Hidden hotspots of amphibian biodiversity in China.

Identifying and protecting hotspots of endemism and species richness is crucial for mitigating the global biodiversity crisis. However, our understanding of spatial diversity patterns is far from complete, which severely limits our ability to conserve biodiversity hotspots. Here, we report a comprehensive analysis of amphibian species diversity in China, one of the most species-rich countries on Earth. Our study combines 20 y of field surveys with new molecular analyses of 521 described species and also identifies 100 potential cryptic species. We identify 10 hotspots of amphibian diversity in China, each with exceptional species richness and endemism and with exceptional phylogenetic diversity and phylogenetic endemism (based on a new time-calibrated, species-level phylogeny for Chinese amphibians). These 10 hotspots encompass 59.6% of China's described amphibian species, 49.0% of cryptic species, and 55.6% of species endemic to China. Only four of these 10 hotspots correspond to previously recognized biodiversity hotspots. The six new hotspots include the Nanling Mountains and other mountain ranges in South China. Among the 186 species in the six new hotspots, only 9.7% are well covered by protected areas and most (88.2%) are exposed to high human impacts. Five of the six new hotspots are under very high human pressure and are in urgent need of protection. We also find that patterns of richness in cryptic species are significantly related to those in described species but are not identical.

A new species of Hemiphyllodactylus (Squamata, Gekkonidae) from southwest Yunnan, China.

A new species of gekkonid, Hemiphyllodactylusgengmaensissp. nov., is described based on six specimens from Gengma Dai and Wa Autonomous County, Yunnan, China. The new species can be distinguished from its congeners by a significant genetic divergence of greater than 9.7% in the mitochondrial ND2 gene and a combination of the following characters: a maximum SVL of 43.24mm; 8 or 9 chin scales; six circumnasal scales; 2 or 3 internasal scales; 9-11 supralabial scales; 8 or 9 infralabial scales; 11-18 dorsal scales; 8-10 ventral scales; a manual lamellar formula of 5-5-5-4 or 5-6-5-4 and a pedal lamellar formula of 5-5-6-5; 20-25 precloacal and femoral pore-bearing scales contiguous in males; dark postorbital stripes or striping on body; dark dorsal transverse blotches present; and a brown postsacral mark bearing anteriorly projecting arms. The discovery of this new species brings the number of Hemiphyllodactylus species in China to 15.

Description of a New Species of the Pareas hamptoni Complex from Yunnan, China, with Confirmation of P. hamptoni Sensu Stricto in China (Squamata, Pareidae).

We describe a new species of the genus Pareas, based on three specimens collected from Guanyinshan Provincial Nature Reserve in Yuanyang County, Honghe Prefecture, Yunnan Province, China. The new species is distinguished from its congeners by one preocular, one postocular or postocular fused with subocular, loreal not bordering the orbit, one row enlarged vertebral scales, five rows keeled mid-dorsal scales at the middle of the body, 189-192 ventral scales and 72-89 subcaudal scales. The dorsal surfaces of the head and body are yellowish red or yellowish brown, and the belly and ventral surfaces of the head and tail are pinkish yellow or yellow with more or less small black spots. Phylogenetic analyses of mitochondrial DNA recovered the new species being the sister taxon to P. hamptoni sensu stricto. The genetic divergences between the new species and P. hamptoni sensu stricto were 4.2% in the Cyt b sequences and 5.0% in the ND4 sequences. In addition, based on specimens collected from Honghe and Wenshan prefectures, we confirmed that P. hamptoni sensu stricto is distributed in China.

Four new species of the genus Diploderma Hallowell, 1861 (Squamata, Agamidae) from China.

