Abstract:Achalinus ningshanensis was first described as a new species of the genus Achalinus in 2022. It consists of A. ningshanensis ningshanensis and A. ningshanensis occidentalis. This study provides morphological and molecular information of A. n. ningshanensis collected from Funiu Mountain, Henan Province, China and determines the taxonomic status of A. n. ningshanensis in Henan.In this study, 22 morphological indexes were selected for morphological analysis. Length was measured by a vernier caliper, and the number of scales was described manually. Genomic DNA was extracted from the liver tissue preserved in 95% alcohol by the FastPure Gel DNA Extraction Mini Kit, and cytochrome c oxidase subunit I (COI) was selected as the target fragment, which was amplified with primers Chmf4 and Chmr4. COI gene sequences of 26 species of Achalinus and three outgroups (Parafimbrios lao, Fimbrios klossi, Xenodermus javanicus) were downloaded from GenBank (Appendix 1). Sequence alignment and calculation of uncorrected pairwise distances (p-distance) among congeners were performed in MEGA 7.0 software. A phylogenetic tree was constructed by maximum likelihood estimation (MLE) analysis via PhyloSuite v1.2.3 software. Morphological comparisons showed that the specimen from Funiu Mountain had two pairs of chin shields with two small scales between the second pair, 181 ventral scales, and dorsal scales of 25-23-23, while A. n.ningshanensis lacked small scales between the second pair of chin shields and had 161 ~ 165 ventral scales and dorsal scales of 23-23-23 (Table 1). The phylogenetic tree indicated that the specimen closely clustered with the topotype of A. n. ningshanensis (Ningshan, Shaanxi, China) (Fig. 3). The genetic distance between them was 0.4%, significantly smaller than the interspecific genetic distance of Achalinus (2.8% ~ 18.8%) (Appendix 2).Combining morphological and molecular phylogenetic methods, we identified the specimen of Achalinus collected from Funiu Mountain, Henan Province as A. n.ningshanensis, representing a new provincial record of snake in Henan.