How little is known about “the little brown frogs”:description of three new species of the genus Leptobrachella(Anura:Megophryidae)from Yunnan Province, China

Asian leaf-litter toads of the genus Leptobrachella represent a great anuran diversification in Asia.Previous studies have suggested that the diversity of this genus is still underestimated. During herpetological surveys from 2013 to 2018, a series of Leptobrachella specimens were collected from the international border areas in the southern and western parts of Yunnan Province, China.Subsequent analyses based on morphological and molecular data revealed three distinct and previously unknown lineages, which we formally describe as three new species herein. Among them, we describe a new species that occurs at the highest known elevation for Leptobrachella in China. Four species of Leptobrachella, including two new species, are found in the same reserve. Furthermore, our results suggest that the population from Longchuan County,Yunnan, may represent an additional new species of Leptobrachella, although we tentatively assigned it to Leptobrachella cf. yingjiangensis due to the small sample size examined. Lastly, we provide the first description of females of L. yingjiangensis. Our results further highlight that both micro-endemism and sympatric distributions of species are common patterns in Leptobrachella, that contribute to taxonomic and conservation challenges in these frogs. We provide an identification key for Leptobrachella known to occur in Yunnan. Given the lack of knowledge on species diversity of Leptobrachella along international border areas, we recommend that future studies include transboundary collaborative surveys.

A new species of the genus Raorchestes(Anura:Rhacophoridae) from Yunnan Province, China

DEAR EDITOR,A new bush frog species, Raorchestes cangyuanensis sp. nov., from Cangyuan, Yunnan Province, China, is described based on morphological and molecular analyses. It differs from all known congeners by a combination of the following characters: body size small, adult snout-vent length (SVL) 16.1–20.0 mm in males (n=3);tympanum indistinct;tips of all fingers and toes expanded into discs with circummarginal grooves;rudimentary webbing between toes;fingers and toes with lateral dermal fringes;inner and outer metacarpal tubercles present;heels meeting when limbs held at right angles to body;crotch with a distinct black patch;discs of fingers and toes orange;male with external single subgular vocal sac and reddish nuptial pad at the base of first finger.

Taxonomic revision of genus Rohanixalus(Anura:Rhacophoridae)in China with description of one new species

DEAR EDITOR,Based upon morphological and molecular evidence,the authors revised the genus Rohanixalus Biju,Garg,Gokulakrishnan,Chandrakasan,Thammachoti,Ren,Gopika,Bisht,Hamidy and Shouche,2020(Anura:Rhacophoridae)in China through describing one new species,adding one species to the fauna(R.shyamrupus)and supplementing data on one species(Rohanixalus hansenae;Supplementary Materials).

First Record of the Genus Wijayarana (Anura, Ranidae) from Yunnan, China

Dear Editor,e family Ranidae is one of the largest and most families of amphibians(Che et al.,2007),with 64 recognized genera and429 species(Frost,2022).However,the systematics and natural history of some ranid groups have not been well-studied.One such example is the taxonomic relationship between Sylvirana,H ylarana and other closely-related taxa(Che et al.,2007;Pyron and Wiens,2011;Oliver et al.,2015).China has a large diversity of ranid frogs,with 146 speciesbelonging to 7 genera,and48 species belonging to 7 genera in the southern province of Yunnan(AmphibiaChina,2022).

辣木叶水提物成分分析及其对小鼠力竭游泳时间的影响

目的:检测辣木叶水提物的活性成分并观察该提取物对小鼠力竭游泳时间的影响。方法:采用超声辅助水浸提法制备辣木叶水提取物,分别采用福林酚法、铝离子显色法、硫酸苯酚法分析其总多酚、总黄酮、总多糖含量;采用FRAP法和试剂盒法评价该提取物的体外抗氧化活性;并用UPLC-MS/MS测定辣木叶水提物中主要的多酚类物质;观察水提物对小鼠力竭游泳时间的影响。结果:分析结果显示,辣木叶水提物中总多酚、总黄酮、总多糖含量分别为(34.6±3.8)mg GAE/g、(39.9±2.0)mg RE/g、(239.6±12.4)mg GE/g;体外抗氧化能力分析表明,总抗氧化力为(285.7±13.9)U/g,羟自由基清除率为(27.7±5.7)%,DPPH自由基清除率为(56.6±3.4)%。UPLC-MS/MS检测结果显示,多酚类物质中木犀草素、槲皮素、山奈酚、绿原酸的相对含量较高。在力竭游泳实验中,与对照组相比,水提物组游泳时间显著延长(P<0.05)。结论:辣木叶水提取物中富含多酚、类黄酮和多糖,具有抗氧化活性和一定的抗疲劳作用。

