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.

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.

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.

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.

Tipping point of plant functional traits of Leymus chinensis to nitrogen addition in a temperate grassland.

It has widely been documented that nitrogen (N) enrichment stimulates plant growth and modifies plant functional traits in the terrestrial ecosystem. However, it remains unclear whether there are critical transitions or tipping points for the response of plant growth or traits to N enrichment, and how these responses differ to different N forms. We chose the native, perennial clonal grass, Leymus chinensis in Inner Mongolia steppe, and conducted a field experiment, in which six N addition rates (0, 2, 5, 10, 20, and 50 g N m-2 year-1) and five N compound types [NH4NO3, (NH4)2SO4, NH4HCO3, CO(NH2)2, slow-release CO(NH2)2] are considered. Here, we found that the different N compound types had no significant effect on the growth of L. chinensis. N addition rate significantly increased plant aboveground biomass and leaf nitrogen concentration, whereas decreased leaf dry matter content. The tipping point for N-induced aboveground biomass increase was at 10 g N m-2 year-1, and the changes in functional traits were at N addition rates of 20 g N m-2 year-1. Our findings suggested that the responses of aboveground biomass and functional traits to N addition were asymmetric, in which responses in aboveground biomass were more sensitive than that in functional traits. The differential sensitivity of aboveground biomass and functional traits of L. chinensis occurred to N deposition highlights the importance of functional traits in mediating ecosystem functioning in the face of N deposition, regardless of which chemical forms dominate in the deposited N.

Nitrogen deposition experiment mimicked with NH4NO3 overestimates the effect on soil microbial community composition and functional potential in the Eurasian steppe.

BACKGROUND: The nitrogenous compound deposited from the atmosphere to the soil is complex, but most field experiments mimic nitrogen deposition with the acid NH4NO3 alone. Thus, whether the acid and non-acid nitrogenous compounds have similar effects on biodiversity and ecosystem functions remains understudied. We mimicked nitrogen deposition with acidic NH4NO3 and (NH4)2SO4, and non-acidic urea, slow-released urea and NH4HCO3 in a temperate steppe, and quantified soil microbial taxonomic and functional gene composition with amplicon sequencing and shotgun metagenomics, respectively. RESULTS: While NH4NO3 and (NH4)2SO4 significantly altered the soil microbial taxonomic and functional composition as well as their carbon decomposition potential, the other three compounds had smaller effects. CONCLUSION: Our results suggested that previous nitrogen deposition experiments mimicked with NH4NO3 or (NH4)2SO4 alone may have overestimated the effect on biodiversity and ecosystem functions in the Eurasian steppe and similar ecosystems affected by mainly nonacidic nitrogen deposition.

Mowing Did Not Alleviate the Negative Effect of Nitrogen Addition on the Arbuscular Mycorrhizal Fungal Community in a Temperate Meadow Grassland.

As nitrogen deposition intensifies under global climate change, understanding the responses of arbuscular mycorrhizal (AM) fungi to nitrogen deposition and the associated mechanisms are critical for terrestrial ecosystems. In this study, the effects of nitrogen addition and mowing on AM fungal communities in soil and mixed roots were investigated in an Inner Mongolia grassland. The results showed that nitrogen addition reduced the α-diversity of AM fungi in soil rather than that of root. Besides, nitrogen addition altered the composition of AM fungal community in soil. Soil pH and inorganic nitrogen content were the main causes of changes in AM fungal communities affected by nitrogen addition. Mowing and the interaction of nitrogen addition and mowing had no significant effect on AM fungal community diversity. In contrast, while mowing may reduce the negative effects of nitrogen addition on the richness and diversity of plants by alleviating light limitation, it could not do so with the negative effects on AM fungal communities. Furthermore, AM fungal communities clustered phylogenetically in all treatments in both soil and roots, indicating that environmental filtering was the main driving force for AM fungal community assembly. Our results highlight the different responses of AM fungi in the soil and roots of a grassland ecosystem to nitrogen addition and mowing. The study will improve our understanding of the effects of nitrogen deposition on the function of ecosystem.

Leaf Functional Traits of Two Species Affected by Nitrogen Addition Rate and Period Not Nitrogen Compound Type in a Meadow Grassland.

Plasticity of plant functional traits plays an important role in plant growth and survival under changing climate. However, knowledge about how leaf functional traits respond to the multi-level N addition rates, multiple N compound and duration of N application remains lacking. This study investigated the effects of 2-year and 7-year N addition on the leaf functional traits of Leymus chinensis and Thermopsis lanceolata in a meadow grassland. The results showed that the type of N compounds had no significant effect on leaf functional traits regardless of duration of N application. N addition significantly increased the leaf total N content (LN) and specific leaf area (SLA), and decreased the leaf total P content (LP) and leaf dry matter content (LDMC) of the two species. Compared with short-term N addition, long-term N addition increased LN, LP, SLA, and plant height, but decreased the LDMC. In addition, the traits of the two species were differentially responsive to N addition, LN and LP of T. lanceolata were consistently higher than those of L. chinensis. N addition would make L. chinensis and T. lanceolata tend to "quick investment-return" strategy. Our results provide more robust and comprehensive predictions of the effects of N deposition on leaf traits.

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.

Effects of nitrogen addition on plant-soil micronutrients vary with nitrogen form and mowing management in a meadow steppe.

