Manipulative Field Experiment Research Articles

Traits estimated when grown alone may underestimate the competitive advantage and invasiveness of exotic species.

Functional differences between native and exotic species, estimated when species are grown alone or in mixtures, are often used to predict the invasion risk of exotic species. However, it remains elusive whether the functional differences estimated by the two methods and their ability to predict species invasiveness (e.g. high abundance) are consistent. We compiled data from two common garden experiments, in which specific leaf area, height, and aboveground biomass of 64 common native and exotic invasive species in China were estimated when grown individually (pot) or in mixtures (field). Exotic species accumulated higher aboveground biomass than natives, but only when grown in field mixtures. Moreover, aboveground biomass and functional distinctiveness estimated in mixtures were more predictive of species persistence and relative abundance in the field mixtures in the second year than those estimated when grown alone. These findings suggest that assessing species traits while grown alone may underestimate the competitive advantage for some exotic species, highlighting the importance of trait-by-environment interactions in shaping species invasion. Therefore, we propose that integrating multi-site or multi-year field surveys and manipulative experiments is required to best identify the key trait(s) and environment(s) that interactively shape species invasion and community dynamics.

Precipitation changes alter plant dominant species and functional groups by changing soil salinity in a coastal salt marsh

Specific mechanisms of precipitation change due to global climate variability on plant communities in coastal salt marsh ecosystems remain unknown. Hence, a field manipulative precipitation experiment was established in 2014 and 5 years of field surveys of vegetation from 2017 to 2021 to explore the effects of precipitation changes on plant community composition. The results showed that changes in plant community composition were driven by dominant species, and that the dominance of key species changed significantly with precipitation gradient and time, and that these changes ultimately altered plant community traits (i.e., community density, height, and species richness). Community height increased but community density decreased with more precipitation averaged five years. Furthermore, changes in precipitation altered dominant species composition and functional groups mainly by influencing soil salinity. Salinity stress caused by decreased precipitation shifted species composition from a dominance of taller perennials and grasses to dwarf annuals and forbs, while the species richness decreased. Conversely, soil desalination caused by increased precipitation increased species richness, especially increasing in the dominance of grasses and perennials. Specifically, Apocynaceae became dominance from rare while Amaranthaceae decreased in response to increased precipitation, but Poaceae was always in a position of dominance. Meanwhile, the dominance of grasses and perennials has the cumulative effect of years and their proportion increased under the increased 60% of ambient precipitation throughout the years. However, the annual forb Suaeda glauca was gradually losing its dominance or even becoming extinct over years. Our study highlights that the differences in plant salinity tolerance are key to the effects of precipitation changes on plant communities in coastal salt marsh. These findings aim to provide a theoretical basis for predicting vegetation dynamics and developing ecological management strategies to adapt to future precipitation changes.

Niche Theory and Species Range Limits along Elevational Gradients: Perspectives and Future Directions

Despite two centuries of research, the mechanisms underlying the formation of species’ elevational range limits remain poorly understood. The climatic variability hypothesis highlights the role of climatic conditions in shaping species’ thermal tolerance and distribution ranges, while the species interactions–abiotic stress hypothesis underscores the relative importance of biotic factors and abiotic stress along environmental gradients. We emphasize Darwin's perspective on the ubiquity of interspecific competition across climatic gradients and the importance of understanding how climate modulates biotic interactions to shape species distributions. Niche theory provides a comprehensive framework, combined with empirical research, to explore how environmental gradients influence species traits, leading to context-dependent species interactions that constrain distributions. In particular, the application of the concept of environmentally weighted performance can further elucidate these complex ecological mechanisms. Future research should integrate multiple approaches, including field and laboratory manipulative experiments, theoretical modeling, and interdisciplinary collaboration, to improve our understanding of species distributions in mountain regions and to inform biodiversity conservation strategies in the face of rapid environmental change.

