Isotopologue ratios of N2O emitted from microcosms with NH4 fertilized arable soils under conditions favoring nitrification

Soils represent the major source of the atmospheric greenhouse gas nitrous oxide (N₂O) and there is a need to better constrain the total global flux and the relative contribution of the microbial source processes. The aim of our study was to determine variability and control of the isotopic fingerprint of N₂O fluxes following NH₄⁺-fertilization and dominated by nitrification. We conducted a microcosm study with three arable soils fertilized with 0–140 mg NH₄⁺–N kg⁻¹. Fractions of N₂O derived from nitrification and denitrification were determined in parallel experiments using the ¹⁵N tracer and acetylene inhibition techniques or by comparison with unfertilized treatments. Soils were incubated for 3–10 days at low moisture (30–55% water-filled pore space) in order to establish conditions favoring nitrification. Dual isotope and isotopomer ratios of emitted N₂O were determined by mass spectrometric analysis of δ¹⁸O, average δ¹⁵N (δ¹⁵N^bulk) and ¹⁵N site preference (SP = difference in δ¹⁵N between the central and peripheral N positions of the asymmetric N₂O molecule). N₂O originated mainly from nitrification (>80%) in all treatments and the proportion of NH₄⁺ nitrified that was lost as N₂O ranged between 0.07 and 0.45%. δ¹⁸O and SP of N₂O fluxes ranged from 15 to 28.4‰ and from 13.9 to 29.8‰, respectively. These ranges overlapped with isotopic signatures of N₂O from denitrification reported previously. There was a negative correlation between SP and δ¹⁸O which is opposite to reported trends in N₂O from denitrification. Variation of average ¹⁵N signatures of N₂O (δ¹⁵N^bulk) did not supply process information, apparently because a strong shift in precursor signatures masked process-specific effects on δ¹⁵N^bulk. Maximum SP of total N₂O fluxes and of nitrification fluxes was close to reported SP of N₂O from NH₄⁺ or NH₂OH conversion by autotrophic nitrifiers, suggesting that SP close to 30‰ is typical for autotrophic nitrification in soils following NH₄⁺-fertilization. The results suggest that the δ¹⁸O/SP fingerprint of N₂O might be used as a new indicator of the dominant source process of N₂O fluxes in soils.     

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.     

Isotopomer signatures of soil-emitted N2O under different moisture conditions—A microcosm study with arable loess soil

Soils represent the major source of the atmospheric greenhouse gas nitrous oxide (N₂O) and there is a need to better constrain the total global flux and the relative contribution of the microbial source processes. The aim of our study was to evaluate isotopomer analysis of N₂O (intramolecular distribution of ¹⁵N) as well as conventional nitrogen and oxygen isotope ratios (i) as a tool to identify N₂O production processes in soils and (ii) to constrain the isotopic fingerprint of soil-derived N₂O. We conducted a microcosm study with arable loess soil fertilized with 20 mg N kg⁻¹ of ¹⁵NO₃⁻-labeled or non-labeled ammonium nitrate. Soils were incubated for 16 d at varying moisture (55%, 75% and 85% water-filled pore space (WFPS)) in order to establish different levels of nitrification and denitrification. Dual isotope and isotopomer ratios of emitted N₂O were determined by mass spectrometric analysis of δ¹⁸O, average δ¹⁵N (δ¹⁵N^bulk) and ¹⁵N site preference (SP=difference in δ¹⁵N between the central and peripheral N-positions of the asymmetric N₂O molecule). Total rates and N₂O emission of denitrification and nitrification were determined by ¹⁵N analysis of headspace gases and soil extracts of the ¹⁵NO₃⁻ treatment. N₂O emission and denitrification increased with moisture whereas gross nitrification was almost constant. In the 55% WFPS treatment, more than half of the N₂O flux was derived from nitrification, whereas denitrification was the dominant N₂O source in the 75% WFPS and 85% WFPS treatments. Moisture conditions were reflected by the isotopic signatures since highly significant differences were observed for average δ¹⁵N^bulk, SP and δ¹⁸O. Experiment means of the 75% WFPS and 85% WFPS treatments gave negative δ¹⁵N^bulk (−18.0‰ and −34.8‰, respectively) and positive SP (8.6‰ and 15.3‰, respectively), which we explained by the fractionation during N₂O production and partial reduction to N₂. In the 55% WFPS treatment, mean SP was relatively low (1.9‰), which suggests that nitrification produced N₂O with low or negative SP. The observed influence of process condition on isotopomer signatures suggests that the isotopomer approach might be suitable for identifying N₂O source processes. However, more research is needed to determine the impact from process rates and microbial community structure. Isotopomer signatures were within the range reported from previous soil studies which supports the assumption that SP of soil-derived N₂O is lower than SP of tropospheric N₂O.     

