A method of selective inhibition to distinguish between nitrification and denitrification as sources of nitrous oxide in soil

Summary Nitrapyrin and C₂H₂ were evaluated as nitrification inhibitors in soil to determine the relative contributions of denitrification and nitrification to total N₂O production. In laboratory experiments nitrapyrin, or its solvent xylene, stimulated denitrification directly or indirectly and was therefore considered unsuitable. Low partial pressures of C₂H₂ (2.5–5.0 Pa) inhibited nitrification and had only a small effect on denitrification, which made it possible to estimate the contribution of denitrification. The contribution of nitrification was estimated by subtracting the denitrification value from total N₂O production (samples without C₂H₂). The critical C₂H₂ concentrations needed to achieve inhibition of nitrification, without affecting the N₂O reductase in denitrifiers, must be individually determined for each set of experimental conditions.     

Gaseous nitrogen losses from fertilized soil during nitrification and denitrification using the acetylene blockage technique

Summary A sandy soil amended with different forms and amounts of fertilizer nitrogen (urea, ammonium sulphate and potassium nitrate) was investigated in model experiments for N₂O emission, which may be evolved during both oxidation of ammonia to nitrate and anaerobic respiration of nitrate. Since C₂H₂ inhibits both nitrification and the reduction of N₂O to N₂ during denitrification, the amount of N₂O evolved in the presence and absence of C₂H₂ represents the nitrogen released through nitrification and denitrification.Results show that amounts of N₂O-N lost from soils incubated anaerobically with 0.1% C₂H₂ and treated with potassium nitrate (23.1 µg N-NO ₃ ^– /g dry soil) exceeded those from soils incubated in the presence of 20% oxygen and treated with even larger amounts of nitrogen as urea and ammonium sulphate. This indicates that nitrogen losses by denitrification may potentially be higher than those occurring through nitrification.     

Contribution of denitrification and autotrophic and heterotrophic nitrification to nitrous oxide production in andosols

To quantify the contribution of denitrification and autotrophic and heterotrophic nitrification to N₂O production in Andosols with a relatively high organic matter content, we first examined the effect of C₂H₂ concentrations on N₂O production and on changes in mineral N contents. The optimum C₂H₂ concentration for inhibiting autotrophic nitrification was 10 Pa. Secondly, and Andosol taken from an arable field was incubated for 32 days at 30°C at 60, 80, and 100% water-holding capacity with or without the addition of NH ₄ ⁺ or NO _inf3 ^sup- (200 mg N kg^-1), and subsamples collected every 4–8 days were further incubated for 24 h with or without C₂H₂ (10 Pa). At 60 and 80% water-holding capacity with NH ₄ ⁺ added, 87–92% of N₂O produced (200–250 g N₂O–N kg^-1) was derived from autotrophic nitrification. In contrast, at 100% water-holding capacity with or without added NO _inf3 ^sup- , enormous amounts of N₂O (29–90 mg N₂O–N kg^-1) were produced rapidly, mostly by denitrification (96–98% of total production). Thirdly, to examine N₂O production by heterotrophic nitrification, the Andosol was amended with peptone or NH ₄ ⁺ (both 1000 mg N kg^-1)+citric acid (20 g C kg^-1) and with or without dicyandiamide (200 mg N kg^-1). Treatment with citric acid alone or with citric acid+dicyandiamide suppressed N₂O production. In contrast, peptone increased N₂O production (5.66 mg N₂O–N kg^-1) mainly by denitrification (80% of total production). However, dicyandiamide reduced N₂O production to 1.1 mg N₂O–N kg^-1. These results indicate that autotrophic nitrification was the main process for N₂O production except at 100% water-holding capacity where denitrification became dominant and that heterotrophic nitrification had a lesser importance in the soils examine.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Contributions from different microbial processes to N2O emission from soil under different moisture regimes

