Linking nutrient resorption stoichiometry with plant growth under long-term nitrogen addition

Increased nitrogen(N)input can potentially lead to secondary phosphorus(P)limitation;however,it remains unclear whether differences in the plant's ability to cope with this P deficiency are related to their growth responses.Using a long-term experiment of N addition in a boreal forest,we explored the potential role of plant nutrient resorption efficiency and its stoichiometry in mediating plant growth responses to increased N input.We recorded the cover and measured the concentration and resorption efficiency of leaf N and P as well as the photosynthesis of a grass Deyeuxia angustifolia and a shrub Vaccinium vitis-idaea.The cover of the grass D.angustifolia increased with increasing N addition,while that of the shrub V.vitis-idaea decreased with N addition rate and almost disappeared from the high-level N addition over time.P resorption efficiency(PRE)increased in D.angustifolia but decreased in V.vitis-idaea with increasing leaf N:P which was increased by N addition for both species.In addition,photosynthesis increased linearly with N resorption efficiency(NRE)and PRE but was better explained by NRE:PRE,changing nonlinearly with the ratio in a hump-shaped trend.Furthermore,the variance(CV)of NRE:PRE for V.vitis-idaea(123%)was considerably higher than that for D.angustifolia(29%),indicating a more stable nutrient resorption stoichiometry of the grass.Taken together,these results highlight that efficient P acquisition and use strategy through nutrient resorption processes could be a pivotal underlying mechanism driving plant growth and community composition shifts under N enrichment.

Effects of nutrient limitations on the sinking velocity of Thalassiosira weissflogii

The sinking of diatoms is critic al to the formation of oceanic biological pumps and coastal hypoxic zones.However,little is known about the effects of different nutrient restrictions on diatom sinking.In this study,we measured the sinking velocity(SV) of Thalassiosira weissflogii using a new phytoplankton video observation instrument and analyzed major biochemical components under varying nutrient conditions.Our results showed that the SV of T.weissflogii under different nutrient limitation conditions varied substantially.The highest SV of(1.77±0.02) m/d was obtained under nitrate limitation,signific antly surpassing that under phosphate limitation at(0.98±0.13) m/d.As the nutrient limitation was released,the SV steadily decreased to(0.32±0.03) m/d and(0.15±0.05) m/d,respectively.Notably;under conditions with limited nitrate and phosphate concentrations,the SV values of T.weissflogii significantly positively correlated with the lipid content(P <0.001),with R^(2) values of 0.86 and 0.69,respectively.The change of the phytoplankton SV was primarily related to the intracellular compo sition,which is controlled by nutrient conditions but did not significantly correlate with transparent extracellular polymer and biosilica contents.The results of this study help to understand the regulation of the vertical sinking process of diatoms by nutrient restriction and provide new insights into phytoplankton dynamics and their relationship with the marine nutrient structure.

Nutrient Limiting Factor and Agronomic Efficiency of Wheat in Fluvo-aquic Soil District in Northwest of Shandong Province

[Objective] This study aimed to provide basis for rational fertilizer application of wheat in fluvo-aquic soil in the northwest of Shandong Province.[Method] In this paper,the treatments of reduced N,P and K were set in order to explore the effects of fertilizer recommendation based on ASI systematic approach on wheat yield,agronomic efficiency and recovery rate of nutrients.[Result] Nitrogen was the main limiting factor for wheat production in that area,followed by phosphorus,and the third was potassium.Compared with the optimum treatment （OPT）,the reduction of N,P and K reduced the grain yield obviously,which came up to 22.4％,14.4％ and 13.4％ respectively.There were no obvious differences in grain yield among Farmer＇s Fertilization Practice （FP）,60％ OPT-N and OPT treatment.[Conclusion] Agronomic efficiency of N,P and K was 6.3,12.9 and 10 kg/kg respectively.The recovery rates of N,P and K in wheat season were 16.41％,17.27％ and 27.27％ respectively.

Nutrient Limiting Factors in Acidic Vegetable Soils

Nutrient limiting factors in acidic soils from vegetable fields of the Chongqing suburbs of China were assessed by employing the systematic approach developed by Agro Services International (ASI) including soil testing, nutrient adsorption study, and pot and field experiments to verify the results of soil testing, with a conventional soil test (CST) used for comparison. The ASI method found the moderately acidic soil (W01) to be N and P deficient; the strongly acidic soil (W04) to be N, K and S deficient; and the slightly acidic soil (W09) to be N, K, S, Cu, Mn, and Zn deficient. The CST method showed that W01 had P, B and Cu deficiencies; W04 had N, P and S deficiencies; and W09 had N, P, S, B, Cu, and Zn deficiencies. There were differences between the two methods. Among the two indicator plants selected, the response of sorghum on the three representative acidic soils was more closely related to the ASI results than that of sweet pepper.

