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Soil enzyme activity can be affected by both production and degradation processes, as enzymes can be degraded by proteases. However, the impact of nutrient addition on enzyme activity is often solely attributed to changes in enzyme production without fully considering degradation. In this study, we demonstrate that the activities of β-1,4-glucosidase (BG), β-D-cellobiohydrolase (CBH), β-1,4-xylosidase (BX), and β-1,4-N-acetyl-glucosaminidase (NAG) in two tropical plantations exhibited comparable levels between nitrogen (N)- and phosphorus (P)-fertilized soils and the unfertilized control under field conditions. However, it was observed that the reduction in enzymatic activity was significantly higher in the fertilized soils during short-term laboratory incubation in the acacia plantation. Additionally, the eucalyptus plantation exhibited a similar tendency, although statistical significance was not achieved due to the high variance of the data. The results show that the interruption of the natural, continuous supply of organic matter or non-soil microbial-derived enzymes, which typically occurs under field conditions, leads to a more significant reduction in soil enzyme activities in fertilized soils compared to unfertilized control. This may be attributed to the higher abundance of protease in fertilized soils, resulting in faster enzyme degradation. Interestingly, P fertilization alone did not have a similar effect, indicating that N fertilization is likely the main cause of the larger decreases in enzyme activity during incubation in fertilized soils compared to unfertilized control soils, despite our study site being poor in P and rich in N. These findings highlight the importance of considering enzyme degradation when investigating material dynamics in forest ecosystems, including the impact of nutrient addition on enzyme activity, as enzyme production alone may not fully explain changes in soil enzyme activity.

Soil fungi play critical roles in ecosystem processes and are sensitive to global changes. Elevated atmospheric nitrogen (N) deposition has been well documented to impact on fungal diversity and community composition, but how the fungal community assembly responds to the duration effects of experimental N addition remains poorly understood. Here, we aimed to investigate the soil fungal community variations and assembly processes under short- (2 years) versus long-term (13 years) exogenous N addition (∼100 kg N ha–1 yr–1) in a N-rich tropical forest of China. We observed that short-term N addition significantly increased fungal taxonomic and phylogenetic α-diversity and shifted fungal community composition with significant increases in the relative abundance of Ascomycota and decreases in that of Basidiomycota. Short-term N addition also significantly increased the relative abundance of saprotrophic fungi and decreased that of ectomycorrhizal fungi. However, unremarkable effects on these indices were found under long-term N addition. The variations of fungal α-diversity, community composition, and the relative abundance of major phyla, genera, and functional guilds were mainly correlated with soil pH and NO3––N concentration, and these correlations were much stronger under short-term than long-term N addition. The results of null, neutral community models and the normalized stochasticity ratio (NST) index consistently revealed that stochastic processes played predominant roles in the assembly of soil fungal community in the tropical forest, and the relative contribution of stochastic processes was significantly increased by short-term N addition. These findings highlighted that the responses of fungal community to N addition were duration-dependent, i.e., fungal community structure and assembly would be sensitive to short-term N addition but become adaptive to long-term N enrichment.

Anthropogenic elevated nitrogen (N) deposition has an accelerated terrestrial N cycle, shaping soil carbon dynamics and storage through altering soil organic carbon mineralization processes. However, it remains unclear how long-term high N deposition affects soil carbon mineralization in tropical forests. To address this question, we established a long-term N deposition experiment in an N-rich lowland tropical forest of Southern China with N additions such as NH4NO3 of 0 (Control), 50 (Low-N), 100 (Medium-N) and 150 (High-N) kg N ha−1 yr−1, and laboratory incubation experiment, used to explore the response of soil carbon mineralization to the N additions therein. The results showed that 15 years of N additions significantly decreased soil carbon mineralization rates. During the incubation period from the 14th day to 56th day, the average decreases in soil CO2 emission rates were 18%, 33% and 47% in the low-N, medium-N and high-N treatments, respectively, compared with the Control. These negative effects were primarily aroused by the reduced soil microbial biomass and modified microbial functions (e.g., a decrease in bacteria relative abundance), which could be attributed to N-addition-induced soil acidification and potential phosphorus limitation in this forest. We further found that N additions greatly increased soil-dissolved organic carbon (DOC), and there were significantly negative relationships between microbial biomass and soil DOC, indicating that microbial consumption on soil-soluble carbon pool may decrease. These results suggests that long-term N deposition can increase soil carbon stability and benefit carbon sequestration through decreased carbon mineralization in N-rich tropical forests. This study can help us understand how microbes control soil carbon cycling and carbon sink in the tropics under both elevated N deposition and carbon dioxide in the future.