Isotopomer analysis of nitrous oxide accumulated in soil cultivated with tea (Camellia sinensis) in Shizuoka,central Japan

Nitrous oxide (N₂O) is a greenhouse gas that is destroying the stratospheric ozone to an increasing degree. Because of nitrogenous fertilizer application, agricultural soil is an important contributor of global N₂O. In Japan, tea fields are amended with the highest level of N fertilizers among agricultural soils, causing soil acidification and large N₂O flux. In soil, microbes play key roles in producing and consuming N₂O. A previous study investigated net N₂O production in tea fields using N₂O flux measurement and soil incubation, which are indirect methods to analyze relevant processes of N₂O production and consumption in soil. In the present study, to analyze N₂O concentrations and isotopomer ratios (bulk nitrogen and oxygen isotope ratios, δ¹⁵N^bulk and δ¹⁸O, and intramolecular ¹⁵N site preference, SP) and to reveal most probable microbial production processes and consumption (N₂O reduction to N₂) process of N₂O, soil gas was collected from a tea field (pH 3.1–4.5) at 10–50 cm depths using a silicone tube. The combination of fertilization, precipitation, and temperature rise produced significantly high N₂O concentrations. During the period of high N₂O concentration (above 5.7 ppmv), SP, the difference in ¹⁵N/¹⁴N ratio between central (α) and terminal (β) nitrogen position in the linear N₂O molecule (^βN^αNO) showed low values of 1.4‰–9.8‰, suggesting that the contribution of bacterial denitrification (nitrifier-denitrification and bacterial denitrifier-denitrification) was greater than that of bacterial nitrification or fungal denitrification. High SP values of 15.0‰–20.1‰ obtained at 10, 35, and 50 cm depths on 31 May 2011 (after one of fertilizations) during which soil temperatures were 15.8 °C–17.9 °C and water-filled pore space (WFPS) was 0.73–0.89 suggest that fungal denitrification and bacterial nitrification contributed to N₂O production to a degree equivalent to that of bacterial denitrification.     

Temporal and spatial patterns of denitrification enzyme activity and nitrous oxide fluxes in three adjacent vegetated riparian buffer zones

The aim of this study was to investigate temporal and spatial patterns of denitrification enzyme activity (DEA) and nitrous oxide (N₂O) fluxes in three adjacent riparian sites (mixed vegetation, forest and grass). The highest DEA was found in the surface (0–30 cm) soil and varied between 0.7±0.1 mg N kg^–1 day^–1 at 5°C and 5.9±0.4 mg N kg^–1 day^–1 at 15°C. There was no significant difference (P >0.05) between the DEA in the uppermost (0–30 cm and 60–90 cm) soil depths under different vegetation covers. In the two deepest (120–150 cm and 180–210 cm) soil depths the DEA varied between 0.0±0.0 mg N kg^–1 day^–1 at 5°C and 4.4±0.9 mg N kg^–1 day^–1 at 15°C and was clearly associated with the accumulation of buried organic carbon (OC). Two threshold values of OC were observed before DEA started to increase significantly, namely 5 and 25 g OC kg^–1 soil at 10–15°C and 5°C, respectively. In the three riparian sites N₂O fluxes varied between a net N₂O uptake of –0.6±0.4 mg N₂O-N m^–2 day^–1 and a net N₂O emission of 2.5±0.3 mg N₂O-N m^–2 day^–1. The observed N₂O emission did not lead to an important pollution swapping (from water pollution to greenhouse gas emission). Especially in the mixed vegetation and forest riparian site highest N₂O fluxes were observed upslope of the riparian site. The N₂O fluxes showed no clear temporal trend.     

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.     

