Species belonging to the genus Microhyla are small-sized frogs that are widely distributed in southern, eastern, and south-eastern Asia. In China, the genus harbors many cryptic species, on two of which—M. beilunensis and M. fanjinshanensis—studies were recently published. In this study, we collected specimens from the Dabie Mountain range, which is at the junction of Anhui, Henan and Hubei Provinces, East China; these specimens belonged to a species previously identified as M. mixtura. Based on phylogenetic analyses, species delimitation analyses, morphological comparisons and advertisement calls comparisons, we found they were significantly different from other known congeners, and thus we describe them as a new species. This study enriches the diversity of Microhylidae, and clarifies the species of the genus Microhyla in the Dabie Mountains.

The Longdong Stream SalamanderBatrachuperus londongensis, living in a mountain stream environment at Mt. Emei in Sichuan Province, China, represents a rare species that is facultatively neotenic in the family Hynobiidae. Although the species has been known to science for some 40 years since its initial discovery in the late 1970s, anatomical details of its osteology remain poorly understood and developmental information is still lacking for the species. This study (1) provides a detailed osteological account ofB. londongensisbased on micro-CT scanning and clearing and staining of multiple specimens from the type locality; (2) provides a discussion of intraspecific variation related to life-history differences; and (3) presents a discussion on limb features related to morphological evolution of limb patterns correlative with ecological adaptation to mountain stream environments. Osteological comparisons with congeneric species has led to recognition of several diagnostic features that are unique toB. londongensis, including: vomers widely separated from one another, lacking a midline contact; presence of uncommon perichondral ossification of the ascending process of the palatoquadrate as part of the suspensorium; and presence of a prominent posterodorsal process of the scapular blade, which serves as a ligamentous insertion of the levator muscle of the scapula. In addition, some but not all neotenic individuals retain the palatine as a discrete element, indicative of its delayed absorption after sexual maturity. Postmetamorphic and neotenic individuals are strikingly different in the complexity of hyobranchial structures. Neotenes display a high degree of ossification of hyobranchial elements, tend to increase ossification of both hypobranchial I and ceratobranchial I during aging, and retain fully ossified ceratobranchial III and IV; in contrast, these elements remain entirely cartilaginous or are totally lost by resorption in postmetamorphic individuals. In addition, all postmetamorphic forms display an inverted “T”-shaped basibranchial II, whereas neotenes show transformation from a “fork”-shaped to the “T”-shaped configuration after sexual maturity.B. londongensisdisplays a mosaic of apomorphic and plesiomorphic states in its limb ossifications: presence of a single centrale element in both the manus and pes is a derived condition in Hynobiidae and other families as well, whereas retention of a postminimus in the pes is obviously plesiomorphic within Urodela. Reduction in number of digits from five to four in the pes and possession of a cornified sheath covering the terminal phalanges are also derived features shared with some but not all mountain stream salamanders that are adapted to a similar type of environment.

Comprehending the determinants of host-associated microbiota is pivotal in microbial ecology. Yet, the links between climatic factors and variations in host-associated microbiota necessitate further clarification. Mountain-dwelling amphibians, with limited dispersal abilities, serve as valuable models for addressing these questions. Our study, using 126 amphibian-associated microbial samples (64 gut and 62 skin) and 101 environmental microbial samples (51 soil and 50 water) from the eastern Tibetan Plateau, revealed host factors as primary drivers of the variations in host-associated microbiota. However, climatic factors contributed to additional variations in gut microbial beta-diversity and skin microbial function. Water microbiota were identified as a significant contributor to the amphibian-associated microbiomes, with their climate-driven variations mediating an indirect association between the variations in climatic factors and host-associated microbiota. These findings extend our understanding of the assembly of host-associated microbiota in amphibians, emphasizing the significance of microbiota in evaluating the impact of climate change on animals.

Most anurans must undergo metamorphosis to adapt to terrestrial life. This process enhances the air-breathing ability of the lungs to cope with the change in oxygen medium from water to air. Revealing the structural construction and molecular switches of lung organogenesis is essential to understanding the realization of the air-breathing function. In this study, histology and transcriptomics were conducted in combination to explore these issues in Microhyla fissipes’ lungs during metamorphosis. During the pro-metamorphic phase, histological structural improvement of the alveolar wall is accompanied by robust substrate metabolism and protein turnover. The lungs, at the metamorphic climax phase, are characterized by an increased number of cilia in the alveolar epithelial cells and collagenous fibers in the connective tissues, corresponding to the transcriptional upregulation of cilia and extracellular matrix-related genes. Post-metamorphic lungs strengthen their contracting function, as suggested by the thickened muscle layer and the upregulated expression of genes involved in muscle contraction. The blood–gas barrier is fully developed in adult lungs, the transcriptional features of which are tissue growth and regulation of differentiation and immunity. Importantly, significant transcriptional switches of pulmonary surfactant protein and hemoglobin facilitate air breathing. Our results illuminated four key steps of lung development for amphibians to transition from water to land.

Genetic adaptation to extremes is a fascinating topic. Nevertheless, few studies have explored the genetic adaptation of closely related species respectively inhabiting distinct extremes. With deep transcriptome sequencing, we attempt to detect the genetic architectures of tadpoles of five closely related toad species adapted to the Tibetan Plateau, middle-altitude mountains and karst caves. Molecular evolution analyses indicated that not only the number of fast evolving genes (FEGs), but also the functioning coverage of FEGs, increased with elevation. Enrichment analyses correspondingly revealed that the highland species had most of the FEGs involved in high-elevation adaptation, for example, amino acid substitutions of XRCC6 in its binding domains might improve the capacity of DNA repair of the toad. Yet, few FEGs and positively selected genes (PSGs) involved in high-elevation adaptation were identified in the cave species, and none of which potentially contributed to cave adaptation. Accordingly, it is speculated that in the closely related toad tadpoles, genetic selection pressures increased with elevation, and cave adaptation was most likely derived from other factors (e.g., gene loss, pseudogenization or deletion), which could not be detected by our analyses. The findings supply a foundation for understanding the genetic adaptations of amphibians inhabiting extremes.