Significance Forest soil carbon (C) storage plays a central role in sequestrating atmospheric CO2 on timescales from centuries to millennia. However, our current understanding of soil C sequestration in response to N deposition mainly focuses on mid-to-high latitudes in the Northern Hemisphere, where N supply typically constrains forest growth. We lack data about changes in soil C stocks in tropical forests, where most ecosystems are N-rich or N-saturated. Using more than a decade of continuous N addition experiment and a meta-analysis, we found that excess N deposition can significantly increase soil C in N-rich tropical forests. However, enhanced C sequestration in tropical soils is not a good reason to justify excess N emissions to the atmosphere., Terrestrial ecosystem carbon (C) sequestration plays an important role in ameliorating global climate change. While tropical forests exert a disproportionately large influence on global C cycling, there remains an open question on changes in below-ground soil C stocks with global increases in nitrogen (N) deposition, because N supply often does not constrain the growth of tropical forests. We quantified soil C sequestration through more than a decade of continuous N addition experiment in an N-rich primary tropical forest. Results showed that long-term N additions increased soil C stocks by 7 to 21%, mainly arising from decreased C output fluxes and physical protection mechanisms without changes in the chemical composition of organic matter. A meta-analysis further verified that soil C sequestration induced by excess N inputs is a general phenomenon in tropical forests. Notably, soil N sequestration can keep pace with soil C, based on consistent C/N ratios under N additions. These findings provide empirical evidence that below-ground C sequestration can be stimulated in mature tropical forests under excess N deposition, which has important implications for predicting future terrestrial sinks for both elevated anthropogenic CO2 and N deposition. We further developed a conceptual model hypothesis depicting how soil C sequestration happens under chronic N deposition in N-limited and N-rich ecosystems, suggesting a direction to incorporate N deposition and N cycling into terrestrial C cycle models to improve the predictability on C sink strength as enhanced N deposition spreads from temperate into tropical systems.

Nitrogen (N) deposition is a component of global change that has considerable impact on belowground carbon (C) dynamics. Plant growth stimulation and alterations of fungal community composition and functions are the main mechanisms driving soil C gains following N deposition in N-limited temperate forests. In N-rich tropical forests, however, N deposition generally has minor effects on plant growth; consequently, C storage in soil may strongly depend on the microbial processes that drive litter and soil organic matter decomposition. Here, we investigated how microbial functions in old-growth tropical forest soil responded to 13 years of N addition at four rates: 0 (Control), 50 (Low-N), 100 (Medium-N), and 150 (High-N) kg N ha-1 year-1 . Soil organic carbon (SOC) content increased under High-N, corresponding to a 33% decrease in CO2 efflux, and reductions in relative abundances of bacteria as well as genes responsible for cellulose and chitin degradation. A 113% increase in N2 O emission was positively correlated with soil acidification and an increase in the relative abundances of denitrification genes (narG and norB). Soil acidification induced by N addition decreased available P concentrations, and was associated with reductions in the relative abundance of phytase. The decreased relative abundance of bacteria and key functional gene groups for C degradation were related to slower SOC decomposition, indicating the key mechanisms driving SOC accumulation in the tropical forest soil subjected to High-N addition. However, changes in microbial functional groups associated with N and P cycling led to coincidentally large increases in N2 O emissions, and exacerbated soil P deficiency. These two factors partially offset the perceived beneficial effects of N addition on SOC storage in tropical forest soils. These findings suggest a potential to incorporate microbial community and functions into Earth system models considering their effects on greenhouse gas emission, biogeochemical processes, and biodiversity of tropical ecosystems.

Soil ecoenzymes play an important role in the decomposition and formation of soil organic matter, and are often considered to be derived from soil microbial and plant root activities. This study found another potentially-important source, transfer of ecoenzymes in phyllosphere to soil via throughfall. Throughfall samples were collected in five subtropical forests, and six ecoenzymes were determined therein. Data from this study showed that the ecoenzyme activity in throughfall was significantly higher than that in rainfall. Although this study has the limitation of a single measurement, it is the first report of the potential importance of ecoenzymes in throughfall in forest ecosystems.

