Six small frogs were collected from location of Xiangsikeng in Mangshan National Nature Reserve (24°56′57.51″N, 112°59′18.21″E, 1 342 m), Hunan Province in April 2016 and May 2017, which were identified as Anura, Rhacophoridae, Rhacophorus, R. minimus, and they are new to Hunan Province. Fourteen morphological measurements of these specimens are presented and morphological characteristics were also described in this study. Then, total DNA extraction, PCR, and16S rRNA DNA sequences were carried out. Compared with other eight tree frogs sequences from GenBank, the phylogenetic tree supported the morphological characteristics. These specimens are preserved in the College of Life Sciences, Hunan Normal University.