Clear-cutting of forests affects the nitrogen cycle and the nitrogen isotopic composition of bioavailable ammonium and nitrate in the soil. Here, we have used nitrogen isotopic variations of tree-rings in red oak (Quercus rubra) and white oak (Quercus alba) as indicators of changes in the nitrogen cycle on a local scale. The delta15N values of late-wood from trees at two remnant forest stands in Ontario, Canada, that underwent large-scale tree-clearing and permanent land-use change at different times were measured. Trees from the perimeter of each stand record a marked 1.5-2.5 per thousand increase in the delta15N values of their tree-rings relative to the values in trees from the center of the stand, with the shift synchronous with the tree-clearing and land-use change. This shift was most likely due to increased rates of nitrification and nitrate leaching in the soil as a result of tree-clearing combined with permanent changes in hydrology and probable fertilizer use accompanying the change in land-use. Nitrogen concentration in tree-rings was not affected bytree-clearing and the associated change in land-use. These results indicate that changes in nitrogen cycling in forest ecosystems, whether due to climate change, land-use change, or other environmental changes (increased O3, other atmospheric pollutants, insects, etc.), can be faithfully monitored with nitrogen isotopic compositions of tree-rings and that dendrogeochemical analysis can be incorporated into studies of the effects of long-term anthropogenic effects on forest ecosystems.

The natural abundance of ¹⁵N (δ¹⁵N) in plants and soils is an ideal tool for assessing ecosystem N dynamics. However, many of the mechanisms driving the variability of foliar and soil δ¹⁵N values within and across ecosystems are still unclear. In this study, we analyzed the patterns of N concentrations and δ¹⁵N values in leaves, bulk soils and soil mineral N as well as soil N turnover rates across four temperate and boreal forest ecosystems along a mountain transect. The results showed that plant species and soil properties directly controlled soil δ¹⁵N patterns and climate factors (air temperature and precipitation) indirectly affected foliar δ¹⁵N patterns. Foliar N concentrations varied consistently with the concentrations of soil available N and soil NO₃⁻-N, whereas foliar δ¹⁵N was most closely associated with the δ¹⁵N of soil NH₄⁺, the most abundant form of N in soil solution. ¹⁵N enrichment in surface mineral soil in high elevation forests was mainly attributed to ¹⁵N-enriched organic N accumulation. Furthermore, the foliar enrichment factor (ε_p/s = δ¹⁵N_foliage−δ¹⁵N_soil) was significantly correlated with N transformation and loss rates, and was negatively correlated with the ratio of NH₄⁺ to total inorganic N. These results suggest that foliar δ¹⁵N value and foliar N concentration together accurately reflect the N availability of forest ecosystems. Foliar ε_p/s can act as an integrated proxy to reflect the status of N cycling within or across forest ecosystems. Soil nitrification and species’ NH₄⁺ to NO₃⁻ uptake ratios are key processes controlling foliar δ¹⁵N patterns in N-limited forest ecosystems. Our findings improve the mechanistic understanding of the commonly observed variability in foliar and soil δ¹⁵N within and across forest ecosystems.

Understanding forest carbon cycling responses to atmospheric N deposition is critical to evaluating ecosystem N dynamics. The natural abundance of N-15 (delta N-15) has been suggested as an efficient and non-invasive tool to monitor N pools and fluxes. In this study, three successional forests in southern China were treated with four levels of N addition. In each treatment, we measured rates of soil N mineralization, nitrification, N2O emission and inorganic N leaching as well as N concentration and delta N-15 of leaves, litters and soils. We found that foliar N concentration and delta N-15 were higher in the mature broadleaf forest than in the successional pine or mixed forests. Three-year continuous N addition did not change foliar N concentration, but significantly increased foliar delta N-15 (p < 0.05). Also, N addition decreased the delta N-15 of top soil in the N-poor pine and mixed forests and significantly increased that of organic and mineral soils in N-rich broadleaf forests (p < 0.05). In addition, the soil N2O emission flux and inorganic N leaching rate increased with increasing N addition and were positively correlated with the N-15 enrichment factor (epsilon (p/s)) of forest ecosystems. Our study indicates that delta N-15 of leaf, litter and soil integrates various information on plant species, forest stand age, exogenous N input and soil N transformation and loss, which can be used to monitor N availability and N dynamics in forest ecosystems caused by increasing N deposition in the future.

