Effectiveness of acetylene inhibition of N2O reduction for measuring denitrification in soils of varying wetness

Abstract

The use of acetylene (C₂H₂) in the inhibition of N₂O to N₂ is widely used for measuring denitrification. The objective of this study was to determine the effectiveness of acetylene inhibition of N₂O reduction for short‐term and prolonged incubation studies in soils of varying water saturation, and to find out the possible reasons for lower N₂O recovery in continuously sealed incubations. Two experiments carried out in the laboratory reconfirmed that acetylene was very effective in inhibiting the reduction of N₂O in denitrification even for the prolonged incubation period (up to 96 h) under moist to saturated soil water contents. With 90 and 120% water‐filled pore space (WFPS), the accumulated N₂O in containers kept sealed throughout the study period, was 28 to 41% less than total headspace N₂O produced in containers that were opened, flushed and fresh C₂H₂ added every 24 h. Interpretation of our results suggest the lower N₂O amount recovered from continuously sealed containers at high WFPS, as compared to short‐term incubations (flushed containers), resulted primarily from delayed N₂O release from soil and not greater N₂O dissolved in soil solution, lower rates of denitrification, or decomposition/loss of C₂H₂ during prolonged incubation. Reduction of N₂O diffusion from soil cores showed direct relationship with head space concentration‐of N₂O and soil WFPS. From these results it is concluded that to obtain quantitative recovery of N₂O produced via denitrification, especially from soil with high WFPS soil cores should be vigorously shaken before head‐space N₂O analysis.     

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.     

Nitrous Oxide Production and Consumption Potential in an Agricultural and a Forest Soil

Both a laboratory incubation experiment using soils from an agricultural field and a forest and field measurements at the same locations were conducted to determine nitrous oxide (N₂O) production and consumption (reduction) potentials using the acetylene (C₂H₂) inhibition technique. Results from the laboratory experiment show that the agricultural soil had a stronger N₂O reduction potential than the forest soil, as indicated by the N₂O/N₂ ratio in denitrification products. Without C₂H₂ inhibition, N₂O could reach a maximum concentration of 51 and 296 ppmv in headspace of the agricultural and forest soil slurries, respectively. Addition of glucose decreased the maximum N₂O concentration to 22 ppmv in headspace of the agricultural soil slurries, but increased to 520 ppmv in the forest soil slurries. Addition of exogenous N₂O did not change such N₂O accumulation maxima during the incubations. The field measurements show that average N₂O emission rates were 0.56 and 0.59 kg N ha^?1 in the agricultural field and forest, respectively. When C₂H₂ was provided in the field measurements, N₂O emission rates from the agricultural field and forest increased by 38 and 51%, respectively. Nitrous oxide consumption under elevated N₂O condition (about 300 ppmv) was found in all five agricultural field measurements, but only in three of the six forest measurements under the same conditions. Field measurements agreed with the laboratory experiment that N₂O reduction activity, which plays a critical role in abating N₂O emissions from soils, largely depended on soil characteristics associated with land use.     

Verification of an in‐situ method for measuring denitr1f1cation

Abstract

The most common direct in‐situ method for the measurement of soil denitrification requires many acetylene (C₂H₂) supply probes and airflow lines to measure nitrous oxide (N₂O) flux from the soil under a sealed cover. A modification to this method simplified C₂H₂ supply by placing a single acetylene supply probe 30 cm deep into the soil and measured soil N₂O emission flux over a 0.11 m² area. Acetylene concentrations ranging from 0.1–10.0% were readily and predictably established by radial diffusion from the supply probe. Over 94% of the N₂O released into the enclosed air space of the soil cover was recovered at an air flow rate of 21 L/h. Recovery decreased rapidly with increased flow rates of 31 and 37 L/h.     

Comparison of two versions of the acetylene inhibition/soil core method for measuring denitrification loss from an irrigated wheat field

