Effects of nitrogen addition on the abundance and composition of soil ammonia oxidizers in Inner Mongolia Grassland

Nitrogen accumulation in soil is increasing in Inner Mongolia which is resulted mainly from fertilization accompanied by conversion of large area of grasslands to croplands. Ammonia-oxidation is the key step of nitrification which is driven by ammonia-oxidizing microorganisms, and study on the response of ammonia-oxidizing microorganisms is necessary for understanding the effects of nitrogen fertilization on ecosystem functions. In this study, the abundance and community structure of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) under long-term N addition of different rates (0, 1.75, 5.25, 10.5, 17.5, and 28 g N m^?2 yr^?1) in a typical steppe of the Inner Mongolia Grassland were investigated using quantitative real-time PCR, cloning and sequencing based on amoA gene. In addition, soil potential ammonia oxidation rate was analyzed. Our results demonstrated that, with the increase of nitrogen addition rate, soil pH declined gradually from 6.6 to 4.9, and potential ammonia oxidation rate also declined which was positively correlated with soil pH (P < 0.01), while the copy number of bacterial amoA gene increased and positively (P < 0.01) correlated with ammonia concentration in soil. The archaeal amoA gene copy number did not change a lot with N nitrogen addition rate below 10.5 g N/m², but significantly decreased with addition of 17.5 and 28 g N m^?2 yr^?1. Sequencing of clone libraries of treatments revealed that in the treatment without N addition, AOB was dominated by Cluster 3a1 of Nitrosospira with a proportion of 87%, while in the treatment with N addition of 28 g N m^?2 yr^?1, proportion of Cluster 2 increased significantly to 41%. All archaeal amoA sequences were affiliated with the soil/sediment clade, and no significant variation of community structure was found between the treatments without N addition and with 28 g N m^?2 yr^?1 addition rate. In conclusion, this study demonstrated significant effects of nitrogen addition on potential ammonia oxidation rate and compositions of ammonia-oxidation microorganisms, which may have important implications for evaluating the impacts of N accumulation on ecosystem functioning. Further, the effects of pH and ammonia concentration on the ammonia oxidation rate and compositions of ammonia-oxidation microorganisms were discussed.     

辉腾锡勒草原干涸湖泊中氨氧化微生物群落结构分析

【目的】以内蒙古辉腾锡勒草原九十九泉湿地为对象,研究湖泊干涸过程中氨氧化微生物的群落结构及其变化。【方法】通过MPN-PCR定量测定氨氧化古菌(AOA)和氨氧化细菌(AOB)的数量;构建amoA基因克隆文库,进行系统发育分析;结合土壤环境因子,探讨湿地退化过程中影响氨氧化微生物的潜在因素。【结果】依湖泊湿地退水梯度的不同样点中,有75%的样点AOB的数量高于AOA,AOB与AOA的数量比率为0.3-18.1。从湖心到湖岸草原带,AOA和AOB的数量有明显增加,但生物多样性呈降低趋势,二者没有呈现正相关。研究发现,AOB的数量与土壤中NH 4+-N的变化存在良好响应。系统发育分析显示,退化湖泊湿地AOA克隆序列均来自于泉古菌门(Crenarchaeota);AOB的amoA基因的克隆序列大部分与亚硝化单胞菌属(Nitrosomonas)有一定同源性,较少部分与亚硝化螺菌属(Nitrosospira)有一定同源性。【结论】湖泊退水过程增加了湿地土壤氨氧化微生物的数量,而氨氧化微生物的种群丰度有所降低。AOA和AOB群落对湖泊湿地的退化过程做出了响应,其中AOB的响应较为明显,氧化条件和土壤铵浓度的改变可能是促成这种响应的重要原因。     

氨氧化古菌及其对氮循环贡献的研究进展

硝化作用先将氨氮氧化为亚硝酸盐氮并进一步氧化为硝酸盐氮,这一过程是氮进行全球生物化学循环的重要环节。随着氨氧化古菌(Ammonia-oxidizing archaea,AOA)基因组序列中氨单加氧酶编码基因(amoA)的发现以及AOA在实验室条件下的成功培养(包括分离纯化和富集培养),基于分子生物学的研究表明AOA在各种环境广泛存在,且多数生境中它的数量远远超过氨氧化细菌(Ammonia-oxidizing bacteria,AOB)。AOA相对于AOB在氮循环中的贡献也引起了多方面的论证和争论。本文就氨氧化古菌的生态分布、系统进化、生境存在丰度及参与硝化作用等进行综述,指出不同生境AOA的活性及其对氮循环的重要性仍需做进一步的研究。     