This paper contrasts the natural and anthropogenic controls on the conversion of unreactive N₂ to more reactive forms of nitrogen (Nr). A variety of data sets are used to construct global N budgets for 1860 and the early 1990s and to make projections for the global N budget in 2050. Regional N budgets for Asia, North America, and other major regions for the early 1990s, as well as the marine N budget, are presented to Highlight the dominant fluxes of nitrogen in each region. Important findings are that human activities increasingly dominate the N budget at the global and at most regional scales, the terrestrial and open ocean N budgets are essentially disconnected, and the fixed forms of N are accumulating in most environmental reservoirs. The largest uncertainties in our understanding of the N budget at most scales are the rates of natural biological nitrogen fixation, the amount of Nr storage in most environmental reservoirs, and the production rates of N₂ by denitrification.

Humans continue to transform the global nitrogen cycle at a record pace, reflecting an increased combustion of fossil fuels, growing demand for nitrogen in agriculture and industry, and pervasive inefficiencies in its use. Much anthropogenic nitrogen is lost to air, water, and land to cause a cascade of environmental and human health problems. Simultaneously, food production in some parts of the world is nitrogen-deficient, highlighting inequities in the distribution of nitrogen-containing fertilizers. Optimizing the need for a key human resource while minimizing its negative consequences requires an integrated interdisciplinary approach and the development of strategies to decrease nitrogen-containing waste.

The nitrogen cycle provides essential nutrients to the biosphere, but its antiquity in modern form is unclear. In a drill core though homogeneous organic-rich shale in the 2.5-billion-year-old Mount McRae Shale, Australia, nitrogen isotope values vary from +1.0 to +7.5 per mil (per thousand) and back to +2.5 per thousand over approximately 30 meters. These changes evidently record a transient departure from a largely anaerobic to an aerobic nitrogen cycle complete with nitrification and denitrification. Complementary molybdenum abundance and sulfur isotopic values suggest that nitrification occurred in response to a small increase in surface-ocean oxygenation. These data imply that nitrifying and denitrifying microbes had already evolved by the late Archean and were present before oxygen first began to accumulate in the atmosphere.

A strong, unambiguous negative trend is found in the nitrogen isotopic composition (delta15N) of nitrate over the industrial period, on the basis of a 100-meter ice core from Summit, Greenland. This record indicates that ice-core nitrate reflects changes in nitrogen oxide (NOx) source emissions and that anthropogenic emissions of NOx have resulted in a 12 per mil decline in delta15N of atmospheric nitrate from preindustrial values to present. Variations in the isotopic composition of nitrate may affect the interpretation of other records of environmental change that are affected by atmospheric nitrate.

Direct or indirect anthropogenic effects on ecosystem nitrogen cycles are important components of global change. Recent research has shown that N isotopes in tree rings reflect changes in ecosystem nitrogen sources or cycles and can be used to study past changes. We analyzed trends in two tree species from a remote and pristine tropical rainforest in Brazil, using trees of different ages to distinguish between the effect of tree age and long-term trends. Because sapwood differed from heartwood in delta(15)N and N concentration and N can be translocated between living sapwood cells, long-term trends are best seen in dead heartwood. Heartwood delta(15)N in Spanish cedar (Cedrela odorata) and big-leaf mahogany (Swietenia macrophylla) increased with tree age, and N concentrations increased with age in Cedrela. Controlling for tree age, delta(15)N increased significantly during the past century even when analyzing only heartwood and after removing labile N compounds. In contrast to northern temperate and boreal forests where wood delta(15)N often decreased, the delta(15)N increase in a remote rainforest is unlikely to be a direct signal of changed N deposition. More plausibly, the change in N isotopic composition indicates a more open N cycle, i.e., higher N losses relative to internal N cycling in the forest, which could be the result of changed forest dynamics.

Deposition of reactive nitrogen (N) from human activities has large effects on temperate forests where low natural N availability limits productivity but is not known to affect tropical forests where natural N availability is often much greater. Leaf N and the ratio of N isotopes (delta(15)N) increased substantially in a moist forest in Panama between ~1968 and 2007, as did tree-ring delta(15)N in a dry forest in Thailand over the past century. A decade of fertilization of a nearby Panamanian forest with N caused similar increases in leaf N and delta(15)N. Therefore, our results indicate regional increases in N availability due to anthropogenic N deposition. Atmospheric nitrogen dioxide measurements and increased emissions of anthropogenic reactive N over tropical land areas suggest that these changes are widespread in tropical forests.

