Preparation of Bispecific IgY-scFvs Inhibition Adherences of Enterotoxigenic Escherichia coli (K88 and F18) to Porcine IPEC-J2 Cell.

Enterotoxigenic Escherichia coli (ETEC) strains are significant contributors to postweaning diarrhea in piglets. Of the ETEC causing diarrhea, K88 and F18 accounted for 92.7%. Despite the prevalence of ETEC K88 and F18, there is currently no effective vaccine available due to the diversity of these strains. This study presents an innovative approach by isolating chicken-derived single-chain variable fragment antibodies (scFvs) specific to K88 and F18 fimbrial antigens from chickens immunized against these ETEC virulence factors. These scFvs effectively inhibited adhesion of K88 and F18 to porcine intestinal epithelial cells (IPEC-J2), with the inhibitory effect demonstrating a dose-dependent increase. Furthermore, a bispecific scFv was designed and expressed in Pichia pastoris. This engineered construct displayed remarkable potency; at a concentration of 25.08 µg, it significantly reduced the adhesion rate of ETEC strains to IPEC-J2 cells by 72.10% and 69.11% when challenged with either K88 or F18 alone. Even in the presence of both antigens, the adhesion rate was notably decreased by 57.92%. By targeting and impeding the initial adhesion step of ETEC pathogenesis, this antibody-based intervention holds promise as a potential alternative to antibiotics, thereby mitigating the risks associated with antibiotic resistance and residual drug contamination in livestock production. Overall, this study lays the groundwork for the development of innovative treatments against ETEC infections in piglets.

Experimental warming altered plant functional traits and their coordination in a permafrost ecosystem.

Knowledge about changes in plant functional traits is valuable for the mechanistic understanding of warming effects on ecosystem functions. However, observations have tended to focus on aboveground plant traits, and there is little information about changes in belowground plant traits or the coordination of above- and belowground traits under climate warming, particularly in permafrost ecosystems. Based on a 7-yr field warming experiment, we measured 26 above- and belowground plant traits of four dominant species, and explored community functional composition and trait networks in response to experimental warming in a permafrost ecosystem on the Tibetan Plateau. Experimental warming shifted community-level functional traits toward more acquisitive values, with earlier green-up, greater plant height, larger leaves, higher photosynthetic resource-use efficiency, thinner roots, and greater specific root length and root nutrient concentrations. However, warming had a negligible effect in terms of functional diversity. In addition, warming shifted hub traits which have the highest centrality in the network from specific root area to leaf area. These results demonstrate that above- and belowground traits exhibit consistent adaptive strategies, with more acquisitive traits in warmer environments. Such changes could provide an adaptive advantage for plants in response to environmental change.

Microbial nitrogen and phosphorus co-limitation across permafrost region.

The status of plant and microbial nutrient limitation have profound impacts on ecosystem carbon cycle in permafrost areas, which store large amounts of carbon and experience pronounced climatic warming. Despite the long-term standing paradigm assumes that cold ecosystems primarily have nitrogen deficiency, large-scale empirical tests of microbial nutrient limitation are lacking. Here we assessed the potential microbial nutrient limitation across the Tibetan alpine permafrost region, using the combination of enzymatic and elemental stoichiometry, genes abundance and fertilization method. In contrast with the traditional view, the four independent approaches congruently detected widespread microbial nitrogen and phosphorus co-limitation in both the surface soil and deep permafrost deposits, with stronger limitation in the topsoil. Further analysis revealed that soil resources stoichiometry and microbial community composition were the two best predictors of the magnitude of microbial nutrient limitation. High ratio of available soil carbon to nutrient and low fungal/bacterial ratio corresponded to strong microbial nutrient limitation. These findings suggest that warming-induced enhancement in soil nutrient availability could stimulate microbial activity, and probably amplify soil carbon losses from permafrost areas.

Spatially explicit estimate of nitrogen effects on soil respiration across the globe.