Relative contributions of archaea and bacteria to microbial ammonia oxidation differ under different conditions during agricultural waste composting

The aim of this study was to compare the relative contribution of ammonia-oxidizing archaea (AOA) and bacteria (AOB) to nitrification during agricultural waste composting. The AOA and AOB amoA gene abundance and composition were determined by quantitative PCR and denaturing gradient gel electrophoresis (DGGE), respectively. The results showed that the archaeal amoA gene was abundant throughout the composting process, while the bacterial amoA gene abundance decreased to undetectable level during the thermophilic and cooling stages. DGGE showed more diverse archaeal amoA gene composition when the potential ammonia oxidation (PAO) rate reached peak values. A significant positive relationship was observed between the PAO rate and the archaeal amoA gene abundance (R2=0.554; P<0.001), indicating that archaea dominated ammonia oxidation during the thermophilic and cooling stages. Bacteria were also related to ammonia oxidation activity (R2=0.503; P=0.03) especially during the mesophilic and maturation stages.     

Ammonia‐oxidizing archaea: important players in paddy rhizosphere soil?

The diversity (richness and community composition) of ammonia-oxidizing archaea (AOA) and bacteria (AOB) in paddy soil with different nitrogen (N) fertilizer amendments for 5 weeks were investigated using quantitative real-time polymerase chain reaction, denaturing gradient gel electrophoresis (DGGE) jand clone library analysis based on the ammonia monooxygenase α-subunit ( amoA ) gene. Ammonia-oxidizing archaea predominated among ammonia-oxidizing prokaryotes in the paddy soil, and the AOA:AOB DNA-targeted amoA gene ratios ranged from 1.2 to 69.3. Ammonia-oxidizing archaea were more abundant in the rhizosphere than in bulk soil. Rice cultivation led to greater abundance of AOA than AOB amoA gene copies and to differences in AOA and AOB community composition. These results show that AOA is dominant in the rhizosphere paddy soil in this study, and we assume that AOA were influenced more by exudation from rice root (e.g. oxygen, carbon dioxide) than AOB.     

Effect of Different Ammonia Concentrations on Community Succession of Ammonia-oxidizing Microorganisms in a Simulated Paddy Soil Column

Ammonia oxidation is performed by both ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA). To explore the effect of ammonia concentration on the population dynamic changes of ammonia-oxidizing microorganisms, we examined changes in the abundance and community composition of AOA and AOB in different layers. Most of the archaeal amoA sequences were Nitrosotalea-related and the proportion that Nitrosotalea cluster occupied decreased in the surface layer and increased in the deep layer during the cultivation process. Nitrosopumilus-related sequences were only detected in the deep layer in the first stage and disappeared later. Both phylogenetic and quantitative analysis showed that there were increased Nitrosomonas-related sequences appeared in the surface layer where the ammonia concentration was the highest. Both AOA and AOB OTU numbers in different layers decreased under selective pressure and then recovered. The potential nitrification rates were 25.06 μg·N·L(-1)·g(-1) dry soil·h(-1) in the mid layer which was higher than the other two layers. In general, obvious population dynamic changes were found for both AOA and AOB under the selective pressure of exogenous ammonia and the changes were different in three layers of the soil column.     

Simazine application inhibits nitrification and changes the ammonia-oxidizing bacterial communities in a fertilized agricultural soil

s-Triazine herbicides are widely used for weed control, and are persistent in soils. Nitrification is an essential process in the global nitrogen cycle in soil, and involves ammonia-oxidizing Bacteria (AOB) and ammonia-oxidizing Archaea (AOA). In this study, we evaluated the effect of the s-triazine herbicide simazine on the nitrification and on the structure of ammonia-oxidizing microbial communities in a fertilized agricultural soil. The effect of simazine on AOB and AOA were studied by PCR-amplification of amoA genes of nitrifying Bacteria and Archaea in soil microcosms and denaturing gradient gel electrophoresis (DGGE) analyses. Simazine [50?μg g(-1) dry weight soil (d.w.s)] completely inhibited the nitrification processes in the fertilized agricultural soil. The inhibition by simazine of ammonia oxidation observed was similar to the reduction of ammonia oxidation by the nitrification inhibitor acetylene. The application of simazine-affected AOB community DGGE patterns in the agricultural soil amended with ammonium, whereas no significant changes in the AOA community were observed. The DGGE analyses strongly suggest that simazine inhibited Nitrosobacteria and specifically Nitrosospira species. In conclusion, our results suggest that the s-triazine herbicide not only inhibits the target susceptible plants but also inhibits the ammonia oxidation and the AOB in fertilized soils.     

