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.     

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.     

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.     

How important is N2O production in removing atmospherically deposited nitrogen from UK moorland catchments?

Nitrate (NO₃⁻) leaching due to anthropogenic nitrogen (N) deposition is an environmental problem in many parts of the UK uplands, associated with surface water acidification and affecting lake nutrient balances. It is often assumed that gaseous return of deposited N to the atmosphere as N₂O through denitrification may provide an important sink for N. This assumption was tested for four moorland catchments (Allt a’Mharcaidh in the Cairngorms, Afon Gwy in mid-Wales, Scoat Tarn in the English Lake District and River Etherow in the southern Pennines), covering gradients of atmospheric N deposition and surface water NO₃⁻ leaching, through a combination of field and laboratory experiments. Field measurements of N₂O fluxes from static chambers with and without additions of NH₄NO₃ solution were carried out every 4 weeks over 1 yr. Wetted soil cores from the same field plots were used in experimental laboratory incubations at 5 and 15 °C with and without additions of NH₄NO₃ solution, followed by measurement of N₂O fluxes. Field measurements showed that significant N₂O fluxes occurred in only a very small number of plots with most showing zero values for much of the year. The maximum fluxes were 0.24 kg-N/ha/yr from unamended plots at the River Etherow and 0.49 kg-N/ha/yr from plots with NH₄NO₃ additions at the Allt a’Mharcaidh. Laboratory incubation experiments demonstrated that large N₂O fluxes could be induced by warming and NH₄NO₃ additions, with the top 5 cm of soil cores responsible for the largest fluxes, reaching 11.8 kg-N/ha/yr from a podsol at Scoat Tarn. Acetylene block experiments showed that while N₂ was not likely to be a significant denitrification product in these soils, reduced N₂O fluxes indicated that nitrification was an important source of N₂O in many cases. A simple model of denitrification suggesting that 10-80% of net N inputs may be denitrified from non-agricultural soils was found to greatly over-estimate fluxes in the UK uplands. The proportion of deposition denitrified was found to be much closer to the IPCC suggested value of 1% with an upper limit of 10%. Interception of N deposition by vegetation may greatly reduce the net supply of N from this source, while soil acidification or other factors limiting carbon supply to soil microbes may prevent large denitrification fluxes even where NO₃⁻ supply is not limiting.     

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.     

The N2O product ratio of nitrification and its dependence on long-term changes in soil pH

The contribution of nitrification to the emission of nitrous oxide (N₂O) from soils may be large, but its regulation is not well understood. The soil pH appears to play a central role for controlling N₂O emissions from soil, partly by affecting the N₂O product ratios of both denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O/(NO₂⁻+NO₃⁻). Mechanisms responsible for apparently high N₂O product ratios of nitrification in acid soils are uncertain. We have investigated the pH regulation of the N₂O product ratio of nitrification in a series of experiments with slurries of soils from long-term liming experiments, spanning a pH range from 4.1 to 7.8. ¹⁵N labelled nitrate (NO₃⁻) was added to assess nitrification rates by pool dilution and to distinguish between N₂O from NO₃⁻ reduction and NH₃ oxidation. Sterilized soil slurries were used to determine the rates of chemodenitrification (i.e. the production of nitric oxide (NO) and N₂O from the chemical decomposition of nitrite (NO₂⁻)) as a function of NO₂⁻ concentrations. Additions of NO₂⁻ to aerobic soil slurries (with ¹⁵N labelled NO₃⁻ added) were used to assess its potential for inducing denitrification at aerobic conditions. For soils with pH?5, we found that the N₂O product ratios for nitrification were low (0.2-0.9‰) and comparable to values found in pure cultures of ammonia-oxidizing bacteria. In mineral soils we found only a minor increase in the N₂O product ratio with increasing soil pH, but the effect was so weak that it justifies a constant N₂O product ratio of nitrification for N₂O emission models. For the soils with pH 4.1 and 4.2, the apparent N₂O product ratio of nitrification was 2 orders of magnitude higher than above pH 5 (76‰ and 14‰). This could partly be accounted for by the rates of chemodenitrification of NO₂⁻. We further found convincing evidence for NO₂⁻-induction of aerobic denitrification in acid soils. The study underlines the role of NO₂⁻, both for regulating denitrification and for the apparent nitrifier-derived N₂O emission.     

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.     