叶酸对1,3-丁二烯诱发的小鼠DNA低甲基化和染色体损伤的影响

目的:探讨叶酸(FA)对1,3-丁二烯(BD)诱发的小鼠遗传损伤和DNA甲基化改变的保护作用。方法:雄性昆明小鼠喂养相应饲料3周建立叶酸正常组(FA-C)、叶酸缺乏组(FA-D)以及叶酸补充组(FA-S)动物模型,每组小鼠18只,再随机分为对照组、BD低剂量和高剂量染毒组(0、13.82、1 382.14 mg/m^3),染毒组小鼠每天吸入染毒6 h,每周5 d,连续染毒2周。采用酶联免疫吸附测定法(ELISA)检测小鼠血清中FA浓度和肝组织基因组DNA总体甲基化水平,荧光定量PCR检测DNA甲基转移酶DNMT1和DNMT3a mRNA的表达,胞质分裂阻滞微核试验分析外周血淋巴细胞的染色体损伤。结果:动物喂养3周后,与FA-C组相比,FA-D组血清FA浓度显著降低,FA-S组显著升高。在相同FA处理条件下,随BD染毒剂量升高,小鼠肝组织基因组DNA甲基化水平及甲基转移酶表达出现降低趋势,微核、核质桥及核芽突比率显著升高。在相同BD染毒条件下,与FA-C组比较,FA-D组的DNA甲基化水平和DNMT1、DNMT3a的mRNA表达出现下降趋势,而微核率等染色体损伤出现升高趋势;相反,FA-S组的DNA甲基化水平及DNMT1、DNMT3a的mRNA表达出现升高趋势,染色体损伤出现降低趋势,尤其在高剂量BD染毒条件下,叶酸补充对上述指标的影响均存在显著性差异(P<0.05或0.01)。结论:BD可诱导小鼠的DNA低甲基化改变及染色体损伤,叶酸缺乏可加重BD所致DNA低甲基化和遗传损伤,而补充叶酸则对BD诱导的甲基化改变和遗传损伤具有保护效应。

Contribution to the taxonomy of the genus Lycodon H.Boie in Fitzinger,1827(Reptilia:Squamata:Colubridae)in China,with description of two new species and resurrection and elevation of Dinodon septentrionale chapaense Angel,Bourret,1933

While considerable progress has been made in the taxonomic studies of the genus Lycodon in Asia,questions remain to be clarified regarding the taxonomy of certain groups,particularly those containing species in China.Not only do many regions in China remain overlooked by herpetologists,resulting in the possibility of undiscovered new species,but the surveyed areas also have suspicious records of recognized congeners that require taxonomic confirmations.Combining both morphological and genetic data,we tackle these outstanding issues in the taxonomy of Lycodon in China.In particular,we discover two new species of Lycodon:one from the previously neglected hot-dry valley in the northern Hengduan Mountain Region close to Tibet,and another recluse and cryptic species from the L.fasciatus complex in the downtown park of a major city in southern Sichuan Province.Additionally,we clarify the distribution of L.septentrionalis in China and resurrect and elevate its junior synonym subspecies,Dinodon septentrionale chapaense,as a full,valid species,and we synonymize the recently described L.namdongensis to the resurrected L.chapaensis comb.nov..Lycodon chapaensis comb.nov.thus represents a new national record of reptilian fauna of China.Lastly,based on literature review,we also correct some of the erroneous records of L.fasciatus and L.ruhstrati in China,point out remaining taxonomic issues of the genus for future research,and update the dichotomous key and distribution of the 20 species of Lycodon currently recorded from China.

Discovery of Japalura chapaensis Bourret, 1937 (Reptilia： Squamata： Agamidae) from Southeast Yunnan Province, China

Due to a paucity of surveys in northern Indochina and lack of international collaborations among neighboring countries, recognized distributional ranges for many amphibian and reptile species end at the political borders for some countries, despite seemingly continuous suitable habitat spanning the region. Combining both morphological and genetic data, we report the first discovery of Japalura chapaensis, a rare agamid lizard believed previously to be endemic to northern Vietnam only, along the border region of southeastern Yunnan Province, China. To facilitate future research on the genus Japalura sensu lato in Indochina, we provide detailed descriptions of additional specimens of this rare species, including the first description of coloration in life and an expanded diagnosis, and discuss the species boundary of J. chapaensis with respect to its congeners.