Soil N<Subscript>2</Subscript>O emissions and N<Subscript>2</Subscript>O/(N<Subscript>2</Subscript>O+N<Subscript>2</Subscript>) ratio as affected by different fertilization practices and soil moisture

The objective of this work was to evaluate the effect of the chemical nature and application frequency of N fertilizers at different moisture contents on soil N₂O emissions and N₂O/(N₂O+N₂) ratio. The research was based on five fertilization treatments: unfertilized control, a single application of 80 kg ha⁻¹ N-urea, five split applications of 16 kg ha⁻¹ N-urea, a single application of 80 kg ha⁻¹ N–KNO₃, five split applications of 16 kg ha⁻¹ N–KNO₃. Cumulative N₂O emissions for 22 days were unaffected by fertilization treatments at 32% water-filled pore space (WFPS). At 100% and 120% WFPS, cumulative N₂O emissions were highest from soil fertilized with KNO₃. The split application of N fertilizers decreased N₂O emissions compared to a single initial application only when KNO₃ was applied to a saturated soil, at 100% WFPS. Emissions of N₂O were very low after the application of urea, similar to those found at unfertilized soil. Average N₂O/(N₂O+N₂) ratio values were significantly affected by moisture levels (p = 0.015), being the lowest at 120% WFPS. The N₂O/(N₂O+N₂) ratio averaged 0.2 in unfertilized soil and 0.5 in fertilized soil, although these differences were not statistically significant.     

Dinitrogen and N2O emissions in arable soils: Effect of tillage, N source and soil moisture

A laboratory investigation was performed to compare the fluxes of dinitrogen (N₂), N₂O and carbon dioxide (CO₂) from no-till (NT) and conventional till (CT) soils under the same water, mineral nitrogen and temperature status. Intact soil cores (0-10 cm) were incubated for 2 weeks at 25 °C at either 75% or 60% water-filled pore space (WFPS) with ¹⁵N-labeled fertilizers (100 mg N kg⁻¹ soil). Gas and soil samples were collected at 1-4 day intervals during the incubation period. The N₂O and CO₂ fluxes were measured by a gas chromatography (GC) system while total N₂ and N₂O losses and their ¹⁵N mole fractions in the soil mineral N pool were determined by a mass spectrometer. The daily accumulative fluxes of N₂ and N₂O were significantly affected by tillage, N source and soil moisture. We observed higher (P<0.05) fluxes of N₂+N₂O, N₂O and CO₂ from the NT soils than from the CT soils. Compared with the addition of nitrate (NO₃⁻), the addition of ammonium (NH₄⁺) enhanced the emissions of these N and C gases in the CT and NT soils, but the effect of NH₄⁺ on the N₂ and/or N₂O fluxes was evident only at 60% WFPS, indicating that nitrification and subsequent denitrification contributed largely to the gaseous N losses and N₂O emission under the lower moisture condition. Total and fertilizer-induced emissions of N₂ and/or N₂O were higher (P<0.05) at 75% WFPS than with 60% WFPS, while CO₂ fluxes were not influenced by the two moisture levels. These laboratory results indicate that there is greater potential for N₂O loss from NT soils than CT soils. Avoiding wet soil conditions (>60% WFPS) and applying a NO₃⁻ form of N fertilizer would reduce potential N₂O emissions from arable soils.     

Influence of pH on the release of NO and N2O from fertilized and unfertilized soil

Summary NO and N₂O release rates were measured in an acidic forest soil (pH 4.0) and a slightly alkaline agricultural soil (pH 7.8) after the pH was adjusted to values ranging from pH 4.0 to 7.8. The total release of NO and N₂O during 20 h of incubation was determined together with the net changes in the concentrations of NH ₄ ⁺ , NO ₂ ^– and NO ₃ ^– in the soil. The release of NO and N₂O increased after fertilization with NH ₄ ⁺ and/or NO ₃ ^– ; it strongly decreased with increasing pH in the acidic forest soil; and it increased when the pH of the alkaline agricultural soil was decreased to pH 6.5. However, there was no simple correlation between NO and N₂O release or between these compounds and activities such as the NO ₂ ^– accumulation, NO ₃ ^– reduction, or NH ₄ ⁺ oxidation. We suggest that soil pH exerts complex controls, e.g., on microbial populations or enzyme activities involved in nitrification and denitrification.     