Nitrous oxide emissions from a sandy-loam textured soil wetted to matric potentials of either-1.0 or-0.1 kPa were determined in laboratory experiments in which the soil was incubated in air (control), air plus 10 Pa C₂H₂ (to inhibit nitrification), 100 kPa O₂ (to suppress denitrification), 10 kPa C₂H₂ (to inhibit N₂O reduction to N₂ in denitrification) or following autoclaving. The total N₂O production, consumption and net N₂O emission from the soils together with the contributions to N₂O emission from different processes of N₂O production were estimated. The rate of N₂O production was significantly greater in the wetter soil (282 pmol N₂O g^-1 soil h^-1) than in the drier soil (192 pmol N₂O g^-1 soil h^-1), but because N₂O consumption by denitrifiers was also greater in the wetter soil, the net N₂O emissions from the wetter and the drier soils did not differ significantly. Non-biological sources made no significant contribution to N₂O emission under either moisture regime and biological processes other than denitrification and nitrification made only a small contribution (1% of the total N₂O production) in the wetter soil. Denitrifying nitrifiers were the predominant source of N₂O emitted from the drier soil and other (non-nitrifying) denitrifiers were the predominant source of N₂O emitted from the wetter soil.     

Nitrifier denitrification as a distinct and significant source of nitrous oxide from soil

Soils are the major source of the greenhouse gas nitrous oxide (N₂O) to our atmosphere. A thorough understanding of terrestrial N₂O production is therefore essential. N₂O can be produced by nitrifiers, denitrifiers, and by nitrifiers paradoxically denitrifying. The latter pathway, though well-known in pure culture, has only recently been demonstrated in soils. Moreover, nitrifier denitrification appeared to be much less important than classical nitrate-driven denitrification. Here we studied a poor sandy soil, and show that when moisture conditions are sub-optimal for denitrification, nitrifier denitrification can be a major contributor to N₂O emission from this soil. We conclude that the relative importance of classical and nitrifier denitrification in N₂O emitted from soil is a function of the soil moisture content, and likely of other environmental conditions as well. Accordingly, we suggest that nitrifier denitrification should be routinely considered as a major source of N₂O from soil.     

Nitrous oxide production at different soil moisture contents in an arable soil in China

Abstract

Laboratory incubations were conducted to investigate nitrous oxide (N₂O) production from a subtropical arable soil (Typic Plinthodults) incubated at different soil moisture contents (SMC) and with different nitrogen sources using a 10% (v/v) acetylene (C₂H₂) inhibitory technique at 25°C. The production of N₂O and CO₂ was monitored during the incubations and changes in the contents of KCl-extractable NO^? ₃-N and NH⁺ ₄-N were determined. The production of N₂O increased slightly with an increase in SMC from 40% water-holding capacity (WHC) to 70% WHC, but increased dramatically at 100% WHC. After incubation the NO^? ₃-N content increased even at a SMC of 100% WHC. At a SMC of 100% WHC, the addition of NH⁺ ₄-N promoted the production of N₂O and CO₂, whereas the addition of NO^? ₃-N decreased N₂O production. Compared with the incubation without C₂H₂, the presence of C₂H₂ increased NH⁺ ₄-N content, but decreased NO^? ₃-N content, and there was no significant difference in N₂O production. These results indicate that heterotrophic nitrification contributes to N₂O production in the soil.     

Contribution of Heterotrophic Nitrification to Nitrous Oxide Production in a Long-Term N-Fertilized Arable Black Soil

To understand the contribution of key microbial processes to nitrous oxide (N₂O) emission in intensively cultivated black soil, laboratory incubation were conducted at 70% water-holding capacity (WHC) and 25 °C, using different gases (air, oxygen, or argon) within the headspace of the incubation chambers to evaluate gas inhibition effects. Arable black soil was sampled from an experimental field that has received urea since October 1979. Nitrification contributed to 57% of total N₂O emission, of which as much as 67% resulted from heterotrophic nitrification. These data strongly suggest that high soil organic carbon concentrations and low pH values are more favorable to N₂O production through heterotrophic, rather than autotrophic, nitrification. Nitrous oxide produced by denitrification accounted for 28% of the total N₂O emission, and the nitrifier denitrification accounted for 15% of the N₂O emitted from the tested soil. These findings indicate that heterotrophic nitrification was the primary N₂O production process in the tested soil.     

