Alpine grasslands are vital in regulating carbon balance on the Qinghai-Tibetan Plateau (QTP) because of the large soil organic carbon (SOC) stocks, while persistent disturbance from the endemic small semifossorial herbivore, plateau pika (Ochotona curzoniae, hereafter pika), may break this balance. Pika affect the soil microclimate by creating a heterogeneous underlying surface, which is expected to alter soil microbial communities and eventually SOC stocks. However, our knowledge regarding the potential influence mechanism is still limited. Here, we investigated vegetation biomass, soil properties and soil microbes among 4 different surfaces (i.e., original vegetation, new pika pile, old pika pile and bare patch) of typical alpine grasslands to reveal soil microbial communities and the associated effect on SOC in response to pika bioturbation. Our results showed that pika bioturbation increased both bacterial and fungal diversity and their phyla abundance for SOC decomposition. Vegetation biomass, electrical conductivity and NH4+-N accounted for the variation in both bacterial and fungal community compositions and diversity. SOC stocks were 15-30% lower in pika piles and bare patches than in the original vegetation, which was mainly attributed to the reduced soil organic matter input from vegetation and the enhanced SOC consumption by soil microbial communities. Overall, we conclude that pika bioturbation altered the diversity and composition of soil microbial communities, which was associated with SOC loss and positive carbon feedback in alpine grasslands. Our findings provide insights into the role of small semifossorial herbivores in the carbon cycle of global grasslands.

The freezing-thawing cycle is a basic feature of a frozen soil ecosystem, and it affects the growth of alpine vegetation both directly and indirectly. As the climate changes, the freezing-thawing mode, along with its impact on frozen soil ecosystems, also changes. In this research, the freezing-thawing cycle of the Nagqu River Basin in the Qinghai-Tibet Plateau was studied. Vegetation growth characteristics and microbial abundance were analyzed under different freezing-thawing modes. The direct and indirect effects of the freezing-thawing cycle mode on alpine vegetation in the Nagqu River Basin are presented, and the changing trends and hazards of the freezing-thawing cycle mode due to climate change are discussed. The results highlight two major findings. First, the freezing-thawing cycle in the Nagqu River Basin has a high-frequency mode (HFM) and a low-frequency mode (LFM). With the influence of climate change, the LFM is gradually shifting to the HFM. Second, the alpine vegetation biomass in the HFM is lower than that in the LFM. Frequent freezing-thawing cycles reduce root cell activity and can even lead to root cell death. On the other hand, frequent freezing-thawing cycles increase microbial (Bradyrhizobium, Mesorhizobium, and Pseudomonas) death, weaken symbiotic nitrogen fixation and the disease resistance of vegetation, accelerate soil nutrient loss, reduce the soil water holding capacity and soil moisture, and hinder root growth. This study provides a complete response mechanism of alpine vegetation to the freezing-thawing cycle frequency while providing a theoretical basis for studying the change direction and impact on the frozen soil ecosystem due to climate change.

Recent global warming models project a significant change in winter climate over the next few decades. The decrease in snowpack in the winter will decrease the heat insulation function of the snowpack, resulting in increased soil freeze-thaw cycles. Here, we examined the impact of winter freeze-thaw cycles on year-round dissolved nitrogen (N) and carbon (C) dynamics and their relationship with dissolved organic matter and microbial biomass in soil by conducting an in situ experimental reduction in snowpack. We investigated dissolved inorganic N (NH4+ and NO3-), dissolved organic N (DON), dissolved organic carbon (DOC), inorganic N leaching, soil microbial biomass, and microbial activities (mineralization and nitrification) in the surface soil of a northern hardwood forest located in Japan. Experimental snowpack reduction significantly increased the number of soil freeze-thaw cycles and soil frost depth. The NH4+ content of the surface soil was significantly increased by the amplified soil freeze-thaw cycles due to decreased snowpack, while the soil NO3- content was unchanged or decreased slightly. The gravimetric soil moisture, DON and DOC contents in soil and soil microbial biomass significantly increased by the snowpack removal in winter. Our results suggest that the amplified freeze-thaw cycles in soil increase the availability of DON and DOC for soil microbes due to an increase in soil freezing. The increases in both DON and DOC in winter contributed to the enhanced growth of soil microbes, resulting in the increased availability of NH4+ in winter from net mineralization following an increase in soil freeze-thaw cycles. Our study clearly indicated that snow reduction significantly increased the availability of dissolved nitrogen and carbon during winter, caused by increased soil water content due to freeze-thaw cycles in winter.