Nitrous oxide production in turfgrass systems: Effects of soil properties and grass clipping recycling

Soil N₂O emissions can affect global environments because N₂O is a potent greenhouse gas and ozone depletion substance. In the context of global warming, there is increasing concern over the emissions of N₂O from turfgrass systems. It is possible that management practices could be tailored to reduce emissions, but this would require a better understanding of factors controlling N₂O production. In the present study we evaluated the spatial variability of soil N₂O production and its correlation with soil physical, chemical and microbial properties. The impacts of grass clipping addition on soil N₂O production were also examined. Soil samples were collected from a chronosequence of three golf courses (10, 30, and 100-year-old) and incubated for 60 days at either 60% or 90% water filled-pore space (WFPS) with or without the addition of grass clippings or wheat straw. Both soil N₂O flux and soil inorganic N were measured periodically throughout the incubation. For unamended soils, cumulative soil N₂O production during the incubation ranged from 75 to 972 ng N g⁻¹ soil at 60% WFPS and from 76 to 8842 ng N g⁻¹ soil at 90% WFPS. Among all the soil physical, chemical and microbial properties examined, soil N₂O production showed the largest spatial variability with the coefficient of variation ~110% and 207% for 60% and 90% WFPS, respectively. At 60% WFPS, soil N₂O production was positively correlated with soil clay fraction (Pearson's r = 0.91, P < 0.01) and soil NH₄⁺–N (Pearson's r = 0.82, P < 0.01). At 90% WFPS, however, soil N₂O production appeared to be positively related to total soil C and N, but negatively related to soil pH. Addition of grass clippings and wheat straw did not consistently affect soil N₂O production across moisture treatments. Soil N₂O production at 60% WFPS was enhanced by the addition of grass clippings and unaffected by wheat straw (P < 0.05). In contrast, soil N₂O production at 90% WFPS was inhibited by the addition of wheat straw and little influenced by glass clippings (P < 0.05), except for soil samples with >2.5% organic C. Net N mineralization in soil samples with >2.5% organic C was similar between the two moisture regimes, suggesting that O₂ availability was greater than expected from 90% WFPS. Nonetheless, small and moderate changes in the percentage of clay fraction, soil organic matter content, and soil pH were found to be associated with large variations in soil N₂O production. Our study suggested that managing soil acidity via liming could substantially control soil N₂O production in turfgrass systems.     

Anaerobic digestates lower N2O emissions compared to cattle slurry by affecting rate and product stoichiometry of denitrification – An N2O isotopomer case study