 Two versions of the acetylene inhibition (AI)/soil core method were compared for the measurement of denitrification loss from an irrigated wheat field receiving urea-N at a rate of 100 kg ha^–1. With AI/soil core method A, the denitrification rate was measured by analysing the headspace N₂O, followed by estimation of N₂O dissolved in the solution phase using Bunsen absorption coefficients. With AI/soil core method B, N₂O entrapped in the soil was measured in addition to that released from soil cores into the headspace of incubation vessels. In addition, the two methods were also compared for measurement of the soil respiration rate. Of the total N₂O produced, 6–77% (average 40%) remained entrapped in the soil, whereas for CO₂, the corresponding figures ranged from 12–65% (average 44%). The amount of the entrapped N₂O was significantly correlated with the water-filled pore space (WFPS) and with the N₂O concentration in the headspace, whereas CO₂ entrapment was dependent on the headspace CO₂ concentration but not on the WFPS. Due to the entrapment of N₂O and CO₂ in soil, the denitrification rate on several (18 of the 41) sampling dates, and soil respiration rate on almost all (27 of the 30) sampling dates were significantly higher with method B compared to method A. Averaged across sampling dates, the denitrification rate measured with method B (0.30 kg N ha^–1 day^–1) was twice the rate measured with method A, whereas the soil respiration rate measured with method B (34.9 kg C ha^–1 day^–1) was 1.6 times the rate measured with method A. Results of this study suggest that the N₂O and CO₂ entrapped in soil should also be measured to ensure the recovery of the gaseous products of denitrification by the soil core method. Received: 12 May 1998     

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.     

A gas-flow soil core method to measure field denitrification rates

A method was developed for rapid measurement of soil denitrification under conditions where natural soil structure and aeration status is maintained. Air was continuously recirculated by means of a membrane pump through a soil core and a sample loop of a gas chromatograph equipped with an electron capture detector. Addition of acetylene to the recirculating air permitted measurement of denitrification in the soil core. Because of the rapid distribution of C₂H₂ and removal of N₂O provided by the gas flow, denitrification rates could usually be determined in less than 2 h. By means of external 6-way and 8-way valves, four soil cores could be simultaneously analyzed on one gas Chromatograph equipped with dual detectors. Soil cores could also be stored at 4°C for later analysis without affecting the denitrification rate. The detection limit for denitrification rate measurements was 0.5 ngN g^?1 soil day^?1 or approximately 2.6 g N ha^?1 day^?1. Coefficients of variation for repeated measurements on the same soil core were usually less than 15%, but coefficients of variation for repacked or natural cores of the same soil were much higher (70–90%) Disruption of the natural soil structure by sieving increased the denitrification rate in an aggregated clay loam soil, but decreased the rate in a non-aggregated sandy soil. These results illustrate the importance of maintaining natural soil structure during denitrification measurements. The effect of pumping gas through soil was evaluated by comparing denitrification rates in soil cores where C₂H₂ was allowed to distribute into the soil by passive diffusion with rates obtained by pumping. Lower denitrification rates were observed in the static incubation presumably due to limited diffusion of C₂H₂ into or N₂O out of the denitrifying sites in the soil. This diffusion limitation could be overcome in the static incubations if C₂H₂ was initially distributed through the soil by pumping. This gas flow method is well suited to the study of soil denitrification rates under nearly natural conditions because the indigenous substrates and anaerobic microsites are preserved, the rapidity in which denitrification rates can be measured, and the high sensitivity and relatively low analytical variability of the method.     

Relationships between soil moisture content and nitrous oxide production during nitrification and denitrification

Summary The effect of soil water content [60%–100% water-holding capacity (WHC)] on N₂O production during autotrophic nitrification and denitrification in a loam soil was studied in a laboratory experiment by selectively inhibiting nitrification with a low C₂H₂ concentration (2.1 Pa). Nitrifiers usually produced more N₂O than denitrifiers. During an initial experimental period of 0–6 days the nitrifiers produced more N₂O than the denitrifiers by a factor ranging from 1.4 to 16.5, depending on the water content and length of incubation. The highest N₂O production rate by nitrifiers was observed at 90% WHC, when the soil had become partly anaerobic, as indicated by the high denitrification rate. At 100% WHC there were large gaseous losses from denitrification, while nitrification losses were smaller except for the first period of measurement, when there was still some O₂ remaining in the soil. The use of 10 kPa C₂H₂ to inhibit reduction of N₂O to N₂ stimulated the denitrification process during prolonged incubation over several days; thus the method is unsuitable for long-term studies.     

Quantifying the underestimation of soil denitrification potential as determined by the acetylene inhibition method

The acetylene inhibition technique (AIT) is commonly used for the determination of soil denitrification potential (SDP). However, the results obtained with this technique have intrinsic uncertainties. This study aimed at quantifying these uncertainties. A silt loam topsoil from an experimental station was incubated under standard conditions for SDP measurements, with and without acetylene additions. Changes in the concentrations of N₂O and N₂ in the headspace of incubation vials were monitored during the incubation, using a robotized sampling and analyzing system. Our results showed that the AIT did not completely inhibit the reduction of N₂O to N₂. As much as 11.7% of the produced N₂O was reduced into N₂ in the presence of acetylene. We conclude that SDP has to be corrected when determined by the AIT. The procedures used in this study will allow the estimation of correction factors, possibly as function of soil type and environmental conditions.     

