Ambient climate determines the directional trend of community stability under warming and grazing.

Changes in ecological processes over time in ambient treatments are often larger than the responses to manipulative treatments in climate change experiments. However, the impacts of human-driven environmental changes on the stability of natural grasslands have been typically assessed by comparing differences between manipulative plots and reference plots. Little is known about whether or how ambient climate regulates the effects of manipulative treatments and their underlying mechanisms. We collected two datasets, one a 36-year long-term observational dataset from 1983 to 2018, and the other a 10-year manipulative asymmetric warming and grazing experiment using infrared heaters with moderate grazing from 2006 to 2015 in an alpine meadow on the Tibetan Plateau. The 36-year observational dataset shows that there was a nonlinear response of community stability to ambient temperature with a positive relationship between them due to an increase in ambient temperature in the first 25 years and then a decrease in ambient temperature thereafter. Warming and grazing decreased community stability with experiment duration through an increase in legume cover and a decrease in species asynchrony, which was due to the decreasing background temperature through time during the 10-year experiment period. Moreover, the temperature sensitivity of community stability was higher under the ambient treatment than under the manipulative treatments. Therefore, our results suggested that ambient climate may control the directional trend of community stability while manipulative treatments may determine the temperature sensitivity of the response of community stability to climate relative to the ambient treatment. Our study emphasizes the importance of the context dependency of the response of community stability to human-driven environmental changes.

Annual ecosystem respiration is resistant to changes in freeze-thaw periods in semi-arid permafrost.

Warming in cold regions alters freezing and thawing (F-T) of soil in winter, exposing soil organic carbon to decomposition. Carbon-rich permafrost is expected to release more CO2 to the atmosphere through ecosystem respiration (Re) under future climate scenarios. However, the mechanisms of the responses of freeze-thaw periods to climate change and their coupling with Re in situ are poorly understood. Here, using 2 years of continuous data, we test how changes in F-T events relate to annual Re under four warming levels and precipitation addition in a semi-arid grassland with discontinuous alpine permafrost. Warming shortened the entire F-T period because the frozen period shortened more than the extended freezing period. It decreased total Re during the F-T period mainly due to decrease in mean Re rate. However, warming did not alter annual Re because of reduced soil water content and the small contribution of total Re during the F-T period to annual Re. Although there were no effects of precipitation addition alone or interactions with warming on F-T events, precipitation addition increased total Re during the F-T period and the whole year. This decoupling between changes in soil freeze-thaw events and annual Re could result from their different driving factors. Our results suggest that annual Re could be mainly determined by soil water content rather than by change in freeze-thaw periods induced by warming in semi-arid alpine permafrost.

Microbial community responses reduce soil carbon loss in Tibetan alpine grasslands under short-term warming.

Changes in labile carbon (LC) pools and microbial communities are the primary factors controlling soil heterotrophic respiration (Rh ) in warming experiments. Warming is expected to initially increase Rh but studies show this increase may not be continuous or sustained. Specifically, LC and soil microbiome have been shown to contribute to the effect of extended warming on Rh . However, their relative contribution is unclear and this gap in knowledge causes considerable uncertainty in the prediction of carbon cycle feedbacks to climate change. In this study, we used a two-step incubation approach to reveal the relative contribution of LC limitation and soil microbial community responses in attenuating the effect that extended warming has on Rh . Soil samples from three Tibetan ecosystems-an alpine meadow (AM), alpine steppe (AS), and desert steppe (DS)-were exposed to a temperature gradient of 5-25°C. After an initial incubation period, soils were processed in one of two methods: (a) soils were sterilized then inoculated with parent soil microbes to assess the LC limitation effects, while controlling for microbial community responses; or (b) soil microbes from the incubations were used to inoculate sterilized parent soils to assess the microbial community effects, while controlling for LC limitation. We found both LC limitation and microbial community responses led to significant declines in Rh by 37% and 30%, respectively, but their relative contributions were ecosystem specific. LC limitation alone caused a greater Rh decrease for DS soils than AMs or ASs. Our study demonstrates that soil carbon loss due to Rh in Tibetan alpine soils-especially in copiotrophic soils-will be weakened by microbial community responses under short-term warming.

Fungal pathogens pose a potential threat to animal and plant health in desertified and pika-burrowed alpine meadows on the Tibetan Plateau.