Fertilization for High Yields in Corn-Sweet Potato-Wheat Rotation: A Systematic Approach to Nutrient Limiting Factors of Soils in Chongqing, China

A systematic approach was adopted to investigate the nutrient limiting factors in gray-brown purple soils and yellow soils derived from limestone in Chongqing, China, to study balanced fertilization for corn, sweet potato and wheat in rotation. The results showed that N, P and K were deficient in both soils, Cu, Mn, S andZn in the gray-brown purple soils and Ca, Mg, Mo and Zn for the yellow soils. Balanced fertilizer application increased yields of corn, sweet potato and wheat by 28.4%, 28.7% and 4.4%, respectively, as compared to the local farmers' practice. The systematic approach can be considered as one of the most efficient and reliable methods in fertility study.

Nutrient retranslocation strategies associated with dieback of Pinus species in a semiarid sandy region of Northeast China

In the semiarid sandy region of Northeast China,Mongolian pine(Pinus sylvestris var.mongolica)suffers dieback after the age of 35,while Japanese red pine(Pinus densiflora)and Chinese pine(Pinus tabuliformis)stay healthy.Foliar nutrient retranslocation reflects the nutrient conservation and utilization mechanism of plants in response to their habitats.However,the nutrient retranslocation strategies employed by three Pinus tree species to cope with nutrient limitations remain largely unknown.For this study,we investigated the seasonal variations in nitrogen(N)and phosphorus(P)concentrations of Mongolian pine,Japanese red pine,and Chinese pine plantations in terms of the green needles of all ages,senesced needles,and soil.Further,the N retranslocation efficiency(NRE),and P retranslocation efficiency(PRE),and correlations between the N:P ratios of needles and soil were analyzed.The results showed that,except for the spring NRE in 1-year-old needles of Mongolian pine,the spring NRE and PRE in 1-and 2-year-old needles of the three tree species were greater than zero.The autumn PRE was higher than zero for Mongolian pine,but lower than zero for Japanese red pine and Chinese pine.Among the three Pinus species,Mongolian pine showed greater spring PRE in 2-year-old needles,and PRE from 1-to 2-yearold needles,and from 2-year-old needles to litter.However,Japanese red pine had higher P concentrations and lower N:P ratios in senesced needles,while greater PRE was found in Chinese pine litter.Significant relationships between the N:P ratios were found in the current year and 1-year-old needles and soil in the Mongolian pine plantation,while there was an insignificant relationship between the N:P ratios of the needles and soil in the Chinese pine plantation.Thus,for Mongolian pine,the removal of P from needles in autumn,and higher P translocation from older needles under P-deficient soil may have contributed to the tree dieback.In contrast,Japanese red pine and Chinese pine stored P in their needles during autumn.Japanese red pine returned more P to the soil via litter,while Chinese pine maintained N:P homeostasis and increased P withdrawal prior to needle abscission.

Concentrations of foliar and surface soil in nutrients Pinus spp. plantations in relation to species and stand age in Zhanggutai sandy land, northeast China

The concentrations of the foliar and surface soil nutrients and the variation with species and stand age were studied inPinus spp. plantations in Zhanggutai area, northeast China. The results showed that the total N, total P and C: N ratio of the soil inP. sylvestris var.mongolica stands were significantly higher in comparison with those inP. tabulaeformis andP. densiflora stands. ForP. sylvestris var.mongolica, the foliar P concentration appeared to decrease with age, and the foliar N and K concentrations did not show a consistent change with age. As for the different tree species of the similar age, the foliar N and P concentrations were significantly different (p<0.05), being withP. sylvestris var.mongolica>P. densiflora>P. tabulaeformis. The foliar N: P ratio ofP. densiflora significantly was higher thanP. sylvestris var.mongolica andP. tabulaeformis, while the foliar K was no obvious difference between the three tree species. There were significant correlation (p<0.05) between soil total N and P, soil organic matter and total P, foliar N and P, but it did not show significant correlations between soil and foliar nutrient concentrations, which might attribute to the excessive litter raking, overgrazing and low soil moisture in this area. Based on the foliar N: P ratio, we introduced a combination threshold index of N: P ratio with their absolute foliar nutrient concentrations to determine the possible limiting nutrient. According to the critical N: P ratio and their absolute foliar N, P concentrations, theP. sylvestris var.mongolica stands showed a decreased N limitation degree with age, theP. densiflora stands showed unlimited by N and P in the whole, and theP. tabulaeformis stands showed co-limited by N and P. No significant difference in soil nutrient concentrations of the surface soils was found between 45, 29, 20-yr-oldPinus sylvestris var.mongolica plantation stands. Keywords coniferous trees - foliar nutrient concentration - limiting nutrients - N - P ratio - Zhanggutai sandy land CLC number S718.55 Document code A Article ID 1007-662X(2004)01-0011-08 Foundation item: This research was supported by Key Knowledge Innovation Project (KZCX3-SW-418) of Chinese Academy of Sciences.Biography: CHEN Guang-sheng (1978-), male, master candidate in Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, P. R. ChinaResponsible editor: Song Funan