灌溉和降水对旱地土壤N2O气态损失的影响

利用土壤探头法和密闭气室法相结合 ,就黄土高原旱地土壤玉米生长期灌溉和降水对N2O气态损失的影响进行了研究 ;并采用乙炔抑制原状土柱培养法 ,对土壤由湿变干和由干变湿过程中N2O变化进行了模拟。试验结果表明 ,在旱地土壤上 ,N2O的变化一般较小 ,但在降雨或者灌溉后无论是土壤N2O通量或者土壤剖面中N2O的浓度均呈现上升趋势 ,且这种变化趋势与同时期降雨量的变化趋势相同。培养结果说明 ,在相同的土壤孔隙水含量 (WFPS)条件下 ,土壤由湿变干过程产生的N2O通量高于土壤由干变湿过程中的产生量 ;在土壤由干变湿过程中N2O通量随土壤WFPS含量的增加而上升 ,但在土壤由湿变干过程中土壤N2O通量在WFPS含量为 70%时达到最大 ,而后随土壤WFPS含量的减少而下降。施肥处理与对照相比两者的变化趋势相同 ,但不施肥处理的变化幅度较小     

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.     

Acetylene inhibition of nitrous oxide reduction and measurement of denitrification and nitrogen fixation in soil

Reduction of N₂O in moist soil was inhibited completely by 10^?2 atm C₂H₂ and partially by 10^?5 atm C₂H₂. The effect of C₂H₄ was 10⁴ times less than that of C₂H₂. Denitrification of NO^?₃ occurred in anaerobically or aerobically incubated waterlogged soil and in anaerobic but not in aerobic moist soil. In the absence of C₂H₂ there was transient accumulation of N₂O. In the presence of C₂H₂ there was stoichiometric conversion of NO^?₃ to N₂O. Some kinetics of the reduction of N₂O and of NO^?₃ to N₂O are presented. Denitrification of 1 μg added NO^?₃-N.g^? could be measured within 1 h. Stoichiometries of production of N₂O from NO^?₂ and NO^?₃, respectively, and production of CO₂ attributable to denitrification were consistent with reported energy yields. Reduction of C₂H₂ to C₂H₄ occurred immediately following complete denitrification of added NO^?₃. The incubation of soil in the presence and in the absence of C₂H₂ thus permits assay of both denitrification and N₂ fixation and provides information on the mole fraction of N₂O in the products of denitrification.     

Small-scale heterogeneity in carbon dioxide, nitrous oxide and methane production from aggregates of a cultivated sandy-loam soil

Spatial variability in carbon dioxide (CO₂), nitrous oxide (N₂O) and methane (CH₄) emissions from soil is related to the distribution of microsites where these gases are produced. Porous soil aggregates may possess aerobic and anaerobic microsites, depending on the water content of pores. The purpose of this study was to determine how production of CO₂, N₂O and CH₄ was affected by aggregate size and soil water content. An air-dry sandy loam soil was sieved to generate three aggregate fractions (<0.25 mm, 0.25–2 mm and 2–6 mm) and bulk soil (<2 mm). Aggregate fractions and bulk soil were moistened (60% water-filled pore space, WFPS) and pre-incubated to restore microbial activity, then gradually dried or moistened to 20%, 40%, 60% or 80% WFPS and incubated at 25 °C for 48 h. Soil respiration peaked at 40% WFPS, presumably because this was the optimum level for heterotrophic microorganisms, and at 80% WFPS, which corresponded to the peak N₂O production. More CO₂ was produced by microaggregates (<0.25 mm) than macroaggregate (>0.25 mm) fractions. Incubation of aggregate fractions and soil at 80% WFPS with acetylene (10 Pa and 10 kPa) and without acetylene showed that denitrification was responsible for 95% of N₂O production from microaggregates, while nitrification accounted for 97–99% of the N₂O produced by macroaggregates and bulk soil. This suggests that oxygen (O₂) diffusion into and around microaggregates was constrained, whereas macroaggregates remained aerobic at 80% WFPS. Methane consumption and production were measured in aggregates, reaching 1.1–6.4 ng CH₄–C kg⁻¹ soil h⁻¹ as aggregate fractions and soil became wetter. For the sandy-loam soil studied, we conclude that nitrification in aerobic microsites contributed importantly to total N₂O production, even when the soil water content permitted denitrification and CH₄ production in anaerobic microsites. The relevance of these findings to microbial processes controlling N₂O production at the field scale remains to be confirmed.     

