Classification Of Odorrana Schmackeri Holopyte And Morphological Variation Genetic Differentiation Studies Of Odorrana Schmackeri Species Complex In Yichang, Hubei Province

In 1892,A French biologist Boettger based on 1 male specimens(SMF6241)which was collected from Yichang Hubei to establish a new species Rana schmackeri,because of the lack of a complete form description of holotype and pictures alive,the identification characteristics and morphological descriptions of this odorous frog have been based on the samples of non-type locality in a very long time.In recent years,some studies have shown that Odorrana schmackeri is probably contains three species complex,so which kind of O.schmackeri species complex does the holotype belongs to? What are the morphological and genetic variations of O.schmackeri in type locality? These problems have great importance to renamed the known species of O.schmackeri species complex,besides,supplimental description of O.schmackeri in type locality can further improve the basic information of O.schmackeri as an effective species.This have provided valuable guidance and reference for find the identification characteristics and new cyptic species in the O.schmackeri species complex.This study is based on 3 O.schmackeri species complex 149 morphological specimens(O.schmackeri 36? 27?,O.hejiangensis 35? 23?,O.sp 23? 5?)and 96 molecular materials(O.schmackeri n=52,O.hejiangensis n=21,O.sp n=23)which were collected from Gaojiayan Yichang Hubei(30 � 39.154 'N;56.665 '110 � E).Morphological specimens were used to morphological comparison,typical discriminant analysis,ANOVA and ANCOVA,96 molecular materials were studied by genetic distance calculation,system genetic relationship and genetic structure analysis based on mitochondrial 12 S,16S rRNA genetic markers and 10 microsatellite locis.This can reveal the morphological variation and genetic differentiation of O.schmackeri species complex in Gaojiayan area.The results show that:(1)3 species of O.schmackeri species complex in type locality,all female were significantly larger than males in ANOVA results(P<0.01).Corrected by snout-vent lenght.ANCOVA results show that 9 morphological indexes: head width,nose-eye space,internasal space,interorbital space,inner corner of eye space,tibia length,tibia width,length of foot and tarsus,inner metatarsal tubercles length are significant difference among them.In the indexes of head width,internasal space,interorbital space,inner corner of eye space,only O.schmackeri male shows significantly larger than female.This indicates that the head of O.schmackeri males are relatively wide compared to O.hejiangensis and O.sp.;In the two indexes of tibia length and tibia width,O.schmackeri shows a relatively larger than O.hejiangensis and O.sp.Moreover,nose-eye space,length of foot and tarsus in O.hejiangensis and inner metatarsal tubercles length in O.sp.are all different from the other two species.(2)The typical discriminant analysis results support the O.schmackeri species complex in Yichang,Hubei province into three groups.The accuracy of male and female is 96.8% and 100%.In this identification model,the square of Mahalanobis distance from O.schmackeri holotype's location to the group center of O.schmackeri O.hejiangensis and O.sp.are 15.622,29.592 and 93.150.(3)The phylogenetic relationship according to O.schmackeri species complex in Yichang,Hubei obviously divided into three lineages(lineageA,lineageC and lineageD),each lineage have the significant support(0.83? PP ?1).The genetic distance between the three lineages is 3.7% to 6.7%,and 0.01% within each lineage.(4)Based on microsatellite locis,Structure clustering analysis shows that the genetic structure of O.schmackeri species complex in Yichang,Hubei are divided into three groups significantly(Cluster 1,Cluster 2 and Cluster 3),and the level of gene flow between each group is very low.Conclusion:(1)The holotype(SMF6241)and O.schmackeri are the same species.(2)The Odorrana schmackeri species complex in Yichang,Hubei included two described species,i.e.,Odorrana schmackeri,O.hejiangensis and one cyptic species O.sp.(3)Significant difference of sexual dimorphism modle and high genetic differentiation level of Odorrana schmackeri,O.hejiangensis and O.sp.in Yichang,Hubei,Indicates their niche might been separated.(4)Significant differences of genetic structure among three species in Yichang,Hubei,and there was little gene flow among the population.