Processes responsible for the nitrous oxide emission from a Costa Rican Andosol under a coffee agroforestry plantation

We used the inhibitor acetylene (C₂H₂) at partial pressures of 10 Pa and 10 kPa to inhibit autotrophic nitrification and the reduction of nitrous oxide (N₂O) to N₂, respectively. Soils (Andosol) from a Coffea arabica plantation shaded by Inga densiflora in Costa Rica were adjusted to 39, 58, 76 and 87% water-filled pore space (WFPS) and incubated for 6 days in the absence or presence of C₂H₂. Soil respiration, nitrification rates and N₂O emissions by both processes were measured in relation to soil moisture conditions. At all WFPS studied, rates of N₂O and N₂ productions were small (4.8; 14.7; 23 and 239.6 ng N–N₂O g⁻¹ d.w. d⁻¹ at 39, 58, 76 and 87% WFPS, respectively), and despite a low soil pH (4.7), N₂O was mainly produced by nitrification, which was responsible for 85, 91, 84 and 87% of the total N₂O emissions at 39, 58, 76 and 87% WFPS, respectively. At the three smaller values of WFPS, a linear relationship was established between WFPS, soil respiration, nitrification and N₂O released by nitrification; no N₂ was produced by denitrification. At more anaerobic conditions achieved by a WFPS of 87%, a large rate of N₂O production was measured during nitrification, and N₂ production accounted for 84% of the gaseous N fluxes caused by denitrification.     

The effect of some carbon substrates on denitrification rates and carbon utilization in soil

Summary Soil was amended with a variety of carbon sources, including four soluble compounds (glucose, sucrose, glycerol and mannitol) and two plant residues (straw and alfalfa).. Potential denitrification rates, measured both as N₂O accumulation and NO₃ ^– disappearance, were compared, and the predicted values of available C, measured as CO₂ production and water-extractable C, were assessed.The two measures of denitrification agreed well although N₂O accumulation was, found to be most sensitive. Soil treated with the four soluble C compounds resulted in the same rate of denitrification although glycerol was not as rapidly oxidized. Alfalfa-amended soil produced a significantly higher rate of denitrification than the same amount of added straw. CO₂ evolution was found to be a good predictor of denitrification over the first 2 days of sampling, but neither measure of available substrate C correlated well with denitrification rate beyond 4 days, when NO₃ ^– was depleted in most treatments. The data with alfalfa-amended soil suggested that denitrifiers used water-extractable C. materials produced by other organisms under anaerobic conditions.     

Role of nitrifier denitrification in the production of nitrous oxide

Nitrifier denitrification is the pathway of nitrification in which ammonia (NH₃) is oxidized to nitrite (NO₂⁻) followed by the reduction of NO₂⁻ to nitric oxide (NO), nitrous oxide (N₂O) and molecular nitrogen (N₂). The transformations are carried out by autotrophic nitrifiers. Thus, nitrifier denitrification differs from coupled nitrification–denitrification, where denitrifiers reduce NO₂⁻ or nitrate (NO₃⁻) that was produced by nitrifiers. Nitrifier denitrification contributes to the development of the greenhouse gas N₂O and also causes losses of fertilizer nitrogen in agricultural soils. In this review article, present knowledge about nitrifier denitrification is summarized in order to give an exact definition, to spread awareness of its pathway and controlling factors and to identify areas of research needed to improve global N₂O budgets. Due to experimental difficulties and a lack of awareness of nitrifier denitrification, not much is known about this mechanism of N₂O production. The few measurements carried out so far attribute up to 30% of the total N₂O production to nitrifier denitrification. Low oxygen conditions coupled with low organic carbon contents of soils favour this pathway as might low pH. As nitrifier denitrification can lead to substantial N₂O emissions, there is a need to quantify this pathway in different soils under different conditions. New insights attained through quantification experiments should be used in the improvement of computer models to define sets of conditions that show where and when nitrifier denitrification is a significant source of N₂O. This may subsequently render the development of guidelines for low-emission farming practices necessary.     