Assessing effects of organic fertilizer applications on N₂O emissions is of great interest because they can cause higher N₂O emissions compared to inorganic fertilizers for a given amount of added nitrogen (N). But there are also reports about enhanced N₂O reduction to climate-neutral elemental N₂ after application of organic manures to soils. Factors controlling the N₂O/(N₂O + N₂) product ratio of denitrification are interrelated, and also the ratio is difficult to study because of limitations in N₂ flux measurements. In this study, we investigated N₂O and N₂ emissions from soil treated with organic fertilizers with different C/N ratios. An N₂O isotopomer approach combined with conventional N₂O and N₂ flux measurements was employed to study underlying microbial pathways.A grassland soil was amended with anaerobic digestate (AD) from food waste digestion (low C/N ratio) or cattle slurry (CS; high C/N ratio), respectively, adjusted to 90% WFPS, and incubated for 52 days under helium–oxygen atmosphere (10% O₂) using a soil incubation system capable of automated N₂O, N₂, and CO₂ measurements. N₂O isotopomer signatures, i.e. the δ¹⁸O and SP values (site preference between ¹⁵N at the central and the peripheral position in the N₂O molecule), were determined by Isotope Ratio Mass Spectrometry and used to model and subsequently estimate the contribution of bacterial denitrification and autotrophic nitrification to N₂O production. For this approach the direct determination of emitted N₂ is essential to take isotope effects during N₂O reduction to N₂ into account by correcting the measured isotope signatures for isotope effects during N₂O reduction using previously determined fractionation factor ranges.The addition of both organic fertilizers to soil drastically increased the rate of gaseous N emissions (N₂O + N₂), probably due to the effects of concurrent presence of nitrate and labile C on the denitrification rate. In the initial phase of the experiment (day 1 to ∼15), gaseous N emissions were dominated by N₂ fluxes in soils amended with organic manures; meanwhile, N₂O emissions were lower compared to untreated Control soils, but increased after 15–20 days relative to the initial fluxes, especially with CS. Extremely low N₂O, but high N₂ emissions in the initial phase suggest that reduction of N₂O to N₂ via denitrification was triggered when the soil was amended with organic fertilizers. In contrast in the untreated Control, N₂O release was highest during the initial phase. Total N₂O release from AD treated soil was similar to Control, while N₂O from CS treated soil was considerably higher, indicating that denitrification was triggered more by the high labile carbon content in CS, while the cumulative N₂O/(N₂O + N₂) product ratio and thus N₂O reduction were similar with both organic fertilizers.The results of the N₂O source partitioning based on the isotopomer data suggest that about 8–25% (AD) and 33–43% (CS) of the cumulated N₂O emission was due to nitrification in organically amended soil, while in the untreated Control nitrification accounted for about 5–16%. The remaining N₂O production was attributed mainly to denitrification, while the poor model fit for other source pathways like fungal denitrification suggested their contribution to be of minor importance. The observed rather distinct phases with predominance first of denitrification and later of nitrification may help developing mitigation measures by addressing N₂O source processes individually with appropriate management options. The observation of relatively large shares of nitrification-derived N₂O is surprising, but may possibly be related to the low soil pH and will require further investigation.The determination of N₂ production is essential for this isotopomer-based source partitioning approach, but so far only applicable under laboratory conditions. The results of this study indicate that the combination of N₂O δ¹⁸O and SP values is very useful in obtaining more robust source estimates as compared to using SP values alone.     

Nitrous oxide emissions from animal urine application on a New Zealand pasture

Animal excreta-nitrogen (N) deposited onto pastoral soils during grazing has been identified as an important source of nitrous oxide (N₂O). Understanding the extent and seasonal variation of N₂O emissions from animal urine is important for the development of best management practices for reducing N₂O losses. The aim of this study was to determine N₂O emissions from cow urine after application onto a pastoral soil in different seasons between 2003 and 2005. A closed soil chamber technique was used to measure the N₂O emissions from a poorly drained silt loam soil which received either 0 (control) or 1,000 kg N ha⁻¹ (as real cow urine) per application. Application of cow urine to soil increased N₂O fluxes above those from the control site for up to 6 weeks, but the duration for which N₂O levels were elevated depended on the season. Nitrous oxide emissions were higher during the winter and spring measurement periods when the soil water-filled pore space (WFPS) was mostly above field capacity, and the emissions were lower during the summer and autumn measurement periods when the soil WFPS was below field capacity. The N₂O emission factor for urine ranged from 0.02 to 1.52% of N applied. This seasonal effect suggests that a reduction in urine return to soil (e.g., through use of standoff pads or animal housing) under wet conditions in New Zealand can potentially reduce N₂O emissions from pastoral soils.     

Isotopologue data reveal bacterial denitrification as the primary source of N2O during a high flux event following cultivation of a native temperate grassland

The source of N₂O in terrestrial ecosystems has long been debated. Both nitrification and denitrification produce N₂O but their relative importance remains uncertain. Here we apply site preference, SP (the difference in δ¹⁵N between the central and outer N atom in N₂O), to estimate the relative importance of bacterial denitrification (including nitrifier denitrification) to total N₂O production from soil. We measured SP over a diurnal cycle following the third year of tillage of a previously uncultivated grassland soil at the Kellogg Biological Station (KBS) in southwestern Michigan. Fluxes of N₂O in our study ranged between 7.8 and 12.1 g N₂O-N ha⁻¹ d⁻¹ and were approximately 3 and 10 times greater than fluxes observed in managed agricultural and successional fields, respectively, at KBS. Consequently, our study captured a period of high flux resulting from the cultivation of a historically never-tilled soil. Concentration weighted SP values decreased from 12.9‰ in the morning to a minimum value of −0.1‰ in the afternoon.Based on SP values reported for bacterial denitrification (−5 to 0‰; Toyoda et al., 2005; Sutka et al., 2006), hydroxylamine oxidation (nitrification) and fungal denitrification (33-37‰; Sutka et al., 2006) we found that production attributable to bacterial denitrification increased from between 52.9 and 60.9% in the morning to between 87.5 and 100% in the afternoon. Further, we observed diurnal variation in flux and SP that is consistent with increased production from bacterial denitrification associated with temperature-driven increases in respiration.     