Asymbiotic nitrogen (N) fixation (ANF) is an important source of N in pristine forests and is predicted to decrease with N deposition. Previous studies revealing N fixation in response to N deposition have mostly applied understory N addition approaches, neglecting the key processes (for example, N retention and uptake) occurring in forest canopy. This study evaluated the effects of N deposition on N fixation in the soil, forest floor, mosses, and canopy leaves in a temperate forest (in central China) and a tropical forest (in southern China) with different treatments: control, understory N addition, and canopy N addition. Results showed that total ANF rates were higher in the temperate forest (2.57 ± 0.19 mg N m−2 d−1) than in the tropical forest (1.34 ± 0.09 mg N m−2 d−1). N addition inhibited the soil, forest floor, moss, and foliar N fixation in the temperate forest, whereas it inhibited only the soil N fixation in the tropical forest. Compared to canopy N addition, understory N addition overestimated the inhibitory effects of N deposition on total ANF slightly in the tropical forest (by 35%) but severely in the temperate forest (by 375–472%) due to neglecting canopy retention of N. In summary, our findings indicate that ANF has different rates and sensitivities to N addition between tropical and temperate forests and that understory N addition overestimates the N deposition effects on ANF in forests, particularly in the temperate forest. These findings are important for our accurate understanding and estimate of terrestrial N fixation under N deposition scenarios.

Because potassium (K) is a rock-derived essential element that can be depleted in highly-weathered tropical soils, K availability may limit some portion of soil microbial activity in tropical forest ecosystems. In this paper we tested if K limits microbial activity in the condition of sufficient labile C supply. An incubation experiment was performed using surface soil samples (0–10 cm depth) obtained from four permanent ecological research plots in a natural sub-tropical forest in southern China. Soil samples were taken in September 2016. Heterotrophic soil respiration rates and microbial biomass were measured after the addition of glucose (both D and L) with and without K (potassium chloride). We did not observe any effects of K addition on soil microbial respiration, suggesting that K does not limit the microbial activity in the condition of sufficient labile C supply. The lack of microbial response to added K can be attributed to the high mobility of K in forest ecosystems, which may have provided sufficient K to microbes in our soil samples (already provided at the beginning of the incubation). However, at the present stage, we cannot conclude that K is not a limiting factor of soil microbial activity in other tropical forest ecosystems because of the heterogeneity of tropical forest ecosystems and few observations. The hypothesis needs to be tested in larger numbers of tropical forests.

The ratio of β-1,4-glucosidase (BG) to β-1,4-N-acetylglucosaminidase (NAG) activity (BG:NAG ratio) is often used as an indicator of the relative resource allocation of soil microbes to C acquisition compared with N. An increasing number of recent studies have used this index to assess the nutrient status of microbes. However, the validity of this index for assessing the nutrient status of microbes is not well tested. In this study, we collected published data and tested that validity by investigating whether N fertilization elevated the BG:NAG ratio, assuming that microbes reduce their allocation to the N-acquiring enzyme (NAG) under N-enriched conditions. Of the data points, 54% (82/151) did not support the hypothesis because those studies showed lower BG:NAG ratios in N-enriched soils than under ambient conditions, especially when the ambient BG:NAG ratio was higher than 2.0 (77%, 59/77). This suggests that the BG:NAG ratio does not always indicate the microbial status for C or N limitation. Rather, we hypothesized that the decomposition stage explained the variation in BG:NAG because N addition accelerates decomposition, and the BG:NAG ratio is lower at later stages of decomposition due to the dominance of NAG-targeting C (chitin or peptidoglycan). A negative correlation of BG:NAG ratio with polyphenol oxidase activity, which increases with decomposition, supported our hypothesis.

Background: Soil fungi play critical roles in ecosystem processes and are sensitive to global changes. Elevated atmospheric nitrogen (N) deposition has been well documented to impact on fungal diversity and community composition, but how fungal community assembly respond to short- and long-term simulative N deposition remains poorly understood. Here, we carried out two field experiments to investigate the soil fungal community variations and assembly processes under short- (2 years) versus long-term (13 years) exogenous nitrogen addition (100 kg N ha-1 yr-1) in a N-rich tropical forest of China. Results: We observed that short-term N addition significantly increased fungal taxonomic and phylogenetic α-diversity, and shifted fungal community composition with significant increases in the relative abundance of Ascomycota and saprotrophic fungi, and decreases in the relative abundance of Basidiomycota and ectomycorrhizal (EcM) fungi. However, unremarkable effects were found under long-term N addition. The variations of fungal α-diversity, community composition, the relative abundance of major phyla, genera and functional guilds were mainly correlated with soil pH and the concentrations of NO3--N, and these correlations were much stronger under short- than long-term N addition. The results of null, neutral community models and the 39 normalized stochasticity ratio (NST) index consistently revealed that stochastic processes played predominant roles in the assembly of soil fungal community under N addition in the tropical forest, and that the relative contributions of stochastic processes were higher at short-term site. Furthermore, both short- and long-term N addition slightly loosened the co-occurrence networks of the fungal community. Conclusions: These findings highlighted that the responses of fungal community structure to N addition were duration-dependent, i.e., the fungal community was sensitive to the short-term N addition but become acclimatized to long-term N enrichment.

