Manipulative Field Experiment Research Articles

Evaluating the role of moonlight-darkness dynamics as proximate spawning cues in an Acropora coral

Abstract For sessile broadcast spawning marine invertebrates, such as corals, successful sexual reproduction depends on conspecifics spawning synchronously. The precise monthly, lunar, and diel timing and the extent of synchrony, i.e., proportion of population reproducing at the same time, are likely to play a key role in coral population recovery, persistence, and adaptation. Despite its importance, the mechanisms by which different environmental factors trigger corals to spawn on specific dates within the lunar cycle remain poorly understood. Periods of darkness post-sunset around full moon of the spawning month have been shown to induce spawning in merulinid corals, whereas for Acropora, moonlight is considered the main determinant driver of night of spawning. Here, we conducted two manipulative field experiments around full moon in Palau using the common table coral Acropora aff. hyacinthus to disentangle the role of moonlight and darkness post-sunset as proximate cues. Coral fragments were assigned to three treatments providing different post-sunset darkness conditions, versus control and procedural control fragments exposed to natural conditions. In contrast to previous studies on Acropora, we found that Acropora aff. hyacinthus can spawn synchronously in the absence of moonlight during the nights leading to spawning. Corals exposed to darkness post-sunset for at least two to three consecutive nights advanced their spawning compared to controls. This finding indicates that periods of darkness post-sunset can act as an inducer for spawning in Acropora as well as in merulinid corals, suggesting that this mechanism may be more widespread than previously thought.

Strategic planting and nutrient amendments to accelerate the revegetation of rapidly retreating coastal dunes

Abstract The increasing frequency and severity of disturbances to coastal dune ecosystems requires developing and implementing restoration strategies that rapidly accelerate re‐establishment of vegetation, enhance dune accretion, and ultimately preserve dune ecosystem services. To assess how to rapidly re‐establish vegetation to counter ecosystem losses, we conducted a manipulative field experiment on a created foredune crest in Northeast Florida, USA to determine what combinations of planting density, outplant species composition, and nutrient addition maximize dune revegetation rate. Above‐ground biomass in densely planted plots with added nutrients was 4.7–7.3 times that of plots planted at conventional restoration densities after only 3 months. Critically, thickly vegetated, high‐density plots with nutrient addition accreted 1–5 cm more sediment after 3 months and 7.5–22 cm more sediment after 15 months, demonstrating that this planting method can rapidly kickstart dune‐building processes. Additionally, above‐ and below‐ground biomass in densely planted, fertilized plots containing bitter panicum (Panicum amarum) were consistently higher and accreted more sand than similar plots planted with sea oats (Uniola paniculata), suggesting that planting schemes involving this early successional species may be most effective for rapid dune‐building. Synthesis and applications. Because coupling nutrient addition with dense planting can trigger self‐sustaining, reinforcing plant growth and dune‐building feedbacks within only months, this planting approach may help to enhance the long‐term success of dune restoration projects.

Conspecific interactions between corals mediate the effect of submarine groundwater discharge on coral physiology.

Land-based inputs, such as runoff, rivers, and submarine groundwater, can alter biologic processes on coral reefs. While the abiotic factors associated with land-based inputs have strong effects on corals, corals are also affected by biotic interactions, including other neighboring corals. The biologic responses of corals to changing environmental conditions and their neighbors are likely interactive; however, few studies address both biotic and abiotic interactions in concert. In a manipulative field experiment, we tested how the natural environmental gradient created by submarine groundwater discharge (SGD) affected holobiont and symbiont metabolic rates and endosymbiont physiology of Porites rus. We further tested how the effect of SGD on the coral was mediated by intra and interspecific interactions. SGD is a natural land-sea connection that delivers nutrients, inorganic carbon, and other solutes to coastal ecosystems worldwide. Our results show that a natural gradient of nutrient enrichment and pH variability as a result of acute SGD exposure generally benefited P. rus, increasing gross photosynthesis, respiration, endosymbiont densities, and chlorophyll a content. Conspecifics in direct contact with the a neighboring coral, however, altered the relationship between coral physiology and SGD, lowering the photosynthetic and respiration rates from expected values when the coral had no neighbor. We show that the response of corals to environmental change is dependent on the types of nearby neighbor corals and how neighbors alter the chemical or physical environment around the coral. Our study underscores the importance of considering biotic interactions when predicting the physiologic responses of corals to the environment.