Diffusion of N-labelled N2O into soil columns: a promising method to examine the fate of N2O in subsoils

Nitrous oxide research has generally focused directly on measuring fluxes of N₂O from the soil surface. The fate of N₂O in the subsoil has often been placed in the ‘too hard’ basket. However, determining the production, fate and movement of N₂O in the subsoil is vital in fully understanding the sources of surface fluxes and in compiling accurate inventories for N₂O emissions. The aim of this study was to generate and introduce into soil columns ¹⁵N labelled N₂O, and to try and determine the consumption of the ¹⁵N₂O and production of ambient N₂O. Columns, 100 cm long by 15 cm diameter, were repacked with sieved soil (sampled from 0 to 5 cm depth) and instrumented with silicone rubber gas sampling ports. Nitrous oxide enriched with ¹⁵N was generated using a thermal decomposition process at 300 °C and then transferred to 2 l flasks. After equilibrating with SF₆ tracer gas the ¹⁵N₂O was introduced into the soil columns via passive diffusion. Gas samples from the soil profile and headspace flux were taken over a 12-day period. A watering event was simulated to perturb the ¹⁵N₂O gas composition in the soil profile. Using the measured ¹⁵N enriched fluxes and the rate of decline in ¹⁵N in the N₂O reservoir, from which the N₂O diffused into the soil, we calculated an N₂O sink (consumption plus absorption by water) equal to 0.48 ng N₂O g⁻¹ soil h⁻¹. The decrease in the ¹⁵N enrichment between successive soil depths indicated N₂O production in the soil profile and we calculated a net N₂O production rate of 0.88 ng N₂O g⁻¹ soil h⁻¹. This pilot study demonstrated the potential for simultaneously measuring both N₂O consumption and production rates, using the ¹⁵N enrichment of the N₂O measured. Further potential refinements of the methodology are discussed.     

Nitrous oxide emission from an irrigated cotton field under semiarid subtropical conditions

In a 1-year study, quantification of nitrous oxide (N₂O) emission was made from a flood-irrigated cotton field fertilized with urea at 100kg N ha⁻¹ a⁻¹. Measurements were made during the cotton-growing season (May–November) and the fallow period (December–April). Of the total 95 sampling dates, 77 showed positive N₂O fluxes (range, 0.1 to 33.3g N ha⁻¹ d⁻¹), whereas negative fluxes (i.e., N₂O sink activity) were recorded on 18 occasions (range, −0.1 to −2.2g N ha⁻¹ d⁻¹). Nitrous oxide sink activity was more frequently observed during the growing season (15 out of 57 sampling dates) as compared to the fallow period (3 out of 38 sampling dates). During the growing season, contribution of N₂O to the denitrification gaseous N products was much less (average, 4%) as compared to that during the fallow period (average, 21%). Nitrous oxide emission integrated over the 6-month growing period amounted 324g N ha⁻¹, whereas the corresponding figure for the 6-month fallow period was 648g N ha⁻¹. Subtracting the N₂O sink activity (30.3g N ha⁻¹ and 3.8g N ha⁻¹ during the growing season and fallow period, respectively), the net N₂O emission amounted 938g N ha⁻¹ a⁻¹. Results suggested that high soil moisture and temperature prevailing under flood-irrigated cotton in the Central Punjab region of Pakistan though favor high denitrification rates, but are also conducive to N₂O reduction thus leading to relatively low N₂O emission.     