Soil respiration (Rs), as the second largest flux of carbon dioxide (CO2 ) between terrestrial ecosystems and the atmosphere, is vulnerable to global nitrogen (N) enrichment. However, the global distribution of the N effects on Rs remains uncertain. Here, we compiled a new database containing 1282 observations of Rs and its heterotrophic component (Rh) in field N manipulative experiments from 317 published papers. Using this up-to-date database, we first performed a formal meta-analysis to explore the responses of Rs and Rh to N addition, and then presented a global spatially explicit quantification of the N effects using a Random Forest model. Our results showed that experimental N addition significantly increased Rs but had a minimal impact on Rh, not supporting the prevailing view that N enrichment inhibits soil microbial respiration. For the major biomes, the magnitude of N input was the main determinant of the spatial variation in Rs response, while the most important predictors for Rh response were biome specific. Based on the key predictors, global mapping visually demonstrated a positive N effect in the regions with higher anthropogenic N inputs (i.e., atmospheric N deposition and agricultural fertilization). Overall, our analysis not only provides novel insight into the N effects on soil CO2 fluxes, but also presents a spatially explicit assessment of the N effects at the global scale, which are pivotal for understanding ecosystem carbon dynamics in future scenarios with more frequent anthropogenic activities.

Priming effect stimulates carbon release from thawed permafrost.

Climate warming leads to widespread permafrost thaw with a fraction of the thawed permafrost carbon (C) being released as carbon dioxide (CO2 ), thus triggering a positive permafrost C-climate feedback. However, large uncertainty exists in the size of this model-projected feedback, partly owing to the limited understanding of permafrost CO2 release through the priming effect (i.e., the stimulation of soil organic matter decomposition by external C inputs) upon thaw. By combining permafrost sampling from 24 sites on the Tibetan Plateau and laboratory incubation, we detected an overall positive priming effect (an increase in soil C decomposition by up to 31%) upon permafrost thaw, which increased with permafrost C density (C storage per area). We then assessed the magnitude of thawed permafrost C under future climate scenarios by coupling increases in active layer thickness over half a century with spatial and vertical distributions of soil C density. The thawed C stocks in the top 3 m of soils from the present (2000-2015) to the future period (2061-2080) were estimated at 1.0 (95% confidence interval (CI): 0.8-1.2) and 1.3 (95% CI: 1.0-1.7) Pg (1 Pg = 1015 g) C under moderate and high Representative Concentration Pathway (RCP) scenarios 4.5 and 8.5, respectively. We further predicted permafrost priming effect potential (priming intensity under optimal conditions) based on the thawed C and the empirical relationship between the priming effect and permafrost C density. By the period 2061-2080, the regional priming potentials could be 8.8 (95% CI: 7.4-10.2) and 10.0 (95% CI: 8.3-11.6) Tg (1 Tg = 1012 g) C year-1 under the RCP 4.5 and RCP 8.5 scenarios, respectively. This large CO2 emission potential induced by the priming effect highlights the complex permafrost C dynamics upon thaw, potentially reinforcing permafrost C-climate feedback.

Accelerated organic matter decomposition in thermokarst lakes upon carbon and phosphorus inputs.