(15)N natural abundances of soil total N, roots and mycorrhizas were studied in surface soil profiles in coniferous and broadleaved forests along a transect from central to northern Europe. Under conditions of N limitation in Sweden, there was an increase in delta(15)N of soil total N of up to 9% from the uppermost horizon of the organic mor layer down to the upper 0-5 cm of the mineral soil. The delta(15)N of roots was only slightly lower than that of soil total N in the upper organic horizon, but further down roots were up to 5% depleted under such conditions. In experimentally N-enriched forest in Sweden, i.e. in plots which have received an average of c. 100 kg N ha(-1) year(-1) for 20 years and which retain less than 50% of this added N in the stand and the soil down to 20 cm depth, and in some forests in central Europe, the increase in delta(15)N with depth in soil total N was smaller. An increase in delta(15)N of the surface soil was even observed on experimentally N-enriched plots, although other data suggest that the N fertilizer added was depleted in(15)N. In such cases roots could be enriched in(15)N relative to soil total N, suggesting that labelling of the surface soil is via the pathway: - available pools of N-plant N-litter N. Under N-limiting conditions roots of different species sampled from the same soil horizon showed similar delta(15)N. By contrast, in experimentally N-enriched forest delta(15)N of roots increased in the sequence: ericaceous dwarf shrubs

稳定性同位素¹⁵N自然丰度法近年来在生态系统氮素循环研究中发挥了和正在发挥着极为重要的作用。在森林生态系统中，自然¹⁵N丰度值是氮循环转换的综合结果，被有的研究者用作生态系统氮饱和状态的指示指标。在本研究中，我们测定了我国西南三个典型森林小流域的植物和土壤的自然¹⁵N丰度值，并进行了比较。植物的自然¹⁵N丰度值的变化范围为-5.0‰~2.7‰,土壤自然¹⁵N丰度值的变化范围为从有机层的 -6.9‰增加到矿质土壤的6.9‰，这些值的变化范围和温带森林生态系统的自然¹⁵N值的变化范围没有明显的区别。在土壤表层，鹿冲关和铁山坪的自然¹⁵N丰度值是明显低于雷公山流域的。在缺氮的鹿冲关流域，土壤表层低的¹⁵N丰度值表明了该流域比较封闭的氮循环状态，而在氮相对饱和的铁山坪流域表层土壤并没有出现¹⁵N富化，这可能是由于大量¹⁵N贫化的氮沉降输入的结果。这些结果表明，土壤的¹⁵N丰度值在我国西南部并不一定能单独作为森林生态系统氮饱和程度的指示指标, 但土壤和植被的δ¹⁵N值对于理解当地森林生态系统的氮循环具有一定的意义。

Variation in stable nitrogen isotope ratios (¹⁵N) was assessed for plants comprising two wetland communities, a bog-fen system and a flood plain, in central Japan. ¹⁵N of 12 species from the bog-fen system and six species from the flood plain were remarkably variable, ranging from –5.9 to +1.1 and from +3.1 to +8.7, respectively. Phragmites australis exhibited the highest ¹⁵N value at both sites. Rooting depth also differed greatly with plant species, ranging from 5cm to over 200cm in the bog-fen system. There was a tendency for plants having deeper root systems to exhibit higher ¹⁵N values; plant ¹⁵N was positively associated with rooting depth. Moreover, an increasing gradient of peat ¹⁵N was found along with depth. This evidence, together with the fact that inorganic nitrogen was depleted under a deep-rooted Phragmites australis stand, strongly suggests that deep-rooted plants actually absorb nitrogen from the deep peat layer. Thus, we successfully demonstrated the diverse traits of nitrogen nutrition among mire plants using stable isotope analysis. The ecological significance of deep rooting in mire plants is that it enables those plants to monopolize nutrients in deep substratum layers. This advantage should compensate for any consequential structural and/or physiological costs. Good evidence of the benefits of deep rooting is provided by the fact that Phragmites australis dominates as a tall mire grass.

