Meta-analysis identifying gut microbial biomarkers of Qinghai-Tibet Plateau populations and the functionality of microbiota-derived butyrate in high-altitude adaptation.

The extreme environmental conditions of a plateau seriously threaten human health. The relationship between gut microbiota and human health at high altitudes has been extensively investigated. However, no universal gut microbiota biomarkers have been identified in the plateau population, limiting research into gut microbiota and high-altitude adaptation. 668 16s rRNA samples were analyzed using meta-analysis to reduce batch effects and uncover microbiota biomarkers in the plateau population. Furthermore, the robustness of these biomarkers was validated. Mendelian randomization (MR) results indicated that Tibetan gut microbiota may mediate a reduced erythropoietic response. Functional analysis and qPCR revealed that butyrate may be a functional metabolite in high-altitude adaptation. A high-altitude rat model showed that butyrate reduced intestinal damage caused by high altitudes. According to cell experiments, butyrate may downregulate hypoxia-inducible factor-1α (HIF-1α) expression and blunt cellular responses to hypoxic stress. Our research found universally applicable biomarkers and investigated their potential roles in promoting human health at high altitudes.

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.

Genome diversity and highland-adaptative variation in Tibet barley landrace population of China.

Barley landraces accumulated variation in adapting to extreme highland environments during long-term domestication in Tibet, but little is known about their population structure and genomic selection traces. In this study, tGBS (tunable genotyping by sequencing) sequencing, molecular marker and phenotypic analyses were conducted on 1,308 highland and 58 inland barley landraces in China. The accessions were divided into six sub-populations and clearly distinguished most six-rowed, naked barley accessions (Qingke in Tibet) from inland barley. Genome-wide differentiation was observed in all five sub-populations of Qingke and inland barley accessions. High genetic differentiation in the pericentric regions of chromosomes 2H and 3H contributed to formation of five types of Qingke. Ten haplotypes of the pericentric regions of 2H, 3H, 6H and 7H were further identified as associated with ecological diversification of these sub-populations. There was genetic exchange between eastern and western Qingke but they shared the same progenitor. The identification of 20 inland barley types indicated multiple origins of Qingke in Tibet. The distribution of the five types of Qingke corresponded to specific environments. Two predominant highland-adaptative variations were identified for low temperature tolerance and grain color. Our results provide new insights into the origin, genome differentiation, population structure and highland adaptation in highland barley which will benefit both germplasm enhancement and breeding of naked barley.

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.

Observation-based global soil heterotrophic respiration indicates underestimated turnover and sequestration of soil carbon by terrestrial ecosystem models.

Soil heterotrophic respiration (Rh ) refers to the flux of CO2 released from soil to atmosphere as a result of organic matter decomposition by soil microbes and fauna. As one of the major fluxes in the global carbon cycle, large uncertainties still exist in the estimation of global Rh , which further limits our current understanding of carbon accumulation in soils. Here, we applied a Random Forest algorithm to create a global data set of soil Rh , by linking 761 field observations with both abiotic and biotic predictors. We estimated that global Rh was 48.8 ± 0.9 Pg C year-1 for 1982-2018, which was 16% less than the ensemble mean (58.6 ± 9.9 Pg C year-1 ) of 16 terrestrial ecosystem models. By integrating our observational Rh with independent soil carbon stock data sets, we obtained a global mean soil carbon turnover time of 38.3 ± 11 year. Using observation-based turnover times as a constraint, we found that terrestrial ecosystem models simulated faster carbon turnovers, leading to a 30% (74 Pg C) underestimation of terrestrial ecosystem carbon accumulation for the past century, which was especially pronounced at high latitudes. This underestimation is equivalent to 45% of the total carbon emissions (164 Pg C) caused by global land-use change at the same time. Our analyses highlight the need to constrain ecosystem models using observation-based and locally adapted Rh values to obtain reliable projections of the carbon sink capacity of terrestrial ecosystems.

The glacial-terrestrial-fluvial pathway: A multiparametrical analysis of spatiotemporal dissolved organic matter variation in three catchments of Lake Nam Co, Tibetan Plateau.

The Tibetan Plateau (TP) is a sensitive alpine environment of global importance, being Asia's water tower, featuring vast ice masses and comprising the world's largest alpine grasslands. Intensified land-use and pronounced global climate change have put pressure on the environment of the TP. We studied the tempo-spatial variability of dissolved organic matter (DOM) to better understand the fluxes of nutrients and energy from terrestrial to aquatic ecosystems in the TP. We used a multiparametrical approach, based on inorganic water chemistry, dissolved organic carbon (DOC) concentration, dissolved organic matter (DOM) characteristics (chromophoric DOM, fluorescence DOM and δ13C of DOM) in stream samples of three catchments of the Nam Co watershed and the lake itself. Satellite based plant cover estimates were used to link biogeochemical data to the structure and degradation of vegetation zones in the catchments. Catchment streams showed site-specific DOM signatures inherited from glaciers, wetlands, groundwater, and Kobresia pygmaea pastures. By comparing stream and lake samples, we found DOM processing and unification by loss of chromophoric DOM signatures and a change towards an autochthonous source of lake DOM. DOM diversity was largest in the headwaters of the catchments and heavily modified in terminal aquatic systems. Seasonality was characterized by a minor influence of freshet and by a very strong impact of the Indian summer monsoon on DOM composition, with more microbial DOM sources. The DOM of Lake Nam Co differed chemically from stream water samples, indicating the lake to be a quasi-marine environment in regards to the degree of chemical modification and sources of DOM. DOM proved to be a powerful marker to elucidate consequences of land use and climatic change on biogeochemical processes in High Asian alpine ecosystems.