Mineralization of dissolved organic matter (DOM) in thermokarst lakes plays a non-negligible role in the permafrost carbon (C) cycle, but remains poorly understood due to its complex interactions with external C and nutrient inputs (i.e., aquatic priming and nutrient effects). Based on large-scale lake sampling and laboratory incubations, in combination with 13 C-stable-isotope labeling, optical spectroscopy, and high-throughput sequencing, we examined large-scale patterns and dominant drivers of priming and nutrient effects of DOM biodegradation across 30 thermokarst lakes along a 1100-km transect on the Tibetan Plateau. We observed that labile C and phosphorus (P) rather than nitrogen (N) inputs stimulated DOM biodegradation, with the priming and P effects being 172% and 451% over unamended control, respectively. We also detected significant interactive effects of labile C and nutrient supply on DOM biodegradation, with the combined labile C and nutrient additions inducing stronger microbial mineralization than C or nutrient treatment alone, illustrating that microbial activity in alpine thermokarst lakes is co-limited by both C and nutrients. We further found that the aquatic priming was mainly driven by DOM quality, with the priming intensity increasing with DOM recalcitrance, reflecting the limitation of external C as energy sources for microbial activity. Greater priming intensity was also associated with higher community-level ribosomal RNA gene operon (rrn) copy number and bacterial diversity as well as increased background soluble reactive P concentration. In contrast, the P effect decreased with DOM recalcitrance as well as with background soluble reactive P and ammonium concentrations, revealing the declining importance of P availability in mediating DOM biodegradation with enhanced C limitation but reduced nutrient limitation. Overall, the stimulation of external C and P inputs on DOM biodegradation in thermokarst lakes would amplify C-climate feedback in this alpine permafrost region.

Patterns and drivers of anaerobic nitrogen transformations in sediments of thermokarst lakes.

Significant attention has been given to the way in which the soil nitrogen (N) cycle responds to permafrost thaw in recent years, yet little is known about anaerobic N transformations in thermokarst lakes, which account for more than one-third of thermokarst landforms across permafrost regions. Based on the N isotope dilution and tracing technique, combined with qPCR and high-throughput sequencing, we presented large-scale measurements of anaerobic N transformations of sediments across 30 thermokarst lakes over the Tibetan alpine permafrost region. Our results showed that gross N mineralization, ammonium immobilization, and dissimilatory nitrate reduction rates in thermokarst lakes were higher in the eastern part of our study area than in the west. Denitrification dominated in the dissimilatory nitrate reduction processes, being two and one orders of magnitude higher than anaerobic ammonium oxidation (anammox) and dissimilatory nitrate reduction to ammonium (DNRA), respectively. The abundances of the dissimilatory nitrate reduction genes (nirK, nirS, hzsB, and nrfA) exhibited patterns consistent with sediment N transformation rates, while α diversity did not. The inter-lake variability in gross N mineralization and ammonium immobilization was dominantly driven by microbial biomass, while the variability in anammox and DNRA was driven by substrate supply and organic carbon content, respectively. Denitrification was jointly affected by nirS abundance and organic carbon content. Overall, the patterns and drivers of anaerobic N transformation rates detected in this study provide a new perspective on potential N release, retention, and removal upon the formation and development of thermokarst lakes.

Stochastic processes regulate belowground community assembly in alpine grasslands on the Tibetan Plateau.

Understanding biogeographical patterns and underlying processes of belowground community assembly is crucial for predicting soil functions and their responses to global environmental change. However, little is known about potential differences of belowground community assembly among bacteria, fungi, protists and soil animals, particularly for alpine ecosystems. Based on the combination of large-scale field sampling, high-throughput marker-gene sequencing and multiple statistical analyses, we explored patterns and drivers of belowground community assembly in alpine grasslands on the Tibetan Plateau. Our results revealed that the distance-decay rates varied among trophic levels, with organisms of higher trophic level having weaker distance-decay pattern. The spatial and environmental variables explained limited variations of belowground communities. By contrast, the stochastic processes, mainly consisting of dispersal limitation and drift, played a primary role in regulating belowground community assembly. Moreover, the relative importance of stochastic processes varied among trophic levels, with the role of dispersal limitation weakening whereas that of drift enhancing in the order of bacteria, fungi, protists and soil animals. These findings advance our understanding of patterns and mechanisms driving belowground community assembly in alpine ecosystems and provide a reference basis for predicting the dynamics of ecosystem functions under changing environment.

Substantial non-growing season carbon dioxide loss across Tibetan alpine permafrost region.