Effects of Ectomycorrhizae and Hyphae on Soil Fungal Community Characteristics Across Forest Gap Positions

The interactive effects of environmental heterogeneity caused by forest gaps and ectomycorrhizae on fungal community characteristics remain insufficiently explored. To address this knowledge gap, we established a three-year field manipulation experiment in a Picea asperata (Picea asperata Mast.) plantation located in the subalpine region of western Sichuan, China. Growth bags with three mesh sizes—1000 μm (allowing ectomycorrhizae and hyphae), 48 μm (excluding ectomycorrhizae), and 1 μm (excluding both)—were placed across forest gaps (closed canopy, gap edge, and gap center) to investigate how gap disturbances influence soil fungal communities via changes in ectomycorrhizal and hyphal turnover alongside soil physicochemical properties. Soil fungal α-diversity was significantly lower under closed-canopy conditions than at forest gap centers and remained unaffected by ectomycorrhizal and hyphal treatments. Particularly, species diversity increased by 9%, and phylogenetic diversity increased by 10% in forest gap centers compared to the closed canopy. In contrast, soil fungal β-diversity responded to both ectomycorrhizal/hyphal treatments (R2 = 0.061; p = 0.001) and forest gap positions (R2 = 0.033; p = 0.003). Pairwise comparative analyses revealed significant distinctions between treatments, concurrently excluding ectomycorrhizal and hyphal treatments versus other experimental treatments, as well as between closed-canopy conditions and forest gap centers. The fungal community was dominated by four major phyla: Ascomycota (25.6%–71.0%), Basidiomycota (17.7%–43.7%), Mortierellomycota (1.4%–24.5%), and Rozellomycota (0.4%–2.9%), the relative abundances of which were unaffected by either ectomycorrhizal/hyphal treatments or forest gap positions. The biomass of ectomycorrhizal and saprotrophic fungi showed no significant response to ectomycorrhizal/hyphal treatments. Notably, the exclusion of ectomycorrhizae and hyphae enhanced the significant correlations between fungal community characteristics and soil physicochemical properties. Hierarchical partitioning analysis revealed that the soil water content (SWC) and dissolved organic carbon content were the key determinants of soil fungal community characteristics beneath closed-canopy conditions. In contrast, at forest gap edges and centers, the fungal communities were predominantly shaped by the SWC and dissolved carbon and nitrogen contents. This study highlights the impacts of forest gap disturbances and ectomycorrhizal treatments on soil fungal communities, offering valuable insights for the sustainable management and biodiversity conservation of subalpine forest ecosystems.

Advanced precipitation peak offsets middle growing-season drought in impacting grassland C sink.

Redistribution of precipitation across seasons is a widespread phenomenon affecting dryland ecosystems globally. However, the impacts of shifting seasonal precipitation patterns on carbon (C) cycling and sequestration in dryland ecosystems remain poorly understood. In this study, we conducted a 10-yr (2013-2022) field manipulative experiment that altered the timing of growing-season precipitation peaks in a semi-arid grassland. We found that the delayed precipitation peak suppressed plant growth and thus reduced gross ecosystem productivity, ecosystem respiration, and net ecosystem productivity due to middle growing-season water stress. Surprisingly, shifting more precipitation to the early growing season can advance plant development, increase the dominance of drought-tolerant forbs, and thus compensate for the negative impacts of middle growing-season water stress on ecosystem C cycling, leading to a neutral change in grassland C sink. Our findings indicate that greater precipitation and plant development in spring could act as a crucial mechanism, maintaining plant growth and stabilizing ecosystem C sink. This underscores the urgent need to incorporate precipitation seasonality into Earth system models, which is crucial for improving projections of terrestrial C cycling and sequestration under future climate change scenarios.

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.