Two new species of Japalura(Squamata:Agamidae)from the Hengduan Mountain Range,China

Until recently, the agamid species, Japalura flaviceps was recognized to have the widest geographic dis- tribution among members of the genus occurring in China, from eastern Tibet to Shaanxi Province. However, recent studies restricted the distribution of J. flaviceps to the Dadu River valley only in north- western Sichuan Province, suggesting that records of J. flaviceps outside the Dadu River valley likely represent undescribed diversity. During two herpeto- faunal surveys in 2013 and 2015, eight and 12 specimens of lizards of the genus Japalura were collected from the upper Nujiang （=Salween） Valley in eastern Tibet, China, and upper Lancang （=Mekong） Valley in northwestern Yunnan, China, respectively. These specimens display a unique suite of diagnostic morphological characters. Our robust comparisons of phenotype reveal that these populations can be distinguished readily from J. flaviceps and all other recognized congeners. Herein, we describe the two Japalura lineages as new spe- cies, Japalura laeviventris sp. nov. and Japalura iadina sp. nov.. In addition, we provide updated con- servation assessments for the new species as well as imperiled congeners according to the IUCN crite- ria for classification, discuss the importance of color patterns in the diagnosis and description of species in the genus Japalura, and discuss directions for future taxonomic studies of the group.

Matrilineal History of the Rana longicrus Species Group(Rana,Ranidae,Anura) and the Description of a New Species from Hunan,southern China

Fieldwork in Jiemuxi National Nature Reserve, Hunan, China, discovered morphologically indis- tinguishable groups of brown frogs that bred at different, exclusive times of the year. A principal components analysis of morphometric data, molecular analyses, and the exclusive breeding season suggested the occurrence of two species. The population that breeds during the winter was found to be an undescribed species and was subsequently described as Rana jiemuxiensis sp. nov. The new cryptic species can be identified from its congeners at the same locality by having a different breeding season and a divergent DNA barcode, Six major lineages of brown frogs were resolved with high nodal support. Rana japonica, R. chaochiaoensis, R. omeimontis, and R. jiemuxiensis have independent maternal lineages. Rana hanlunica and R. maoershanensis have essentially identical maternal lineages and they appear to represent the same species. The validity of the species R. longicrus, R. zhenhaiensis, and R. culaiensis and their genealogical relationships are not resolved and deserve further study. The genealogy indicates that sympatric cryptic lineages probably widely exist in the R. longicrus group. This highlights the importance of future more fine-scaled samplings and the inclusion of nuclear genes.

AmphibiaChina： an online database of Chinese Amphibians

AmphibiaChina, an open-access, web-based database, is designed to provide comprehensive and up-to-date information on Chinese amphibians. It offers an integrated module with six major sections. Compared to other known databases including AmphibiaWeb and Amphibian Species of the World, AmphibiaChina has the following new functions： （1） online species identification based on DNA barcode sequences; （2） comparisons and discussions of different major taxonomic systems; and （3） phylogenetic progress on Chinese amphibians. This database offers a window for the world to access available information of Chinese amphibians. AmphibiaChina with its Chinese version can be accessed at http：// www.amphibiachina.org.

A new species of the genus Amolops （Amphibia： Ranidae） from southeastern Tibet, China

A new species of the genus Amolops Cope, 1865 is described from Nyingchi, southeastern Tibet, China, based on morphological and molecular data. The new species, Amolops nyingchiensis sp. nov. is assigned to the Amolops monticola group based on its skin smooth, dorsolateral fold distinct, lateral side of head black, upper lip stripe white extending to the shoulder. Amolops nyingchiensis sp. nov. is distinguished from all other species of Amolops by the following combination of characters： （1） medium body size, SVL 48.5-58.3 mm in males, and 57.6-70.7 mm in females; （2） tympanum distinct, slightly larger than one third of the eye diameter; （3） a small tooth-like projection on anteromedial edge of mandible; （4） the absence of white spine on dorsal surface of body; （5） the presence of circummarginal groove on all fingers; （6） the presence of vomerine teeth; （7） background coloration of dorsal surface brown, lateral body gray with yellow; （8） the presence of transverse bands on the dorsal limbs; （9） the presence of nuptial pad on the first finger in males; （10） the absence of vocal sac in males. Taxonomic status of the populations that were previously identified to A. monticola from Tibet is also discussed.