有机无机肥配施对酸性菜地土壤硝化作用的影响

通过室内培养和田间试验, 研究了有机无机肥配施对酸性菜地土硝化作用的影响。培养试验条件为60%土壤最大持水量和25 ℃。 结果表明,土壤硝化作用模式为指数方程,延滞期10天。与纯化肥处理（NPK）相比,鲜猪粪配施无机肥（FPM+NPK）和猪粪堆肥配施无机肥（CPM+NPK）均能降低土壤硝化势和氨氧化潜势,猪粪堆肥配施无机肥还能增加土壤微生物量碳、 氮。鲜猪粪配施无机肥和猪粪堆肥配施无机肥处理在硝化培养和田间试验期间N2O释放量均没有差异,但硝化培养期间鲜猪粪配施无机肥的N2O释放量显著低于纯化肥处理,田间试验期间猪粪堆肥配施无机肥的N2O释放量显著低于纯化肥处理。培养试验结束后的土壤pH值与土壤硝化势间,以及硝化培养期间N2O累积释放量与土壤硝化势间均存在显著正相关关系。本研究表明, 有机无机肥配施显著影响土壤硝化作用以及硝化培养期间和田间N2O释放。     

Nitrous oxide production in grassland soils: assessing the contribution of nitrifier denitrification

Nitrifier denitrification is the reduction of NO₂⁻ to N₂ by nitrifiers. It leads to the production of the greenhouse gas nitrous oxide (N₂O) as an intermediate and possible end product. It is not known how important nitrifier denitrification is for the production of N₂O in soils. We explored N₂O production by nitrifier denitrification in relation to other N₂O producing processes such as nitrification and denitrification under different soil conditions. The influence of aeration of the soil, different N sources, and pH were tested in four experiments. To differentiate between sources of N₂O, an incubation method with inhibitors was used [Biol. Fertil. Soils 22 (1996) 331]. Sets of four incubations included controls without addition of inhibitors, incubations with addition of small concentrations of C₂H₂ (0.01-0.1 kPa), large concentrations of O₂ (100 kPa), or a combination of C₂H₂ and O₂. The results indicate that the availability of NO₂⁻ stimulated the apparent N₂O production by nitrifier denitrification. A decreasing O₂ content increased the total N₂O production, but decreased N₂O production by nitrifier denitrification. No significant effect of pH could be found. The study revealed problems concerning the use of the inhibitors C₂H₂ and O₂. Almost one-third of all incubations with inhibitors produced more N₂O than the controls. Possible reasons for the problems are discussed. The inhibitors C₂H₂ and O₂ need to be tested thoroughly for their effects on different N₂O producing processes before further application.     

Effects of acetylene on nitrification and denitrification in two soils during incubation with ammonium nitrate

A loam from the Frilsham and one from the Wickham Series were incubated at 50 and 90 per cent of their water contents at saturation with 100 μg NH₄NO₃-N_g^?1 soil in the presence and absence of C₂H₂ (0.5 per cent, v/v). Acetylene inhibited nitrification in both soils, but had no effect on mineralization of N. No denitrification (measured as the production of N₂O in the presence of C₂H₂) occurred during incubation at 50 per cent saturation. At 90 per cent saturation, denitrification resulted in a loss of 28.4 and 36.7 μg N_g^?1 after 48 h from the Frilsham and Wickham soils, respectively. The concurrent inhibition of nitrification had no effect on the extent of denitrification at this time. In the Wickham soil, NO₃^? was exhausted after 168 h incubation in the presence of C₂H₂ and denitrification was underestimated by 13 μg N_g^?. The data suggested that concurrent inhibition of nitrification during measurement of denitrification using the C₂H₂ inhibition technique is most likely to affect the estimate of denitrification loss when NO₃^?supply is limited by the inhibition of nitrification.     