Nitrogen addition alleviates the negative effects of reduction in precipitation on soil multifunctionality in a typical steppe

The increasing frequency of drought events and nitrogen deposition have fundamentally changed soil microbial functions in terrestrial ecosystems. However, most studies have mainly concentrated on the impact of a single environmental factor on ecosystem functions; how drought and nitrogen enrichment interactively affect soil multifunctionality remains largely unknown. In this study, the effects of different drought scenarios [intense drought (ID), chronic drought (CD), and reduced rainfall frequency (RF)] and nitrogen addition on soil microbial biomass, and soil multifunctionality (determined using soil enzyme activity) were examined in the fourth year of a field manipulative experiment conducted in a typical steppe in northern China. The results demonstrated that both ID and CD significantly reduced soil multifunctionality, microbial biomass carbon (MBC) and microbial biomass nitrogen (MBN). The CD treatment also decreased the ratio of MBC to MBN, while RF had less impacts on soil multifunctionality and the biomass of soil microbes. In contrast, nitrogen addition enhanced soil multifunctionality, MBC and MBN. Structural equation modeling analysis demonstrated that drought decreased soil multifunctionality directly and indirectly by reducing MBN and soil water content, whereas nitrogen addition increased soil multifunctionality mainly by increasing MBN and soil inorganic nitrogen. This study provides the first experimental evidence of the opposing impacts of reduction in precipitation and nitrogen enrichment on soil microbial biomass and soil multifunctionality in a semiarid typical steppe, and suggests that nitrogen fertilization could be an effective measure to alleviate the negative effects of climate drought on soil functions in nitrogen- and water-limited grassland ecosystems.

Rising temperatures may increase fungal epizootics in northern populations of the invasive spongy moth in North America

Insect pest species are generally expected to become more destructive with climate change because of factors such as weakened host tree defences during droughts and increased voltinism under rising temperatures; however, responses will vary by species due to a variety of factors, including altered interactions with their natural enemies. Entomopathogens are a substantial source of mortality in insects, but the likelihood of epizootics can depend strongly on climatic conditions. Previous research indicates that rates of infection of the spongy moth (Lymantria dispar) by its host-specific fungal pathogen, Entomophaga maimaiga, increase with environmental moisture and decrease as temperatures rise. High temperatures may have direct and indirect (due to the associated drying) effects on the fungus, but the interactive effects between temperature and moisture level on larval infection are unclear. Here, we test the hypothesis that warmer, drier conditions will decrease rates of infection of spongy moth larvae by E. maimaiga. We evaluated the effects of precipitation and temperature on larval mortality caused by E. maimaiga with a manipulative field experiment, conducted in one of the northernmost and coldest parts of the spongy moth’s non-native range in North America. We caged laboratory-reared spongy moth larvae in experimentally warmed open-air forest plots, exposing the larvae to soil inoculated with E. maimaiga resting spores during two consecutive trials. Caged larvae were exposed to three temperature treatments — ambient, 1.7 °C above ambient and 3.4 °C above ambient — and either supplemental precipitation (+173 mm per trial) or ambient precipitation. Opposite to our hypothesis, there was no significant effect of supplemental precipitation, nor an interaction between precipitation and temperature. There was, however, a significant positive effect of increasing temperature on the number of larvae infected. On average, in each respective trial, larval infection increased by 44% and 50% under the elevated temperature treatments compared to ambient temperature. Experimental warming may have increased infections because ambient temperatures at the field site were suboptimal for fungal germination. The results from this experiment suggest that, in colder portions of the spongy moth’s invasive range, increasing temperatures due to climate change may enhance the ability of E. maimaiga to help control populations of the spongy moth.

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.

Behavioural responses to mammalian grazing expose insect herbivores to elevated risk of avian predation.

Large mammalian herbivores (LMH) are important functional components and drivers of biodiversity and ecosystem functioning in grasslands. Yet their role in regulating food-web dynamics and trophic cascades remains poorly understood. In the temperate grasslands of northern China, we explored whether and how grazing domestic cattle (Bos taurus) alter the predator-prey interactions between a dominant grasshopper (Euchorthippus unicolor) and its avian predator the barn swallow (Hirundo rustica). Using two large manipulative field experiments, we found that in the presence of cattle, grasshoppers increased their jumping frequency threefold, swallows increased foraging visits to these fields sixfold, and grasshopper density was reduced by about 50%. By manipulatively controlling the grasshoppers' ability to jump, we showed that jumping enables grasshoppers to avoid being incidentally consumed or trampled by cattle. However, jumping behaviour increased their consumption rates by swallows 37-fold compared with grasshoppers that were unable to jump. Our findings illustrate how LMH can indirectly alter predator-prey interactions by affecting behaviour of avian predators and herbivorous insects. These non-plant-mediated effects of LMH may influence trophic interactions in other grazing ecosystems and shape community structure and dynamics. We highlight that convoluted multispecies interactions may better explain how LMH control food-web dynamics in grasslands.