Warming and rainfall reduction alter soil microbial diversity and co-occurrence networks and enhance pathogenic fungi in dryland soils

In this 9-year manipulative field experiment, we examined the impacts of experimental warming (2 °C, W), rainfall reduction (30 % decrease in annual rainfall, RR), and their combination (W + RR) on soil microbial communities and native vegetation in a semi-arid shrubland in south-eastern Spain. Warming had strong negative effects on plant performance across five coexisting native shrub species, consistently reducing their aboveground biomass growth and long-term survival. The impacts of rainfall reduction on plant growth and survival were species-specific and more variable. Warming strongly altered the soil microbial community alpha-diversity and changed the co-occurrence network structure. The relative abundance of symbiotic arbuscular mycorrhizal fungi (AMF) increased under W and W + RR, which could help buffer the direct negative impacts of climate change on their host plants nutrition and enhance their resistance to heat and drought stress. Indicator microbial taxa analyses evidenced that the marked sequence abundance of many plant pathogenic fungi, such as Phaeoacremonium, Cyberlindnera, Acremonium, Occultifur, Neodevriesia and Stagonosporopsis, increased significantly in the W and W + RR treatments. Moreover, the relative abundance of fungal animal pathogens and mycoparasites in soil also increased significantly under climate warming. Our findings indicate that warmer and drier conditions sustained over several years can alter the soil microbial community structure, composition, and network topology. The projected warmer and drier climate favours pathogenic fungi, which could offset the benefits of increased AMF abundance under warming and further aggravate the severe detrimental impacts of increased abiotic stress on native vegetation performance and ecosystem services in drylands.

Effect of Nitrogen Application Rate on the Relationships between Multidimensional Plant Diversity and Ecosystem Production in a Temperate Steppe.

Nitrogen (N) deposition, as one of the global change drivers, can alter terrestrial plant diversity and ecosystem function. However, the response of the plant diversity-ecosystem function relationship to N deposition remains unclear. On one hand, in the previous studies, taxonomic diversity (i.e., species richness, SR) was solely considered the common metric of plant diversity, compared to other diversity metrics such as phylogenetic and functional diversity. On the other hand, most previous studies simulating N deposition only included two levels of control versus N enrichment. How various N deposition rates affect multidimensional plant diversity-ecosystem function relationships is poorly understood. Here, a field manipulative experiment with a N addition gradient (0, 1, 2, 4, 8, 16, 32, and 64 g N m-2 yr-1) was carried out to examine the effects of N addition rates on the relationships between plant diversity metrics (taxonomic, phylogenetic, and functional diversity) and ecosystem production in a temperate steppe. Production initially increased and reached the maximum value at the N addition rate of 47 g m-2 yr-1, then decreased along the N-addition gradient in the steppe. SR, functional diversity calculated using plant height (FDis-Height) and leaf chlorophyll content (FDis-Chlorophyll), and phylogenetic diversity (net relatedness index, NRI) were reduced, whereas community-weighted means of plant height (CWMHeight) and leaf chlorophyll content (CWMChlorophyll) were enhanced by N addition. N addition did not affect the relationships of SR, NRI, and FDis-Height with production but significantly affected the strength of the correlation between FDis-Chlorophyll, CWMHeight, and CWMChlorophyll with biomass production across the eight levels of N addition. The findings indicate the robust relationships of taxonomic and phylogenetic diversity and production and the varying correlations between functional diversity and production under increased N deposition in the temperate steppe, highlighting the importance of a trait-based approach in studying the plant diversity-ecosystem function under global change scenarios.

Nitrogen deposition differentially regulates the sensitivity of gross primary productivity to extreme drought versus wetness.