Four new species of Diploderma are described from Sichuan and Yunnan provinces, southwestern China, based on an integrative taxonomic approach, combining morphological and genetic data. The first new species from Danba County, Sichuan Province, is morphologically most similar and phylogenetically closely related to D.flaviceps, but it can be diagnosed from the latter by having a relatively much shorter tail and by a genetic distance of 4.4% in the ND2 gene; the second new species from Muli County, Sichuan Province, is phylogenetically closely related to D.daochengense, D.yongshengense, and D.yulongense, but it can be diagnosed from the latter three species by having a pale yellow gular spot and by genetic distances of 5.6-6.7% in the ND2 gene; the third new species from Jiulong County, Sichuan Province, is morphologically most similar and phylogenetically closely related to D.angustelinea, but it can be diagnosed from the latter by having a relatively much longer tail and by a genetic distance of 2.8% in the ND2 gene; and the last new species from Weixi County, Yunnan Province, is phylogenetically closely related to D.aorun, but it can be diagnosed from the latter by having a pale yellow gular spot and by a genetic distance of 2.9% in the ND2 gene. Our work brings the number of species within the genus Diploderma to 46.

Description of a New Cobra (Naja Laurenti, 1768; Squamata, Elapidae) from China with Designation of a Neotype for Naja atra.

Taxonomic frameworks for medically important species such as cobras (genus Naja Laurenti, 1768; Squamata, Elapidae) are essential for the medical treatment of snake bites and accurate antivenin development. In this paper, we described the former N. kaouthia populations recorded from China as a new species and designated a neotype for N. atra-based morphological and mitochondrial phylogenetic analysis. The new species N. fuxisp. nov. was morphologically diagnosed from N. kaouthia by (1) regular single narrow crossband present on the middle and posterior parts of the dorsum (3-15, 7.9 ± 2.7, n = 32) and the dorsal surface of the tail (1-6, 4.2 ± 1.1, n = 32) of both adults and juveniles, buff-colored with dark fringes on both edges, vs. South Asian populations (n = 39) and Southeast Asian populations (n = 35) without cross bands, with irregular cross bands or multiple light-colored crossbands pairs, or densely woven lines; (2) small scales between the posterior chin shields, usually three (40%) or two (37%), rarely four (13%), or one (10%) (n = 30) vs. mostly one (81%) and rarely two (19%) (n = 28); (3) ventrals 179-205 (195.4 ± 6.7, n = 33) vs. South Asian populations 179-199 (188.7 ± 5.9, n = 12); Southeast Asian populations 168-186 (177.8 ± 4.9, n = 18). Phylogenetically, the new species forms an independent sister clade to the clade including N. atra, N. kaouthia, N. oxiana and N. sagittifera. Furthermore, the subspecies N. naja polyocellata should be resurrected and recognized as a full species, N. polyocellatacomb. nov., and the subspecies N. sumatrana miolepis should be resurrected.

Complete Mitogenome of Oreolalax Omeimontis Reveals Phylogenetic Status and Novel Gene Arrangement of Archaeobatrachia.

Species of the genus Oreolalax displayed crucial morphological characteristics of vertebrates transitioning from aquatic to terrestrial habitats; thus, they can be regarded as a representative vertebrate genus for this landing phenomenon. But the present phylogenetic status of Oreolalax omeimontis has been controversial with morphological and molecular approaches, and specific gene rearrangements were discovered in all six published Oreolalax mitogenomes, which are rarely observed in Archaeobatrachia. Therefore, this study determined the complete mitogenome of O. omeimontis with the aim of identifying its precise phylogenetic position and novel gene arrangement in Archaeobatrachia. Phylogenetic analysis with Bayesian inference and maximum likelihood indicates O. omeimontis is a sister group to O. lichuanensis, which is consistent with previous phylogenetic analysis based on morphological characteristics, but contrasts with other studies using multiple gene fragments. Moreover, although the duplication of trnM occurred in all seven Oreolalax species, the translocation of trnQ and trnM occurred differently in O. omeimontis to the other six, and this unique rearrangement would happen after the speciation of O. omeimontis. In general, this study sheds new light on the phylogenetic relationships and gene rearrangements of Archaeobatrachia.

A new species of Cyrtodactylus Gray, 1827 (Squamata, Gekkonidae) from southwestern Yunnan, China.

