Analysis On The Distribution About The Species In Genus Rana In Dabieshan Area And The Distribution Of Polypedates Braueri In Anhui

In recent years,the records of new species and new distributuion of amphibians have been reported ceaselessly.These new findings are mainly due to the fact that early classification mostly relies on morphological metrics.In the process of comparison,there are many misjudgments in species identification and classification because of the differences among individuals and the inconsistency of measurement standards.Nowadays,because of the develop of molecular markers,species identification and classification have reached a new height.This study is divided into two aspects:(1)Through morphological comparison and phylogenetic relationship analysis,we discuss species belonging to Rana in Dabie Mountains;(2)Through the phylogenetic relationships,we discuss the species belonging to Polypedates in Dabie Mountain area,besides,the microsatellite locis of the species were developed.There has been considerable controversy on the taxonomy of Rana species in the Dabie Mountains.30 individuals belong to genus Rana were collected to make a correct judgment on the attribution of species of Rana in the Dabie Mountains.In the comparison of morphological features,it was found that in addition to the existence of the identified Rana chensinensis in the area,there was also found a species from genus Rana different from all known Rana species.The characteristics of the suspected new species of Rana differing from other Rana species in morphology are as follow:(1)large body size(males SVL 50.9-62.8 mm,n = 8 and females 53.0-68.3 mm,n = 2);(2)snout obtusely pointed,projecting beyond lower jaw;(3)distinct canthus rostralis;(4)smooth dorsal skin,lacking warts;(5)relative finger lengths?>?>?>?,without finger webbing,the toe lengths ?<?<?<?<? with two third webbed,web type ?2-1-?2+-1+ ?3-? 2-2 2+ ?;(6)no external vocal sac;(7)presence of a straight dorsal fold from temporal to crotch;(8)the color of the dorsum varies with the surrounding environment in life,from golden to brown.In addition to morphological comparisons,the study was also based on the phylogenetic relationships of multiple genes(12S rRNA,16S rRNA,Cyt b,ND2,BDNF,Tyr and RAG1).The species within Rana are mainly divided into seven large branches.The suspected new species in the study clustered within the group of Rana longicrus group frogs,far away from other species in the Rana.Combined the morphological comparison and molecular identification,the species found in this area were defined as new species and named as Rana dabieshanensis sp.nov.Because the species in the Polypedates and Rhacophorus are not easily to distinguish,different scholars have their own opinions on the attribution of species within these two genus.According to historical records,there are four species of tree frogs in the Dabie Mountains:Rhacophorus.nigropunctatus,Polypedates megacephalus,Rhacophus dennysi,and Rhacophorus anhuiensis.Considering that some species of the genus Rhacophorus and Polypedates are similar,so,this experiment aims to classify Rhacophoridae species in the Dabie Mountains by partial mitochondrion sequences of 12S rRNA and 16S rRNA.Combined available homologous sequence of Polypedates,genetic analyses suggest that the sample from the Dabie Mountains is P.braueri rather than P.megacephalus as previously recorded.It is the first time to report the Dabie Mountains is a new distribution area of P.braueri.In order to further study the conservation genetics of the species,the 3 base microsatellite loci of the species were isolated and screened in this study.A total of 13 pairs of microsatellite primers were used for the 21 individuals.The size of the amplified fragment was 112-267bp,and the allele(Na)range was 2-5.The expected heterozygosity(HE)and the observed heterozygosity(HO)range were 0.463?0.712 and 0.174-0.957 respectively.