We investigated the impacts of 14-year continuous N addition on activities of arylsulfatase (AS) and phosphodiesterase (PDE), which catalyze soil organic sulfur (S) and phosphorus (P), respectively. The response of AS to N addition was compared with that of C- and N-acquiring enzymes, i.e., β-1,4-glucosidase (BG), β-D-cellobiohydrolase (CBH), β-1,4-Xylosidase (BX), β-1,4-N-acetyl-glucosaminnidase (NAG), and leucine amino peptidase (LAP). We also compared the impact of N addition on PDE activity with that on phosphomonoesterase (PME) activity. The results showed that N addition clearly decreased soil AS activity, whereas activities of C- and N-acquiring enzymes did not exhibit similar changes. The inconsistent response of AS activity with the enzymes was attributed to soil acidification induced by long-term N addition, which shifts the pH condition to a more optimal condition for AS activity and accelerates the accumulation of S in soils. The ratio of PME to PDE were significantly elevated by N addition, implying that the microbial and plant resource allocation to PME relative to PDE increased after N addition at our study site. The shift in resource allocation may have occurred because (i) the types of phosphatases secreted by biota shifted toward phosphatases that require one-step reaction to obtain P (such as PME) in the more-severely P-depleted condition, or (ii) soil acidification provided a more optimal condition for PDE than PME, which resulted in lower PDE production appearing as lower enzyme activity at the same pH condition in the laboratory. Overall, our results indicated different responses of exoenzyme activities to external N input. This highlights the importance of atmospheric N deposition on microbial activity and collateral C and nutrient dynamics in tropical natural forests.

Lowland tropical forests with chronic nitrogen (N) deposition and/or abundant N-fixing organisms are commonly rich in N relative to other nutrients. The tropical N richness introduces a paradoxical relationship in which many tropical forests sustain high rates of asymbiotic N fixation despite the soil N richness and the higher energy cost of N fixation than of soil N uptake. However, the mechanism underlying this phenomenon remains unclear. Our study aims to test this phenomenon and examine potential mechanisms of nutrient concentrations vs. substrate stoichiometry in regulating N fixation using multiple linear regression models. We hypothesized that the rates of asymbiotic N fixation would be low in an N-rich forest under N deposition and substrate stoichiometry would explain the variation in N fixation better than nutrient concentrations. We conducted a chronic N-addition experiment in an N-saturated tropical forest in southern China and measured the N fixation rates, carbon (C), N, and phosphorus (P) concentrations, and stoichiometry in different substrates (soil, forest floor, mosses, and canopy leaves). Total N fixation rates were high (10.35-12.43 kg N·ha-1 ·yr-1 ) in this N-saturated forest because of the high substrate C:N and N:P stoichiometry (which explained 13-52% of the variation in N fixation, P 0.05). Atmospheric N deposition (34-50 kg N·ha-1 ·yr-1 ) failed to down-regulate asymbiotic N fixation in this forest possibly because the N deposition rate was insufficient to inhibit N fixation or N deposition maintained high N fixation rates by increasing C sequestration in the substrates. Our N-addition experiment showed the insensitivity of N fixation in all the tested substrates to low N addition (50 kg N·ha-1 ·yr-1 ); however, medium and high N addition (100-150 kg N·ha-1 ·yr-1 ) stimulated the moss and foliar N fixation because of the increases in substrate C:N stoichiometry (which explained 30-34% of the variation in N fixation, P < 0.001). Overall, our results emphasize the importance of substrate (particularly mosses and foliage) stoichiometry as a driver of asymbiotic N fixation and sustained N richness in lowland tropical forests.

Nitrogen (N) deposition induces soil acidification in natural forests; however, whether it increases soil acidity in tropical plantations with simple tree structures compared with natural forests remains unclear. This study aimed to investigate the effects of N deposition on the soil acidity of tropical broadleaf plantations dominated by Acacia auriculiformis and Eucalyptus urophylla in South China, which has been enduring N deposition for over 30 years, and investigate the reasons for the changes in soil acidity. Long-term N addition did not affect soil acidity in the two plantations, with no significant changes in soil pH values, and exchangeable non-acidic and acidic cation concentrations. Long-term N deposition did not significantly affect the plant and total soil N concentrations, but significantly increased the soil nitrous oxide emission rates and total dissolved N concentrations in the soil solutions. Our findings indicate that most of the added N was lost via leaching and emissions, such that long-term N addition did not exacerbate soil acidification in broadleaf plantations, thereby providing novel insight into the effects of atmospheric N deposition on forest ecosystems. Overall, our study indicates that long-term N deposition does not always lead to soil acidification in tropical forests, as previously expected.