Global hydroclimatic variability is increasing with more frequent extreme dry and wet years, severely destabilizing terrestrial ecosystem productivity. However, what regulates the consequence of precipitation extremes on productivity remains unclear. Based on a 9-year field manipulation experiment on the Qinghai-Tibetan Plateau, we found that the responses of gross primary productivity (GPP) to extreme drought and wetness were differentially regulated by nitrogen (N) deposition. Over increasing N deposition, extreme dry events reduced GPP more. Among the 12 biotic and abiotic factors examined, this was mostly explained by the increased plant canopy height and proportion of drought-sensitive species under N deposition, making photosynthesis more sensitive to hydraulic stress. While extreme wet events increased GPP, their effect did not shift over N deposition. These site observations were complemented by a global synthesis derived from the GOSIF GPP dataset, which showed that GPP sensitivity to extreme drought was larger in ecosystems with higher N deposition, but GPP sensitivity to extreme wetness did not change with N deposition. Our findings indicate that intensified hydroclimatic variability would lead to a greater loss of land carbon sinks in the context of increasing N deposition, due to that GPP losses during extreme dry years are more pronounced, yet without a synchronous increase in GPP gains during extreme wet years. The study implies that the conservation and management against climate extremes merit particular attention in ecosystems subject to N deposition.

Effects of Canopy Nitrogen Addition and Understory Vegetation Removal on Nitrogen Transformations in a Subtropical Forest

The management of understory vegetation and anthropogenic nitrogen (N) deposition has significantly resulted in a nutrient imbalance in forest ecosystems. However, the effects of canopy nitrogen addition and understory vegetation removal on N transformation processes (mineralization, nitrification, ammonification, and leaching) along with seasonal variations (spring, summer, autumn, and winter) remain unclear in subtropical forests. To fill this research gap, a field manipulation experiment was conducted with four treatments, including: (i) CK, control; (ii) CN, canopy nitrogen addition (25 kg N ha−1 year−1); (iii) UR, understory vegetation removal; and (iv) CN+UR, canopy nitrogen addition plus understory vegetation removal. The results revealed that CN increased net mineralization and nitrification by 294 mg N m−2 month−1 in the spring and 126 mg N m−2 month−1 in the winter, respectively. UR increased N mineralization and nitrification rates by 618 mg N m−2 month−1 in the summer. In addition, CN effectively reduced N leaching in the spring, winter, and autumn, while UR increased it in the spring and winter. UR increased annual nitrification rates by 93.4%, 90.3%, and 38.9% in the winter, spring, and summer, respectively. Additionally, both net N ammonification and annual nitrification rates responded positively to phosphorus availability during the autumn. Overall, UR potentially boosted nitrification rates in the summer and ammonification in the spring and winter, while CN reduced N leaching in the spring, winter, and autumn. Future research should integrate canopy nitrogen addition, understory vegetation removal, and phosphorus availability to address the global N deposition challenges in forest ecosystems.

Effects of microclimate on disease prevalence across an urbanization gradient.

Increased temperatures associated with urbanization (the "urban heat island" effect) have been shown to impact a wide range of traits across diverse taxa. At the same time, climatic conditions vary at fine spatial scales within habitats due to factors including shade from shrubs, trees, and built structures. Patches of shade may function as microclimate refugia that allow species to occur in habitats where high temperatures and/or exposure to ultraviolet radiation would otherwise be prohibitive. However, the importance of shaded microhabitats for interactions between species across urbanized landscapes remains poorly understood. Weedy plants and their foliar pathogens are a tractable system for studying how multiple scales of climatic variation influence infection prevalence. Powdery mildew pathogens are particularly well suited to this work, as these fungi can be visibly diagnosed on leaf surfaces. We studied the effects of shaded microclimates on rates of powdery mildew infection on Plantago host species in (1) "pandemic pivot" surveys in which undergraduate students recorded shade and infection status of thousands of plants along road verges in urban and suburban residential neighborhoods, (2) monthly surveys of plant populations in 22 parks along an urbanization gradient, and (3) a manipulative field experiment directly testing the effects of shade on the growth and transmission of powdery mildew. Together, our field survey results show strong positive effects of shade on mildew infection in wild Plantago populations across urban, suburban, and rural habitats. Our experiment suggests that this relationship is causal, where microclimate conditions associated with shade promote pathogen growth. Overall, infection prevalence increased with urbanization despite a negative association between urbanization and tree cover at the landscape scale. These findings highlight the importance of taking microclimate heterogeneity into account when establishing links between macroclimate or land use context and prevalence of disease.