Evaluation of nutrient limitation in aquatic ecosystems with nitrogen fixing bacteria

There has always been a great need for simple and accurate bioassays for evaluating nutrient limitation in aquatic ecosystems. Whereas organic carbon is usually considered to be the limiting nutrient for microbial growth in many aquatic ecosystems, there are, however, many water sources that are limited by phosphorus or nitrogen. A method named ＂nitrogen fixing bacterial growth potential＂ （NFBGP） test, which is based on pre-culturing of autochthonous （target） microorganisms was described. The method was applied to evaluate phosphorus or nitrogen nutrient limitation in lake and sewage water samples using an isolate of the nitrogen fixing bacterium, Azorhizobium sp. WS6. The results corresponded well to those from the traditional algal growth potential （AGP） test and the bacterial regrowth potential （BRP） test, suggesting that the NFBGP test is a useful supplementary method for evaluating the limiting nutrient, especially phosphorus, in an aquatic environment.

Nutrient composition and distributions in coastal waters impacted by the Changjiang plume

Four cruises were conducted during 2002--2003 in the Changjiang Estuary and adjacent coastal areas. The data presented show a clear coast to open sea gradient in nutrients related to the river inputs. Maximum values of chlorophyll a were typically observed at intermediate salinities at surface water and coincided with non-conservative decreases in nutrients along the salinity gradient, indicating that removal of nutrients was related to phytoplankton uptake. The seasonal variations of nutrient concentrations were just opposite to those of chlorophyll a, indicating that the seasonal variations of nutrients were mainly controlled by phytoplankton uptake, whereas riverine inputs merely weakened or balanced its extent. During the estuarine mixing, phosphate demonstrated some remobilization during all the four cruises; whereas both conservative and non-conservative behaviors for dissolved inorganic nitrogen and silicate were observed in the study area, indicating that both biotic and abiotic events may affect their behaviors during the estuarine mixing. Under the influence of freshwater inputs with high value of ratio of nitrogen to phosphorus, the estuarine and coastal waters impacted by the Changjiang plume were high （ 〉 30） in ratio of nitrogen to phosphorus, but rates of primary production were apparently not constrained by any kind of nutrient elements. However, the low （ 〈 1 ） ratio of silicate to nitrogen in most of the study area might be linked with the rapidly increasing frequency of harmful algal bloom （HAB） incidents in recent years in the coastal waters impacted by the Changjiang plume.

Model study on Bohai ecosystem 2. Annual cycle of nutrient-phytoplankton dynamics

Using the coupled bio-physical model described in the first paper of this series of studies, the annual variations of algae biomass and nutrient concentration in the Bohai Sea are simulated. Modeled results show that the onset of spring bloom is induced by high nutrient stocks stored in winter, though the initial time is earlier in shallow waters than in deep waters, for which the evolution of the vertical stratification in deep waters plays an important role; on the other hand, newly added river-berne nutrients and resuspending sediment-berne nutrients are responsible for the outburst of autumn blooms. On the basis of modeled results, it is also found that the BS ecosystem, as a whole, is limited by nitrogen all the year round, though the phosphorus limitation is apparent in the Laizhou Bay where the ratio of nitrogen concentration to phosphorus concentration is higher than 16 due to the contribution of newly added nutrient species from Huanghe River discharges.

