Study On Taxonomic Status Of Murina Rongjiangensis(Chiroptera:Vesertilionidae)

Genus Murina is one of the controversial taxa in the taxonomic study of Chiroptera and relatively least studied.In recent decade,ten new species of the genus have been discovered in China due to intensified field surveys using harp trap.However,some new species are reported based on limited specimens and single molecular marker,which leads to controversies in determining the species of Murina.Hence,detailed taxonomic studies confirming the species determination has become necessary.Murina rongjiangensis is one of 10 new species of the genus Murina,discovered by Chen et al.from Rongjiang County,Guizhou Province,China in 2017,which showed similar external and skull morphology with M.shuifuensis.In this study,it was found that limited samplings and single mitochondrial marker were used for the species determination of M.rongjiangensis,and the cited literature data and evidence are insufficient.To further clarify the taxonomic status of M.rongjiangensis,55 specimens were collected from the type localities and surrounding area of the two new species mentioned above.The taxonomic status of M.rongjiangensisand the relationship between M.shuipuensis and M.rongjiangensis were verified by integrating evidence from morphology,phylogeography and acoustic characteristics.The results of the study are as follows:1)The results of phylogeographic analysis based on mitochondrial COI,Cytb genes and nuclear gene RAG2 showed that M.rongjiangensis and M.shuipuensis are closely related and do not support the former as an independent viable species.Although mitochondrial markers suggested significant genetic differentiation between Guizhou and outer Guizhou populations,both nuclear gene and microsatellite markers presented mixed results.In the absence of more evidences,this study tends to classify it into the M.shuipuensiscomplex.Furthermore,some haplotypes in Shuifu village,Libo county,Guizhou province(the type locality of M.shuipuensis)are embedded in clade of M.rongjiangensis,suggesting the sympatric distribution of the two species.2)phylogeographic analysis showed that the results of mitochondrialand nuclear gene markers were inconsistent.phylogeographic analysis based on microsatellite showed that males havehigher genetic relationship among the seven populations and females do the opposite,suggesting that male-biased dispersal exists in this group.This behavior and male-mediated nuclear gene introgression led to the conflict between nuclear gene and mitochondrial gene markers in this study.3)The results of multivariate statistical analysis of external,skull morphology and echolocation call were unable to distinguish M.rongjiangensis from M.shuipuensis,suggesting that M.rongjiangensis should not be considered as a valid species.However,the result of analysis of morphology and ultrasonic frequency characteristics show that there was some differentiation between Guizhou and outer Guizhou populations which conforms to the boundaries between central China,east China,south China and southwest China,but further studies are needed to verify this geographical variation.4)Coat color is one of the key indicators of M.shuipuensisand M.rongjiangensisfor species publication(For example,throat hair is bright orange),However,this study found that there were color variations in the throat and abdominal hair of the complex based on a larger range of specimens.The results of multivariate statistical analysis showed that the saturation was significantly correlated with the wear rate of teeth,suggesting that age may be the main factor leading to the color variation.This finding further suggests that hair color should be used with caution for classification based on limited specimens.In overall,M.shuipuensis and M.rongjiangensis are sympatrically distributed,with little genetic differentiation,and no obvious differentiation in appearance,skull morphology,ultrasonic and phylogeography analysis of nuclear gene.Therefore,the taxonomic status of M.rongjiangensis as an independent valid species is not valid,so M.shuipuensis and M.rongjiangensis should be synonym.Although there were geographical variations in skull morphology,echolocation call and mitochondrial genes betweenouter Guizhou and Guizhou populations,the nuclear gene markers were mixed and indistinguishable.Therefore,the outer Guizhou population was temporarily divided into M.shuipuensis complex.