动物对极端环境的适应是动物生理生态学研究的核心内容之一。高山倭蛙作为青藏高原特有物种，是世界上海拔分布最高的两栖动物，因其独特的高原适应能力而备受关注。高海拔种群栖息地冬季的月平均气温在0°C以下，这必然对其生存是一大挑战。高山倭蛙如何度过并适应高海拔地区寒冷漫长的冬季是一个非常值得关注的问题，但目前关于其低温生存策略及机制知之甚少。前期研究证实了冬眠期高山倭蛙具有耐受结冰的能力，通过积累小分子代谢物及组织脱水来保护机体。本项目拟选择冬眠期高山倭蛙作为研究对象，围绕耐受结冰的四个方面：结冰存活能力、抗冻保护剂、冰成核剂、基因表达调控，结合代谢组学、转录组学和16S扩增子测序技术，从生理生化、分子及微生物群落水平系统地探究冬眠期高山倭蛙耐受结冰的分子机制。本项目的研究成果将有助于揭示两栖动物适应高原极端环境的机制，丰富低温生物学、动物生理生态学研究的理论体系，为保护高原生物多样性提供参考。

The adaptation of animals to an extreme environment is a major concern in animal physiological ecology. Nanorana parkeri is a frog species endemic to the Tibetan Plateau and figures among the highest altitude amphibians in the world. This species has attracted much attention due to its unique plateau adaptability. The monthly mean air temperature of high-altitude habitat is below 0°C in winter, which is a great challenge for this species' survival. How to adapt to the cold and long winter for this species is a very important issue. However, the survival strategy and underlying mechanisms have been poorly understood. Our previous results showed that hibernating N. parkeri living at high altitude has the ability to tolerate extracellular freezing with accumulation of low molecular weight metabolites and tissue dehydration in winter. In the current project, we systematically investigate four aspects of freeze tolerance: freezing survival ability, cryoprotectants, ice nucleating agents, and gene expression regulation in hibernating N. parkeri to explore the underlying molecular mechanisms from the physiological, biochemical, molecular, and microbial community level using a combination of metabonomics, transcriptomics, and 16S rRNA amplicon sequencing. These results will contribute to revealing the mechanism of amphibians adapting to the extreme environment of the Tibetan Plateau, enriching the theory of cryobiology and animal physiological ecology, and providing reference for the conservation of plateau biodiversity.