Effect of urease and nitrification inhibitors on ammonia volatilization and abundance of N‐cycling genes in an agricultural soil

The effect of the combined application of urease and nitrification inhibitors on ammonia volatilization and the abundance of nitrifier and denitrifier communities is largely unknown. Here, in a mesocosm experiment, ammonia volatilization was monitored in an agricultural soil treated with urea and either or both of the urease inhibitor N‐(n‐butyl) thiophosphoric triamide (NBPT) and the nitrification inhibitor 3,4‐dimethylpyrazole phosphate (DMPP), with 50% and 80% water‐filled pore space (WFPS). The effect of the treatments on the abundance of bacteria and archaea was estimated by quantitative PCR (qPCR) amplification of their respective 16S rRNA gene, that of nitrifiers using amoA genes, and that of denitrifiers by qPCR of the norB and nosZI denitrification genes. After application of urea, N losses due to NH₃ volatilization accounted for 23.0% and 9.2% at 50% and 80% WFPS, respectively. NBPT reduced NH₃ volatilization to 2.0% and 2.4%, whereas DMPP increased N losses by up to 36.8% and 26.0% at 50% and 80% WFPS, respectively. The combined application of NBPT and DMPP also increased NH₃ emissions, albeit to a lesser extent than DMPP alone. As compared with unfertilized control soil, both at 50% and 80% WFPS, NBPT strongly affected the abundance of bacteria and archaea, but not that of nitrifiers, and decreased that of denitrifiers at 80% WFPS. Regardless of moisture conditions, treatment with DMPP increased the abundance of denitrifiers. DMPP, both in single and in combined application with NBPT, increased the abundance of nitrification and denitrification genes.     

Effect of the inhibitors nitrapyrin and sodium chlorate on nitrification and N2O formation in an acid forest soil

An acid forest soil from beech forest gaps, which were either limed or unlimed, and the undisturbed forest was investigated for the type of nitrifying populations and the process of N₂O evolution. To see whether nitrifiers were of heterotrophic or autotrophic origin, the nitrification inhibitors nitrapyrin and sodium chlorate were applied to disturbed soil samples which underwent laboratory incubations. Nitrapyrin inhibits autotrophic nitrification. In different studies, sodium chlorate has been identified as an inhibitor either of autotrophic or of heterotrophic nitrification. In the samples investigated only nitrapyrin inhibited the autotrophic nitrification occurring in the limed soil. Sodium chlorate effectively inhibited heterotrophic nitrification. In the limed forest floor samples, where most autotrophic nitrification occured, sodium chlorate showed no inhibitory effect. In another laboratory incubation experiment, N₂O evolution from undisturbed soil columns, to which the above inhibitors were applied, was investigated. In those samples, in which nitrification had been reduced, neither inhibitor significantly reduced N₂O evolution. Thus it was concluded that the contribution of nitrification to N₂O losses is negligible, and that N₂O evolution arises from the activity of denitrifying organisms. Microbial biomass and respiration measurements showed that the inhibitors did not affect microflora negatively.     