Denitrification in waterlogged soils: In situ temperature-dependent variations

A method to measure nitrous oxide (N₂O) produced by denitrification in the field is described. The system shows a relationship between N₂O emission and the daily temperature variations of the surface soil. We compare the results with those obtained by complementary use of acetylene. A conversion factor between emitted N₂O (without C₂H₂) and reduced NO^?₃ is discussed.     

Effects of phosphorus addition on N2O and NO emissions from soils of an Acacia mangium plantation

Abstract

An incubation experiment was conducted to examine the effects of the phosphorus (P) application on nitrous oxide (N₂O) and nitric oxide (NO) emissions from soils of an Acacia mangium plantation in Indonesia. The soils were incubated with and without the addition of P (Ca[H₂PO₄]₂; 2 mg P g soil)^?1) after adjusting the water-filled pore space (WFPS) to 75% or 100%. The P addition increased N₂O emissions under both WFPS conditions and NO emissions at 75% WFPS. Some possible mechanisms are considered. First, the P addition stimulated nitrogen (N) cycling, and N used for nitrification and/or denitrification also increased. Second, the P addition could have relieved the P shortage for nitrifying and/or denitrifying bacteria, producing N₂O and NO. Our results suggest that the application of P fertilizer has the potential to stimulate N₂O and NO emissions from Acacia mangium plantations, at least when soils are under relatively wet conditions.     

土壤温度、水分和NH4+-N浓度对土壤硝化反应速度及N2O排放的影响

硝化反应是土壤、特别是干旱半干旱地区农业土壤N2O产生的重要途径之一。但是,目前环境条件对硝化反应中N2O排放的影响研究较少,而在国内外通用的几个模型中均用固定比例估算硝化反应过程中N2O的排放。本文通过砂壤土培养试验,研究了土壤温度、水分和NH4+-N浓度对硝化反应速度及硝化反应中N2O排放的影响,并用数学模型定量表示了各因素对硝化反应的作用,用最小二乘法最优拟合求得该土壤的最大硝化反应速度及N2O最大排放比例。结果表明,随着温度升高,硝化反应速度呈指数增长;水分含量由20%充水孔隙度(WFPS)增加到40%WFPS时,反应速度增加,水分含量增加到60%WFPS时反应速度略有降低;NH4+-N浓度增加对硝化反应速度起抑制作用。用米氏方程描述该土壤的硝化反应过程,其最大硝化反应速度为6.67mg·kg?1·d?1。硝化反应中N2O排放比例随温度升高而降低;随NH4+-N浓度增加而略有增加;20%和40%WFPS水分含量时,硝化反应中N2O排放比例为0.43%～1.50%,最小二乘法求得的最大比例为3.03%,60%WFPS时可能由于反硝化作用,N2O排放比例急剧增加,还需进一步研究水分对硝化反应中N2O排放的影响。     

Influence of air porosity on distribution of gases in soil under assay for denitrification

 There has been concern that the measurement of gas emissions from a soil surface may not accurately reflect gas production within the soil profile. But, there have been few direct assessments of the error associated with the use of surface emissions for estimating gas production within soil profiles at different water contents. To determine the influence of air porosity on the distribution of gases within soil profiles, denitrification assays were performed using soil columns incubated with different water contents to provide air porosities of 18%, 13%, and 0% (equivalent to 62%, 73%, and 100% water-filled pore space, respectively). The soil columns were formed by packing sieved soil into cylinders which could be sealed at the top to form a headspace for the measurement of surface emissions of soil gases. Gas-permeable silicone tubing was placed at three depths (4.5, 9, and 13.5 cm) within each soil core to permit the measurement of gas concentration gradients within the soil core. Assays for denitrification were initiated by the addition of acetylene (5 kPa) to the soil column, and gas samples were taken from both the headspace and gas-permeable tubing at various times during a 46-h incubation. The results showed that at 18% air porosity, the headspace gases were well equilibrated with pore-space gases, and that gas emissions from the soil could provide good estimates of N₂O and CO₂ production. At air porosities of 13% and 0%, however, substantial storage of these gases occurred within the soil profiles, and measurements of surface emissions of gas from the soils greatly underestimated gas production. For example, the sole use of N₂O emission measurements caused three to five fold underestimates of N₂O production in soil maintained at 13% air porosity. It was concluded that the confounding influence of soil moisture on gas production and transport in soil greatly limits the use of surface emissions as a reliable indicator of gas production. This is particularly pertinent when assessing processes such as denitrification in which N gas production is greatly promoted by the conditions that limit O₂ influx and concurrently limit N gas efflux. Received: 15 January 1999     