Response of nutrients and the surface phytoplankton community to ice melting in the central Arctic Ocean

During the fourth Chinese National Arctic Research Expedition cruise in summer 2010, a time-series observation was carried out to examine the response of nutrients and phytoplankton community in the ice-water interface to the ice melting ice in the central Arctic Ocean. Phosphate and silicate in the ice-water interface were rich relative to dissolved inorganic nitrogen （DIN）, based on the Redfield ratio （16N：1P：16Si）, suggesting that DIN was the potential limiting nutrient. DIN concentrations in the sea ice were about 3-4 times that in the surface seawater, indicating that melting ice delivered DIN to the surface water. Pigment analysis showed that fucoxanthin and chlorophyll a contribute to carotenoids and chlorophylls in particles. The mean concentrations of chlorophyll c, diatoxanthin, diadinoxanthin and fucoxanthin from 15 August to 18 August were 6 ug.m-3, 22 ug.m-a, 73 ug.m-3 and 922 ug.m-3, respectively, suggesting that diatoms dominated in the phytoplankton community composition. Furthermore, a notable enhancement in fucoxanthin and chlorophyll a during a large-scale ice melting was likely attributed to senescent diatoms released from the bottom sea-ice as well as phytoplankton diatoms growth in the water column due to the input of nutrients （i.e., DIN） and reducing light limitation from melting ice. Temporal distribution patterns of prasinoxanthin and lutein differed from fucoxanthin, indicating that the response of green algae and diatoms to ice melting were different.

Study on Nutrient Limitation of Phytoplankton in the Field Experiment of the Yangtze River Estuary in Summer

From July 23rd to August 15th, 2001, a field cultivation experiment was carried out to determine the limitation factors of phytoplankton in the Yangtze River estuary and the adjacent areas. The results indicated that the potential limiting nutrient was phosphorus in the Yangtze River diluted water area, nitrogen in the offshore of the Yangtze River estuary and the conversion of phosphorus to nitrogen in the middle area. Iron and silicon were not the potential limiting factors. If there were some kinds of limiting factors in the water, the growth of phytoplankton would be limited obviously. In case of disappearance of the limiting factor, the phytoplankton would grow fast. When the Noctiluca scintiuans bloom occurred, the phytoplankton biomass level was very low in a short time due to the grazing pressure. When the grazing pressure disappeared, the phytoplankton would grow quickly in abundant nutrients condition.

Species divergence in seedling leaf traits and tree growth response to nitrogen and phosphorus additions in an evergreen broadleaved forest of subtropical China

Tree competitiveness generally depends on trait plasticity in response to environmental change.The effects of nitrogen(N)and phosphorus(P)on leaf trait variability by species is poorly understood,especially in China’s subtropical forests.This study examined the seedling leaf traits and net primary productivity of all trees>5 cm DBH of two dominant species,Schima superba and Castanopsis carlesii,in an evergreen broadleaved forest fertilized with nitrogen(+N),phosphorus(+P),and nitrogen plus phosphorus(N+P).The effect of N on seedling leaf traits was stronger than P,while fertilization in general was species dependent.Leaf mass per unit area decreased with N for S.superba seedlings but not for C.carlesii.Leaf N,P,and N/P ratios changed with N addition for both species.All four N fractions of carboxylation,bioenergetics,cell wall,and other N metabolites in C.carlesii leaves responded significantly to fertilization,while only the cell wall in S.superb a leaves responded.Other leaf functional traits,including light-saturated photosynthetic rates,water,N,and P use efficiencies,chlorophyll and non structural carbohydrate contents increased with N addition in S.superb a and by P addition in C.carlesii.Canopy closure at the stand-level increased due to N.Litter biomass and relative growth rate of S.superb a was not affected by any treatments,while both for C.carlesii significantly decreased with N+P addition.Collectively,nutrient limitation may vary at a small scale among species in a subtropical forest based on their responses of seedling traits and net primary productivity to fertilization.Seedling traits are not correlated with the net primary productivity of larger trees except for N fractions,because low light conditions induced by fertilization reduces the proportion of N allocated to photosynthesis in seedlings.In addition,acclimation differences of tree species may increase the uncertainty of community succession.

Application of N: P Stoichiometry to Ecology Studies

Stoichiometry has long been addressed in the studies of ecosystem ecology, but it was almost ignored for a long time. Until recently, ecologists have become aware that stoichiometry could provide a new tool to study ecology from genes to the biosphere. Among this trend, N:P stoichiometry is used actively in ecological interactions since nitrogen (N) and phosphorus (P) are the two most important elements in most ecosystems. This article reviews the application of N:P stoichiometry to the studies of ecological problems at different levels, including ecosystem, community and species. Meanwhile, we also provide the cellular basis of N:P stoichiometry, identify the shortages in the use of N:P stoichiometry theory, and put forward some perspectives for future research to be conducted.

