The wood frog (Rana sylvatica) is one of only a few vertebrate species that can survive extensive freezing of its body fluids during the winter. The mechanisms of natural freeze tolerance include metabolic rate depression to conserve energy and the implementation of cryoprotective strategies, especially the synthesis of huge amounts of glucose as a cryoprotectant. Liver is the main source of glucose production/export (and other cryoprotective actions) and plays a central role in freezing survival of the whole animal. Freezing is a multi-component stress that includes anoxia/ischemia due to the cessation of blood flow and dehydration of cells caused by ice accumulation in extracellular spaces. To help endure these stresses, cells need to suppress and reprioritize ATP-expensive cell functions. One of these is cell growth and proliferation, and we hypothesized that cell cycle arrest would be key to freezing survival. The present study examines the responses by key cell cycle components to freezing, anoxia and dehydration stresses in wood frog liver. Immunoblotting was used to investigate protein expression of Cdc 2, Cdks (2, 4, 6), and cyclins (A, B1, D1, E) as well as the phosphorylation states of Cdks (Thr14/Tyr15), the phosphatases Cdc25a (Ser76) and Cdc25c (Ser216) and the CIP/KIP Cdk inhibitors p21 (Thr145) and p27 (Thr187). Responses to 24 h freezing, 24 h anoxia and 40% dehydration as well as recovery from these stresses were analyzed. The results showed very similar responses by cell cycle components to anoxia or dehydration and were consistent with cell cycle suppression under stress and reversal during recovery. Freezing showed elements of cell cycle suppression, including reduced protein levels of Cdks and cyclins A and B1, but also showed unique responses by cyclin D1, Cdc25 phosphatases and p21/p27. These may be linked with alternative actions by these proteins that contribute to cryoprotection; e.g., an alternative action of cyclin D1 as a transcription factor may contribute to the upregulation of glucose-6-phosphatase, a key enzyme needed for the export of glucose cryoprotectant.

Plant diversity plays an important role in maintaining the stability of ecosystem functioning. Based on field surveys and indoor analyses, this study investigated the relationship between species diversity and community stability at different stages of bare patch succession in degraded alpine meadow ecosystems. Results show that: (1) Using the ICV (the Inverse of the Coefficient of Variation) method to analyze changes in plant community stability, community stability was generally ranked as follows: Long-term recovered patches > Healthy alpine meadow > Degraded alpine meadow > Short-term recovered patch > Bare Patches. (2) Using factor analysis to construct an evaluation system, the stability ranking based on species diversity was as follows: Healthy alpine meadow > Long-term recovered patches > Degraded alpine meadow > Short-term recovered patches > Bare Patches. (3) The community stability index was significantly positively correlated with vegetation coverage, height, biomass, species richness, Shannon–Wiener diversity index, species evenness, and Simpson’s diversity index (p < 0.05). Therefore, a positive correlation exists between plant diversity and community stability, such that plant communities with a higher species diversity tend to be more stable. To maintain the plant diversity and community stability of alpine meadow ecosystems, it is necessary to consider the characteristics of grassland plant composition and community structure, as well as their influencing factors, and promote the positive succession process of grasslands.

Identifying the relative contributions of climate change and human activities to alpine grassland dynamics is critical for understanding grassland degradation mechanisms. In this study, first, the actual NPP (NPP_a) was obtained by MOD17A3. Second, we used the Zhou Guangsheng model to simulate the potential met net primary productivity (NPP_p). Finally, the NPP generated by anthropogenic activities (NPP_h) was estimated by calculating the difference between NPP_p and NPP_a. Then, the relative contributions of climate change and human activities to NPP changes in grasslands were quantitatively assessed by analyzing trends in NPP_p and NPP_a. Thereby, the drivers of NPP change in the Yellow River source grassland were identified. The results showed that the temperature and precipitation in the study area showed a warm-humid climate trend from 2000 to 2020. The NPP_p and NPP_a increased at a rate of 1.07 g C/m² and 1.51 g C/m² per year, respectively, while the NPP_h decreased at a rate of 0.46 g C/m² per year. It can be seen that human activities had a positive effect on the change of NPP in the Yellow River source grassland from the change rate. The relative contribution analysis showed that 55.90% of grassland NPP increased due to climate change, 40.16% of grassland NPP increased due to human activities, and the grassland degradation was not significant. The research results can provide a theoretical basis and technical support for the next step of the Yellow River source grassland ecological protection project.