Microbial functional changes mark irreversible course of Tibetan grassland degradation.

The Tibetan Plateau's Kobresia pastures store 2.5% of the world's soil organic carbon (SOC). Climate change and overgrazing render their topsoils vulnerable to degradation, with SOC stocks declining by 42% and nitrogen (N) by 33% at severely degraded sites. We resolved these losses into erosion accounting for two-thirds, and decreased carbon (C) input and increased SOC mineralization accounting for the other third, and confirmed these results by comparison with a meta-analysis of 594 observations. The microbial community responded to the degradation through altered taxonomic composition and enzymatic activities. Hydrolytic enzyme activities were reduced, while degradation of the remaining recalcitrant soil organic matter by oxidative enzymes was accelerated, demonstrating a severe shift in microbial functioning. This may irreversibly alter the world´s largest alpine pastoral ecosystem by diminishing its C sink function and nutrient cycling dynamics, negatively impacting local food security, regional water quality and climate.

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.

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.

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.

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.

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.

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.

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.

Warming and land use change concurrently erode ecosystem services in Tibet.

Alpine meadows on the Tibetan Plateau comprise the largest alpine ecosystem in the world and provide critical ecosystem services, including forage production and carbon sequestration, on which people depend from local to global scales. However, the provision of these services may be threatened by climate warming combined with land use policies that are altering if and how pastoralists can continue to graze livestock, the dominant livelihood practice in this region for millennia. We synthesized findings from a climate warming and yak grazing experiment with landscape-level observations in central Tibet to gain insight into the trajectories of change that Tibet's alpine meadows will undergo in response to expected changes in climate and land use. We show that within 5 years, experimental warming drove an alpine community with intact, sedge-dominated turfs into a degraded state. With removal of livestock, consistent with policy intended to reverse degradation, a longer-term shift to a more shrub-dominated community will likely occur. Neither degraded nor shrub meadows produce forage or sequester carbon to the same degree as intact meadows, indicating that climate warming and drying will reduce the ability of Tibet's alpine meadows to provide key ecosystem services, and that livestock reduction policies intended to counteract trajectories of land degradation instead endanger contemporary livelihoods on the Tibetan Plateau.

Responses of sequential and hierarchical phenological events to warming and cooling in alpine meadows.

Organisms' life cycles consist of hierarchical stages, from a single phenological stage (for example, flowering within a season), to vegetative and reproductive phases, to the total lifespan of the individual. Yet phenological events are typically studied in isolation, limiting our understanding of life history responses to climate change. Here, we reciprocally transfer plant communities along an elevation gradient to investigate plastic changes in the duration of sequential phenological events for six alpine species. We show that prolonged flowering leads to longer reproductive phases and activity periods when plants are moved to warmer locations. In contrast, shorter post-fruiting leaf and flowering stages led to shorter vegetative and reproductive phases, respectively, which resulted in shorter activity periods when plants were moved to cooler conditions. Therefore, phenological responses to warming and cooling do not simply mirror one another in the opposite direction, and low temperature may limit reproductive allocation in the alpine region.

Plant functional traits mediate reproductive phenology and success in response to experimental warming and snow addition in Tibet.

Global climate change is predicted to have large impacts on the phenology and reproduction of alpine plants, which will have important implications for plant demography and community interactions, trophic dynamics, ecosystem energy balance, and human livelihoods. In this article we report results of a 3-year, fully factorial experimental study exploring how warming, snow addition, and their combination affect reproductive phenology, effort, and success of four alpine plant species belonging to three different life forms in a semiarid, alpine meadow ecosystem on the central Tibetan Plateau. Our results indicate that warming and snow addition change reproductive phenology and success, but responses are not uniform across species. Moreover, traits associated with resource acquisition, such as rooting depth and life history (early vs. late flowering), mediate plant phenology, and reproductive responses to changing climatic conditions. Specifically, we found that warming delayed the reproductive phenology and decreased number of inflorescences of Kobresia pygmaea C. B. Clarke, a shallow-rooted, early-flowering plant, which may be mainly constrained by upper-soil moisture availability. Because K. pygmaea is the dominant species in the alpine meadow ecosystem, these results may have important implications for ecosystem dynamics and for pastoralists and wildlife in the region.