大紫蛱蝶三个地理种群的RAPD遗传多样性分析

RAPD ANALYSIS OF GENETIC DIVERSITY AMONG THREE POPULATIONS OF SASAKIA CHARONDA

采用RAPD技术分析了大紫蛱蝶朝鲜亚种Sasakiacharondacoreana 3个地理种群的遗传多样性 ,并以同属的黑紫蛱蝶指名亚种Sasakiafunebrisfunebris (Leech)及近缘属种黑脉蛱蝶指名亚种Hestinaassimilisassimilis (Lin naeus)为外群 ,探讨了它们之间的亲缘关系。经筛选的 10个随机引物对供试的 5 8只蝶类标本共产生 2 0 0条带 ,扩增谱带具有明显的属、种间多态性。

Random Amplified Polymorphic DNA (RAPD)markers were applied to analyze genetic diversity among three populations of Sasakia charonda coreana, Sasakia funebris funebris (Leech) and Hestina assimilis assimilis (L.) were used as outgroups. Genomic DNA of fifty-eight individuals was extracted from dissected leg muscle using saturated NaCl and then amplified by 10 random primers (10 bp) which were previously selected. A total of 200 clear and reproducible bands were generated. The RAPD patterns showed polymorphic variations among and within different genera and species. Shannon's index showed that higher genetic diversity existed within Hestina assimilis assimilis (Linnaeus), while Sasakia funebris funebris (Leech) had the lowest genetic diversity. Meanwhile, genetic diversity also occurred among different populations of Sasakia charonda coreana, the genetic diversity value estimated by Shannon's index is 18.5%. The amplified fragments were analyzed by the Phyltools 6.0, the molecular phylogenetic tree based on Nei's genetic distance of RAPD markers was constructed using UPGMA and Neighbor-Joining. The dendrogram indicated strong similarities within populations. Among three populations of Sasakia charonda coreana, the individuals of Hubei population and Shanxi population were closely clustered together; and then they were clustered with Liaoning population. The clustering followed the order of, first the individuals within a population, then the different populations of the same species, later the species of the same genus, and finally the species among genera. Sasakia charonda coreana was closely related to Sasakia funebris funebris (Leech), whereas Hestina assimilis assimilis (L.) has far phylogenetic relationship with them, the mean Nei's genetic distance is 0.113 and 0.147 respectively. These data suggested that several implications. Firstly, RAPD is useful as an effective molecular marker to distinguish the divergence in Sasakia and its closed related genera. Secondly, there seems to be a tendency of correlations between the genetic and geographic distance among the 3 different populations of Sasakia charonda coreana from the south to the north. Thirdly, the RAPD similarity levels follows the order of individuals>populations>species>genera. The results of dendrogram based on RAPD markers are consistent with the previous conclusions of morphologic classification.