Abstract:Five specimens (2 males and 3 females) of the Leptobrachella were collected in Pohe Village, Zhelang Township, Longlin County, Guangxi, China at 105°13′27″ E, 24°44′58″ N altitude 826 m during a field survey in May 2021. The specimens were measured with digital calipers for following measurements: snout-vent length, head length, head width, snout length, internasal space, interorbital space, width of upper eyelid, diameter of eye, diameter of tympanum, length of lower arm, hindlimb length, tibia length, foot length, and compared these characters with the L. ventripunctata. All specimens were fixed in 10% buffered formalin, transferred to 95% ethanol, and deposited in Yulin Normal University. Five liver samples attained from euthanasia specimens and then preserved in 95% ethanol and stored at ﹣20 ℃ were used for molecular analysis. All samples were 17 sequences from all known Leptobrachella species, two sequences from the out-group Brachytarsophrys and Leptobrachium were obtained from GenBank and incorporated into our dataset (Detail information of these materials was shown in Table 2). Genomic DNA was extracted from liver tissue samples using DNA extraction kit. Partial sequences of 12S rRNA and 16S rRNA gene were amplified according the method of Lyu et al. (2017). The resulting fragments were sequenced with both forward and reverse primers. The Clustal W algorithm was used in MEGA 7.0 software to compare all the 12S rRNA and 16S rRNA gene sequences and the Kimura two-parameter model was used to calculate the genetic distances between species of the genus Leptobrachella. We assessed phylogenetic and systematic relationships among the species mentioned above of the mitochondrial DNA genes of 12S rRNA and 16S rRNA using maximum likelihood and Bayesian inference methods by PhyloSuit v1.2.2 software. In this study, we found that the specimens were similar with L. ventripunctata in morphological, morphometric measurements (Table 3) and feature (Fig. 1). Bayesian inference tree and maximum likelihood tree showed that the five specimens were clustered with the L. ventripunctata with a high support, which the Bayesian posterior probabilities (PP) were 1.00 and the bootstrap support (BS) was 100 (Fig. 2). The genetic distance between the specimens and L. ventripunctata was 0.7%﹣0.9%, much lower than that between Leptobrachella species (4.4%﹣23.4%) (Table 3). The specimens from Longlin County are identified as L. ventripunctata based on morphological and molecular comparison, and it is a new record of this species in Guangxi, China.