One of the major uncertainties for projecting permafrost carbon (C)-climate feedback is a poor representation of the non-growing season carbon dioxide (CO2 ) emissions under a changing climate. Here, combining in situ field observations, regional synthesis and a random forest model, we assessed contemporary and future soil respired CO2 (i.e., soil respiration, Rs ) across the Tibetan alpine permafrost region, which has received much less attention compared with the Arctic permafrost domain. We estimated the regional mean Rs of 229.8, 72.9 and 302.7 g C m-2  year-1 during growing season, non-growing season and the entire year, respectively; corresponding to the contemporary losses of 296.9, 94.3 and 391.2 Tg C year-1 from this high-altitude permafrost-affected area. The non-growing season Rs accounted for a quarter of the annual soil CO2 efflux. Different from the prevailing view that temperature is the most limiting factor for cold-period CO2 release in Arctic permafrost ecosystems, precipitation determined the spatial pattern of non-growing season Rs on the Tibetan Plateau. Using the key predictors, model extrapolation demonstrated additional losses of 38.8 and 74.5 Tg C from the non-growing season for a moderate mitigation scenario and a business-as-usual emissions scenario, respectively. These results provide a baseline for non-growing season CO2 emissions from high-altitude permafrost areas and help for accurate projection of permafrost C-climate feedback.

Nitrogen input enhances microbial carbon use efficiency by altering plant-microbe-mineral interactions.

Microbial growth and respiration are at the core of the soil carbon (C) cycle, as these microbial physiological performances ultimately determine the fate of soil C. Microbial C use efficiency (CUE), a critical metric to characterize the partitioning of C between microbial growth and respiration, thus controls the sign and magnitude of soil C-climate feedback. Despite its importance, the response of CUE to nitrogen (N) input and the relevant regulatory mechanisms remain poorly understood, leading to large uncertainties in predicting soil C dynamics under continuous N input. By combining a multi-level field N addition experiment with a substrate-independent 18 O-H2 O labelling approach as well as high-throughput sequencing and mineral analysis, here we elucidated how N-induced changes in plant-microbial-mineral interactions drove the responses of microbial CUE to N input. We found that microbial CUE increased significantly as a consequence of enhanced microbial growth after 6-year N addition. In contrast to the prevailing view, the elevated microbial growth and CUE were not mainly driven by the reduced stoichiometric imbalance, but strongly associated with the increased soil C accessibility from weakened mineral protection. Such attenuated organo-mineral association was further linked to the N-induced changes in the plant community and the increased oxalic acid in the soil. These findings provide empirical evidence for the tight linkage between mineral-associated C dynamics and microbial physiology, highlighting the need to disentangle the complex plant-microbe-mineral interactions to improve soil C prediction under anthropogenic N input.

Depth-dependent drivers of soil microbial necromass carbon across Tibetan alpine grasslands.

Microbial necromass carbon (C) has been considered an important contributor to persistent soil C pool. However, there still lacks large-scale systematic observations on microbial necromass C in different soil layers, particularly for alpine ecosystems. Besides, it is still unclear whether the relative importance of biotic and abiotic variables such as plant C input and mineral properties in regulating microbial necromass C would change with soil depth. Based on the combination of large-scale sampling along a ~2200 km transect across Tibetan alpine grasslands and biomarker analysis, together with a global data synthesis across grassland ecosystems, we observed a relatively low proportion of microbial-derived C in Tibetan alpine grasslands compared to global grasslands (topsoil: 45.4% vs. 58.1%; subsoil: 41.7% vs. 53.7%). We also found that major determinants of microbial necromass C depended on soil depth. In topsoil, both plant C input and mineral protection exerted dominant effects on microbial necromass C. However, in subsoil, the physico-chemical protection provided by soil clay particles, iron-aluminum oxides, and exchangeable calcium dominantly facilitated the preservation of microbial necromass C. The differential drivers over microbial necromass C between soil depths should be considered in Earth system models for accurately forecasting soil C dynamics and its potential feedback to global warming.

Storage, patterns and influencing factors for soil organic carbon in coastal wetlands of China.