Nitrous oxide production of heavy metal contaminated soil

Arsenic (As), lead (Pb), copper (Cu) and zinc (Zn) can be found in large concentrations in mine spills of central and northern Mexico. Interest in these heavy metals has increased recently as they contaminate drinking water and aquifers in large parts of the world and severely affect human health, but little is known about how they affect biological functioning of soil. Soils were sampled in seven locations along a gradient of heavy metal contamination with distance from a mine in San Luis Potosí (Mexico), active since about 1800 AD. C mineralization and N₂O production were monitored in an aerobic incubation experiment. Concentrations of As in the top 0-10 cm soil layer ranged from 8 to 22,992 mg kg⁻¹, from 31 to 1845 mg kg⁻¹ for Pb, from 27 to 1620 mg kg⁻¹ for Cu and from 81 to 4218 mg kg⁻¹ for Zn. There was a significant negative correlation between production rates of CO₂ and concentrations of As, Pb, Cu and Zn, and there was a significant positive correlation with pH, water holding capacity (WHC), total N and soil organic C. There was a significant negative correlation (P<0.05) between production rate of nitrous oxide (N₂O) attributed to nitrification by the inhibition method in soil incubated at 50% WHC and total concentrations of Pb and Zn, and there was a significant positive correlation (P<0.05) with pH and total N content. There was a significant negative correlation (P<0.05) between the production rate of N₂O attributed to denitrification by the inhibition method in soil incubated at 100% WHC and total concentrations of Pb, Cu and Zn, and a significant positive correlation (P<0.01) with pH; there was a significant positive correlation (P<0.05) between the production of N₂O attributed to other processes by the inhibition method and WHC, inorganic C and clay content. A negative value for production rate of N₂O attributed to nitrifier denitrification by the inhibition method was obtained at 100% WHC. The large concentrations of heavy metals in soil inhibited microbial activity and the production rate of N₂O attributed to nitrification by the inhibition method when soil was incubated at 50% WHC and denitrification when soil was incubated at 100% WHC. The inhibitor/suppression technique used appeared to be flawed, as negative values for nitrifier denitrification were obtained and as the production rate of N₂O through denitrification increased when soil was incubated with C₂H₂.     

Assessing the potential of ammonia oxidizing bacteria to produce nitrous oxide in soils of a high arctic lowland ecosystem on Devon Island, Canada

The contribution of nitrifiers (ammonia-oxidizing bacteria (AOB)) and denitrifiers to nitrous oxide (N₂O) emission from arctic soils remains inconclusive. Based on preliminary experiments, we hypothesized that AOB are the primary producers of N₂O in a high arctic lowland ecosystem on Devon Island, Nunavut, Canada. In part 1 of the study, flux chambers were installed in a catena to determine in situ fluxes of gases (N₂O and carbon dioxide (CO₂)) from 16 June to 13 July 2004. Although fluxes were low, N₂O production occurred in the wettest area of the landscape when ammonium levels were high. As ammonium, but not nitrate, levels declined in the wet sedge meadow, N₂O emissions correspondingly decreased. In part 2, the contribution of nitrification and denitrification to N₂O production was assessed by Acetylene Inhibition Assay and ¹⁵N isotopically enriched incubations. Ammonium fertilization stimulated N₂O emissions to a greater extent than nitrate, and acetylene had a greater impact on N₂O emissions in ammonium-fertilized soils than in nitrate-amended soils. Stable isotope analysis indicated that at 50-55% water filled pore space, nitrification was the dominant (>80%) N₂O emitting process. In part 3, molecular analyses of the two N₂O producing groups indicated the both nitrifiers and denitrifiers did not differ between landforms. Our results suggest nitrifier denitrification is the dominant process occurring in these arctic soils and that the role of denitrifiers in N₂O release from arctic soils needs to be re-evaluated.     

