Genetic Diversity Of Liaoning Province, Northeast Rana Rana System Development Of The Bohai Sea Region

The genetic diversity and genetic structure among four populations of R. dybowskii distributed in Liaoning were analyzed using RAPD molecular markers. It indicated that there was the correlation between genetic differentiation and geographic distance in R. dybowskii population. An understanding of both genetic diversity and population structure of R. dybowskii provides insight for the conservation and management. We can obtain the phylogenetic relationships and evolution mechanism among species of the rana, and then to provide the basis of geological information from the perspective of biogeography, through the analysis of the mitochondrial 16S rRNA gene sequence of the brown frog from Bohai region. The main results are as followed:(I) The samples of R. dybowskii from Huanren, Benxi, Fushun and Fuxin respectively, were collected. RAPD molecular markers were used to test the genetic diversity and genetic variation. All of the genetic parameters, such as allele frequency, percent of polymorphic loci (PPL), Nei's gene diversity (h), coefficient of gene differentiation (Gst), and Shannon's information index (I) were calculated. The genetic structure was further estimated using analysis of molecular variance (AMOVA). The phylogenetic tree was constructed based on UPGMA. The results and conclusions were as follow:(1) Nine arbitrary primers were used to amplify 110 samples of R. dybowskii. Seven to thirteen distinct bands were observed each primer. All bands expressed polymorphism among 95 bands tested and polymorphic percentage was 100%. However, the data of Shannon's information index and Nei's gene diversity showed lower genetic diversity in R. dybowskii. Huanren populations had the higher genetic diversity than other populations, Shannon's information index and Nei's gene diversity were 0.2337 and 0.1341 respectively.(2) The coefficient of genetic differentiation (Gst= 0.0231) among 4 geographic populations was lower, and the gene flow (Nm) is higher. It indicated that genetic differentiation was less significant among 4 geographic populations. The genetic diversity within populations (Hs= 0.0753) in the proportion of the total genetic diversity (Ht= 0.0771) was 97%. The results of analysis of molecular variance (AMOVA) showed that genetic variation within populations was large part of all.(Ⅱ) Based on partial sequences of 16S rRNA gene, the phylogenetic relationships among the brown frogs from Bohai estimated. The MP and ML trees were constructed using PAUP*4.10 and the analyses of Bayesian inference using MrBayes version 3.1.2. ML/BI-based phylogenetic analyses, the optimal model of sequence evolution was chosen on the basis of Hierarchical Likelihood Ratio Tests (hLRT) as implemented in Modeltest 3.7. The results and conclusions were as follow:(1) Based on the phylogenetic relationships, R. huanrenensis should be classified to the R. chensinesis group.(2) Based on phylogenetic tree and biogeological information, there were the same ancestors in R. japonica R. zhenhaiensis and R. culaiensis.(3) 16S rRNA gene partial sequences of six samples in R. culaiensis shared the same haplotype, in addition to the analysis on morphology of the adult frog and labial tooth row formula of the tadpole, it suggested that R. culaiensis was valid species.