A new species of Mountain Dragon(Reptilia:Agamidae:Diploderma) from the D. dymondi complex in southern Sichuan Province, China

Despite continuous studies on the cryptic diversity of the Diploderma flaviceps complex in Southwest China for the pastdecade, little attention has been given to other widespreadcongeners in China. Combining both morphological and phylogenetic data, we describe a new species of Diplodermafrom populations identified previously as D. dymondi in thelower Yalong River Basin in southern Sichuan Province. Thenew species is morphologically most similar to D. dymondiand D. varcoae, but it can be differentiated by a considerablegenetic divergence and a suite of morphological characters,including having taller nuchal crest scales, smaller tympana,and a distinct oral coloration. Additionally, we discuss otherputative species complexes within the genus Diploderma inChina.

Discovery of a wild,genetically pure Chinese giant salamander creates new conservation opportunities

Effective conservation of threatened biota relies on accurate assessments and scientific guidance.As an unfortunate example,Chinese giant salamanders(Andrias,CGS)remain critically endangered in nature.Misguided conservation efforts,e.g.,commercial propagation and releasing of millions of likely non-indigenous or interspecific hybrids,have further compromised conservation initiatives.Limited information on wild populations of CGS poses a significant conservation challenge.Following 18-month long field monitoring,we now report the discovery of a wild population of CGS in a closed nature reserve in Jiangxi Province,China.Genomic assessments reveal its genetic distinctiveness and do not detect genetic admixture with other species.Based on morphological and molecular evidences,we describe this CGS as a new species Andrias jiangxiensis sp.nov.This is the only known species of CGS today with a genetically pure,reproducing,in situ population.This discovery emphasizes the important role that closed nature reserves play in protecting species,and the necessity of integrating long-term field monitoring and genetic assessments.It sets a new pathway for discovering and conserving endangered species,especially for those biotas that are similarly being extirpated by anthropogenic translocations and overexploitation.

A new genus and species of treefrog from Medog,southeastern Tibet,China(Anura,Rhacophoridae)

A new genus and species of threefrog is described from Medog, southeastern Tibet, China based on morphological and phylogenetic data. The new genus can be distinguished from other treefrog genera by the following combination of characters： （1） body size moderate, 45.0 mm in male; （2） snout rounded; （3） canthus rostralis obtuse and raised prominently, forming a ridge from nostril to anterior corner of eyes; （4） web rudimentary on fingers; （5） web moderately developed on toes; （6） phalange ＂Y＂ shaped, visible from dorsal side of fingers and toes; （7） skin of dorsal surfaces relatively smooth, scatted with small tubercles; （8） iris with a pale yellow, ＂X＂ shaped pattern of pigmentation.

A New Species of Japalura (Squamata: Sauria:Agamidae) from Upper Lancang (Mekong) Valley of Eastern Tibet, China

A new species of the agamid genus Japalura is described based on 15 specimens from the upper Lancang（Mekong） Valley of eastern Tibet, PR China. Populations of the new species, Japalura vela sp. nov., were previously recognized as J. flaviceps. The new species is morphologically most similar to J. batangensis, J. micangshanensis, J. variegata, and J. zhaoermii, but is distinguished from the four species and all remaining congeners by the following combination of morphological characters： 1） small adult size（SVL 56–69 mm in males, 59–66 mm in females）; 2） ratio of tail TAL/SVL 1.85–2.06; 3） ratio of hind limb HLL/SVL 0.72–0.81; 4） T4 S 24 or 25; 5） concealed tympanum; 6） transverse gular fold present; 7） gular pouch present; 8） axillary fold present; 9） a pronounced, continuous, sail-like vertebral crest along length of body in males; 10） ground dorsal coloration black in males; 11） distinct gray transverse streaks on dorsal surface of head; 12） black radiated streaks around eyes; 13） distinct, black vermiculate stripes on ventral surface of head in both sexes; 14） a strongly jagged dorsolateral stripe from neck to base of tail on each side of vertebral crest in males; and 15） absence of gular spots in both sexes. General distribution patterns of the genus in the Hengduan Mountains region are also discussed.

