We described a new species of the genus Leptobrachella from Southern Guangxi, China, based on morphological characteristics, molecular analyses and bioacoustics. This new species, Leptobrachella shiwandashanensis sp. nov., occurs in sympatry with Leptobrachella shangsiensis. However, phylogenetic analyses based on the mitochondrial 16S gene fragment revealed that the new species was clustered in a monophyletic group and a considerable genetic divergence existed between the new species and L. shangsiensis (p-distance: 7.9%) as well as its congeners (minimum p-distance: 7.0%). The new species also differed from its congeners by a combination of the following characteristics: (1) small size (SVL 26.8–29.7 mm in males; 33.7–35.9 mm in females); (2) pale brown dorsal surfaces, with a brown inverse-triangle-shaped marking between the eyes; (3) creamy white ventral surface with brown spots on the lateral margin, with a near immaculate creamy white throat and chest; (4) without webbing and lateral fringes on toes; (5) flanks with irregular black spots; (6) tibia-tarsal articulation reaching the posterior of the eye in males but reaching the shoulder in females; (7) heels that do not meet when the thighs are appressed at right angles to the body; (8) bicolored iris, with the upper half brownish-red, fading to silver in the lower half; and (9) a call consisting of a single note and a dominant frequency of 5.3–5.7 kHz (recorded at 23°C).
A new species of the genus Leptobrachella, L. guinanensis sp. nov., is described in this study based on morphological, molecular, and bioacoustic data. The species was discovered in the Shiwandashan National Nature Reserve in Shangsi County, Guangxi, China. Phylogenetically, L. guinanensis sp. nov. is closely related to L. ventripunctata. However, there are distinct morphological differences between L. guinanensis sp. nov. and L. ventripunctata, as well as three other sympatric species (L. shangsiensis, L. shiwandashanensis, and L. sungi). These differences include body size (SVL 30.5–32.5 mm in males; 38.7–41.8 mm in females in the new species vs 25.5–28.0 mm in males, 31.5–35.0 mm in females in L. ventripunctata), the absence of brown spots on the ventral surface (vs chest and belly creamy white with many scattered brown spots in L. ventripunctata), 1/3 toe webbing and wide toe lateral fringes (vs no toe webbing and no lateral fringes in L. ventripunctata), and distinct dermal ridges under toes (vs absent in L. ventripunctata). Furthermore, the dominant vocal frequencies of the new species range from 7.3 to 8.3 kHz, which is unique compared to other Leptobrachella species and represents the highest dominant frequencies ever recorded. The Shiwandashan National Nature Reserve is now home to four known sympatric species of Leptobrachella.
The black‐spotted tokay and the red‐spotted tokay are morphologically distinct and have largely allopatric distributions. The black‐spotted tokay is characterized by a small body size and dark skin with sundry spots, while the red‐spotted tokay has a relatively large body size and red spots. Based on morphological, karyotypic, genetic, and distribution differences, recent studies suggested their species status; however, their classifications remain controversial, and additional data such as ecological niches are necessary to establish firm hypotheses regarding their taxonomic status. We reconstructed their ecological niches models using climatic and geographic data. We then performed niche similarity tests (niche identity and background tests) and point‐based analyses to explore whether ecological differentiation has occurred, and whether such differences are sufficient to explain the maintenance of their separate segments of environmental ranges. We found that both niche models of the black‐ and the red‐spotted tokay had a good fit and a robust performance, as indicated by the high area under the curve (AUC) values (“black” = 0.982,SD = ± 0.002, “red” = 0.966 ± 0.02). Significant ecological differentiation across the entire geographic range was found, indicating that the involvement of ecological differentiation is important for species differentiation. Divergence along the environmental axes is highly associated with climatic conditions, with isothermality being important for the “black” form, while temperature seasonality, precipitation of warmest quarter, and annual temperature range together being important for the “red” form. These factors are likely important factors in niche differentiation between the two forms, which result in morphological replacement. Overall, beside morphological and genetic differentiation information, our results contribute to additional insights into taxonomic distinction and niche differentiation between the black‐ and the red‐spotted tokay.
