Abstract:The genus Boulenophrys belongs to the family Megophryidae, Anura. This genus includes 56 species, and they are widely distributed in Sichuan, Guizhou, Zhejiang, Fujian, Jiangxi, Guangdong and other regions in China. Currently, 5 species of genus Boulenophrys have been recorded in Sichuan Province, including B. binlingensis, B. minor, B. ombrophila, B. spinata and B. wushanensis. In July 2021, two subadult amphibians were collected from Wulinggou, Daxueshan town (27°53′ N, 104°45′ E, 1380m) in the Daxueshan Scenic Area (Yibin City, Sichuan Province, China). The specimens were measured according to Fe et al.’s method (2001), and their mitochondrial 16S rRNA and COI genes were amplified and sequenced. We gathered all available 16S rRNA and COI sequences of the Boulenophrys genus and the corresponding sequences of Brachytarsophrys carinense from GenBank (Table 2), and used maximum likelihood method to build a phylogenetic tree of Boulenophrys. Genetic distances among species were measured using uncorrected p-distances calculated by MEGA 7. Through morphometric measurements (Table 1) and external morphological (Fig. 2) examination, the two specimens were found to be similar to B. qianbeiensis. Two mitochondrial 16S rRNA and COI gene sequences, length 1094 bp, were obtained. The phylogenetic tree of species in Boulenophrys showed that the two specimens were clustered with the type of B. qianbeiensis (Fig. 3). and the genetic distance of COI gene between the specimen and B. qianbeiensis was 0.000﹣0.004, whereas the genetic distance between the specimen and the B. omeimontis group was 0.038﹣0.113. Phylogenetic analysis based on 16S rRNA and COI genes as well as morphological traits have supported the two specimens were B. qianbeiensis, which should be a new amphibian record of Sichuan Province. Previously, B. qianbeiensis was only found in Huanglian Nature Reserve and Kuankuoshui National Nature Reserve in Guizhou Province. Our discovery not only expanded the distribution range of B. qianbeiensis, but also provided important information for protection and evolutionary research of B. qianbeiensis.