Natural (15)N abundance values were measured in needles, twigs, wood, soil, bulk precipitation, throughfall and soil water in a Douglas fir (Pseudotsuga menziesii (Mirb.) and a Scots pine (Pinus sylvestris L.) stand receiving high loads of nitrogen in throughfall (>50 kg N ha(-1) year(-1)). In the Douglas fir stand delta(15)N values of the vegetation ranged between -5.7 and -4.2 per thousand with little variation between different compartments. The vegetation of the Scots pine stand was less depleted in (15)N and varied from -3.3 to -1.2 per thousanddelta(15)N. At both sites delta(15)N values increased with soil depth, from -5.7 per thousand and -1.2 per thousand in the organic layer to +4.1 per thousand and +4.7 per thousand at 70 cm soil depth in the Douglas fir and Scots pine stand, respectively. The delta(15)N values of inorganic nitrogen in bulk precipitation showed a seasonal variation with a mean in NH4(+)-N of -0.6 per thousand at the Douglas fir stand and +10.8 per thousand at the Scots pine stand. In soil water below the organic layer NH4(+)-N was enriched and NO3(-)-N depleted in (15)N, which was interpreted as being caused by isotope fractionation accompanying high nitrification rates in the organic layers. Mean delta(15)N values of NH4(+) and NO3(-) were very similar in the drainage water at 90 cm soil depth at both sites (-7.1 to -3.8 per thousand). A dynamic N cycling model was used to test the sensitivity of the natural abundance values for the amount of N deposition, the (15)N ratio of atmospheric N deposited and for the intrinsic isotope discrimination factors associated with N transformation processes. Simulated delta(15)N values for the N saturated ecosystems appeared particularly sensitive to the (15)N ratio of atmospheric N inputs and discrimination factors during nitrification and mineralization. The N-saturated coniferous forest ecosystems studied were not characterized by elevated natural (15)N abundance values. The results indicated that the natural (15)N abundance values can only be used as indicators for the stage of nitrogen saturation of an ecosystem if the delta(15)N values of the deposited N and isotope fractionation factors are taken into consideration. Combining dynamic isotope models and natural (15)N abundance values seems a promising technique for interpreting natural (15)N abundance values found in these forest ecosystems.

*Humans are increasing both the deposition of reactive nitrogen (N) and concentrations of atmospheric CO(2) on Earth, but the combined effects on terrestrial ecosystems are not clear. In the absence of historical records, it is difficult to know if N availability is currently increasing or decreasing on regional scales. *To determine the nature and timing of past changes in grassland ecosystem dynamics, we measured the composition of stable carbon (C) and N isotopes in leaf tissue from 545 herbarium specimens of 24 vascular plant species collected in Kansas, USA from 1876 to 2008. We also parameterized a simple model of the terrestrial N cycle coupled with a stable isotope simulator to constrain processes consistent with observed patterns. *A prolonged decline in foliar N concentrations began in 1926, while a prolonged decline in foliar delta(15)N values began in 1940. Changes in the difference between foliar and atmospheric C isotopes reveal slightly increased photosynthetic water use efficiency since 1876. *The declines in foliar N concentrations and foliar delta(15)N suggest declining N availability in these grasslands during the 20th century despite decades of anthropogenic N deposition. Our results are consistent with progressive-nitrogen-limitation-type hypotheses where declines in N availability are driven by increased ecosystem N storage as a result of increased atmospheric CO(2).

The use of stable isotopes of N and O in N2O has been proposed as a way to better constrain the global budget of atmospheric N2O and to better understand the relative contributions of the main microbial processes (nitrification and denitrification) responsible for N2O formation in soil. This study compared the isotopic composition of N2O emitted from soils under different tree species in the Brazilian Amazon. We also compared the effect of tree species with that of soil moisture, as we expected the latter to be the main factor regulating the proportion of nitrifier- and denitrifier-derived N2O and, consequently, isotopic signatures of N2O. Tree species significantly affected delta15N in nitrous oxide. However, there was no evidence that the observed variation in delta15N in N2O was determined by varying proportions of nitrifier- vs. denitrifier-derived N2O. We submit that the large variation in delta15N-N2O is the result of competition between denitrifying and immobilizing microorganisms for NO3(-). In addition to altering delta15N-N2O, tree species affected net rates of N2O emission from soil in laboratory incubations. These results suggest that tree species contribute to the large isotopic variation in N2O observed in a range tropical forest soils. We found that soil water affects both 15N and 18O in N2O, with wetter soils leading to more depleted N2O in both 15N and 18O. This is likely caused by a shift in biological processes for 15N and possible direct exchange of 18O between H2O and N2O.