CO2 enrichment accelerates alpine plant growth via increasing water-use efficiency

Phenological changes in global vegetation are often attributed to climate warming. However, climate warming and elevated atmospheric CO2 concentration (eCO2) are two co-occurring global change factors, and how eCO2 would affect vegetation phenology has received less attention. The partial pressure of atmospheric CO2 on the Tibetan Plateau (TP) is lower than that in regions of lower altitudes. Consequently, the growth and phenology of alpine plants in this region could be more sensitive to eCO2, but this hypothesis is not yet supported by empirical evidence. Here we explored the effect of eCO2 on plant phenology (including phenophases of green-up, budding, and flowering) through a 5-year field manipulation experiment in a high-altitude (4600 m above sea level) alpine grassland on the TP. Our results showed that eCO2 significantly advanced the spring phenology of an early-flowering species (Kobresia pygmaea), while it had no impact on the phenology of two mid-flowering species (Potentilla saundersiana and Potentilla cuneata). Compared to other low-altitude regions, plant phenology on the TP underwent greater alterations under eCO2, which supports our hypothesis that the growth of high-altitude plants is more sensitive to eCO2. Furthermore, we found that eCO2 significantly reduced the overlapping of flowering between contrasting plant species, mainly due to the phenological advancement of the K. pygmaea induced by eCO2. The observed advancement of the spring phenology in K. pygmaea under eCO2 was associated with increasing ecosystem water-use efficiency (WUE), thereby advancing its subsequent phenological development, such as budding and flowering. Our findings provide experimental evidence that atmospheric CO2 enrichment can accelerate plant growth processes in high-altitude regions, and suggest that large-scale model simulations should consider the effects of elevated atmospheric CO2 concentration on plant growth and phenology.

Effects of nitrogen addition and leaf age on needle traits and the relationships among traits in Pinus koraiensis

Leaf functional traits are essential for the growth and adaptation of trees to environmental changes. However, how needle traits and trait–trait relationships respond to different nitrogen (N) addition rates and leaf ages remain unclear for evergreen coniferous species. Our study aimed to investigate the effects of N addition and leaf age on needle traits and their bivariate correlations. Understanding resource utilization and distribution strategies during tree growth under the background of increased N deposition is important. We conducted a six-year manipulative field experiment in a Pinus koraiensis plantation in northeastern China with variable N addition levels: control (no N addition, CK); 20 kg N ha−1yr−1 (low-N, LN); 40 kg N ha−1yr−1 (medium-N, MN); and 80 kg N ha−1yr−1 (high-N, HN). The needle trait data were collected in September. The morphological traits, i.e., specific leaf area (SLA) and leaf dry matter content (LDMC); and chemical traits, i.e., leaf nitrogen concentration (LNC), leaf phosphorus concentration (LPC), and leaf nonstructural carbohydrates (NSC) concentrations, were measured to investigate the changes in needle traits and their bivariate correlations. We found that N addition and needle age significantly affected needle traits, and the effect of needle age was much stronger than that of N addition. N addition and needle age had no significant interaction effects on any of the traits except for the LNC. Due to the dispersal trait values of current-year needles, there were almost no significant bivariate correlations between traits under different N addition rates. The SLA–LDMC and LDMC–LNC relationships were found to differ among the various N addition rates, and other traittrait relationships may be relatively stable because of the weak plasticity of older needles. Additionally, needle age significantly affected the slope of the SLA–LDMC relationship and the elevation of the LDMC–LPC and NSC–LNC relationships, indicating differences in the ecological strategies of needles at different ages. In summary, our findings emphasize that trait–trait relationships in current-year and older needles were inconsistent in response to N addition and that the optimal allocation of resources in needles was based on their functional needs during leaf aging.