Nitrogen (N) addition and mowing can significantly influence micronutrient cycling in grassland ecosystems. It remains largely unknown about how different forms of added N affect micronutrient status in plant-soil systems. We examined the effects of different N compounds of (NH4)2SO4, NH4NO3, and urea with and without mowing on micronutrient Fe, Mn, Cu, and Zn in soil-plant systems in a meadow steppe. The results showed that (NH4)2SO4 addition had a stronger negative effect on soil pH compared with NH4NO3 and urea, resulting in higher increases in soil available Fe and Mn herein. Nitrogen addition decreased plant community-level biomass weighted (hereafter referred to as community-level) Fe concentration but increased Mn concentration, with a greater effect under (NH4)2SO4 addition. Community-level Cu concentration increased with (NH4)2SO4 and NH4NO3 addition only under mowing treatment. Mowing synergistically interacted with urea addition to increase community-level Mn and Zn concentrations even with decreased soil organic matter, possibly because of compensatory plant growth and thus higher plant nutrient uptake intensity under mowing treatment. Overall, responses of plant-soil micronutrients to N addition varied with mowing and different N compounds, which were mainly regulated by soil physicochemical properties and plant growth. Different magnitude of micronutrient responses in plants and soils shed light on the necessity to consider the role of various N compounds in biogeochemical models when projecting the effects of N enrichment on grassland ecosystems.

Correction to: Mowing mitigates the negative impacts of N addition on plant species diversity.

Unfortunately, the panels of (f) in Figures 1, 2, and 4.

The relative contributions of intra- and inter-specific variation in driving community stoichiometric responses to nitrogen deposition and mowing in a grassland.

AIMS: The stoichiometric characteristics of plant communities are important controller for several fundamental ecological processes. The effects of environmental changes on community stoichiometric characteristics are driven by intra- and inter-specific variation. However, the relative importance of both pathways has seldom been empirically examined. METHODS: We quantified the relative contribution of intra- and inter-specific variation to the changes of community nitrogen (N) and phosphorus (P) concentrations after seven-year factorial N addition and mowing treatments in a semi-arid grassland of northern China. RESULTS: Nitrogen addition significantly increased community N and P concentrations and N:P ratio. Mowing significantly increased community N concentration and N:P. Intra-specific variation contributed more than inter-specific variation to the total variability of all the nutritional and stoichiometric characteristics, with intra-specific variation accounting for 68%, 70%, and 75% of the total variation in community-level N, P, and N:P, respectively. Negative covariations between the contribution of intra- and inter-specific variation occurred for community N and P concentrations. Further, N addition and mowing interacted to affect the impacts of intra- and inter-specific variation on community N concentration and N:P stoichiometry. CONCLUSIONS: Our results highlight different ways of trait selection for N addition and mowing treatments. Interactions between those two factors make it more difficult to accurately predict the responses of plant-mediated biogeochemical cycles under co-occurrence of environmental changes.

Mowing mitigates the negative impacts of N addition on plant species diversity.

Increasing availability of reactive nitrogen (N) threatens plant diversity in diverse ecosystems. While there is mounting evidence for the negative impacts of N deposition on one component of diversity, species richness, we know little about its effects on another one, species evenness. It is suspected that ecosystem management practice that removes nitrogen from the ecosystem, such as hay-harvesting by mowing in grasslands, would mitigate the negative impacts of N deposition on plant diversity. However, empirical evidence is scarce. Here, we reported the main and interactive effects of N deposition and mowing on plant diversity in a temperate meadow steppe with 4-year data from a field experiment within which multi-level N addition rates and multiple N compounds are considered. Across all the types of N compounds, species richness and evenness significantly decreased with the increases of N addition rate, which was mainly caused by the growth of a tall rhizomatous grass, Leymus chinensis. Such negative impacts of N addition were accumulating with time. Mowing significantly reduced the dominance of L. chinensis, and mitigated the negative impacts of N deposition on species evenness. We present robust evidence that N deposition threatened biodiversity by reducing both species richness and evenness, a process which could be alleviated by mowing. Our results highlight the changes of species evenness in driving the negative impacts of N deposition on plant diversity and the role of mowing in mediating such negative impacts of N deposition.

The impacts of nitrogen deposition on community N:P stoichiometry do not depend on phosphorus availability in a temperate meadow steppe.

Nitrogen (N) enrichment has great consequences on several fundamental ecological processes through its impacts on plant nutrition traits (i.e. nutrient concentration and stoichiometric ratios); however, the extent to which the effects of N enrichment depend on phosphorus (P) availability are less well understood. While there is mounting evidence for the species-specific responses of plant nutrition traits to nutrient enrichment, we know little about the changes at the community-level. Here, we measured community-level biomass weighted (CWM) and non-weighted (CM) plant N and P concentrations and N:P ratio in a temperate meadow steppe after four years factorial N and P addition, with biomass and nutrition traits of each species in each plot being recorded. Nitrogen addition significantly increased community-level N concentration, decreased P concentration, and enhanced community N:P ratio. Phosphorus addition had no impacts on community-level N concentration, significantly increased P concentration, and reduced community N:P ratio. The impacts of N addition on community nutrition traits were not dependent on P addition and the community-level nutrition trait responses to N and P additions were primarily driven by intraspecific trait variation (ITV) rather than by species turnover. Community-level nutrition traits in the temperate meadow steppe were sensitive to the projected N and P enrichment. While nutrient enrichment had substantially changed community composition, its impacts on community nutrition traits were driven by ITV. Nitrogen deposition would result in imbalance of N and P in plant community, as indicated by the substantial increase in community-level N:P, which was not affected by increased P availability.