Interannual fluctuations in precipitation shape the trajectory of ecosystem respiration along a grazing exclusion chronosequence in a typical steppe in Inner Mongolia.

Grazing exclusion (GE), as an effective strategy for revitalizing degraded grasslands, possesses the potential to increase ecosystem respiration (Re) and significantly influence the capacity of grassland soils to sequester carbon. However, our current grasp of Re dynamics in response to varying durations of GE, particularly in the context of precipitation fluctuations, remains incomplete. To fill this knowledge gap, we conducted a monitoring of Re over a 40-year GE chronosequence within Inner Mongolia temperate typical steppe across two distinct hydrologically years. Overall, Re exhibited a gradual saturation curve and an increasing trend with the duration of GE in the wet year of 2021 and the normal precipitation year of 2022, respectively. The variance primarily stemmed from relatively higher microbial biomass carbon observed in the short-term GE during 2022 in contrast to 2021. Moreover, the impacts of GE on the sensitivities of Re to moisture and temperature were intricately tied to precipitation patterns. increasing significantly with prolonged GE duration in 2022 but not in 2021. Our study highlights the intricate interplay between GE duration, precipitation variability, and Re dynamics. This deeper understanding enhances our ability to predict and manage carbon cycling within typical steppe in Inner Mongolia, offering invaluable insights for effective restoration strategies and climate change mitigation.

The coronavirus proofreading exoribonuclease mediates extensive viral recombination.

Recombination is proposed to be critical for coronavirus (CoV) diversity and emergence of SARS-CoV-2 and other zoonotic CoVs. While RNA recombination is required during normal CoV replication, the mechanisms and determinants of CoV recombination are not known. CoVs encode an RNA proofreading exoribonuclease (nsp14-ExoN) that is distinct from the CoV polymerase and is responsible for high-fidelity RNA synthesis, resistance to nucleoside analogues, immune evasion, and virulence. Here, we demonstrate that CoVs, including SARS-CoV-2, MERS-CoV, and the model CoV murine hepatitis virus (MHV), generate extensive and diverse recombination products during replication in culture. We show that the MHV nsp14-ExoN is required for native recombination, and that inactivation of ExoN results in decreased recombination frequency and altered recombination products. These results add yet another critical function to nsp14-ExoN, highlight the uniqueness of the evolved coronavirus replicase, and further emphasize nsp14-ExoN as a central, completely conserved, and vulnerable target for inhibitors and attenuation of SARS-CoV-2 and future emerging zoonotic CoVs.

Different nitrogen saturation thresholds for above-, below-, and total net primary productivity in a temperate steppe.

Identifying the thresholds for the positive responses of total net primary productivity (NPP) to nitrogen (N) enrichment is an essential prerequisite for predicting the benefits of N deposition on ecosystem carbon sequestration. However, the responses of below-ground NPP (BNPP) to N enrichment are unknown in many ecosystems, which limits our ability to understand the carbon cycling under the scenario of increasing N availability. We examined the changes in above-ground NPP (ANPP), BNPP, and NPP of a temperate meadow steppe across a wide-ranging N addition gradient (0, 2, 5, 10, 20, and 50 g N m-2 year-1 ) during 5 years. Both ANPP and NPP increased nonlinearly with N addition rates. The N saturation threshold for ANPP (TA ) and NPP (TN ) was at the rate of 13.11 and 6.70 g N m-2 year-1 , respectively. BNPP decreased with increasing N addition when N addition rates ˃5 g N m-2 year-1 , resulting in much lower TN than TA . Soil N enrichment played a key role in driving the negative impacts of high N addition rates on BNPP, and consequently on the earlier occurrence of N saturation threshold for NPP. Our results highlight the negative effects of soil N enrichment on NPP in natural grasslands super-saturated with N. Furthermore, by considering ANPP and BNPP simultaneously, our results indicate that previous findings from above-ground might have over-estimated the positive effects of N deposition on primary productivity.

Enhanced foliar 15 N enrichment with increasing nitrogen addition rates: Role of plant species and nitrogen compounds.