Plant species collected from tundra ecosystems located along a north-south transect from central Alaska to the north coast of Alaska showed large and consistent differences in (15)N natural abundances. Foliar delta(15)N values varied by about 10% among species within each of two moist tussock tundra sites. Differences in (15)N contents among species or plant groups were consistent across moist tussock tundra at several other sites and across five other tundra types at a single site. Ericaceous species had the lowest delta(15)N values, ranging between about -8 to -6 per thousand. Foliar (15)N contents increased progressively in birch, willows and sedges to maximum delta(15)N values of about +2 per thousand in sedges. Soil (15)N contents in tundra ecosystems at our two most intensively studied sites increased with depth and delta(15)N values were usually higher for soils than for plants. Isotopic fractionations during soil N transformations and possibly during plant N uptake could lead to observed differences in (15)N contents among plant species and between plants and soils. Patterns of variation in (15)N content among species indicate that tundra plants acquire nitrogen in extremely nutrient-poor environments by competitive partitioning of the overall N pool. Differences in plant N sources, rooting depth, mycorrhizal associations, forms of N taken up, and other factors controlling plant N uptake are possible causes of variations in delta(15)N values of tundra plant species.

Nutrient enrichment is increasingly affecting many tropical ecosystems, but there is no information on how this affects tree biodiversity. To examine dynamics in vegetation structure and tree species biomass and diversity, we annually remeasured tree species before and for six years after repeated additions of nitrogen (N) and phosphorus (P) in permanent plots of abandoned pasture in Amazonia. Nitrogen and, to a lesser extent, phosphorus addition shifted growth among woody species. Nitrogen stimulated growth of two common pioneer tree species and one common tree species adaptable to both high- and low-light environments, while P stimulated growth only of the dominant pioneer tree Rollinia exsucca (Annonaceae). Overall, N or P addition reduced tree assemblage evenness and delayed tree species accrual over time, likely due to competitive monopolization of other resources by the few tree species responding to nutrient enrichment with enhanced establishment and/or growth rates. Absolute tree growth rates were elevated for two years after nutrient addition. However, nutrient-induced shifts in relative tree species growth and reduced assemblage evenness persisted for more than three years after nutrient addition, favoring two nutrient-responsive pioneers and one early-secondary tree species. Surprisingly, N + P effects on tree biomass and species diversity were consistently weaker than N-only and P-only effects, because grass biomass increased dramatically in response to N + P addition. The resulting intensified competition probably prevented an expected positive N + P synergy in the tree assemblage. Thus, N or P enrichment may favor unknown tree functional response types, reduce the diversity of coexisting species, and delay species accrual during structurally and functionally complex tropical rainforest secondary succession.

Seasonal changes in foliage nitrogen (N) and carbon (C) concentrations and δ¹⁵N and δ¹³C ratios were monitored during a year in Erica arborea, Myrtus communis and Juniperus communis co-occurring at a natural CO₂ spring (elevated [CO₂], about 700 μmol mol⁻¹) and at a nearby control site (ambient [CO₂], 360 μmol mol⁻¹) in a Mediterranean environment. Leaf N concentration was lower in elevated [CO₂] than in ambient [CO₂] for M. communis, higher for J. communis, and dependent on the season for E. arborea. Leaf C concentration was negatively affected by atmospheric CO₂ enrichment, regardless of the species. C/N ratio varied concomitantly to N. Leaves in elevated [CO₂] showed lower δ¹³C, and therefore likely lower water use efficiencies than leaves at the control site, regardless of the species, suggesting substantial photosynthetic acclimation under long-term CO₂-enriched atmosphere. Leaves of E. arborea showed lower values of δ¹⁵N under elevated [CO₂], but this was not the case of M. communis and J. communis foliage. The use of the resources and leaf chemical composition are affected by elevated [CO₂], but such an effect varies during the year, and is species-dependent. The seasonal dependency and species specificity suggest that plants are able to exploit different available water and N resources within Mediterranean sites.