Mutual facilitation between foundation species Mytilus edulis and Lanice conchilega promotes habitat heterogeneity on tidal flats

Foundation species that modify their habitat can facilitate other species, including other foundation species. Most studies focus solely on a single foundation species, overlooking such facilitation cascades. In this study, we investigated the interactions between the two coastal foundation species Mytilus edulis (blue mussel) and Lanice conchilega (sand mason worm). We investigated whether these species engage in facilitative interactions or if their association simply reflects a shared ecological niche on the soft-sediment intertidal flats of the Dutch Wadden Sea. To do so, we performed species distribution modeling, manipulative field experiments, and field surveys. We found a positive association between both foundation species, with a 2.45 times higher occurrence of both species compared to a random distribution. In addition, these species partially occupied the same ecological niche. We demonstrated that L. conchilega provided settlement substrate for M. edulis spat, increasing densities by 400 times compared to bare plots. Furthermore, M. edulis reefs facilitated L. conchilega occurrence in the wake of the reef. Biogenic reef development revealed that this interspecific facilitation resulted in spatial habitat heterogeneity. Therefore, we conclude that interspecific facilitation can significantly enhance the occurrence of these two important intertidal foundation species. Acknowledgment of such complex facilitation interactions has an untapped potential for improving the success of restoration and conservation programs.

Linkages among leaf nutrient concentration, resorption efficiency, litter decomposition and their stoichiometry to canopy nitrogen addition and understory removal in subtropical plantation

BackgroundThe prevalence of understory removal and anthropogenic nitrogen (N) deposition has significantly altered the ecological processes of forest ecosystems at both regional and global scales. However, it remains a pressing challenge to understand how N deposition and understory removal affect leaf nutrient dynamics, nutrient resorption, litter decomposition, and their linkages for better managing forest ecosystems under nutrient imbalances induced by N enrichment. To address this research gap, a field manipulation experiment was carried out in a subtropical Cunninghamia lanceolata plantation with four treatments including: control (CK), canopy N addition (CN), understory removal (UR), and canopy N addition plus understory removal (CN × UR). Green and senesced leaf N and phosphorus (P) concentrations, N and P resorption efficiencies, litter decomposition, and their correlations were measured.ResultsThe results revealed that the average N concentrations of green early and late leaves in UR were increased by 6.61 and 18.89% compared to CK. UR had the highest whereas CN had the lowest P concentrations in green leaves across the two sampling seasons. Following this, UR, leaf type, season, and their interactions significantly affected leaf N, P, and N:P (P < 0.05). The highest leaf N resorption (32.68%) and P resorption efficiencies (63.96%) were recorded in UR. Litter decomposition was significantly retarded in UR (P < 0.01) relative to CN. The regression analysis demonstrated that leaf nutrient status was significantly interconnected with leaf nutrient resorption efficiencies. In addition, leaf nutrient dynamics were strongly correlated with litter nutrients, indicating that both were coupled.ConclusionThese findings can deepen our knowledge of biogeochemical cycling and reveal contrasting nutrient-acquisition strategies on N and P limitation in response to UR and CN. Considering the P limitation, it is important to note that P was resorbed more efficiently, illustrating a remarkable nutrient preservation approach for nutrient-limitations. Resorption may be a crucial mechanism for keeping nutrients in these forests, so better understory management practices are required to prevent reliance on external nutrient pools. Overall, this study sheds meaningful insights into the ability of forest adaptation in response to global climatic change.