Intact Tibetan meadows provide significant defense against soil-borne pathogen dispersal. However, dramatic meadow degradation has been observed due to climate change and pika damage, but their impacts on soil-borne pathogens are still unclear. With approximately 40% of the world's population living in Tibetan Plateau and its downstream watersheds, this lack of knowledge should be of great concern. Here, we used Illumina amplicon sequencing to characterize the changes in potential human, domestic animal, plant, and zoonotic bacterial and fungal pathogens in nondegraded, desertified, and pika-burrowed meadows. The relative abundance of bacterial domestic animal pathogens and zoonotic pathogens were significantly increased by desertification. Pika burrowing significantly increased the relative abundance of bacterial human pathogens and zoonotic pathogens. The species richness and relative abundance of fungal pathogens was significantly increased by desertification and pika burrowing. Accordingly, fungal plant and animal pathogens categorized by FUNGuid significantly increased in desertified and pika-burrowed meadows. Soil chemical and plant properties explained 38% and 64% of the bacterial and fungal pathogen community variance, respectively. Therefore, our study indicates for the first time that both alpine meadow desertification and pika burrowing could potentially increase infectious disease risks in the alpine ecosystem, especially for fungal diseases.

Asymmetric warming reduces the strength of selection pressure of moderate grazing on reproductive phenology in alpine plants.

Both warming and grazing already affect the reproductive phenology of alpine plants. However, their effects have mostly been studied in isolation, and their interaction is still unclear. In this study, an asymmetric warming (average + 1.2 °C during daytime and + 1.7 °C during nighttime and + 1.5 °C during summer and + 2.0 °C during winter) with moderate grazing experiment was conducted for four years to determine their individual and interactive effects on the onsets and durations of reproductive phenophases for fifteen alpine plant species on the Qinghai-Tibetan Plateau. Individual warming and grazing simultaneously advanced the average start dates and ending dates of budding, flowering and fruiting by 5.3-6.2 days, and further resulted in smaller effects on their durations for most plant species. The interactions between warming and grazing on them varied with plant species and year, which advanced by average 12.1 days for all plant species. The effects of grazing on the temperature sensitivity of the start dates of reproductive phenophases (average by -8.5 days °C-1) were greater than that of warming alone (average by -3.4 days °C-1) and warming with grazing (average by -5.5 days °C-1) for most of the alpine plant species. There were significant effects of the previous phenological events on subsequent reproductive phenophases. Therefore, our results suggested that both warming and grazing advanced reproductive phenophases through altered soil temperature and soil moisture and carry-over effects of previous phenological events on subsequent phenological events. Warming reduced the temperature sensitivity of the start dates of reproductive phenophases to grazing, suggesting that it depressed strength of selection pressure of grazing on the onsets of reproductive phenology in alpine plants.

Warming delays but grazing advances leaf senescence of five plant species in an alpine meadow.

Leaf senescence is the final stage in the life cycle of leaves and is critical to plants' fitness as well as to ecosystem carbon and nutrient cycling. To date, most understanding about the responses of leaf senescence to environmental changes has derived from research in forests, but the topic has been relatively neglected, especially under grazing conditions, in natural grasslands. We conducted a 3-year manipulative asymmetric warming with moderate grazing experiment to explore the responses of leaf senescence of five main species in an alpine meadow on the Qinghai-Tibetan Plateau. We found that warming prolonged leaf longevity through earlier leaf-out and later leaf senescence, and grazing prolonged it through a greater advance in leaf-out than first leaf coloration for all plants. Warming did not affect leaf nitrogen (N) content or N resorption efficiency (NRE), but grazing increased N content in coloring leaves for P. anserine and P. nivea and decreased NRE for K. humilis, P. anserine under no-warming, and for P. nivea under warming. The interactive effects of warming and grazing on leaf phenology and leaf traits depended on species identity and year. There were positive relationships between leaf-out and leaf senescence mainly derived from grazing, and positive relationships between NRE from old leaves and leaf senescence for three out of five plant species. Therefore, our results indicated that earlier leaf-out could result in earlier leaf senescence only under grazing, but depending on plant species. Delayed leaf coloring increased NRE from old leaves for some plant species measured under warming and grazing. Our results suggested that alpine plants may develop strategies to adapt to warming and grazing to assimilate more carbon through prolonged leaf longevity rather than increased NRE through earlier leaf coloring in the alpine meadow.