Twentieth-century warming could lead to increases in the moisture-holding capacity of the atmosphere, altering the hydrological cycle and the characteristics of precipitation. Such changes in the global rate and distribution of precipitation may have a greater direct effect on human well-being and ecosystem dynamics than changes in temperature itself. Despite the co-variability of both of these climate variables, attention in long-term climate reconstruction has mainly concentrated on temperature changes. Here we present an annually resolved oxygen isotope record from tree-rings, providing a millennial-scale reconstruction of precipitation variability in the high mountains of northern Pakistan. The climatic signal originates mainly from winter precipitation, and is robust over ecologically different sites. Centennial-scale variations reveal dry conditions at the beginning of the past millennium and through the eighteenth and early nineteenth centuries, with precipitation increasing during the late nineteenth and the twentieth centuries to yield the wettest conditions of the past 1,000 years. Comparison with other long-term precipitation reconstructions indicates a large-scale intensification of the hydrological cycle coincident with the onset of industrialization and global warming, and the unprecedented amplitude argues for a human role.

The effect of interspecific competition and element additions (N and P) on four grassland species (Poa pratensis, Lolium perenne, Festuca valida, Taraxacum officinale) grown under field conditions was studied. Two grasses (L. perenne, F. valida) grown in monoculture (absence of competition) showed lower carbon isotope discrimination (¹³C) and enriched ¹⁵N values. Nitrogen addition (as urea) had inconsistent effects on species ¹³C while caused enrichment of ¹⁵N of P. pratensis and F. valida but strong depletion of ¹⁵N of T. officinale. Phosphorous had no significant effect on ¹³C but depleted ¹⁵N of all species.

Nitrogen enrichment is widely thought to be responsible for the loss of plant species from temperate terrestrial ecosystems. This view is based on field surveys and controlled experiments showing that species richness correlates negatively with high productivity and nitrogen enrichment. However, as the type of nutrient limitation has never been examined on a large geographical scale the causality of these relationships is uncertain. We investigated species richness in herbaceous terrestrial ecosystems, sampled along a transect through temperate Eurasia that represented a gradient of declining levels of atmospheric nitrogen deposition--from approximately 50 kg ha(-1) yr(-1) in western Europe to natural background values of less than 5 kg ha(-1) yr(-1) in Siberia. Here we show that many more endangered plant species persist under phosphorus-limited than under nitrogen-limited conditions, and we conclude that enhanced phosphorus is more likely to be the cause of species loss than nitrogen enrichment. Our results highlight the need for a better understanding of the mechanisms of phosphorus enrichment, and for a stronger focus on conservation management to reduce phosphorus availability.

Southern Ontario receives the highest levels of atmospheric nitrogen (N) deposition in Canada and there are concerns that forests in the region may be approaching a state of ‘N saturation’. In order to evaluate whether potential chemical indices provide evidence of N saturation, 23 hardwood plots were sampled along a modeled N-deposition gradient ranging from 9.3 to 12.8 kg/ha/year. All plots were dominated by sugar maple (Acer saccharum Marsh.) and foliar N and foliar δ¹⁵N were positively correlated with modeled N deposition. However, forest floor N content and the C:N ratio were unrelated to N deposition, but were instead related to soil pH and annual temperature; lower C:N ratios and higher N content in the forest floor were found at the most acidic sites in the cooler, northern part of the study region despite lower N deposition. Likewise, δ¹⁵N values in surface mineral soil and the ¹⁵N enrichment factor of foliage (δ¹⁵N foliage − δ¹⁵N soil) are correlated to soil pH and temperature and not N deposition. Further, potential N mineralization, ammonification, and nitrification in Ontario maple stands were highest in the northern part of the region with the lowest modeled N deposition. Nitrogen cycling in soil appears to be primarily influenced by the N status of the forest floor and other soil properties rather than N deposition, indicating that chemical indices in soil in these hardwood plots may not provide an early indicator of N saturation.

In a 12-year experimental study of nitrogen (N) deposition on Minnesota grasslands, plots dominated by native warm-season grasses shifted to low-diversity mixtures dominated by cool-season grasses at all but the lowest N addition rates. This shift was associated with decreased biomass carbon (C):N ratios, increased N mineralization, increased soil nitrate, high N losses, and low C storage. In addition, plots originally dominated by nonnative cool-season grasses retained little added N and stored little C, even at low N input rates. Thus, grasslands with high N retention and C storage rates were the most vulnerable to species losses and major shifts in C and N cycling.