Abstract:[Objectives] The long-standing issue of classification controversy between Zhangixalus zhoukaiyae and Z. lishuiensis has existed since their first publication. To further explore and resolve this controversy, an extensive field monitoring study was conducted at the border of Zhejiang and Anhui Provinces, specifically (Huangshan, Anhui and Chun’an, Zhejiang). Five specimens of the Zhangixalus genus were collected. Morphological comparisons and phylogenetic analysis were performed on these five specimens alongside specimens of Z. zhoukaiyae and Z. lishuiensis obtainedfrom their respective type localities. [Methods] Morphological characteristics of the five specimens including 12 morphological data indicators were measured by a vernier caliper, and analyzed together with the holotype of Z. zhoukaiyae and Z. lishuiensis (Table 2). The 16S rRNA gene fragments were amplified for these five specimens, combined with NCBI GenBank data (Table 1), a Bayesian tree was constructed using Mrbayes 3.1.2 and p-distance model in MEGA 6 was used to calculate the genetic distance between individuals of Zhangixalus. [Results] Our results show no significant morphological differences between the five specimens and the type specimens of Z. zhoukaiyae and Z. lishuiensis. Based on the phylogenetic analysis of the 16S rRNA gene, the five samples clustered into a monophyletic group with the sequences of the type specimen of Z. zhoukaiyae and Z. lishuiensis (Fig. 2). The genetic distance between our five samples and the type specimen of Z. zhoukaiyae is 1.3%, and that of the type specimen of Z. lishuiensis is 0.8%. The genetic distance between the type specimens of Z. zhoukaiyae and Z. lishuiensis is 1.6% (Appendix 1). [Conclusion] Our analysis inferred that Z. lishuiensis is a synonym of Z. zhoukaiyae, our results resolved the existing classification controversy between them. We therefore recommend that Z. lishuiensis and Z. zhoukaiyae should be treated as different geographical populations, as they have not reached the level of interspecific divergence.