PCR profiling of ammonia-oxidizer communities in acidic soils subjected to nitrogen and sulphur deposition

Communities of ammonia-oxidizing bacteria (AOB) were characterized in two acidic soil sites experimentally subjected to varying levels of nitrogen and sulphur deposition. The sites were an acidic spruce forest soil in Deepsyke, Southern Scotland, with low background deposition, and a nitrogen-saturated upland grass heath in Pwllpeiran, North Wales. Betaproteobacterial ammonia-oxidizer 16S rRNA and ammonia monooxygenase (amoA) genes were analysed by cloning, sequencing and denaturing gradient gel electrophoresis (DGGE). DGGE profiles of amoA and 16S rRNA gene fragments from Deepsyke soil in 2002 indicated no effect of nitrogen deposition on AOB communities, which contained both Nitrosomonas europaea and Nitrosospira. In 2003, only Nitrosospira could be detected, and no amoA sequences could be retrieved. These results indicate a decrease in the relative abundance of AOB from the year 2002 to 2003 in Deepsyke soil, which may be the result of the exceptionally low rainfall in spring 2003. Nitrosospira-related sequences from Deepsyke soil grouped in all clusters, including cluster 1, which typically contains only sequences from marine environments. In Pwllpeiran soil, 16S rRNA gene libraries were dominated by nonammonia oxidizers and no amoA sequences were detectable. This indicates that autotrophic AOB play only a minor role in these soils even at high nitrogen deposition.     

Simultaneous effect of temperature,cyanide and ammonia-oxidizing bacteria concentrations on ammonia oxidation

For biological nitrification, a set of experiments were carried out to approximate the response of lag period along with ammonia oxidation rate with respect to different concentrations of cyanide (CN⁻) and ammonia-oxidizing bacteria (AOB), and temperature variation in laboratory-scale batch reactors. The effects of simultaneous changes in these three factors on ammonia oxidation were quantitatively estimated and modeled using response surface analysis. The lag period and the ammonia oxidation rate responded differently to changes in the three factors. The lag period and the ammonia oxidation rate were significantly affected by the CN⁻ and AOB concentrations, while temperature changes only affected the ammonia oxidation rate. The increase of AOB concentration and temperature alleviated the inhibition effect of cyanide on ammonia oxidation. The statistical method used in this study can be extended to estimate the quantitative effects of other environmental factors that can change simultaneously.     

Acclimation and activity of ammonia-oxidizing bacteria with respect to variations in zinc concentration, temperature, and microbial population

Activity of ammonia-oxidizing bacteria (AOB) to simultaneous variation in Zn²⁺ concentration (0.01-3.5 mg/L), temperature (23-33 °C), and AOB concentration (3-30 × 10⁶ gene copies/mL) in a steel industry wastewater treatment plant was evaluated. Two equations were developed to describe the lag period (i.e., AOB acclimation) and ammonia oxidation rate (i.e., growth of the AOB) depending on the variables. AOB concentration and temperature both had significant effects on lag period and the ammonia oxidation rate. Zn²⁺ concentration only had a significant effect on ammonia oxidation rate at 5% α-level. There was a significant interaction between AOB concentration and temperature for both lag period and ammonia oxidation rate. The effects of the variables were not significant when AOB concentration was higher than 2.0 × 10⁷ copies/mL. There was no visible shift or changes in AOB communities based on DGGE analysis with amoA gene primers.     

Dynamics of ammonia-oxidizing communities in barley-planted bulk soil and rhizosphere following nitrate and ammonium fertilizer amendment