A new species of the Cyrtodactyluschauquangensis species group is described based on four specimens collected from the karst formations of Menglian County, Puer City, Yunnan Province, China. The new species can be separated from all other congeners by having a unique combination of morphological characters: a medium-sized body; ventrolateral folds present with interspersed small tubercles; seven precloacal pores in a continuous series in males, absent in females; enlarged femoral scales and femoral pores absent; two postcloacal tubercles on each side; and one or two rows of enlarged subcaudals. Genetically, the new species most closely related to C.wayakonei and the uncorrected sequence divergences of the ND2 gene and its flanking tRNAs between the new species and investigated congeners range from 7.2% to 18.4%.

First record of Calotesvindumbarbatus Wagner, Ihlow, Hartmann, Flecks, Schmitz & Böhme, 2021 (Squamata: Agamidae) from China, with revised diagnosis of this species.

BACKGROUND: Three new species were recently described from the Calotesmystaceus Duméril & Bibron, 1837 complex. Of the three new species, C.vindumbarbatus Wagner, Ihlow, Hartmann, Flecks, Schmitz & Böhme, 2021 was known only from northern Myanmar. NEW INFORMATION: Seven specimens of lizard were collected from Tongbiguan Nature Reserve, western Yunnan, China. Phylogenetically, these specimens clustered with the type specimens of Calotesvindumbarbatus from Myanmar with strong support and showed inappreciable genetic divergence from the type specimens of C.vindumbarbatus. We report the first country record of C.vindumbarbatus from China. In addition, a supplementary description, based on the newly-collected specimens and revised diagnosis of this species, was provided.

Genomic insights into the evolution of the critically endangered soft-shelled turtle Rafetus swinhoei.

The 2019 death of the female Swinhoes soft-shelled turtle (Rafetus swinhoei) individual in China put this species in a dire conservation state. To avoid the loss of genetic information from the female individual and facilitate the protection and preservation of R. swinhoei, we sequenced and assembled its draft chromosome-level genome by combining BGISEQ short reads, Nanopore long reads, and Hi-C sequencing reads. We characterized its potential ZZ/ZW sex determination system. Phylogeny and divergence time analyses indicated that R. swinhoei has a close relationship with Pelodiscus sinensis, and they shared a common ancestor ~54.4 Ma. Population history analyses indicated that R. swinhoei has the lowest heterozygosity among all turtles examined, and its population size has declined dramatically over the past 10 million years. Many core genes involved in autophagy and DNA damage response, such as Rab5a and Parp1, were under positive selection and/or evolving rapidly, and these genes may make important contributions to the long lifespan and evolutionary adaptation of R. swinhoei. The loss of several tooth-related genes in turtle genomes explains the genetic basis of their toothless phenotype. This study not only provides a genomic resource for scientists to study the species using the strategy of comparative genomics, but also provides a data basis for us to find more potential R. swinhoei individuals in the wild.

Cryptic diversity and phylogeography of the Rhabdophis nuchalis group (Squamata: Colubridae).

Previous studies, have found that the rapid uplift of the Tibetan plateau accelerated the diversification of species. However, there are few relevant biogeographic data for the Colubridae in this region. We conducted a comprehensive study of the Rhabdophis nuchalis Group, which presently contains four nominal species, R. nuchalis, R. pentasupralabialis, R. leonardi, and R. chiwen. Building upon previous studies with specimens we have recently examined, greater interspecific and intraspecific diversity has been revealed. Here we address three questions: (1) Do the intraspecific differences represent only geographic variation within lineages, or are there cryptic species? (2) What are the interspecific relationships among members of the R. nuchalis Group? (3) What has been the biogeographic history of this species group? To resolve these questions we used four mitochondrial gene sequences and one nuclear sequence to investigate the molecular phylogenetic and geographic relationships among populations. Our molecular analysis reveals cryptic species diversity within the R. nuchalis Group, and seven clades were identified in the analysis. Ancestral area estimation suggests that the R. nuchalis Group originated in the Hengduan Mountains approximately 6.24 Mya and expanded its range northward to the Qinling-Daba Mountains. The Sichuan Basin appears to have been a barrier to migration. Species divergence seems to have been related to the rapid uplift of the Qinghai-Tibet Plateau.