Natural abundance of 15N (δ15N) in plants and soils can provide time-integrated information related to nitrogen (N) cycling within ecosystems, but it has not been well tested in warm and humid subtropical forests. In this study, we used ecosystem δ15N to assess effects of increased N deposition on N cycling in an old-growth broad-leaved forest and a secondary pine forest in a high-N-deposition area in southern China. We measured δ15N of inorganic N in input and output fluxes under ambient N deposition, and we measured N concentration (%N) and δ15N of major ecosystem compartments under ambient deposition and after decadal N addition at 50 kg N ha−1yr−1, which has a δ15N of −0.7 ‰. Our results showed that the total inorganic N in deposition was 15N-depleted (−10 ‰) mainly due to high input of strongly 15N-depleted NH4+-N. Plant leaves in both forests were also 15N-depleted (−4 to −6 ‰). The broad-leaved forest had higher plant and soil %N and was more 15N-enriched in most ecosystem compartments relative to the pine forest. Nitrogen addition did not significantly affect %N in the broad-leaved forest, indicating that the ecosystem pools are already N-rich. However, %N was marginally increased in pine leaves and significantly increased in understory vegetation in the pine forest. Soil δ15N was not changed significantly by the N addition in either forest. However, the N addition significantly increased the δ15N of plants toward the 15N signature of the added N, indicating incorporation of added N into plants. Thus, plant δ15N was more sensitive to ecosystem N input manipulation than %N in these subtropical forests. We interpret the depleted δ15N of plants as an imprint from the high and 15N-depleted N deposition that may dominate the effects of fractionation that are observed in most warm and humid forests. Fractionation during the steps of N cycling could explain the difference between negative δ15N in plants and positive δ15N in soils, and the increase in soil δ15N with depths. Nevertheless, interpretation of ecosystem δ15N from high-N-deposition regions needs to include data on the deposition 15N signal.

The prevailing paradigm for soil microbial activity in tropical forests is that microbial activity is limited by phosphorus (P) availability. Thus, exogenous P addition should increase rates of organic matter decomposition. Studies have also confirmed that soil respiration is accelerated when P is added experimentally. However, we hypothesize that the increased rates of soil microbial respiration could be due to the release of organic material from the surface of soil minerals when P is added, because P is more successful at binding to soil particles than organic compounds. In this study, we demonstrate that P addition to soil is associated with significantly higher dissolved organic carbon (DOC) content in a tropical evergreen forest in southern China. Our results indicate that P fertilization stimulated soil respiration but suppressed litter decomposition. Results from a second sorption experiment revealed that the recovery ratio of added DOC in the soil of a plot fertilized with P for 9 years was larger than the ratio in the soil of a non-fertilized plot, although the difference was small. We also conducted a literature review on the effects of P fertilization on the decomposition rates of litter and soil organic matter at our study site. Previous studies have consistently reported that P addition led to higher response ratios of soil microbial respiration than litter decomposition. Therefore, experiments based on P addition cannot be used to test whether microbial activity is P-limited in tropical forest soils, because organic carbon desorption occurs when P is added. Our findings suggest that the prevailing paradigm on the relationship between P and microbial activity in tropical forest soils should be re-evaluated.

Rates of biological nitrogen fixation (BNF) were measured in ecosystem. In this study, we measured rates of BNF in ecosystem compartments (bulk soil, forest floor, rhizosphere soil, and nodule) in two mature tropical plantations in southern China with legume trees (Acacia auriculiformis, AA) and with non-legume trees (Eucalyptus urophylla, EU) after 4 years of nitrogen (N) fertilization (0, 50, and 100 kg N ha−1 year−1). BNF rates of bulk soil were comparable between plantations, while rates of rhizosphere soil were significantly higher in the EU plantation and rates of forest floor were significantly higher in the AA plantation. Thus, total BNF rates were comparable between plantations (AA = 6.04 kg N ha−1 year−1; EU= 6.42 kg N ha−1 year−1). In the AA plantation, N addition significantly decreased BNF rates in all measured compartments and thus the total rates. In the EU plantation, N addition did not change BNF rates of forest floor, but significantly decreased rates of bulk soil and increased rates of rhizosphere soil; thus, total rates did not change. Our findings provide evidence that forest type is an important factor regulating the effects of external N input on BNF, and suggest that elevated atmospheric N deposition in recent decades will suppress total N fixation in mature forests with legume trees but not in those with non-legume trees.