Oxidation of ammonia by nitrifying microorganisms is a major pathway that fertilizer nitrogen (N) may take upon application to agricultural soils, but the relative roles of bacterial (AOB) vs. archaeal (AOA) ammonia oxidizers are controversial. We explored the effects of various forms of mineral N fertilizer on the AOB and AOA community dynamics in two different soils planted with barley. Ammonia oxidizers were monitored via real-time PCR and terminal restriction fragment length polymorphism analysis of bacterial and archaeal amoA genes following the addition of either [NH?]?SO?, NH?NO? or KNO?. AOB and AOA communities were also studied specifically in the rhizospheres of two different barley varieties upon [NH?]?SO? vs. KNO? addition. AOB changed in community composition and increased in abundance upon ammonium amendment in bulk soil and rhizosphere, with changes in bacterial amoA copy numbers lagging behind relative to changes in soil ammonium. In both soils, only T-RFs corresponding to phylotypes related to Nitrosospira clade 3a underwent significant community changes. Increases in AOB abundance were generally stronger in the bulk soil than in the rhizosphere, implying significant ammonia uptake by plant roots. AOA underwent shifts in the community composition over time and fluctuated in abundance in all treatments irrespective of ammonia availability. AOB were thus considered as the main agents responsible for fertilizer ammonium oxidation, while the functions of AOA in soil N cycling remain unresolved.     

酸性土壤氨氧化微生物及其影响因素研究进展

自然条件变化和人类活动不仅加剧了土壤酸化,扩大了酸性土壤面积,而且严重影响了土壤氮循环。氨氧化过程作为硝化作用的限速步骤,是全球氮循环的核心环节,受到国内外研究者的广泛关注。探究酸性土壤氨氧化作用及其功能微生物对完善氮循环机制和促进土壤养分循环具有重要意义。本文主要综述了土壤中氨氧化代谢途径,对比了氨氧化细菌(ammoniaoxidizing bacteria, AOB)、氨氧化古菌(ammonia-oxidizing archaea, AOA)和全程硝化菌(complete ammoniaoxidizers,Comammox)对酸性土壤氨氧化作用的相对贡献,分析了微生物内源功能差异及pH、底物浓度等外部环境因素对氨氧化微生物丰度、活性和群落结构的影响,最后对氨氧化微生物研究进行了展望,以期为酸性土壤氨氧化作用研究和微生物修复技术应用与实践提供科学参考。     

Quantitative analyses of the abundance and composition of ammonia-oxidizing bacteria and ammonia-oxidizing archaea of a Chinese upland red soil under long-term fertilization practices

The abundance and composition of soil ammonia-oxidizing bacteria (AOB) and ammonia-oxidizing archaea (AOA) were investigated by using quantitative real-time polymerase chain reaction, cloning and sequencing approaches based on amoA genes. The soil, classified as agri-udic ferrosols with pH (H(2)O) ranging from 3.7 to 6.0, was sampled in summer and winter from long-term field experimental plots which had received 16 years continuous fertilization treatments, including fallow (CK0), control without fertilizers (CK) and those with combinations of fertilizer nitrogen (N), phosphorus (P) and potassium (K): N, NP, NK, PK, NPK and NPK plus organic manure (OM). Population sizes of AOB and AOA changed greatly in response to the different fertilization treatments. The NPK + OM treatment had the highest copy numbers of AOB and AOA amoA genes among the treatments that received mineral fertilizers, whereas the lowest copy numbers were recorded in the N treatment. Ammonia-oxidizing archaea were more abundant than AOB in all the corresponding treatments, with AOA to AOB ratios ranging from 1.02 to 12.36. Significant positive correlations were observed among the population sizes of AOB and AOA, soil pH and potential nitrification rates, indicating that both AOB and AOA played an important role in ammonia oxidation in the soil. Phylogenetic analyses of the amoA gene fragments showed that all AOB sequences from different treatments were affiliated with Nitrosospira or Nitrosospira-like species and grouped into cluster 3, and little difference in AOB community composition was recorded among different treatments. All AOA sequences fell within cluster S (soil origin) and cluster M (marine and sediment origin). Cluster M dominated exclusively in the N, NP, NK and PK treatments, indicating a pronounced difference in the community composition of AOA in response to the long-term fertilization treatments. These findings could be fundamental to improve our understanding of the importance of both AOB and AOA in the cycling of nitrogen and other nutrients in terrestrial ecosystems.     