Soil organic carbon (SOC) in coastal wetlands, also known as "blue C," is an essential component of the global C cycles. To gain a detailed insight into blue C storage and controlling factors, we studied 142 sites across ca. 5000 km of coastal wetlands, covering temperate, subtropical, and tropical climates in China. The wetlands represented six vegetation types (Phragmites australis, mixed of P. australis and Suaeda, single Suaeda, Spartina alterniflora, mangrove [Kandelia obovata and Avicennia marina], tidal flat) and three vegetation types invaded by S. alterniflora (P. australis, K. obovata, A. marina). Our results revealed large spatial heterogeneity in SOC density of the top 1-m ranging 40-200 Mg C ha-1 , with higher values in mid-latitude regions (25-30° N) compared with those in both low- (20°N) and high-latitude (38-40°N) regions. Vegetation type influenced SOC density, with P. australis and S. alterniflora having the largest SOC density, followed by mangrove, mixed P. australis and Suaeda, single Suaeda and tidal flat. SOC density increased by 6.25 Mg ha-1 following S. alterniflora invasion into P. australis community but decreased by 28.56 and 8.17 Mg ha-1 following invasion into K. obovata and A. marina communities. Based on field measurements and published literature, we calculated a total inventory of 57 × 106 Mg C in the top 1-m soil across China's coastal wetlands. Edaphic variables controlled SOC content, with soil chemical properties explaining the largest variance in SOC content. Climate did not control SOC content but had a strong interactive effect with edaphic variables. Plant biomass and quality traits were a minor contributor in regulating SOC content, highlighting the importance of quantity and quality of OC inputs and the balance between production and degradation within the coastal wetlands. These findings provide new insights into blue C stabilization mechanisms and sequestration capacity in coastal wetlands.

An integrated belowground trait-based understanding of nitrogen-driven plant diversity loss.

Belowground plant traits play important roles in plant diversity loss driven by atmospheric nitrogen (N) deposition. However, the way N enrichment shapes plant microhabitats by patterning belowground traits and finally determines aboveground responses is poorly understood. Here, we investigated the rhizosheath trait of 74 plant species in seven N-addition simulation experiments across multiple grassland ecosystems in China. We found that rhizosheath formation differed among plant functional groups and contributed to changes in plant community composition induced by N enrichment. Compared with forb species, grass and sedge species exhibited distinct rhizosheaths; moreover, grasses and sedges expanded their rhizosheaths with increasing N-addition rate which allowed them to colonize belowground habitats. Grasses also shaped a different microenvironment around their roots compared with forbs by affecting the physicochemical, biological, and stress-avoiding properties of their rhizosphere soil. Rhizosheaths act as a "biofilm-like shield" by the accumulation of protective compounds, carboxylic anions and polysaccharides, determined by both plants and microorganisms. This enhanced the tolerance of grasses and sedges to stresses induced by N enrichment. Conversely, forbs lacked the protective rhizosheaths which renders their roots sensitive to stresses induced by N enrichment, thus contributing to their disappearance under N-enriched conditions. This study uncovers the processes by which belowground facilitation and trait matching affect aboveground responses under conditions of N enrichment, which advances our mechanistic understanding of the contribution of competitive exclusion and environmental tolerance to plant diversity loss caused by N deposition.

Microbial traits determine soil C emission in response to fresh carbon inputs in forests across biomes.

Soil priming is a microbial-driven process, which determines key soil-climate feedbacks in response to fresh carbon inputs. Despite its importance, the microbial traits behind this process are largely undetermined. Knowledge of the role of these traits is integral to advance our understanding of how soil microbes regulate carbon (C) emissions in forests, which support the largest soil carbon stocks globally. Using metagenomic sequencing and 13 C-glucose, we provide unprecedented evidence that microbial traits explain a unique portion of the variation in soil priming across forest biomes from tropical to cold temperature regions. We show that microbial functional profiles associated with the degradation of labile C, especially rapid simple sugar metabolism, drive soil priming in different forests. Genes involved in the degradation of lignin and aromatic compounds were negatively associated with priming effects in temperate forests, whereas the highest level of soil priming was associated with ß-glucosidase genes in tropical/subtropical forests. Moreover, we reconstructed, for the first time, 42 whole bacterial genomes associated with the soil priming effect and found that these organisms support important gene machinery involved in priming effect. Collectively, our work demonstrates the importance of microbial traits to explain soil priming across forest biomes and suggests that rapid carbon metabolism is responsible for priming effects in forests. This knowledge is important because it advances our understanding on the microbial mechanisms mediating soil-climate feedbacks at a continental scale.