Side Effects of Acetylene on the Conversion of Nitrate in Soil

Laboratory studies were conducted to determine if acetylene affects the rate of NO₃^? reduction to N₂O and N₂, if C₂H₂ ist anaerobically metabolized in the presence of nitrate, and if C₂H₂ affects soil carbon metabolism. These studies in water saturated soil, incubated under air or N₂ atmosphere, with or without acetone-free acetylene, show that C₂H₂ can accelerate NO₃^? reduction. Acetylene inhibited carbon mineralization when NO₃^? was limited but accelerated it when sufficient NO₃^? was available. After three days, only two per cent or less of the added C₂H₂ was directly oxidized to CO₂, however, up to 28 % of the added C₂H₂ carbon remained in the soil. The residual C₂H₂ carbon was oxidized aerobically and anaerobically when NO₃^? was added. These data suggest that when C₂H₂ is used in denitrification studies, the results must be carefully scrutinized. Once a soil is exposed to C₂H₂ it should not be used again soon to assess denitrification.     

Emissions of CO2 and N2O from a pasture soil from Madagascar—Simulating conversion to direct‐seeding mulch‐based cropping in incubations with organic and inorganic inputs

In the highlands of Madagascar, agricultural expansion gained on grasslands and cropping systems based on direct seeding with permanent vegetation cover are emerging as a means to sustain upland crop production. The objective of this study was to examine how such agricultural practices affect greenhouse‐gas emissions from a loamy Ferralsol previously used as a pasture. We conducted an experiment under controlled laboratory conditions combining cattle manure, crop residues (rice straw), and mineral fertilizers (urea plus NPK or di‐NH₄‐phosphate) to mimic on‐field inputs and examined soil CO₂ and N₂O emissions during a 28‐d incubation at low and high water‐filled pore space (40% and 90% WFPS). Emissions of N₂O from the control soil, i.e., soil receiving no input, were extremely small (< 5 ng N₂O‐N (g soil)^–1 h^–1) even under anaerobic conditions. Soil moisture did not affect the order of magnitude of CO₂ emissions while N₂O fluxes were up to 46 times larger at high soil WFPS, indicating the potential influence of denitrification under these conditions. Both CO₂ and N₂O emissions were affected by treatments, incubation time, and their interactions. Crop‐residue application resulted in larger fluxes of CO₂ but reduced N₂O emissions probably due to N immobilization. The use of di‐NH₄‐phosphate was a better option than NPK to reduce N₂O emissions without increasing CO₂ fluxes when soil received mineral fertilizers. Further studies are needed to translate the findings to field conditions and relate greenhouse‐gas budgets to crop production.     

Contributions of nitrification and denitrification to N2O emissions from soils at different water-filled pore space

A combination of stable isotope and acetylene (0.01% v/v) inhibition techniques were used for the first time to determine N₂O production during denitrification, autotrophic nitrification and heterotrophic nitrification in a fertilised (200 kg N ha^–1) silt loam soil at contrasting (20–70%) water-filled pore space (WFPS). ¹⁵N-N₂O emissions from ¹⁴NH₄¹⁵NO₃ replicates were attributed to denitrification and ¹⁵N-N₂O from ¹⁵NH₄¹⁵NO₃ minus that from ¹⁴NH₄¹⁵NO₃ replicates was attributed to nitrification and heterotrophic nitrification in the presence of acetylene, as there was no dissimilatory nitrate reduction to ammonium or immobilisation and remineralisation of ¹⁵N-NO₃^–. All of the N₂O emitted at 70% WFPS (31.6 mg N₂O-N m^–2 over 24 days; 1.12 g N₂O-N g dry soil^–1; 0.16% of N applied) was produced during denitrification, but at 35–60% WFPS nitrification was the main process producing N₂O, accounting for 81% of ¹⁵N-N₂O emitted at 60% WFPS, and 7.9 g ¹⁵N-N₂O m^–2 (0.28 ng ¹⁵N-N₂O g dry soil^–1) was estimated to be emitted over 7 days during heterotrophic nitrification in the 50% WFPS treatment and accounted for 20% of ¹⁵N-N₂O from this treatment. Denitrification was the predominant N₂O-producing process at 20% WFPS (2.6 g ¹⁵N-N₂O m^–2 over 7 days; 0.09 ng ¹⁵N-N₂O g dry soil^–1; 85% of ¹⁵N-N₂O from this treatment) and may have been due to the occurrence of aerobic denitrification at this WFPS. Our results demonstrate the usefulness of a combined stable isotope and acetylene approach to quantify N₂O emissions from different processes and to show that several processes may contribute to N₂O emission from agricultural soils depending on soil WFPS.