3种鲍基因组DNA多态性的RAPD分析

RAPD Analysis of the polymorphism of genomic DNA among three abalones

利用随机扩增多态DNA技术检测了中间鲍Haliotismidae、皱纹盘鲍Haliotisdiscushannai和九孔鲍Haliotisdiversicolorsupertexte3种鲍基因组DNA的多态性,初步进行了分子水平上的分类研究。经24个随机引物扩增,共得到376条多态性片段,片段长度为250～2500bp。根据片段的共享度计算出平均遗传距离指数,并采用UPGMA和NJ两种聚类分析方法进行处理得到系统树。在UP GMA聚类中,中间鲍与九孔鲍先聚在一起,有较近的亲缘关系,其次是皱纹盘鲍,而它们之间的遗传距离相差不大,NJ聚类图上就反映出3种鲍聚为3类,所以两种聚类分析结果基本一致,与它们形态上存在明显差异的结果基本吻合,说明RAPD技术可以作为鲍种质鉴定和分类的一种有效、简便的辅助手段。

Polymorphism of genomic DNA and molecular classification of three species of abalones, Haliotis midae, Haliotis discus hannai, Haliotis diversicolor supertexte,were determined by random amplified polymorphic DNA (RAPD). Amplifications with 24 primers gave 376 reproducible amplified fragments ranging between 250 and 2500 bp in length,the amplified fragments were scored as present(1) or absent(0) for each DNA sample and an index of degree of band sharing(F) was calculated by Nei and Lis matching coefficient method. The value of(1-F) was used to evaluate genetic distances between species. The phylogenetic trees were constructed with the methods of UPGMA and NJ on the basis of genetic distances. UPGMA analysis indicated that there was a close relationship between H. midae and H. diversicolor supertexte, followed by H. discus hannai, and no significant genetic distances between the abalones. RAPD approach provides a new method for efficient and simple classification and identification of new strains for abalones.