古尔班通古特沙漠生物土壤结皮对氨氧化微生物生态位的影响

生物结皮作为荒漠地表的重要覆被类型, 在荒漠生态系统的氮素循环中扮演重要角色。融雪期为古尔班通古特沙漠生物结皮的复苏和生长提供了充足的水分, 也成为该沙漠氮素固定和转化的重要时期, 但该时期生物结皮如何影响驱动氨氧化转化的微生物群落动态尚未明确。因此, 我们利用荧光定量PCR (fluorescent quantitative PCR, qPCR)方法分析融雪期生物结皮与去除结皮不同土层(0-2, 2-5, 5-10和10-20 cm)氨氧化菌群丰度特征, 结合潜在硝化速率和土壤理化参数, 探究融雪期生物结皮对荒漠土壤氮素转化作用。结果表明: 氨氧化古菌(ammonia-oxidizing archaea, AOA)是古尔班通古特沙漠土壤优势氨氧化菌, 生物结皮对0-2 cm层土壤中AOA、氨氧化细菌(ammonia-oxidizing bacteria, AOB) amoA基因丰度具有显著抑制作用(P < 0.01), 对10-20 cm层土壤中AOA amoA基因丰度具有显著促进作用(P < 0.01)。冗余分析(redundancy analysis, RDA)表明, AOA、AOB amoA基因丰度主要受土壤含水量和铵态氮含量的影响, 占总条件效应的54.90%。氨氧化速率分析发现, 去除生物结皮显著降低古尔班通古特沙漠土壤硝化作用潜力(P < 0.001), 证实生物结皮对荒漠土壤氮素转化具有重要的调控作用。综上所述, 古尔班通古特沙漠氨氧化微生物的分布规律受环境因子调控, 特别是生物结皮可以通过调节土壤含水量和铵态氮含量影响AOA和AOB的空间生态位分化, 促进沙漠土壤的硝化作用。     

Nitrogen cycling and relationships between ammonia oxidizers and denitrifiers in a clay-loam soil

This study investigated the effect of municipal solid waste (MSW) compost (0, 50, and 100 t/ha) on N cycling and the microorganisms involved in it, in a clay-loam soil. After a release of nitrates (NO₃ ^?-N) in the first 6 days after compost incorporation, soil NO₃ ^?-N content remained constant in all the treatments until day?62, suggesting N immobilization induced by the soil used in this study. Then, soil NO₃ ^?-N content increased in all treatments and especially in the highest compost dose, providing evidence that immobilization effect has been at least partially relieved. amoA gene copies of ammonia-oxidizing archaea (AOA) and bacteria (AOB) followed the overall pattern of soil NO₃ ^?-N content; however, no differences were found in amoA gene copies among treatments, except in the last sampling, an effect attributed to the slight differences in the potential nitrification rate among them. Ammonia oxidizer pattern provided evidence that both groups were involved in ammonia oxidation and changes in their abundance can be used as ‘indicator’ to predict changes in soil nitrification status. Moreover, the strong correlation between AOA and AOB amoA copies (R ²?=?0.94) and the high slope (13) of the curve suggest that AOA had probably an important role on ammonia oxidation. Denitrifying genes (nirS, nirK, nosZ) also followed the general pattern of soil NO₃ ^?-N, and they were strongly correlated with both groups of ammonia oxidizers, and particularly AOA, suggesting strong interrelationships among them. Losses of N through denitrification, as they were estimated by total nitrogen, were inversely related to soil NO₃ ^?-N content. Similar to ammonia oxidizers, denitrifying gene copies did not differ among compost treatments an effect that could be probably explained by the low availability of organic-C in the MSW compost and hence the competition with aerobic heterotrophs.     

Comparative phylogeny of the ammonia monooxygenase subunit A and 16S rRNA genes of ammonia-oxidizing bacteria

A fragment of the ammonia monooxygenase gene (amoA) from 31 strains of ammonia-oxidizing bacteria (AOB) was sequenced and analysed phylogenetically. The results were compared with the phylogeny of 16S rDNA from AOB. For most groups of AOB we found a high consistency between the phylogenetic trees based on the 16S rDNA and amoA sequences. Although it is not a phylogenetic marker, using the amoA as a probe when studying microbial diversity will probably reduce the amount of non-AOB detected, compared to using rDNA based probes. The data presented in this paper extend and improve the basis for application of amoA in studies of AOB in the environment.     

Diversity and abundance of ammonia-oxidizing archaea and bacteria in polluted mangrove sediment