High potential of stable carbon sequestration in phytoliths of China's grasslands.

Phytolith carbon (C) sequestration plays a key role in mitigating global climate change at a centennial to millennial time scale. However, previous estimates of phytolith-occluded carbon (PhytOC) storage and potential in China's grasslands have large uncertainties mainly due to multiple data sources. This contributes to the uncertainty in predicting long-term C sequestration in terrestrial ecosystems using Earth System Models. In this study, we carried out an intensive field investigation (79 sites, 237 soil profiles [0-100 cm], and 61 vegetation assessments) to quantify PhytOC storage in China's grasslands and to better explore the biogeographical patterns and influencing factors. Generally, PhytOC production flux and soil PhytOC density in both the Tibetan Plateau and the Inner Mongolian Plateau had a decreasing trend from the Northeast to the Southwest. The aboveground PhytOC production rate in China's grassland was 0.48 × 106 t CO2 a-1 , and the soil PhytOC storage was 383 × 106 t CO2 . About 45% of soil PhytOC was stored in the deep soil layers (50-100 cm), highlighting the importance of deep soil layers for C stock assessments. Importantly, the Tibetan Plateau had the greatest contribution (more than 70%) to the PhytOC storage in China's grasslands. The results of multiple regression analysis indicated that altitude and soil texture significantly influenced the spatial distribution of soil PhytOC, explaining 78.1% of the total variation. Soil phytolith turnover time in China's grasslands was mainly controlled by climatic conditions, with the turnover time on the Tibetan Plateau being significantly longer than that on the Inner Mongolian Plateau. Our results offer more accurate estimates of the potential for phytolith C sequestration from ecological restoration projects in degraded grassland ecosystems. These estimates are essential to parameterizing and validating global C models.

Divergent Trajectory of Soil Autotrophic and Heterotrophic Respiration upon Permafrost Thaw.

Warming-induced permafrost thaw may stimulate soil respiration (Rs) and thus cause a positive feedback to climate warming. However, due to the limited in situ observations, it remains unclear about how Rs and its autotrophic (Ra) and heterotrophic (Rh) components change upon permafrost thaw. Here we monitored variations in Rs and its components along a permafrost thaw sequence on the Tibetan Plateau, and explored the potential linkage of Rs components (i.e., Ra and Rh) with biotic (e.g., plant functional traits and soil microbial diversity) and abiotic factors (e.g., substrate quality). We found that Ra and Rh exhibited divergent responses to permafrost collapse: Ra increased with the time of thawing, while Rh exhibited a hump-shaped pattern along the thaw sequence. We also observed different drivers of thaw-induced changes in the ratios of Ra:Rs and Rh:Rs. Except for soil water status, plant community structure, diversity, and root properties explained the variation in Ra:Rs ratio, soil substrate quality and microbial diversity were key factors associated with the dynamics of Rh:Rs ratio. Overall, these findings demonstrate divergent patterns and drivers of Rs components as permafrost thaw prolongs, which call for considerations in Earth system models for better forecasting permafrost carbon-climate feedback.

Response to Comment on 'Soil carbon persistence governed by plant input and mineral protection at regional and global scales'.

We demonstrated that ignoring the non-linear relationship between topsoil Δ14 C and plant carbon (C) input in Wu et al.'s analysis was the fundamental reason for the discrepancy between their analysis and ours. By considering such a non-linear relationship, plant C input still predominantly governs the topsoil C turnover.

Soil carbon persistence governed by plant input and mineral protection at regional and global scales.

