Background: Understanding how organisms adapt to high-elevation environments at a genome scale provides novel insights into the process of adaptive evolution. Previous studies have mainly focused on endothermic organisms, while poikilothermic species may have evolved different mechanisms to cope with high-elevation environments. In this context, we sequenced transcriptomes of a pair of closely related anuran species, Rana chensinensis and R. kukunoris, which inhabit respective low- and high-elevation habitats. By comparing the two transcriptomes, we identified candidate genes that may be involved in high-elevation adaption in poikilothermic species. Results: Over 66 million sequence reads from each transcriptome were generated. A total of 41,858 and 39,293 transcripts for each species were obtained by de novo assembly. By comparing the orthologous transcripts, we identified 125 protein-coding genes that have likely experienced strong positive selection (Ka/Ks>1). In addition, 335 genes that may bear a signature of positive selection (1≥Ka/Ks>0.5) were also recognized. By considering their functions, fourteen candidate genes were determined to be likely involved in high-elevation adaptation, including two CYP genes, USP-1, and several others. Conclusions: We identified a set of candidate genes that may have promoted adaptation of R. kukunoris to its high-elevation environment. These include several genes that have previously been associated with oxygen binding, response to UV radiation, and repair of free radical injury. Detailed molecular, physiological, and phenotypic studies are required to better understand the roles of these genes in improving the performance of R. kukunoris in a high-elevation environment. We have evidence for both convergent and divergent evolution between endothermic and poikilothemic species, but additional research across a wide range of organisms will be necessary to comprehend the complexity of high-elevation adaptation.

While crypsis is a prominent antipredator adaptation, the role of the brain in predator-driven evolution remains controversial. Resolving this controversy requires contextualizing the brain with established antipredator traits and predation pressure. We hypothesize that the reduced predation risk through crypsis relaxes predation-driven selection on the brain and provide comparative evidence across 102 Chinese frog species for our hypothesis. Specifically, our phylogenetic path analysis reveals an indirect relationship between predation risk and crypsis that is mediated by brain size. This result suggests that at a low predation risk, frogs can afford to be conspicuous and use their large brain for cognitive predator evasion. This strategy may become less efficient or energetically costlier under higher predation pressure, favoring smaller brains and instead increasing crypsis.

Recent studies have highlighted the underestimated diversity of the genus Diploderma Hallowell, 1861 in the Hengduan Mountain Region in Southwest China, but much of the region remains poorly surveyed for reptile diversity. In this study we describe two new species of Diploderma from the upper Jinsha and middle Yalong River Valley, based on evaluations of morphological, genetic, and distribution data. The two new species are morphologically most similar to D. angustelinea and D. vela, but they can be diagnosed from both recognized taxa and all remaining congeners by a suite of morphological features, particularly the distinct coloration of gular spots. Additionally, both new species either render other recognized species paraphyletic or are allopatric with respect to their morphologically similar congeners. Furthermore, we rediscover D. brevicaudum in the wild for the first time, which was known from historical museum specimens only. We estimate the phylogenetic position of D. brevicaudum within the genus Diploderma based on mitochondrial genealogy, and we provide an expanded diagnosis and comparisons against closely related congeners and provide a detailed description of coloration in life based on newly collected specimens. Our discoveries of the new Diploderma species further highlight the urgent conservation needs of the currently neglected hot-dry valley ecosystems in the Hengduan Mountain Region of China.

High-altitude adaptation provides an excellent system for studying how organisms cope with multiple environmental stressors and interacting genetic modifications. To explore the genetic basis of high-altitude adaptation in poikilothermic animals, we acquired transcriptome sequences from a high-altitude population and a low-altitude population of the Asiatic toad (Bufo gargarizans). Transcriptome data from another high-altitude amphibian, Rana kukunoris and its low-altitude relative R. chensiensis, which are from a previous study, were also incorporated into our comparative analysis. More than 40,000 transcripts were obtained from each transcriptome, and 5107 one-to-one orthologs were identified among the four taxa for comparative analysis. A total of 29 (Bufo) and 33 (Rana) putative positively selected genes were identified for the two high-altitude species, which were mainly concentrated in nutrient metabolism related functions. Using SNP-tagging and FST outlier analysis, we further tested 89 other nutrient metabolism related genes for signatures of natural selection, and found that two genes, CAPN2 and ITPR1, were likely under balancing selection. We did not detect any positively selected genes associated with response to hypoxia. Amphibians clearly employ different genetic mechanisms for high-altitude adaptation compared to endotherms. Modifications of genes associated with nutrient metabolism feature prominently while genes related to hypoxia tolerance appear to be insignificant. Poikilotherms represent the majority of animal diversity, and we hope that our results will provide useful directions for future studies of amphibians as well as other poikilotherms.

Competition between animals for limited resources often involves signaling to establish ownership or dominance. In some species, the defended resource relates to suitable thermal conditions and refuge from predators. This is particularly true of burrow-dwelling lizards such as the Qinghai toad-headed agama (Phrynocephalus vlangalii), which are found on the Tibetan plateau of western China. Male and female lizards occupy separate burrows, which are vital for anti-predator behaviour during warmer months when lizards are active and, crucially, provide shelter from harsh winter conditions. These lizards are readily observed signaling by means of tail displays on the sand dunes they inhabit. Given the selective pressure to hold such a resource, both males and females should exhibit territorial behaviour and we considered this study system to examine in detail how social context influences motion based territorial signaling. We confirmed that territorial signaling was used by both sexes and by adopting a novel strategy that permitted 3D reconstruction of tail displays, we identified significant variation due to social context. However, signal structure was not related to lizard morphology. Clearly, the burrow is a highly valued resource and we suggest that additional variation in signaling behaviour might be mediated by resource quality.