Ammonia oxidation by microorganisms is a critical process in the nitrogen cycle. Recent research results show that ammonia-oxidizing archaea (AOA) are both abundant and diverse in a range of ecosystems. In this study, we examined the abundance and diversity of AOA and ammonia-oxidizing beta-proteobacteria (AOB) in estuarine sediments in Hong Kong for two seasons using the ammonia monooxygenase A subunit gene (amoA) as molecular biomarker. Relationships between diversity and abundance of AOA and AOB and physicochemical parameters were also explored. AOB were more diverse but less abundant than AOA. A few phylogenetically distinct amoA gene clusters were evident for both AOA and AOB from the mangrove sediment. Pearson moment correlation analysis and canonical correspondence analysis (CCA) were used to explore physicochemical parameters potentially important to AOA and AOB. Metal concentrations were proposed to contribute potentially to the distributions of AOA while total phosphorus (TP) was correlated to the distributions of AOB. Quantitative PCR estimates indicated that AOA were more abundant than AOB in all samples, but the ratio of AOA/AOB (from 1.8 to 6.3) was smaller than most other studies by one to two orders. The abundance of AOA or AOB was correlated with pH and temperature while the AOA/AOB ratio was with the concentrations of ammonium. Several physicochemical factors, rather than any single one, affect the distribution patterns suggesting that a combination of factors is involved in shaping the dynamics of AOA and AOB in the mangrove ecosystem.     

Climate warming increases soil erosion,carbon and nitrogen loss with biofuel feedstock harvest in tallgrass prairie

Anthropogenic soil erosion severely affects land ecosystems by reducing plant productivity and stimulating horizontal carbon and nitrogen movement at the surface. Climate warming may accelerate soil erosion by altering soil temperature, moisture, and vegetation coverage. However, no experiments have been carried out to quantify soil erosion with warming. In a long‐term field experiment, we explored how annual clipping for biofuel feedstock production and warming caused soil erosion and accompanying carbon and nitrogen losses in tallgrass prairie in Oklahoma, USA. We measured relative changes in soil surface elevation between clipped and unclipped plots with or without experimental warming. Our results show that average relative erosion depth caused by clipping was 1.65±0.09 and 0.54±0.08 mm yr^?1, respectively, in warmed and control plots from November 21, 1999 to April 21, 2009. The soil erosion rate was 2148±121 g m^?2 yr^?1 in the warmed plots and 693±113 g m^?2 yr^?1 in the control plots. Soil organic carbon was lost at a rate of 69.6±5.6 g m^?2 yr^?1 in the warmed plots and 22.5±2.7 g m^?2 yr^?1 in the control plots. Total nitrogen was lost at a rate of 4.6±0.4 g m^?2 yr^?1 in the warmed plots and 1.4±0.1 g m^?2 yr^?2 in the control plots. The amount of carbon and nitrogen loss caused by clipping is equivalent to or even larger than changes caused by global change factors such as warming and rising atmospheric CO₂ concentration. In addition, soil erosion rates were significantly correlated with clipping‐induced changes in soil moisture. Our results suggest that clipping for biofuel harvest results in significant soil erosion and accompanying losses of soil carbon and nitrogen, which is aggravated by warming.     

Nitrogen addition reduces soil respiration in a mature tropical forest in southern China

Response of soil respiration (CO₂ emission) to simulated nitrogen (N) deposition in a mature tropical forest in southern China was studied from October 2005 to September 2006. The objective was to test the hypothesis that N addition would reduce soil respiration in N saturated tropical forests. Static chamber and gas chromatography techniques were used to quantify the soil respiration, following four‐levels of N treatments (Control, no N addition; Low‐N, 5 g N m^?2 yr^?1; Medium‐N, 10 g N m^?2 yr^?1; and High‐N, 15 g N m^?2 yr^?1 experimental inputs), which had been applied for 26 months before and continued throughout the respiration measurement period. Results showed that soil respiration exhibited a strong seasonal pattern, with the highest rates found in the warm and wet growing season (April–September) and the lowest rates in the dry dormant season (December–February). Soil respiration rates showed a significant positive exponential relationship with soil temperature, whereas soil moisture only affect soil respiration at dry conditions in the dormant season. Annual accumulative soil respiration was 601±30 g CO₂‐C m^?2 yr^?1 in the Controls. Annual mean soil respiration rate in the Control, Low‐N and Medium‐N treatments (69±3, 72±3 and 63±1 mg CO₂‐C m^?2 h^?1, respectively) did not differ significantly, whereas it was 14% lower in the High‐N treatment (58±3 mg CO₂‐C m^?2 h^?1) compared with the Control treatment, also the temperature sensitivity of respiration, Q₁₀ was reduced from 2.6 in the Control with 2.2 in the High‐N treatment. The decrease in soil respiration occurred in the warm and wet growing season and were correlated with a decrease in soil microbial activities and in fine root biomass in the N‐treated plots. Our results suggest that response of soil respiration to atmospheric N deposition in tropical forests is a decline, but it may vary depending on the rate of N deposition.