RAPD Genetic Markers in Six Species of Marine Crab
摘要
The phylogenetic relationship of six marine crabs was studied using \nrandom amplified polymorphic DNA (RAPD) technique.Predetermined optimal \nreaction conditions were as follows:Samples were first heated at \n94°C for 4 min;and followed by 45 cycles of 1 min at 94°C,1 \nmin at 36°C,and 2 min at 72°C then held at 72°C for 10 \nmin.With 16 primers,123 reproducible amplified fragments ranging \nbetween 500 and 14 000 bp were acquired.In all bands,2 bands were \nshared by all crabs,13 shared by Charybdis japonica ,C.Acuta \nand C.Feriatus,16 shared by Portunus sanguinolentus and \nP.Trituberculatus.The amplified fragments were scored as present (1) or \nabsent (0) for each DNA sample and an index of degree of band sharing \n(F) was calculated by using Nei and Li's matching coefficient \nmethod.The degree of F was 0.0606-0.8000.The value of 1-F was used to \nevaluate genetic distances between species.The genetic distances \nbetween species were 0.2000-0.9394.The phylogenetic trees were \nconstructed with the method of UPGMA and NJ on the basis of genetic \ndistances.The results from the two methods of cluster analysis are \nsimilar in general,and the relationships indicated by the phylogenetic \ntrees show the difference between genus species.The results are in good \noverall agreement with classical taxonomy.