Enhanced spring temperature sensitivity of carbon emission links to earlier phenology.

Phenology has a great effect on the carbon cycle. Significant relationships have been well demonstrated between phenology and photosynthesis. However, few studies have been undertaken to characterize relationships between phenology and ecosystem respiration (Re). We conducted a reciprocal transplant experiment among three elevations for two-years to measure Re over six phenological sequences throughout the growing seasons. Our results showed that changes in phenological duration were mainly determined by the onset of phenology, as one day advance of phenological onset could lengthen 0.13 days of phenological duration. Advances in early spring phenophases (i.e., first leaf-out, first bud/boot-set and first flowering) under warming strengthened the temperature sensitivity of Re. However, the late phenophases (i.e., first seeding-set, first post-fruiting vegetation and first leaf-coloring) had non-significant relationships with Re. In total, after pooling all the data, one day advance of phenophases would increase Re by 2.23% under warming. In particular, Re would increase by 29.12% with an advance of phenophases by 8.46 days of under a 1.5 °C warming scenario. Our analysis of the coupling between temperature/moisture-phenology-Re may further supplement evidence that warmer spring temperature increases carbon emission by advancing early phenophases. This points to a faster and easier way to investigate how aboveground functional traits (phenology) affect unseen functional traits (Re) on the Tibetan Plateau.

Opposite effects of winter day and night temperature changes on early phenophases.

Changes in day (maximum temperature, TMAX ) and night temperature (minimum temperature, TMIN ) in the preseason (e.g., winter and spring) may have opposite effects on early phenophases (e.g., leafing and flowering) due to changing requirements of chilling accumulations (CAC) and heating accumulations (HAC), which could cause advance, delay or no change in early phenophases. However, their relative effects on phenology are largely unexplored, especially on the Tibetan Plateau. Here, observations were performed using a warming and cooling experiment in situ through reciprocal transplantation (2008-2010) on the Tibetan Plateau. We found that winter minimum temperature (TMIN ) warming significantly delayed mean early phenophases by 8.60 d/°C, but winter maximum temperature (TMAX ) warming advanced them by 12.06 d/°C across six common species. Thus, winter mean temperature warming resulted in a net advance of 3.46 d/°C in early phenophases. In contrast, winter TMIN cooling, on average, significantly advanced early phenophases by 5.12 d/°C, but winter TMAX cooling delayed them by 7.40 d/°C across six common species, resulting in a net delay of 2.28 d/°C for winter mean temperature cooling. The opposing effects of TMAX and TMIN warming on the early phenophases may be mainly caused by decreased CAC due to TMIN warming (5.29 times greater than TMAX ) and increased HAC due to TMAX warming (3.25 times greater than TMIN ), and similar processes apply to TMAX and TMIN cooling. Therefore, our study provides another insight into why some plant phenophases remain unchanged or delayed under climate change.

Divergent Responses of Community Reproductive and Vegetative Phenology to Warming and Cooling: Asymmetry Versus Symmetry.

Few studies have focused on the response of plant community phenology to temperature change using manipulative experiments. A lack of understanding of whether responses of community reproductive and vegetative phenological sequences to warming and cooling are asymmetrical or symmetrical limits our capacity to predict responses under warming and cooling. A reciprocal transplant experiment was conducted for 3 years to evaluate response patterns of the temperature sensitivities of community phenological sequences to warming (transferred downward) and cooling (transferred upward) along four elevations on the Tibetan Plateau. We found that the temperature sensitivities of flowering stages had asymmetric responses to warming and cooling, whereas symmetric responses to warming and cooling were observed for the vegetative phenological sequences. Our findings showed that coverage changes of flowering functional groups (FFGs; i.e., early-spring FFG, mid-summer FFG, and late-autumn FFG) and their compensation effects combined with required accumulated soil temperatureto codetermined the asymmetric and symmetric responses of community phenological sequences to warming and cooling. These results suggest that coverage change in FFGs on warming and cooling processes can be a primary driver of community phenological variation and may lead to inaccurate phenlogical estimation at large scale, such as based on remote sensing.