Three new species of Diploderma Hallowell, 1861 (Squamata, Agamidae) from the Hengduan Mountain Region, south-western China.

Three new species of Diploderma are described from the Hengduan Mountain Region in south-western China, based on morphological and genetic data. The first new species from Yulong County, Yunnan Province is morphologically most similar and phylogenetically closely related to D.brevicauda, but it can be diagnosed from the latter by having a relatively longer tail; the second new species from Xiangcheng County, Sichuan Province is phylogenetically closely related to D.bowoense, but it can be diagnosed from the latter by the absence of a distinct gular spot; and the third new species from Yongsheng County, Yunnan Province is phylogenetically closely related to D.yulongense, but it can be diagnosed from the latter by having different colourations of the ventral and ventrolateral surfaces of the body. Taxonomy and diversity survey are the basis of species conservation, our discoveries contributing to better conservation of the species of this genus.

Confirmation of Tylototritonziegleri Nishikawa, Matsui & Nguyen, 2013 in China, with discussion on the relationship between T.verrucosus Anderson, 1871 and T.panwaensis Grismer, Wood, Quah, Thura, Espinoza & Murdoch, 2019 (Caudata, Salamandridae).

Background: The distribution of the Ziegler's Crocodile Newt Tylototritonziegleri Nishikawa, Matsui & Nguyen, 2013 in China has been controversial. This species was originally recorded uncertainly from Guangxi Autonomous Region, China. Subsequently, this species was recorded from Yunnan and Guangdong provinces, China. Thereafter, the record from Guangdong was denied and the record from Yunnan was questioned. New information: Two specimens of Tylototriton Anderson, 1871 were collected from Wenshan Prefecture, Yunnan Province, China, in 2020. Phylogenetically, the sequences of these two specimens clustered with the sequences of T.ziegleri (including the holotype) from its type locality with strong support, and morphologically agree well with the original description of T.ziegleri. We confirm the record of T.ziegleri in China and present detailed collection site and morphological description of the specimens from China. In addition, we found that T.panwaensis Grismer, Wood, Quah, Thura, Espinoza & Murdoch, 2019 may be the synonym of T.verrucosus Anderson, 1871. We discussed the relationship between T.verrucosus and T.panwaensis.

A new species of Nanorana Günther, 1896 (Anura, Dicroglossidae) from Yunnan, China.

A new species of Nanorana Günther, 1896 is described from Yunnan Province, China, based on morphological and molecular evidence. Morphologically, Nanorana xuelinensis sp. nov. is distinguished from its congeners by a combination of the following diagnostic characters: body size large; adult males with keratinized spines on chest, belly, lateral body, posterior dorsum, buttocks, outer side of the fore limbs, the inner metacarpal tubercle, fingers I and II, and upper eyelids; no spines on the inner side of the lower and upper arm; forelimbs strongly hypertrophied in adult males; anterior dorsum skin smooth; dorsolateral folds absent; finger I longer than finger II; webbing deeply incurved between tips of toes; present outer metacarpal tubercle and absent outer metatarsal tubercle. The new species is separated from all other congeners by uncorrected genetic distances ranging from 5.2% to 7.3% based on mitochondrial 16S rRNA gene and ranging from 3.9% to 7.6% based on mitochondrial 12S rRNA gene.

A new species of Cyrtodactylus Gray, 1827 (Squamata, Gekkonidae) from Yunnan, China.