The effect of urea and pig slurry fertilization on denitrification, direct nitrous oxide emission, volatile fatty acids, water-soluble carbon and anthrone-reactive carbon in maize-cropped soil from the Po plain (Modena, Italy)

 The use of zootechnical slurries in agriculture can increase N losses as N₂O by direct emission and by denitrification. The aim of this research was to determine the influence of pig slurry, as well as its combination with mineral N, on N₂O emissions in the field and their relationships with some fractions of soil organic matter, with soil moisture and with rainfall. In spite of varying amounts of organic substance applied, the diverse agronomic treatments did not produce substantial differences in N losses due to denitrification. Wide variations between the slurry fertilized and the urea-fertilized plots were not found, whereas the combination of pig slurry with urea usually produced an increase both in N₂O emissions due to denitrification and in direct N₂O emissions (N losses corresponding to about 50% of those due to denitrification). The greatest losses of N₂O-N occurred in the first month following fertilizer administration. N₂O emissions due to denitrification were highest in the days immediately following the administration of fertilizers and lowest in a later period. N₂O emissions due to nitrification occurred later. Therefore, N₂O emission via nitrification differed from N₂O losses via denitrification which, under optimal conditions, presented peaks of activity during the whole growth cycle. The N₂O-N losses were highly influenced by physical parameters, particularly rain. An increase in micropore water creates conditions of scarce oxygenation or of anaerobiosis which influence oxidation-reduction processes and, at the same time, can limit the diffusion of bacteria-produced gas towards the soil surface. Received: 14 January 1998     

Increase in nitrous oxide production in soil induced by ammonium and organic carbon

We observed that soil cores collected in the field containing relatively high NH _inf4 ^sup+ and C substrate levels produced relatively large quantities of N₂O. A series of laboratory experiments confirmed that the addition of NH _inf4 ^sup+ and glucose to soil increase N₂O production under aerobic conditions. Denitrifying enzyme activity was also increased by the addition of NH _inf4 ^sup+ and glucose. Furthermore, NH _inf4 ^sup+ and glocose additions increased the production of N₂O in the presence of C₂H₂. Therefore, we concluded that denitrification was the most likely source of N₂O production. Denitrification was not, however, directly affected by NH _inf4 ^sup+ in anaerobic soil slurries, although the use of C substrate increased. In the presence of a high substrate C concentration, N₂O production by denitrifiers may be affected by NO _inf3 ^sup- supplied from NH _inf4 ^sup+ through nitrification. Alternatively, N₂O may be produced during mixotrophic and heterotrophic growth of nitrifiers. The results indicated that the NH _inf4 ^sup+ concentration, in addition to NO _inf3 ^sup- , C substrate, and O₂ concentrations, is important for predicting N₂O production and denitrification under field conditions.     

高氮和NO2-对中亚热带森林土壤N2O和NO产生的影响

利用15N同位素标记方法,研究在两种水分条件即60％和90％ WHC下,添加硝酸盐(NH4NO3,N 300 mg kg-1)和亚硝酸盐(NaNO2,N 1 mg kg-1)对中亚热带天然森林土壤N2O和NO产生过程及途径的影响.结果表明,在含水量为60％ WHC的情况下,高氮输入显著抑制了N2O和NO的产生(p＜0.01);但当含水量增为90％ WHC后,实验9h内抑制N2O产生,之后转为促进.所有未灭菌处理在添加NO2-后高氮抑制均立即解除并大量产生N2O和NO,与对照成显著差异(p＜0.01),在60％ WHC条件下,这种情况维持时间较短(21 h),但如果含水量高(90％ WHC)这种情况会持续很长时间(2周以上),说明水分有效性的提高和外源NO2-在高氮抑制解除中起到重要作用.本实验中N2O主要来源于土壤反硝化过程,而且加入未标记NO2-后导致杂合的N2O(14N15NO)分子在实验21 h内迅速增加,表明这种森林土壤的反硝化过程可能主要是通过真菌的“共脱氮”来实现,其贡献率可多达80％以上.Spearman秩相关分析表明未灭菌土壤NO的产生速率与N2O产生速率成显著正相关性(p＜0.05),土壤含水量越低二者相关性越高.灭菌土壤添加NO2-能较未灭菌土壤产生更多的NO,但却几乎不产生N2O,表明酸性土壤的化学反硝化对NO的贡献要大于N2O.