摘要
选取日本 (普陀、嵊泗两个自然群体 )、锈斑、锐齿、三疣梭子蟹、红星梭子蟹、青蟹共 6种海产蟹 ,用RAPD方法检测了其基因组DNA的多态性。在事先优化的反应条件下 ,用 16个随机引物共扩增出 12 3条多态性片段 ,片段大小在 5 0 0～ 14 0 0 0bp。在 12 3条扩增片段中 ,6种蟹共享的片段有 2条 ,3种有13条 ,2种梭子蟹有 16条 ;6种蟹之间随机扩增DNA片段的共享度在 0 0 6 0 6～ 0 80 0 0 ,相对遗传距离指数在0 2 0 0 0～ 0 9394。根据获得的相对遗传距离指数 ,用PHYLIP 3 5软件包中的UPGMA和NJ方法进行聚类处理。UPGMA和NJ聚类图均显示三疣梭子蟹和红星梭子蟹先聚在一起 ,再和青蟹聚在一起。
The phylogenetic relationship of six marine crabs was studied using random amplified polymorphic DNA (RAPD) technique.Predetermined optimal reaction conditions were as follows:Samples were first heated at 94?℃ for 4 min;and followed by 45 cycles of 1 min at 94?℃,1 min at 36?℃,and 2 min at 72?℃ then held at 72?℃ for 10 min.With 16 primers,123 reproducible amplified fragments ranging between 500 and 14 000 bp were acquired.In all bands,2 bands were shared by all crabs,13 shared by Charybdis japonica , C.acuta and C.feriatus ,16 shared by Portunus sanguinolentus and P.trituberculatus .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 using Nei and Li's matching coefficient method.The degree of F was 0 0606-0 8000.The value of 1-F was used to evaluate genetic distances between species.The genetic distances between species were 0 2000-0 9394.The phylogenetic trees were constructed with the method of UPGMA and NJ on the basis of genetic distances.The results from the two methods of cluster analysis are similar in general,and the relationships indicated by the phylogenetic trees show the difference between genus species.The results are in good overall agreement with classical taxonomy.
基金
浙江省自然科学基金资助项目 (3 0 2 44 0 )
宁波市青年基金资助项目 (2 0 0 3A62 0 13 )