In view of the problem of sustainable restoration of vegetation in alpine mining areas, vegetation communities and physical and chemical properties of soil under different artificial restoration measures (i.e., grass monoculture, sowing quantity and topsoil replacement) were investigated for five consecutive years (2016–2020) in the sloped eastern area of a northern slag mound in the Jiangcang mining area of the Muli coalfield in Qinghai, China. The results showed that the vegetation characteristics of different sowing treatments with different kinds of grass species, such as Elymus nutans, Poa pratensis cv. Qinghai and Poa crymophila cv. Qinghai, were significantly different (p < 0.05). The content of soil available nitrogen and phosphorus was significantly different among different grass species (p < 0.05). Vegetation coverage and ramet density of sowing treatment five were significantly higher than those of a lower sowing quantity (p < 0.05). There was no significant difference in the vegetation characteristics among different grass species treatments five years after the restoration (p > 0.05). There was no significant difference in vegetation height of different soil covering treatments (p > 0.05). In the early stage of restoration, vegetation coverage and ramet density in TR2 (resurfacing soil 10 cm) and TR3 (resurfacing soil 15 cm) were significantly higher than those without treatment. Soil N, P and organic matter under the treatments of TR1 (resurfacing soil 5 cm), TR2 and TR3 were significantly higher than those in CK (p < 0.05) in the early stage of restoration, but there was no significant difference in soil N, P and organic matter after five years of restoration. Overall, the vegetation and soil characteristics showed a trend of increase first and then decrease during the 5-year restoration period under different artificial measures. There were significant differences in the vegetation and soil characteristics among different treatments in the second year of restoration (p < 0.05), but there was no significant difference between the first year and the fifth year of restoration, which indicated that vegetation and soil began to degrade after five years of restoration, and substrate nutrients in the mining waste soil could only support the short-term restoration of vegetation. Therefore, it is necessary to target matrix nutrients in future vegetation restoration in alpine mining areas.

This study aimed to understand the response of vegetation community characteristics in the degraded alpine meadow of the Source Zone of the Yellow River to exclosure of various lengths. Artificial fences were erected to prevent livestock grazing and let the degraded meadow recover naturally as a means of restoration. The research focused on a typical degraded alpine meadow in which four plots were fenced off for three periods of 1 year (E1), 4 years (E4), and 10 years (E10), plus a freely grazed plot as the control. The study compared and analyzed the differences in plant community characteristics, carbon (C), nitrogen (N), and phosphorus (P) reserves, as well as the stoichiometric characteristics of main functional groups in the alpine meadow over different exclosure durations. The results indicated that E10 long-term exclosure significantly increased the aboveground biomass of gramineous plants but reduced the aboveground biomass of miscellaneous grasses. However, when compared to E4 short-term exclosure, E10 resulted in a reduction in the aboveground biomass of Cyperaceae plants. On the other hand, E4 medium-term exclosure significantly increased the aboveground biomass of Gramineae and Cyperaceae. Exclosure significantly increased the nitrogen (N) and phosphorus (P) reserves of the aboveground plant communities. Among these communities, the plant communities in the E10 long-term exclosure had the highest N and P reserves. However, this exclosure length also led to a significant reduction in plant diversity. Furthermore, except for Cyperaceae, all functional groups were observed in E10 and E4 plots. The carbon–nitrogen ratio and carbon–phosphorus ratio of these groups were significantly lower than those of groups G and E1. Medium-term exclosure (E4) has a positive impact on the aboveground biomass as well as plants’ nitrogen and phosphorus reserves. However, long-term exclosure (E10) has been observed to decrease species diversity and nutrient utilization efficiency of alpine meadow vegetation, which can be detrimental to the sustainable development of the alpine meadow ecosystem. Therefore, it is not recommended to implement long-term exclosure. Instead, a moderate level of grazing should be adopted after 4 years of exclosure.