Phosphorus limitation in biofiltration for drinking water treatment

Bacterial growth potential(BGP) method and two parallel pilot scale biofilters were used to investigate phosphorus limitation and its effect on the removal of organic matters in biofiltration for drinking water treatment. Addition of phosphorus can substantially increase the BGPs of the samples. Its effect was equivalent to that of addition of a mixture of various inorganic nutrients including phosphorus. The biofilter with phosphate added into its influent performed a higher biological stability of the effluent and a higher COD Mn removal than the control filter. These results suggested that phosphorus was the limiting nutrient in the biofiltration and the removal efficiency of organic matters could be improved by adding phosphate into the influent.

Improving Removal Efficiency of Organic Matters by Adding Phosphorus in Drinking Water Biofiltration Treatment

Objective To investigate phosphorus limitation and its effect on the removal efficiency of organic matters in drinking water biological treatment. Methods Bacterial growth potential (BGP) method and a pair of parallel pilot-scale biofilters were used for the two objectives, respectively. Results The addition of phosphorus could substantially increase the BGPs of the water samples and the effect was stronger than that of the addition of carbon. When nothing was added into the influents, both CODMn removals of the parallel biofilters (BF1 and BF2) were about 15%. When phosphate was added into its influent, BF1 performed a CODMn removal, 6.02 percentage points higher than the control filter (BF2) and its effluent had a higher biological stability. When the addition dose was <20μg. L-1, no phosphorus pollution would occur and there was a good linear relationship between the microbial utilization of phosphorus and the removal efficiency of organic matters. Conclusions Phosphorus was a limiting nutrient and its limitation was stronger than that of carbon. The addition of phosphate was a practical way to improve the removal efficiency of organic matters in drinking water biological treatment.

Leaf N and P stoichiometry of 57 plant species in the Karamori Mountain Ungulate Nature Reserve,Xinjiang,China

Nitrogen （N） and phosphorus （P） are the major nutrients that constrain plant growth and development, as well as the structure and function of ecosystems. Hence, leaf N and P patterns can contribute to a deep understanding of plant nutrient status, nutrient limitation type of ecosystems, plant life-history strategy and differentiation of functional groups. However, the status and pattern of leaf N and P stoichiometry in N-deficiency desert ecosystems remain unclear. Under this context, the leaf samples from 57 plant species in the Karamori Mountain Ungulate Nature Reserve, eastern Junggar Desert, China were investigated and the patterns and interrelations of leaf N and P were comparatively analyzed. The results showed that the average leaf N concentration, P concentration, and N：P ratio were 30.81 mg/g, 1.77 mg/g and 17.72, respectively. This study found that the leaf N concentration and N：P ratio were significantly higher than those of studies conducted at global, national and regional scales; however, the leaf P concentration was at moderate level. Leaf N concentration was allometrically correlated with leaf P and N：P ratio across all species. Leaf N, P concentrations and N：P ratio differed to a certain extent among plant functional groups. C4 plants and shrubs, particularly shrubs with assimilative branches, showed an obviously lower P concentration than those of C3 plants, herbs and shrubs without assimilative branches. Shrubs with assimilative branches also had lower N concentration. Fabaceae plants had the highest leaf N, P concentrations （as well as Asteraceae） and N：P ratio; other families had a similar N, P-stoichiometry. The soil in this study was characterized by a lack of N （total N：P ratio was 0.605）, but had high N availability compared with P （i.e. the available N：P ratio was 1.86）. This might explain why plant leaves had high N concentration （leaf N：P ratio〉16）. In conclusion, the desert plants in the extreme environment in this study have formed their intrinsic and special stoichiometric characteristics in relation to their life-history strategy.

