Reevaluation Of The Holotype Of Odorrana Schmackeri Boettger, 1892 (Amphibia: Anura: Ranidae) And Characterization Of One Cryptic Species In O. Schmackeri Sensu Lato Through Integrative Approaches

The Odorrana schmackeri sensu lato is a group of cascade frogs that arc widely distributed in southern China However, their taxonomy has long been unresolved and several cryptic species have been reported in the past decade. Previous analyses revealed the existence of three sympatric species (i.e., O. schmackeri, O. hejiangensis and O. sp2) at the type locality of O. schmackeri (Gaojiayan Town, Yichang City, Hubei Province, China). However, the lack of an original description for the holotype specimen (SMF6241) of O. schmackeri poses a confusing and controversial problem in species nomination within this group. Here, we reevaluate the taxonomic affinities of SMF6241 and examine the distinctness of O. sp2 based on specimens from the type locality. We use integrative approaches that combine morphology with 10 microsatellite loci and bioacoustics. Canonical discriminant analysis of morphological characteristics revealed three distinct groupings (i.e., a schmackeri, O. hejiangensis and O. sp2) and attributed SMF6241 to O. schmackeri. Genetic clustering analysis of nuclear DNA also identified three corresponding clusters. In addition, analysis of interspecific variation in morphometrics and acoustics showed divergence between O. sp2 and its sympatric congeners. All lines of evidence from morphological, molecular and bioacoustic approaches suggest the species status of O. sp2, namely Odorrana ichangensis Chen, sp. nov. Overall, this study resolves the taxonomic assignment of the holotype of O. schmackeri and provides an example for species delimitation using integrative approaches.