Nitrogen (N) deposition in China has increased greatly, but the general impact of elevated N deposition on carbon (C) dynamics in Chinese terrestrial ecosystems is not well documented. In this study we used a meta-analysis method to compile 88 studies on the effects of N deposition C cycling on Chinese terrestrial ecosystems. Our results showed that N addition did not change soil C pools but increased above-ground plant C pool. A large decrease in below-ground plant C pool was observed. Our result also showed that the impacts of N addition on ecosystem C dynamics depend on ecosystem type and rate of N addition. Overall, our findings suggest that 1) decreased below-ground plant C pool may limit long-term soil C sequestration; and 2) it is better to treat N-rich and N-limited ecosystems differently in modeling effects of N deposition on ecosystem C cycle.

Differences in nitrogen (N) acquisition patterns between plant species are often reflected in the natural 15N isotope ratios (&delta, 15N) of the plant tissues, however, such differences are poorly understood for co-occurring plants in tropical and subtropical forests. To evaluate species variation in N acquisition traits, we measured leaf N concentration (%N) and &delta, 15N in tree and understory plant species under ambient N deposition (control) and after a decade of N addition at 50 kg N ha&minus, 1 yr&minus, 1 (N-plots) in an old-growth subtropical forest in southern China. We also measured changes in leaf &delta, 15N after one-year of 15N addition in both the control and N-plots. The results show consistent significant species variation in leaf %N in both control and N-plots, but decadal N addition did not significantly affect leaf %N. Leaf &delta, 15N values were also significantly different among the plant species both in tree and understory layers, and both in control and N-plots, suggesting differences in N acquisition strategies such as variation in N sources and dominant forms of N uptake and dependence on mycorrhizal associations among the co-occurring plant species. Significant differences between the plant species (in both control and N-plots) in changes in leaf &delta, 15N after 15N addition were observed only in the understory plants, indicating difference in access (or use) of deposited N among the plants. Decadal N addition had species-dependent effects on leaf &delta, 15N, suggesting the N acquisition patterns of these plant species are differently affected by N deposition. These results suggest that co-occurring plants in N-rich and subtropical forests vary in their N acquisition traits, these differences need to be accounted for when evaluating the impact of N deposition on N cycling in these ecosystems.

The data presented in this article are referred to the research article “A potential source of soil ecoenzymes: From the phylllosphere to soil via throughfall” (Mori et al., 2019). The data included the activities of β-1,4-glucosidase (BG, EC 3.2.1.21), β- d -cellobiosidase (CBH, EC 3.2.1.91), β-1,4-N-acetyl-glucosaminidase (NAG, EC 3.2.1.52), leucine amino peptidase (LAP, EC 3.4.11.1), polyphenol oxidase (PPO, EC 1.10.3.2), and phosphomonoesterase (PME, EC 3.1.3.2). The informatin of study sites and sampling method are shown in Fig. 1 and 2.

Anthropogenic nitrogen (N) deposition has accelerated terrestrial N cycling at regional and global scales, causing nutrient imbalance in many natural and seminatural ecosystems. How added N affects ecosystems where N is already abundant, and how plants acclimate to chronic N deposition in such circumstances, remains poorly understood. Here, we conducted an experiment employing a decade of N additions to examine ecosystem responses and plant acclimation to added N in an N-rich tropical forest. We found that N additions accelerated soil acidification and reduced biologically available cations (especially Ca and Mg) in soils, but plants maintained foliar nutrient supply at least in part by increasing transpiration while decreasing soil water leaching below the rooting zone. We suggest a hypothesis that cation-deficient plants can adjust to elevated N deposition by increasing transpiration and thereby maintaining nutrient balance. This result suggests that long-term elevated N deposition can alter hydrological cycling in N-rich forest ecosystems.