Effect of soil water status and mulching on N<Subscript>2</Subscript>O and CH<Subscript>4</Subscript> emission from lowland rice field in China

Cultivation of rice in unsaturated soils covered with mulch is receiving more attention in China because of increasingly serious water shortage; however, greenhouse gas emission from this cultivation system is still poorly understood. A field experiment was conducted in 2001 to compare nitrous oxide (N₂O) and methane (CH₄) emission from rice cultivated in unsaturated soil covered with plastic or straw mulch and the traditional waterlogged production system. Trace gas fluxes from the soil were measured weekly throughout the entire growth period using a closed chamber method. Nitrous oxide emissions from unsaturated rice fields were large and varied considerably during the rice season. They were significantly affected by N fertilizer application rate. In contrast, N₂O emission from the waterlogged system was very low with a maximum of 0.28 mg N₂O m^–2 h^–1. However, CH₄ emission from the waterlogged system was significantly higher than from the unsaturated system, with a maximum emission rate of 5.01 mg CH₄ m^–2 h^–1. Our results suggested that unsaturated rice cultivation with straw mulch reduce greenhouse gas emissions.     

CH4 oxidation and N2O emissions at varied soil water-filled pore spaces and headspace CH4 concentrations

Emission of N₂O and CH₄ oxidation rates were measured from soils of contrasting (30-75%) water-filled pore space (WFPS). Oxidation rates of ¹³C-CH₄ were determined after application of 10 μl ¹³C-CH₄ l⁻¹ (10 at. % excess ¹³C) to soil headspace and comparisons made with estimates from changes in net CH₄ emission in these treatments and under ambient CH₄ where no ¹³C-CH₄ had been applied. We found a significant effect of soil WFPS on ¹³C-CH₄ oxidation rates and evidence for oxidation of 2.2 μg ¹³C-CH₄ d⁻¹ occurring in the 75% WFPS soil, which may have been either aerobic oxidation occurring in aerobic microsites in this soil or anaerobic CH₄ oxidation. The lowest ¹³C-CH₄ oxidation rate was measured in the 30% WFPS soil and was attributed to inhibition of methanotroph activity in this dry soil. However, oxidation was lowest in the wetter soils when estimated from changes in concentration of ¹²⁺¹³C-CH₄. Thus, both methanogenesis and CH₄ oxidation may have been occurring simultaneously in these wet soils, indicating the advantage of using a stable isotope approach to determine oxidation rates. Application of ¹³C-CH₄ at 10 μl ¹³C-CH₄ l⁻¹ resulted in more rapid oxidation than under ambient CH₄ conditions, suggesting CH₄ oxidation in this soil was substrate limited, particularly in the wetter soils. Application of and (80 mg N kg soil⁻¹; 9.9 at.% excess ¹⁵N) to different replicates enabled determination of the respective contributions of nitrification and denitrification to N₂O emissions. The highest N₂O emission (119 μg ¹⁴⁺¹⁵N-N₂O kg soil⁻¹ over 72 h) was measured from the 75% WFPS soil and was mostly produced during denitrification (18.1 μg ¹⁵N-N₂O kg soil⁻¹; 90% of ¹⁵N-N₂O from this treatment). Strong negative correlations between ¹⁴⁺¹⁵N-N₂O emissions, denitrified ¹⁵N-N₂O emissions and ¹³C-CH₄ concentrations (r=−0.93 to −0.95, N₂O; r=−0.87 to −0.95, denitrified ¹⁵N-N₂O; P<0.05) suggest a close relationship between CH₄ oxidation and denitrification in our soil, the nature of which requires further investigation.     

Responses of soil organic matter and greenhouse gas fluxes to soil management and land use changes in a humid temperate region of southern Europe

We studied the effects of soil management and changes of land use on soils of three adjacent plots of cropland, pasture and oak (Quercus robur) forest. The pasture and the forest were established in part of the cropland, respectively, 20 and 40 yr before the study began. Soil organic matter (SOM) dynamics, water-filled pore space (WFPS), soil temperature, inorganic N and microbial C, as well as fluxes of CO₂, CH₄ and N₂O were measured in the plots over 25 months. The transformation of the cropland to mowed pasture slightly increased the soil organic and microbial C contents, whereas afforestation significantly increased these variables. The cropland and pasture soils showed low CH₄ uptake rates (<1 kg C ha⁻¹ yr⁻¹) and, coinciding with WFPS values >70%, episodes of CH₄ emission, which could be favoured by soil compaction. In the forest site, possibly because of the changes in soil structure and microbial activity, the soil always acted as a sink for CH₄ (4.7 kg C ha⁻¹ yr⁻¹). The N₂O releases at the cropland and pasture sites (2.7 and 4.8 kg N₂O-N ha⁻¹ yr⁻¹) were, respectively, 3 and 6 times higher than at the forest site (0.8 kg N₂O-N ha⁻¹ yr⁻¹). The highest N₂O emissions in the cultivated soils were related to fertilisation and slurry application, and always occurred when the WFPS >60%. These results show that the changes in soil properties as a consequence of the transformation of cropfield to intensive grassland do not imply substantial changes in SOM or in the dynamics of CH₄ and N₂O. On the contrary, afforestation resulted in increases in SOM content and CH₄ uptake, as well as decreases in N₂O emissions.     