Nighttime warming and nitrogen addition effects on the microclimate of a freshwater wetland dominated by Phragmites australis

The critical impacts of microclimate on carbon (C) cycling have been widely reported. However, the potential effects of global change on wetland microclimate remain unclear, primarily because of the absence of field manipulative experiment in inundated wetland. This study was designed to examine the effects of nighttime warming and nitrogen (N) addition on air, water, and sediment temperature and also reveal the controlling factors in a Phragmites australis dominated freshwater wetland on the North China Plain. Nighttime warming increased daily air, water, and sediment temperature by 0.24 °C, 0.27 °C, and 0.36 °C, respectively. The diurnal temperature range of water was decreased by 0.44 °C under nighttime warming, whereas warming had no effect on diurnal temperature range of air and sediment. In addition, N addition caused a reduction of 0.20 °C and 0.14 °C in daily water and sediment temperature by increasing vegetation coverage. There was a significant interaction between nighttime warming and N addition on water temperature. Furthermore, the vapor pressure deficit is the main factor affecting the extent of the warming-induced increases in air temperature. The changes of height and leaf area index of Phragmites australis are responsible for the cooling effects in the N addition plots. This study provides empirical evidence for the positive climate warming – microclimate feedback in freshwater wetland. However, N deposition leads to decreased water and sediment temperature. Our findings highlight the importance of incorporating the differential impacts of nighttime warming and N addition on air, water, and sediment temperature into the predictions of wetland C cycling responses to climate change.

Forb stability, dwarf shrub stability and species asynchrony regulate ecosystem stability along an experimental precipitation gradient in a semi-arid desert grassland.

Precipitation pattern changes may affect plant biodiversity, which could impact ecosystem stability. However, the effects of changes in precipitation regime on ecosystem stability and their potential mechanisms are still unclear. We conducted a 3-year field manipulation experiment with five precipitation treatments (-40%, -20%, 0% (CK), +20% and +40% of ambient growing season precipitation) in a semi-arid desert grassland to examine the effects of precipitation alterations on functional group stability, species asynchrony, and diversity, and the underlying mchanisms of ecosystem stability using structural equation modelling. Alterations in precipitation had different effects on community biomass and functional group biomass. Moreover, ecosystem stability was mainly driven by forb stability (path coefficient = 0.79). Changes in precipitation had significant effects on soil dissolved inorganic N (P < 0.01) further affecting ecosystem stability through species asynchrony (path coefficient = 0.25). Dwarf shrubs had a stabilizing effect on ecosystem stability (path coefficient = 0.32), mainly via deep roots. Ecosystem stability tended to be lower in the -40% (4.72) and +40% (2.74) precipitation treatments. The common reduction in species asynchrony and stability of forb and dwarf shrub functional groups resulted in lower ecosystem stability under the -40% treatment. The lower stability under the +40% treatment might be ascribed to unimproved dwarf shrub stability. Higher dwarf shrub and forb stability contributed to higher ecosystem stability under normal precipitation changes (±20% treatments) and CK. Species diversity was not a crucial driver of ecosystem stability. Our results indicate that precipitation alteration can regulate ecosystem stability via functional group stability (e.g. forb stability, dwarf shrub stability) and species asynchrony in a semiarid desert grassland.

Artificial Light at Night Reduces Flashing in Photinus and Photuris Fireflies During Courtship and Predation

Artificial light at night (ALAN) affects species-specific communication in a wide range of nocturnal species, including fireflies (Lampyridae). Fireflies rely on bioluminescent signals for communicating. In this study, we conducted two manipulative field experiments to evaluate the effect of artificial light at night on the flashing activity of male and female neotropical fireflies during courtship and predation. Our results showed a significant reduction in the flashing activity of both males and females exposed to ALAN during courtship and predation. Remarkably, the effect of ALAN on male flashing activity seems to be independent of female flashing activity. In conclusion, ALAN disrupts bioluminescent intraspecific (courtship) and interspecific (predation) communication, which in turn could influence mating success, thus negatively affecting neotropical firefly populations in the long term. Our findings contribute to understanding the challenges faced by neotropical firefly communities in the presence of ALAN.