A new species of Cyrtodactylus is described on the basis of five specimens collected from the karst formations of Zhenkang County, Yunnan Province, China. Cyrtodactylus zhenkangensis sp. nov. is recognized by having a unique combination of morphological characters, the most diagnostic being: 12-15 enlarged femoral scales on each thigh; 2-5 femoral pores on each thigh in males, 0-3 pitted scales on each thigh in females; eight or nine precloacal pores in a continuous row or separated by one poreless scale in males, 7-9 pitted scales in females; subcaudals enlarged, arranged alternately as single and double on anterior and mostly single at middle and posterior; dorsal surface of head with obvious reticulations. Phylogenetic analyses show that the new species is a member of the C. wayakonei species group and a sister taxon to a clade consisting of C. wayakonei and C. martini based on Maximum Likelihood analyses and Bayesian Inference and differs from its congeners by at least 12.0% genetic divergence in a fragment of the COI gene.

A new species of the genus Pareas (Squamata, Pareidae) from Yunnan, China.

A new species of Pareas is described from Yunnan Province, China, based on morphological comparisons and molecular data. Genetically, the new species is most closely related to the recently-described Pareas geminatus, for which we present new topotypic findings. The genetic divergence (uncorrected p-distance) of the cytb gene between the new species and congeners ranged from 6.14% to 24.68%. Morphologically, the new species can be distinguished from P. geminatus and all other congeners. Our work brings the total number of species within the genus Pareas to 22.

The complete mitochondrial genome of Amyda cartilaginea (Testudines: Trionychidae).

The Asiatic softshell turtle, also known as the black-rayed softshell turtle (Amyda cartilaginea; Accession no: MT039230), is found in northeastern India (Mizoram), Brunei Darussalam, Indonesia, Malaysia, Singapore, Myanmar, Laos, Vietnam, Cambodia, and Thailand. This turtle is thought to have been introduced into the Sunda Islands, Sulawesi, and Yunnan, China, through the Malay Peninsula to Sumatra, Java, and Borneo. Herein, we determined the complete mitochondrial genome of A. cartilaginea for the first time using next-generation sequencing (NGS). The assembled mitogenome was 16,763 bp in length and encoded 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA genes (12S rRNA and 16S rRNA), and one control region (CR). The PCGs based maximum-likelihood phylogeny discriminated A. cartilaginea from other Testudines and clusters within family Trionychidae with the sister taxa of Nilssonia nigricans.

A new species of the genus Acanthosaura (Squamata, Agamidae) from Yunnan, China, with comments on its conservation status.

A new species of Acanthosaura from Yunnan, China, is described based on morphological and genetic data. The new species can be separated from all other species of the genus by having a different shape of the black eye patch, a different coloration of the postorbital and occipital spines and nuchal crest, and a different color of the gular pouch. Genetically, uncorrected sequence divergences of COI between the new species and investigated congeners ranged from 16.12% to 24.11%. The conservation status of the new species is also discussed.

Author Correction: Review of the genus Brachytarsophrys (Anura: Megophryidae), with revalidation of Brachytarsophrys platyparietus and description of a new species from China.

Following the publication of our paper (Li et al., 2020), it has come to our attention that, due to our oversight, the contributions of some colleagues were not reflected in the "ACKNOWLEDGEMENTS" section. Hereby, we wish to add the following to the "ACKNOWLEDGEMENTS" section as a correction.

Scale microornamentation of five species of Pareas (Serpentes, Pareidae) from China.

Due to the highly conserved morphology of its members, the taxonomy of Pareas remains contentious. Microornamentation was examined on the exposed surfaces of dorsal scales from the midbody region of five species of genus Pareas. The results showed that the ultrastructural characteristics are similar in some respects and different in others. Similarly, all species exhibit arc and pore structures, but the shape, number and arrangement of these arcs and pores differ. In particular, the microornamentation of the dorsal scales of Pareas yunnanensis, thus far regarded as a synonym of P. chinensis, is obviously different from that of P. chinensis. Thus, P. yunnanensis is revalidated. The ultrastructural characteristics of these five species of Pareas are very different from those of reported for P. carinatus. P. carinatus may not belong to the same genus as other Pareas species.