Phylogenetic Relationships of the Genus Paramesotriton(Caudata: Salamandridae) with the Description of a New Species from Qixiling Nature Reserve, Jiangxi, Southeastern China and a Key to the species

The matrilineal genealogy of the genus Paramesotriton is hypothesized based on DNA sequences from mitochondrial NADH subunit two （ND2） and its flanking tRNAs （tRN^Arp and a partial tRNA^Ala）. The genealogy identifies a highly divergent, unnamed lineage from Qixiling Nature Reserve, Jiangxi, China and places it as the sister taxon of P. chinensis. The newly discovered population differs from other congeners by several features of external morphology including having large clusters of dark brown conical warts on the dorsum of the head, lateral surface of the body and dorsolateral ridges. Its intermittent dorsal vertebral ridge is the same color as other parts of the dorsum and tail narrows gradually from the base to the tip. Further, the new population differs from all congeners by an uncorrected P-distance of more than 9.38% in ND2. Consequently, we describe the new species of Asian warty newt （Salamandridae） as Paramesotriton qixilingensis sp. nov.

The Australasian frog family Ceratobatrachidae in China,Myanmar and Thailand:discovery of a new Himalayan forest frog clade

In an effort to study the systematic affinities and species- level phylogenetic relationships of the enigmatic anurans vadably assigned to the genera Ingerana or Limnonectes （family Dicroglossidae）, we collected new molecular sequence data for five species including four Himalayan taxa, Limnonectes xizangensis, Lim. medogensis, Lim. alpine, Ingerana borealis and one southeast Asian species, I. tasanae, and analyzed these together with data from previous studies involving other ostensibly related taxa. Our surprising results demonstrate unequivocally that Lim. xizangensis, Lim. medogensis and Lira. alpine form a strongly supported clade, the sister-group of the family Australasian forest frog family Ceratobatrachidae. This discovery requires an expansion of the definition of Ceratobatrachidae and represents the first record of this family in China. These three species are distinguished from the species of Ingerana and Limnonectes by the： （1） absence of interdigital webbing of the foot, （2） absence of terminal discs on fingers and toes, （3） absence of circumarginal grooves on the fingers and toes, and （4） absence of tarsal folds. Given their phylogenetic and morphological distinctiveness, we assign them to the oldest available generic name for this clade, Liurana Dubois 1987, and transfer Liurana from Dicroglossidae to the family Ceratobatrachidae. In contrast, Ingerana tasanae was found to be clustered with strong support with the recently described genus Alcalus （Ceratobatrachidae）, a small clade of otherwise Sundaic species; this constitutes a new record of the family Ceratobatrachidae for Myanmar and Thailand. Finally, Ingerana borealis clustered with the ＂true＂ Ingerana （family Dicroglossidae）, for which the type species is L tenasserimensis.

A new species of the genus Scutiger （Anura： Mego- phryidae） from Medog of southeastern Tibet, China

A new species of Scutiger Theobald, 1868 is described from Medog, southeastern Tibet, China, based on morphological and molecular data. The new species was previously identified as Scutiger nyingchiensis, but it can be differentiated from the latter and all other congeners by the following combination of characters： （1） medium adult body size, SVL 50.5-55.6 mm in males and 53.8-57.2 mm in females; （2） maxillary teeth absent; （3） web rudimen- tary between toes; （4） prominent, conical-shaped tubercles on dorsal and lateral surfaces of body and limbs; （5） tubercles covered by black spines in both sexes in breeding condition; （6） a pair of pectoral glands and a pair of axillary glands present and covered by black spines in males in breeding condition, width of axillary gland less than 50% of pectoral gland; （7） nuptial spines present on dorsal surface of first and second fingers, and inner side of third finger in males in breeding condition; （8） spines absent on the abdominal region; （9） vocal sac absent. In addition, the distribution and conservation status of the new species are also discussed

Comparative Skin Histology of Frogs Reveals High-elevation Adaptation of the Tibetan Nanorana parkeri

Adaptations to extreme environmental conditions are intriguing. Animal skin, which directly interacts with external environment, plays diverse and important roles in adaptive evolution. The thin and bare skin of amphibians is sensitive to external environmental conditions and, thus, it facilitates investigations into adaptations for living in extreme environments. Herein, we compare the structures of skin in four anuran species living at elevations ranging from 100 m to 4500 m to assess phenotypic innovations in the skin of Nanorana parkeri, which lives at extremely high elevations. Analyses reveal similar basic skin structures, but N. parkeri differs from the other species by having more epidermal capillaries and granular glands, which correlate highly with responses to hypoxia and/or ultraviolet(UV) radiation. Further intraspecific comparisons from frogs taken at ~4500 m and ~2900 m reveal that all of the changes are fixed. Changes occurring only in the higher elevation population, such as possessing more skin pigments, may represent local adaptations to coldness and/or UV radiation. These results provide a morphological basis for understanding further the molecular adaptations of these frogs.