Summary Nitrogen (N) saturation hypothesis suggests that when an ecosystem reaches N-saturation, continued N input will cause increased N leaching, nitrous oxide (N2O) emission, and N mineralization and nitrification rates. It also suggests that a different element will become the main limiting factor when N saturation has been reached. Although this hypothesis has been tested in temperate forests, whether they can be directly applied to N-saturated tropical forests remain poorly addressed. To test this hypothesis, soil inorganic N, soil N mineralization and nitrification rate, soil N2O emission rate and nitrate (NO3−) leaching rate were measured in an N-saturated old-growth tropical forest in southern China, after 6 years of N and P addition. We hypothesized that N addition would stimulate further N saturation, but P addition might alleviate N saturation. As expected, our results showed that six continuous years of experimental N addition did cause further N saturation, which was indicated by significant increases in soil inorganic N concentration, N2O emission and nitrate (NO3−) leaching. However, in contrast to our expectations, N addition significantly decreased in situ rates of net N mineralization and nitrification, which could be related to associated changes in enzyme activity and microbial community composition. On the other hand, P addition mitigated N saturation, as expected. Soil inorganic N concentration, N2O emission and NO3− leaching decreased significantly after P addition, but the net rates of N mineralization and nitrification were significantly increased. Our results provide a new understanding of the N saturation hypothesis, suggesting that the effects of long-term N deposition on net N mineralization and nitrification rates in N-saturated tropical forests can be negative and that P addition can alleviate N saturation in such tropical systems.

Atmospheric nitrogen (N) deposition has accelerated in the last several decades due to anthropogenic activities, such as nitrogen fertilization, N-fixing plants cultivation and fossil fuel and biomass combustion. Increasing N deposition has become one of the important factors regulating N cycle in forest ecosystems. Forest ecosystems can retain part of deposited N in soil by biotic and abiotic mechanisms, but when N inputs exceed the capacity of soil retention, N losses will aggravate in terms of N oxide emission and/or nitrate leaching. The excess N input has threatened ecosystem health via acidification and eutrophication, causing declines in terrestrial biodiversity and forest productivity in forest ecosystems of Europe and North America. Recently, China has become one of the three areas that undergo severe N deposition in the world. Impacts of N deposition on soil N cycle in Chinese forest ecosystems have received increasing concern. In this paper, we reviewed the processes of soil N cycle and their responses to atmospheric N deposition based on available literature. The objective is to enhance our understanding on how N deposition affects soil N cycle in forest ecosystems and provide scientific information for sustainable forest management. The review mainly includes the following four aspects: (1) processes of soil N cycle and their controlling factors. These processes include biological N fixation (BNF), decomposition, mineralization, nitrification, denitrification, N oxide emission and NO3−N leaching. The controlling factors of these processes are complicated and interactional. Only one of these factors altered may affect soil N cycle. For example, C/N is the factor that controls BNF, decomposition, mineralization and NO3−N leaching. (2) Research methods and current results about studies are related to the impact of N deposition on soil N cycle in forest ecosystems. In general, the research methods are long-term simulated N deposition experiment, N deposition gradient method, roof clean rain method and 15N tracing method. Effects of N deposition on soil N cycle vary depending on different initial N statuses and lengths of experiment. In “N-limited” forests, N deposition tended to have positive effect on soil N cycling processes, such as accelerating litter decomposition rate and N mineralization rate. However, such result generally showed in short-term fertilization experiments. In some long-term fertilization experiments, it showed that the negative effects would rise when the forests reached N saturation. Compared to “N-limited” forests in temperate region, N deposition tended to have negative or neutral effects in “N-rich” tropical forest. For example, N deposition promoted nitrification process in tropical forests. (3) Possible mechanisms for the effect of N deposition on soil N cycle: N deposition can affect soil N cycle through altering the chemical characteristic of forest substrates, the biomass and community composition of plant and microorganism. (4) Current problems and future research needs for the study about the effect of N deposition on soil N cycle: What role does regional diversity, changes in forest type, and interaction of carbon (C), N and phosphorus (P) play on the effect of N deposition on soil N cycle in forest ecosystems deserve our further study in the future.

Soil acidification is defined as the process in which exchangeable cations are leaching and soil H+ concentration is raising thereby increases soil acidity. Changes in soil pH value and acid neutralizing capacity are mainly indicators of soil acidification. Soil acidification is considered to be a serious ecological and environmental issue, which not only reduces soil quality, but also decreases biodiversity of forest ecosystem and induces forest decline. With nitrogen (N) deposition rapidly increasing, its contribution to soil acidification becomes a major concern in the world. However, the impact of increased N deposition on soil acidification is not well addressed highlighting the need for further attention to the issue. In this paper, the studies on forest soil acidification induced by N deposition were reviewed. The factors related to soil acidification driven by N deposition were classified and discussed, which included soil acidic buffering capacity, N components in atmospheric N deposition, climate, plant species in forests, and N status in ecosystem. Iron (Fe) buffering phase and the consequent Fe toxicity occurring to the acidified soil caused by high N deposition were concerned. The scarcity of phosphorus (P) element induced by soil acidification was particularly emphasized. The research methods used to study soil acidification driven by N deposition were also evaluated. In the end we stressed the importance of the study on soil acidification especially in tropical and subtropical regions driven by N deposition and its mechanisms. This paper can serve for maintaining sustainable forest and agricultural ecosystems.

