The mitochondrial 16S rRNA of two Hyla gongshanensis samples from Xinyi city in Guizhou province was amplified and sequenced,the phylogenetic trees of various groups of H.gongshanensis was constructed using double monophyletic analyse.The results showed that this specimen was belonged to H.gongshanensis.H.gongshanensis recovered 3 lineages,including the Xinyi lineage,the Quqing lineage and the Sabah (Viet Nam) lineage,but the relationship among the three lineages was still unresolved.