含笑属RAPD反应条件优化及遗传多态性分析

[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、Mg；^2＋和Taq DNA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125ng模板DNA，0．3μmol／L引物，0．5U Taq聚合酶，1．5mmol／L MgCl2，0．1mmol／L dNTP，2μl 10x 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。

Genetic Diversity Analysis and Optimization of RAPD Reaction System of Michelia

[Objective] The optimization of RAPD reaction condition were established to determine the relationship of seven species of Michelia.[Method] The improved CTAB 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 polymerase 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 MgCl2,0.1 mmol/L dNTP and 2 μl 10×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 Linn classification and genetic diversity analysis.