N2O and NO production in various Chinese agricultural soils by nitrification

Eleven types of agricultural soils were collected from Chinese uplands and paddy fields to compare their N₂O and NO production by nitrification under identical laboratory conditions. Before starting the assays, all air-dried soils were preincubated for 4 weeks at 25 °C and 40% WFPS (water-filled pore space). The nitrification activities of soils were determined by adding (NH₄)₂SO₄ (200 mg N kg⁻¹ soil) and incubating for 3 weeks at 25 °C and 60% WFPS. The net nitrification rates obtained fitted one of two types of models, depending on the soil pH: a zero-order reaction model for acidic soils and one neutral soil (Group 0); or a first-order reaction model for one neutral soil and alkaline soils (Group 1). The results suggest that pH is the most important factor in determining the kinetics of soil nitrification from ammonium. In the Group 1 soils, initial emissions (i.e. during the first week) of N₂O and NO were 82.6 and 83.6%, respectively, of the total emissions during 3 weeks of incubation; in the Group 0 soils, initial emissions of N₂O and NO were 54.7 and 59.9%, respectively, of the total emissions. The net nitrification rate in the first week and second-third weeks were highly correlated with the initial and subsequent emissions (i.e. during the second and third weeks), respectively, of N₂O and NO. The average percentages of emitted (N₂O+NO)-N relative to net nitrification N in initial and subsequent periods were 2.76 and 0.59 for Group 0, and 1.47 and 0.44 for the Group 1, respectively. The initial and subsequent emission ratios of NO/N₂O from Group 0 (acidic) soils were 3.77 and 2.52 times, respectively, higher than those from Group 1 soils (P<0.05).     

Seasonal variations of nitrous oxide emission in relation to nitrogen fertilization and energy crop types in sandy soil

Nitrous oxide (N₂O) is a greenhouse gas and agricultural soils are major sources of atmospheric N₂O. Its emissions from soils make up the largest part in the global N₂O budget. Research was carried out at the experimental fields of the Leibniz-Institute of Agricultural Engineering Potsdam-Bornim (ATB). Different types (mineral and wood ash) and levels (0, 75 and 150 kg N ha⁻¹) of fertilization were applied to annual (rape, rye, triticale and hemp) and perennial (poplar and willow) plants every year. N₂O flux measurements were performed 4 times a week by means of gas flux chambers and an automated gas chromatograph between 2003 and 2005. Soil samples were also taken close to the corresponding measuring rings. Soil nitrate and ammonium were measured in soil extracts.N₂O emissions had a peak after N fertilization in spring, after plant harvest in summer and during the freezing–thawing periods in winter. Both fertilization and plant types significantly altered N₂O emission. The maximum N₂O emission rate detected was 1081 μg N₂O m⁻² h⁻¹ in 2004. The mean annual N₂O emissions from the annual plants were more than twofold greater than those of perennial plants (4.3 kg ha⁻¹ vs. 1.9 kg ha⁻¹). During January, N₂O fluxes considerably increased in all treatments due to freezing–thawing cycles. Fertilization together with annual cropping doubled the N₂O emissions compared to perennial crops indicating that N use efficiency was greater for perennial plants. Fertilizer-derived N₂O fluxes constituted about 32% (willow) to 67% (rape/rye) of total soil N₂O flux. Concurrent measurements of soil water content, NO₃ and NH₄ support the conclusion that nitrification is main source of N₂O loss from the study soils. The mean soil NO₃-N values of soils during the study for fertilized soils were 1.6 and 0.9 mg NO₃-N kg⁻¹ for 150 and 75 kg N ha⁻¹ fertilization, respectively. This value reduced to 0.5 mg NO₃-N kg⁻¹ for non-fertilized soils.