Determining the abundance of N isotope (δ15 N) in natural environments is a simple but powerful method for providing integrated information on the N cycling dynamics and status in an ecosystem under exogenous N inputs. However, whether the input of different N compounds could differently impact plant growth and their 15 N signatures remains unclear. Here, the response of 15 N signatures and growth of three dominant plants (Leymus chinensis, Carex duriuscula, and Thermopsis lanceolata) to the addition of three N compounds (NH4 HCO3 , urea, and NH4 NO3 ) at multiple N addition rates were assessed in a meadow steppe in Inner Mongolia. The three plants showed different initial foliar δ15 N values because of differences in their N acquisition strategies. Particularly, T. lanceolata (N2 -fixing species) showed significantly lower 15 N signatures than L. chinensis (associated with arbuscular mycorrhizal fungi [AMF]) and C. duriuscula (associated with AMF). Moreover, the foliar δ15 N of all three species increased with increasing N addition rates, with a sharp increase above an N addition rate of ~10 g N m-2  year-1 . Foliar δ15 N values were significantly higher when NH4 HCO3 and urea were added than when NH4 NO3 was added, suggesting that adding weakly acidifying N compounds could result in a more open N cycle. Overall, our results imply that assessing the N transformation processes in the context of increasing global N deposition necessitates the consideration of N deposition rates, forms of the deposited N compounds, and N utilization strategies of the co-existing plant species in the ecosystem.

Assessing community assembly controls over community-scale nutrient resorption responses to nitrogen deposition.

Nutrient resorption is a fundamental physiological process in plants, with important ecological controls over numerous ecosystem functions. However, the role of community assembly in driving responses of nutrient resorption to perturbation remains largely unknown. Following the Price equation framework and the Community Assembly and Ecosystem Function framework, we quantified the contribution of species loss, species gain, and shared species to the reduction of community-level nutrient resorption efficiency in response to multi-level nitrogen (N) addition in a temperate steppe, after continuous N addition for seven years. Reductions of both N and phosphorus (P) resorption efficiency (NRE and PRE, respectively) were positively correlated with N addition levels. The dissimilarities in species composition between N-enriched and control communities increased with N addition levels, and N-enriched plots showed substantial species losses and gains. Interestingly, the reduction of community-scale NRE and PRE mostly resulted from N-induced decreases in resorption efficiency for the shared species in the control and N-enriched communities. There were negative correlations between the contributions of species richness effect and species identity effect and between the number and identity of species gained for the changes in both NRE and PRE following N enrichment. By simultaneously considering N-induced changes in species composition and in species-level resorption, our work presents a more complete picture of how different community assembly processes contribute to N-induced changes in community-level resorption.

Functional structure mediates the responses of productivity to addition of three nitrogen compounds in a meadow steppe.

Atmospheric nitrogen (N) deposition is altering grassland productivity and community structure worldwide. Deposited N comes in different forms, which can have different consequences for productivity due to differences in their fertilization and acidification effects. We hypothesize that these effects may be mediated by changes in plant functional traits. We investigated the responses of aboveground primary productivity and community functional composition to addition of three nitrogen compounds (NH4NO3, [NH4]2SO4, and CO[NH2]2) at the rates of 0, 5, 10, 20 g N m-2 yr-1. We used structural equation modeling (SEM) to evaluate how functional structure influences the responses of productivity to the three N compounds. Nitrogen addition increased community-level leaf chlorophyll content but decreased leaf dry matter content and phosphorus concentration. These changes were mainly due to intra-specific variation. Functional dispersion of traits was reduced by N addition through changes in species composition. SEM revealed that fertilization effects were more important than soil acidification for the responses of productivity to CO(NH2)2 addition, which enhanced productivity by decreasing functional trait dispersion. In contrast, the effects of (NH4)2SO4 and NH4NO3 were primarily due to soil acidification, influencing productivity via community-weighted means of functional traits. Our results suggest that N forms with different fertilizing and acidifying effects influence productivity via different functional traits pathways. Our study also emphasizes the need for in situ experiments with the relevant N compounds to accurately understand and predict the ecological effects of atmospheric N deposition on ecosystems.

Decoupled responses of above- and below-ground stability of productivity to nitrogen addition at the local and larger spatial scale.

Temporal stability of net primary productivity (NPP) is important for predicting the reliable provisioning of ecosystem services under global changes. Although nitrogen (N) addition is known to affect the temporal stability of aboveground net primary productivity (ANPP), it is unclear how it impacts that of belowground net primary productivity (BNPP) and NPP, and whether such effects are scale dependent. Here, using experimental N addition in a grassland, we found different responses of ANPP and BNPP stability to N addition at the local scale and that these responses propagated to the larger spatial scale. That is, N addition significantly decreased the stability of ANPP but did not affect the stability of BNPP and NPP at the two scales investigated. Additionally, spatial asynchrony of both ANPP and BNPP among communities provided greater stability at the larger scale and was not affected by N addition. Our findings challenge the traditional view that N addition would reduce ecosystem stability based on results from aboveground dynamics, thus highlighting the importance of viewing ecosystem stability from a whole system perspective.