Grazing legacy mediates the diverse responses of grassland multidimensional stability to resource enrichment

Long-term grazing and resource enrichment are known to affect grassland ecosystem stability independently. However, the interactive effects of grazing legacy and resource enrichment on ecosystem multidimensional stability (e.g., temporal stability (TS): the degree of constancy of ecosystem components over time and temporal resistance (TR): the ability of ecosystem components to withstand the environmental change) in grassland ecosystems remain underexplored. In this study, we conducted a field manipulation experiment to assess the impact of 4-yr of resource addition (water and nitrogen) on multidimensional stability of multiple ecosystem components and underlying drivers in Inner Mongolia steppe with 7-yr different experimental grazing history. We found that the positive effects of water + nitrogen addition on multifunctional TS increased with increasing grazing intensity and an opposing trend was observed for multifunctional TR response. However, neither grazing legacy nor resource addition had an effect on community compositional stability. Resource addition enhanced the stability of species evenness, CWM (community weighted mean) species biomass and CWM-traits. Furthermore, water + nitrogen additions stimulated soil temperature TS but reduced its TR in heavy grazing intensity, with high grazing intensity bolstering soil moisture stability. Multifunctional stability was primarily governed by species asynchrony, while stability of species evenness fostered compositional stability. In particular, TS of functional diversity enhanced multifunctional TS in light grazing intensity, while TS of CWM-traits promoted multifunctional TS in heavy grazing intensity. Our study underscores the decoupling responses of functional stability and compositional stability to grazing legacy combined with resource enrichment. These findings highlight that the significance of concurrently considering multiple stability dimensions and components for a comprehensive understanding of grassland ecosystem stability under intensifying land use and global change scenarios.

Effects of simulated precipitation changes on soil respiration:Progress and prospects.

Soil respiration, the main pathway for transferring terrestrial carbon pool to atmospheric carbon pool, is profoundly affected by the intensification in global precipitation variability in the context of climate change. Nowadays, variable controlling methods and field manipulation experiments are two main methods widely used to investigate the effects of simulated precipitation changes on soil respiration. Yet, due to the heterogeneity of soil properties, vegetation types, and the magnitude of precipitation change, there is substantial inconsistency in the conclusions of simulated precipitation change effects on soil respiration. Here, we analyzed data from domestic and foreign literature, and examined the effects of simulated precipitation change on soil respiration. Firstly, we described the response pattern of soil respiration to soil moisture fluctuation and pointed out that the magnitude and direction of the response of soil respiration to simulated precipitation change depended on whether soil moisture was optimally conditioned at different precipitation treatments. Second, we summarized the response patterns of soil respiration to symmetric increase and decrease in precipitation, which mainly included symmetric and asymmetric responses (positive and negative asymmetric). Meanwhile, the adaptation of plants and soil microorganisms to drought stress and soil oxygen limitation, as well as the reduction of organic substrates, were the main mechanisms accounting for the shifts of soil respiration response patterns to simulated precipitation change from symmetric to asymmetric responses. Third, we identified a significant effect of ambient climate on soil respiration in response to precipitation treatments as increasing duration of the experimental treatments. In addition, cumulative or buffering effects of ambient climatic conditions on precipitation treatment could affect the sensitivity of soil respiration along precipitation gradient by altering hydrothermal conditions. Finally, to accurately assess the implications of precipitation changes on soil carbon balance processes, we proposed three aspects of future precipitation effects on soil respiration for attention: 1) focusing on the phenomenon of "threshold effects" in the asymmetric response of soil respiration along precipitation gradients; 2) distinguishing the intrinsic mechanisms of autotrophic and heterotrophic components in soil respiration in response to precipitation changes; and 3) focusing on the impacts of intensified precipitation variability on soil respiration in the context of future climate extremes. In conclusion, with the intensified variability in global precipitation patterns, clarifying the response mechanism of soil respiration to precipitation changes is of great significance for accurately predicting and evaluating the alterations of soil carbon cycle processes and carbon balance in the context of global changes.

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.

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.