Responses of Dodonaea viscosa growth and soil biological properties to nitrogen and phosphorus additions in Yuanmou dry-hot valley

Nitrogen(N) and phosphorus(P) are limited nutrients in terrestrial ecosystems, and their limitation patterns are being changed by the increase in N deposition. However, little information concerns the plant growth and the soil biological responses to N and P additions among different soils simultaneously, and these responses may contribute to understand plant-soil interaction and predict plant performance under global change. Thus, this study aimed to explore how N and P limitation changes in different soil types, and reveal the relationship between plant and soil biological responses to nutrient additions. We planted Dodonaea viscosa, a globally distributed species in three soil types(Lixisols, Regosols and Luvisols) in Yuanmou dry-hot valley in Southwest China and fertilized them factorially with N and P. The growth and biomass characters of D. viscosa, soil organic matter, available N, P contents and soil carbon(C), N, P-related enzyme activities were quantified. N addition promoted the growth and leaf N concentration of D. viscosa in Lixisols; N limitation in Lixisols was demonstrated by lower soil available N with higher urease activity. P addition promoted the growth and leaf P concentration of D. viscosa in Luvisols; severe P limitation in Luvisols was demonstrated by a higher soil available N: P ratio with higher phosphatase activity. Urease activity was negatively correlated with soil available N in Nlimited Lixisols, and phosphatase activity was negatively correlated with soil available P in P-limited Luvisols. Besides, the aboveground biomass and leaf N concentration of D. viscosa were positively correlated with soil available N in Lixisols, but the aboveground biomass was negatively correlated with soil available P. Our results show similar nutrient limitation patterns between plant and soil microorganism in the condition of enough C, and the nutrient limitations differ across soil types. With the continued N deposition, N limitation of the Lixisols in dry hot valleys is expected to be alleviated, while P limitation of the Luvisols in the mountaintop may be worse in the future, which should be considered when restoring vegetation.

Leaf stoichiometry of Leontopodium lentopodioides at high altitudes on the northeastern Qinghai-Tibetan Plateau,China

Altitude affects leaf stoichiometry by regulating temperature and precipitation,and influencing soil properties in mountain ecosystems.Leaf carbon concentration(C),leaf nitrogen concentration(N),leaf phosphorous concentration(P),and their stoichiometric ratios of Leontopodium lentopodioides(Willd.)Beauv.,a widespread species in degraded grasslands,were investigated to explore its response and adaptation strategy to environmental changes along four altitude gradients(2500,3000,3500,and 3800 m a.s.l.)on the northeastern Qinghai-Tibetan Plateau(QTP),China.The leaf C significantly varied but without any clear trend with increasing altitude.Leaf N showed an increasing trend,and leaf P showed a little change with increasing altitude,with a lower value of leaf P at 3500 m than those at other altitudes.Similarity,leaf C:P and N:P exhibited a little change with increasing altitude,which both had greater values at 3500 m than those at other altitudes.However,leaf C:N exhibited a decreasing trend with increasing altitude.Soil NH^(+)_(4)-N,soil pH,soil total phosphorus(STP),mean annual temperature(MAT),and mean annual precipitation(MAP)were identified as the main factors driving the variations in leaf stoichiometry of L.lentopodioides across all altitudes,with NH^(+)_(4)-N alone accounting for 50.8%of its total variation.Specifically,leaf C and N were mainly controlled by MAT,soil pH,and NH^(+)_(4)-N,while leaf P by MAP and STP.In the study area,it seems that the growth of L.lentopodioides may be mainly limited by STP.The results could help to strengthen our understanding of the plasticity of plant growth to environmental changes and provide new information on global grassland management and restoration.

Topography modulates effects of nitrogen deposition on microbial resource limitation in a nitrogen-saturated subtropical forest

Background:Nitrogen(N)saturation theory proposes that an ecosystem might switch from N limitation to carbon(C),phosphorus(P),or other nutrient limitations if it receives continuous N input.Yet,after N limitation is removed,which nutrient is the most limited and whether topography modulates such change is rarely tested at a microbial level.Here,we conducted a two-year N addition experiment under two different topography positions(i.e.a slope and a valley)in a N-saturated subtropical forest.Soil enzyme activity was measured,and ecoenzymatic stoichiometry indexes were calculated as indicators of microbial resource limitation.Results:In the valley,two-year N addition changed the activity of all studied enzymes to various degrees.As a result,microbial C limitation was aggravated in the valley,and consequently microbial decomposition of soil labile organic C increased,but microbial P limitation was alleviated due to the stoichiometry balance.On the slope,however,N addition did not significantly change the activity of the studied enzymes,and did not alter the status of microbial resource limitation.Conclusions:These results indicate that C is a more limited element for microbial growth than P after removing N limitation,but we also highlight that topography can regulate the effect of N deposition on soil microbial resource limitation in subtropical forests.These findings provide useful supplements to the N saturation theory.