Leguminous tree plantations at phosphorus (P) limited sites may result in excess nitrogen (N) and higher rates of nitrous oxide (N2O) emissions. However, the effects of N and P applications on soil N2O emissions from plantations with N-fixing vs. non-N-fixing tree species have rarely been studied in the field. We conducted an experimental manipulation of N and/or P additions in two plantations with Acacia auriculiformis (AA, N-fixing) and Eucalyptus urophylla (EU, non-N-fixing) in South China. The objective was to determine the effects of N or P addition alone, as well as NP application together on soil N2O emissions from these tropical plantations. We found that the average N2O emission from control was greater in the AA (2.3 ± 0.1 kg N2O–N ha−1 yr−1) than in EU plantation (1.9 ± 0.1 kg N2O–N ha−1 yr−1). For the AA plantation, N addition stimulated N2O emission from the soil while P addition did not. Applications of N with P together significantly decreased N2O emission compared to N addition alone, especially in the high-level treatments (decreased by 18%). In the EU plantation, N2O emissions significantly decreased in P-addition plots compared with the controls; however, N and NP additions did not. The different response of N2O emission to N or P addition was attributed to the higher initial soil N status in the AA than that of EU plantation, due to symbiotic N fixation in the former. Our result suggests that atmospheric N deposition potentially stimulates N2O emissions from leguminous tree plantations in the tropics, whereas P fertilization has the potential to mitigate N-deposition-induced N2O emissions from such plantations.

Elevated anthropogenic nitrogen (N) deposition has greatly altered terrestrial ecosystem functioning, threatening ecosystem health via acidification and eutrophication in temperate and boreal forests across the northern hemisphere. However, response of forest soil acidification to N deposition has been less studied in humid tropics compared to other forest types. This study was designed to explore impacts of long-term N deposition on soil acidification processes in tropical forests. We have established a long-term N-deposition experiment in an N-rich lowland tropical forest of Southern China since 2002 with N addition as NH4 NO3 of 0, 50, 100 and 150 kg N ha(-1) yr(-1) . We measured soil acidification status and element leaching in soil drainage solution after 6-year N addition. Results showed that our study site has been experiencing serious soil acidification and was quite acid-sensitive showing high acidification (pH(H2O)

Aims Elevated nitrogen (N) deposition in tropical regions may accelerate ecosystem phosphorus (P) limitation. However, it is not explicitly addressed that how changes in soil N and P availability affect foliar nutrients and photosynthesis of plants in tropical forests. In this study, we examined the effects of N and P additions on foliar nutrients and net photosynthesis of two dominant understory species, Randia canthioides (R. canthioides) and Cryptocarya concinna (C. concinna) in an N-saturated old-growth tropical forest (>400-year-old) in south ern China. Methods A full factorial NP addition experiment (2

Natural abundance of 15N (δ15N) in plants and soils can provide integrated information on ecosystem nitrogen (N) cycling, but it has not been well tested in warm and humid sub-tropical forests. In this study, we examined the measurement of δ15N for its ability to assess changes in N cycling due to increased N deposition in an old-growth broadleaved forest and a secondary pine forest in a high N deposition area in southern China. We measured δ15N of inorganic N in input and output fluxes under ambient N deposition, and N concentration (N %) and δ15N of major ecosystem compartments under ambient and after decadal N addition at 50 kg N ha−1 yr−1. Our results showed that the N deposition was δ15N-depleted (−12 ‰) mainly due to high input of depleted NH4&plus;-N. Plant leafs in both forest were also δ15N-depleted (−4 to −6 ‰). The old-growth forest had higher plant and soil N %, and was more 15N-enriched in most ecosystem compartments relative to the pine forest. Nitrogen addition did not significantly affect N % in both forests, indicating that the ecosystem pools are already N-rich. Soil δ15N was not changed significantly by the N addition in both forests. However, the N addition significantly increased the δ15N of plants toward the 15N signature of the added N (~ 0 ‰), indicating incorporation of added N into plants. Thus, plant δ15N was sensitive to ecosystem N input manipulation although N % was unchanged in these N-rich sub-tropical forests. We interpret the depleted δ15N values of plants as an imprint from the high and δ15N-depleted N deposition. The signal from the input (deposition or N addition) may override the enrichment effects of fractionation during the steps of N cycling that are observed in most warm and humid forests. Thus, interpretation of ecosystem δ15N values from high N deposition regions need to include data on the deposition δ15N signal.