Elucidating the processes underlying the persistence of soil organic matter (SOM) is a prerequisite for projecting soil carbon feedback to climate change. However, the potential role of plant carbon input in regulating the multi-layer SOM preservation over broad geographic scales remains unclear. Based on large-scale soil radiocarbon (∆14 C) measurements on the Tibetan Plateau, we found that plant carbon input was the major contributor to topsoil carbon destabilisation despite the significant associations of topsoil ∆14 C with climatic and mineral variables as well as SOM chemical composition. By contrast, mineral protection by iron-aluminium oxides and cations became more important in preserving SOM in deep soils. These regional observations were confirmed by a global synthesis derived from the International Soil Radiocarbon Database (ISRaD). Our findings illustrate different effects of plant carbon input on SOM persistence across soil layers, providing new insights for models to better predict multi-layer soil carbon dynamics under changing environments.

Large-scale evidence for microbial response and associated carbon release after permafrost thaw.

Permafrost thaw could trigger the release of greenhouse gases through microbial decomposition of the large quantities of carbon (C) stored within frozen soils. However, accurate evaluation of soil C emissions from thawing permafrost is still a big challenge, partly due to our inadequate understanding about the response of microbial communities and their linkage with soil C release upon permafrost thaw. Based on a large-scale permafrost sampling across 24 sites on the Tibetan Plateau, we employed meta-genomic technologies (GeoChip and Illumina MiSeq sequencing) to explore the impacts of permafrost thaw (permafrost samples were incubated for 11 days at 5°C) on microbial taxonomic and functional communities, and then conducted a laboratory incubation to investigate the linkage of microbial taxonomic and functional diversity with soil C release after permafrost thaw. We found that bacterial and fungal α diversity decreased, but functional gene diversity and the normalized relative abundance of C degradation genes increased after permafrost thaw, reflecting the rapid microbial response to permafrost thaw. Moreover, both the microbial taxonomic and functional community structures differed between the thawed permafrost and formerly frozen soils. Furthermore, soil C release rate over five month incubation was associated with microbial functional diversity and C degradation gene abundances. By contrast, neither microbial taxonomic diversity nor community structure exhibited any significant effects on soil C release over the incubation period. These findings demonstrate that permafrost thaw could accelerate C emissions by altering the function potentials of microbial communities rather than taxonomic diversity, highlighting the crucial role of microbial functional genes in mediating the responses of permafrost C cycle to climate warming.

Altered microbial structure and function after thermokarst formation.

Permafrost thaw could induce substantial carbon (C) emissions to the atmosphere, and thus trigger a positive feedback to climate warming. As the engine of biogeochemical cycling, soil microorganisms exert a critical role in mediating the direction and strength of permafrost C-climate feedback. However, our understanding about the impacts of thermokarst (abrupt permafrost thaw) on microbial structure and function remains limited. Here we employed metagenomic sequencing to analyze changes in topsoil (0-15 cm) microbial communities and functional genes along a permafrost thaw sequence (1, 10, and 16 years since permafrost collapse) on the Tibetan Plateau. By combining laboratory incubation and a two-pool model, we then explored changes in soil labile and stable C decomposition along the thaw sequence. Our results showed that topsoil microbial α-diversity decreased, while the community structure and functional gene abundance did not exhibit any significant change at the early stage of collapse (1 year since collapse) relative to non-collapsed control. However, as the time since the collapse increased, both the topsoil microbial community structure and functional genes differed from the control. Abundances of functional genes involved in labile C degradation decreased while those for stable C degradation increased at the late stage of collapse (16 years since collapse), largely driven by changes in substrate properties along the thaw sequence. Accordingly, faster stable C decomposition occurred at the late stage of collapse compared to the control, which was associated with the increase in relative abundance of functional genes for stable C degradation. These results suggest that upland thermokarst alters microbial structure and function, particularly enhances soil stable C decomposition by modulating microbial functional genes, which could reinforce a warmer climate over the decadal timescale.