Bioturbating megafauna limits the expansion of Cymodocea nodosa seagrass meadows

In seagrass systems, there is a realization that biotic interactions affecting sediment stability might be as important as abiotic stressors in shaping the growth and distribution patterns of seagrass vegetation. However, few studies have looked at the interaction between seagrass and large (mega) bioturbating species, although through sediment reworking activity they may have the capacity to cause large scale impact on seagrass establishment. We assessed the effect of bioturbation by mobile megabioturbators (species lager than 5 cm) on subtidal seagrass (Cymodocea nodosa) using a manipulative field experiment where access by megabioturbators was restricted for 4 months during the growth season. There was a negative effect of megabioturbators on both the biomass and density of young seagrass ramets extending outwards from a mature seagrass meadow, but megabioturbators did not have a notable effect on seagrass growth in the meadows. Interestingly, the exclusion of megabioturbators in all tested habitats significantly modified the infaunal community composition, indicating the presence of complexity in biotic interactions and the possibility of indirect effects caused by the megabioturbators. Overall, megabioturbators have direct and indirect impact in seagrass habitats, playing a significant role in seagrass distribution patterns.

Moderate increase of precipitation stimulates CO2 production by regulating soil organic carbon in a saltmarsh.

Saltmarsh is widely recognized as a blue carbon ecosystem with great carbon storage potential. Yet soil respiration with a major contributor of atmospheric CO2 can offset its carbon sink function. Up to date, mechanisms ruling CO2 emissions from saltmarsh soil remain unclear. In particular, the effect of precipitation on soil CO2 emissions is unclear in coastal wetlands, due the lack of outdoor data in real situations. We conducted a 7-year field manipulation experiment in a saltmarsh in the Yellow River Delta, China. Soil respiration in five treatments (-60%, -40%, +0%, +40%, and + 60% of precipitation) was measured in the field. Topsoils from the last 3 years (2019-2021) were analyzed for CO2 production potential by microcosm experiments. Furthermore, quality and quantity of soil organic carbon and microbial function were tested. Results show that only the moderate precipitation rise of +40% induced a 66.2% increase of CO2 production potential for the microcosm experiments, whereas other data showed a weak impact. Consistently, soil respiration was also found to be strongest at +40%. The CO2 production potential is positively correlated with soil organic carbon, including carbon quantity and quality. But microbial diversity did not show any positive response to precipitation sizes. r-/K-strategy seemed to be a plausible explanation for biological factors. Overall, our finding reveal that a moderate precipitation increase, not decrease or a robust increase, in a saltmarsh is likely to improve soil organic carbon quality and quantity, and bacterial oligotroph:copiotroph ratio, ultimately leading to an enhanced CO2 production.

Increased precipitation alters the effects of nitrogen deposition on soil bacterial and fungal communities in a temperate forest

Anthropogenic nitrogen (N) deposition and increased precipitation are known to alter soil microbial communities. However, the combined effects of elevated N deposition and increased precipitation on soil microbial community dynamics and co-occurrence networks in temperate forests remain elusive. In this study, we conducted a field manipulation experiment by applying N solution and water to the forest canopy to simulate natural N deposition and increased precipitation in a temperate forest. We collected samples in the litter layer, organic soil layer, and mineral soil layer in 2018–2019 after 6–7 years of N and water treatments, and explored how elevated N deposition and increased precipitation regulate soil microbial diversity, community composition, and co-occurrence networks in different soil layers and at different sampling times. We found that the effects of N deposition and increased precipitation on soil microbial communities varied with soil layers and sampling times. Compared to the ambient environment, single canopy N addition (CN) or single canopy water addition (CW) did not affect bacterial Shannon diversity in the mineral soil layer in 2018, but the combined canopy N and water additions (CNW) decreased it in this layer at this time. CN increased fungal OTU richness in the organic and mineral soil layers in 2018; however, CW and CNW did not have an effect on it in the same layer at the same time. CW and CNW, but not CN, significantly affected bacterial and fungal community compositions in the litter layer in 2018 and in the organic soil layer in 2019. In contrast, CN, but not CW or CNW, significantly affected fungal community composition in the litter layer in 2019. CNW exhibited higher complexities of bacterial and fungal co-occurrence networks than CN and the ambient environment, indicating increased precipitation can strengthen the effect of N deposition on the complexity of bacterial and fungal co-occurrence networks. Our findings suggest that increased precipitation alters the effects of atmospheric N deposition on soil bacterial and fungal communities in this temperate forest, depending on soil layer and sampling time. Moreover, both bacterial and fungal community compositions are sensitive to increased precipitation, but the bacterial community composition is more sensitive to N deposition than the fungal community composition in the organic and mineral soil layers in this forest.