Soil aggregate microbiomes steer plant community overyielding in ungrazed and intensively grazed grassland soils.

Plant and soil microbial community composition play a central role in maintaining ecosystem functioning. Most studies have focused on soil microbes in the bulk soil, the rhizosphere and inside plant roots, however, less is known about the soil community that exists within soil aggregates, and how these soil communities influence plant biomass production. Here, using field-conditioned soil collected from experimental ungrazed and grazed grasslands in Inner Mongolia, China, we examined the composition of microbiomes inside soil aggregates of various size classes, and determined their roles in plant-soil feedbacks (PSFs), diversity-productivity relationships, and diversity-dependent overyielding. We found that grazing induced significantly positive PSF effects, which appeared to be mediated by mycorrhizal fungi, particularly under plant monocultures. Despite this, non-additive effects of microbiomes within different soil aggregates enhanced the strength of PSF under ungrazed grassland, but decreased PSF strength under intensively grazed grassland. Plant mixture-related increases in PSF effects markedly enhanced diversity-dependent overyielding, primarily due to complementary effects. Selection effects played far less of a role. Our work suggests that PSF contributes to diversity-dependent overyielding in grasslands via non-additive effects of microbiomes within different soil aggregates. The implication of our work is that assessing the effectiveness of sustainable grassland restoration and management on soil properties requires inspection of soil aggregate size-specific microbiomes, as these are relevant determinants of the feedback interactions between soil and plant performance.

Increasing rates of long-term nitrogen deposition consistently increased litter decomposition in a semi-arid grassland.

The continuing nitrogen (N) deposition observed worldwide alters ecosystem nutrient cycling and ecosystem functioning. Litter decomposition is a key process contributing to these changes, but the numerous mechanisms for altered decomposition remain poorly identified. We assessed these different mechanisms with a decomposition experiment using litter from four abundant species (Achnatherum sibiricum, Agropyron cristatum, Leymus chinensis and Stipa grandis) and litter mixtures representing treatment-specific community composition in a semi-arid grassland under long-term simulation of six different rates of N deposition. Decomposition increased consistently with increasing rates of N addition in all litter types. Higher soil manganese (Mn) availability, which apparently was a consequence of N addition-induced lower soil pH, was the most important factor for faster decomposition. Soil C : N ratios were lower with N addition that subsequently led to markedly higher bacterial to fungal ratios, which also stimulated litter decomposition. Several factors contributed jointly to higher rates of litter decomposition in response to N deposition. Shifts in plant species composition and litter quality played a minor role compared to N-driven reductions in soil pH and C : N, which increased soil Mn availability and altered microbial community structure. The soil-driven effect on decomposition reported here may have long-lasting impacts on nutrient cycling, soil organic matter dynamics and ecosystem functioning.

Carbon limitation overrides acidification in mediating soil microbial activity to nitrogen enrichment in a temperate grassland.

Higher ecosystem nitrogen (N) inputs resulting from human activities often suppress soil microbial biomass and respiration, thereby altering biogeochemical cycling. Soil acidification and carbon (C) limitation may drive these microbial responses, yet their relative importance remains elusive, which limits our understanding of the longer term effects of increasing N inputs. In a field experiment with continuous N addition at seven different rates from 0 to 50 g N m-2  year-1 over 6 years in a temperate grassland of Inner Mongolia, China, we examined the responses of soil microbial biomass and respiration to changes in soil acidity and C availability by adding lime and/or glucose to soil samples. Soil microbial biomass and respiration did only weakly respond to increasing soil pH, but increased strongly in response to higher C availability with increasing N addition rates. Soil net N immobilization increased in response to glucose addition, and soil microbial biomass increased at higher rates than microbial respiration along the gradient of previous N addition rates, both suggesting increasingly reinforced microbial C limitation with increasing N addition. Our results provide clear evidence for strong N-induced microbial C limitation, but only little support for soil acidity effects within the initial pH range of 4.73-7.86 covered by our study. Field data support this conclusion by showing reduced plant C allocation belowground in response to N addition, resulting in soil microbial C starvation over the long term. In conclusion, soil microbial biomass and respiration under N addition were strongly dependent on C availability, most likely originating from plant belowground C inputs, and was much less affected by changes in soil pH. Our data help clarify a long-standing debate about how increasing N input rates affect soil microbial biomass and respiration, and improve the mechanistic understanding of the linkages between ecosystem N enrichment and C cycling.