Calotes wangisp. nov., a new species of the agamid genus Calotes Cuvier, 1817, from southern China and northern Vietnam, is described. This species can be distinguished from all known congeners by a combination of morphological characteristics and genetic divergence in the mitochondrial tRNA, ND2, and CO1 genes. Molecular phylogenetic analysis revealed that the new species was formed as a monophyletic group and that considerable genetic divergence existed between its congeners (minimum p-distance, 4.6%). Calotes wangisp. nov. is distinguished by a combination of the following characteristics: average SVL < 90 mm for adult males; 10–14 dorsal eyelid scales; scales on side of neck and adjacent shoulder area pointing obliquely upward; keels on neck scales weakly to strongly developed; fold in front of the shoulder absent; pair of dark triangular patches extending from the front of the shoulder to the jaw angles; and orange coloration of the tongue. Calotes wangisp. nov. is similar to C. irawadi but differs in having scales between the nasal shield and the orbit and a fourth toe with a claw that can reach between the eyes and tympanum (even to the snout when hind the limbs are adpressed forward). Phylogenetic analyses revealed two well-supported subspecies, Lineages A and B in C. wangisp. nov., with mean uncorrected p-distances between them of 2%. We propose that Lineage A, which is mainly from the central and southern Wuzhi Mountains on Hainan Island, is a subspecies, C. w. hainanensisssp. nov. Lineage B mainly comprises individuals from other sites on the island plus the adjacent mainland, and is described as subspecies, C. w. wangissp. nov. A diagnostic key to all Calotes species of China is also provided.
Patterns of geographic variation in body size are predicted to evolve as adaptations to local environmental gradients. However, many of these clinal patterns in body size, such as Bergmann's rule, are controversial and require further investigation into ectotherms such as reptiles on a regional scale. To examine the environmental variables (temperature, precipitation, topography and primary productivity) that shaped patterns of geographic variation in body size in the reptileCalotes versicolor, we sampled 180 adult specimens (91 males and 89 females) at 40 locations across the species range in China. TheMANOVAresults suggest significant sexual size dimorphism inC. versicolor(F23,124 = 11.32,p < .001). Our results showed thatC. versicolorfailed to fit the Bergmann's rule. We found that the most important predictors of variation in body size ofC. versicolordiffered for males and females, but mechanisms related to heat balance and water availability hypotheses were involved in both sexes. Temperature seasonality, precipitation of the driest month, precipitation seasonality, and precipitation of the driest quarter were the most important predictors of variation in body size in males, whereas mean precipitation of the warmest quarter, mean temperature of the wettest quarter, precipitation seasonality, and precipitation of the wettest month were most important for body size variation in females. The discrepancy between patterns of association between the sexes suggested that different selection pressures may be acting in males and females.
Authentication of traditional Chinese medicine (TCM) is challenging due to DNA degradation in Chinese medicinal materials, which are usually processed and stored dry. The standard DNA barcoding length (648 bp) or longer are difficult to amplify, which makes it difficult to identify adulterants in Chinese medicinal materials. In this study, we used the mitochondrial 16S rRNA gene (< 200bp) as a barcode to differentiate black-spotted tokay geckoes (Gekko reevesii) from the related species used as counterfeits. We collected 63 specimens from 17 species of G. reevesii and their counterfeits, and each specimen generated a 189 bp 16S rRNA gene sequence. The average uncorrected p-distances within genuine G. reevesii was 0.9%, while the average uncorrected p-distances between G. reevesii and their counterfeits was 6.3% (at a minimum). According to phylogenetic analysis and genetic distances, the genuine G. reevesii samples collected in this study constitute a monophyly that can be distinguished from its counterfeits in TCM formulations, including G. gecko (red-spotted tokay geckos), which have very similar morphology. Thus, the short 16S rRNA barcode provides an effective tool for distinguishing G. reevesii from its counterfeits, ensuring the safety and efficacy of clinical medications containing components from G. reevesii in TCM.