Warming differentially affects above- and belowground ecosystem functioning of the semi-arid alpine grasslands

Rapid climate warming is altering multiple ecosystem functions of alpine grasslands. However, the responses of the above- and belowground ecosystem multifunctionality (EMF) to climate warming might exhibit difference in semi-arid alpine grasslands. Based on manipulative field experiments at an alpine meadow and an alpine steppe, we explored warming effects on the functioning of alpine grassland ecosystems on the Tibetan Plateau. Warming significantly decreased plant diversity and aboveground biomass, but tended to increase belowground biomass, soil carbon, and soil nutrient contents. Experimental warming generally had neutral effects on the EMF of both alpine grasslands. Nevertheless, warming differentially affects the above- and belowground ecosystem functioning of Tibetan semi-arid alpine grasslands, with the aboveground EMF (AEMF) deceased but the belowground EMF (BEMF) increased under warmer conditions. Our results further showed that the negative effect of experimental warming on AEMF was mainly regulated by the changes of plant and soil biodiversity. However, plant productivity had a pivotal role in propelling the positive effect of warming on BEMF. Our results emphasized the potential impacts of plant and soil biodiversity, productivity, and soil nutrients in maintaining the EMF of alpine grasslands, which could offer novel views for sustainable management of Tibetan semi-arid alpine ecosystems.

Soil extracellular enzyme stoichiometry reveals the nutrient limitations in soil microbial metabolism under different carbon input manipulations

Changes in the quality and quantity of litter and root inputs due to climate change and human activities can influence below-ground biogeochemical processes in forest ecosystems. However, it is unclear whether and how much aboveground litter and root inputs affect soil microbial metabolism and nutrient limitation mechanisms. In this study, according to a 4-years field manipulation experiment, litter and root manipulations (control (CK), double litter input (DL), no litter (NL), no root (NR), and no inputs (NI)) were set up to analyze the extracellular enzyme activities and stoichiometric ratios characteristics of 0–10 cm and 10–20 cm soils, explore the metabolic limitations of microorganisms, and clarify the main driving factors restricting nutrient limitation. The results showed that the enzyme activities associated with the C cycling (β-1,4-glucosidase (BG), cellulose disaccharide hydrolase (CBH)) and N cycling (β-1,4-N-acetylglucosaminidase (NAG), leucine aminopeptidase (LAP)) in DL treatment were significantly higher than those in NR treatment. Moreover, enzyme activities related to P cycling are significantly higher in comparison to other treatments. The acid phosphatase (AP), which is related to the P cycle, showed the highest activity under NR treatment. In addition, there was no significant difference in soil microbial metabolic limitation by the different carbon inputs, which did not change the original nutrient limitation pattern. The main drivers of microbial nutrient metabolic limitation included soil physicochemical properties, soil total nutrients, and available nutrients, among which soil SWC and pH presented the greatest influence on microbial C limitation and soil total nutrients showed the greatest influence on microbial N limitation. Changes in soil carbon input altered soil extracellular enzyme activities and their stoichiometric ratios by affecting soil physicochemical properties, total nutrients. This study provides data for the understanding of material cycling in forest ecosystems under environmental change.