 词：含笑属 反应体系 RAPD 聚类分析 

分 类 号：Q943[生物学—植物学]