Gaseous and leaching nitrogen losses from no-tillage and conventional tillage systems following surface application of cattle manure

Previous studies have demonstrated inconsistent results on the impact of tillage systems on nitrogen (N) losses from field-applied manure. This study assessed the impact of no-tillage (NT) and conventional tillage (CT) systems on gaseous N losses, N₂O:N₂O + N₂ ratios and NO₃⁻-N leaching following surface application of cattle manure. The study was undertaken during the 2003/2004 and 2004/2005 seasons at two field sites in Nova Scotia namely, Streets Ridge (SR) in Cumberland County and the Bio-environmental Engineering Centre (BEEC) in Truro. Results showed that the NT system had higher (p < 0.05) NH₃ losses than CT. Over the two seasons, manure incorporation in CT reduced NH₃ losses on average by 86% at SR and 78% at BEEC relative to NT. At both sites and during both seasons, denitrification rates and N₂O fluxes in NT were generally higher than in CT plots, presumably due to higher soil water and organic matter content in NT. Over the two seasons, mean denitrification rates at SR were 239 and 119 g N ha⁻¹ d⁻¹, while N₂O fluxes were 120 and 64 g N ha⁻¹ d⁻¹ under NT and CT, respectively. At BEEC mean denitrification rates were 114 and 71 g N ha⁻¹ d⁻¹, while N₂O fluxes were 52 and 27 g N ha⁻¹ d⁻¹ under NT and CT, respectively. Conversely, N₂O:N₂O + N₂ ratios were lower in NT than CT suggesting more complete reduction of N₂O to N₂ under NT. When averaged across all soil depths, NO₃⁻-N was higher (p < 0.05) in CT than NT. Nitrate-N decreased with depth at both sites regardless of tillage. In most cases, NO₃⁻-N was higher under CT than NT at all soil depths. Similarly, flow-weighted average NO₃⁻-N concentrations in drainage water were generally higher under CT. This may be partly attributed to higher denitrification rates under NT. Therefore, NT may be a viable strategy to remove NO₃⁻-N from the soil, and thus, reduce NO₃⁻-N contamination of groundwater. However, it should be noted that while the use of NT reduces NO₃⁻-N leaching it may come with unintended environmental tradeoffs, including increased NH₃ and N₂O emissions.     

Oxygen exchange between nitrogen oxides and H2O can occur during nitrifier pathways

Interpretation of the oxygen isotopic signature of soil-derived N₂O may be flawed when it is based on reaction stoichiometry and fractionation alone. In fact, oxygen (O) exchange between H₂O and intermediates of N₂O production pathways may largely determine this O isotopic signature. Although in our previous work we conclusively proved the occurrence of O exchange during N₂O production by denitrification of NO₃⁻, its occurrence in N₂O production pathways by nitrifiers remains unclear. The aim of this study was to examine the likeliness of O exchange during various stages of N₂O production in soil via nitrification, nitrifier denitrification and denitrification. We evaluated a set of scenarios on the presence of such exchange using data from a series of ¹⁸O and ¹⁵N tracing experiments. The measured actual O incorporation from H₂O into N₂O (AOI) was compared with the theoretical maximum O incorporation (MOI) from various scenarios that differed in their assumptions on the presence of O exchange. We found that scenarios where O exchange was assumed to occur exclusively during denitrification could not explain the observed AOI, as it exceeded the MOI for 9 out of 10 soils. This demonstrates that additional O exchange must have occurred in N₂O production through nitrifier pathways. It remains to be determined in which steps of these pathways O exchange can take place. We conclude that O exchange is likely to be mediated by ammonia oxidizers during NO₂⁻ reduction (nitrifier denitrification), and that it could possibly occur during NO₂⁻ oxidation to NO₃⁻ by nitrite oxidizers as well.     

Nitrous oxide emissions from fertilized,irrigated cotton (Gossypium hirsutum L.) in the Aral Sea Basin,Uzbekistan: Influence of nitrogen applications and irrigation practices

Nitrous oxide emissions were monitored at three sites over a 2-year period in irrigated cotton fields in Khorezm, Uzbekistan, a region located in the arid deserts of the Aral Sea Basin. The fields were managed using different fertilizer management strategies and irrigation water regimes. N₂O emissions varied widely between years, within 1 year throughout the vegetation season, and between the sites. The amount of irrigation water applied, the amount and type of N fertilizer used, and topsoil temperature had the greatest effect on these emissions.Very high N₂O emissions of up to 3000 μg N₂O-N m^?2 h^?1 were measured in periods following N-fertilizer application in combination with irrigation events. These “emission pulses” accounted for 80–95% of the total N₂O emissions between April and September and varied from 0.9 to 6.5 kg N₂O-N ha^?1.. Emission factors (EF), uncorrected for background emission, ranged from 0.4% to 2.6% of total N applied, corresponding to an average EF of 1.48% of applied N fertilizer lost as N₂O-N. This is in line with the default global average value of 1.25% of applied N used in calculations of N₂O emissions by the Intergovernmental Panel on Climate Change.During the emission pulses, which were triggered by high soil moisture and high availability of mineral N, a clear diurnal pattern of N₂O emissions was observed, driven by daily changes in topsoil temperature. For these periods, air sampling from 8:00 to 10:00 and from 18:00 to 20:00 was found to best represent the mean daily N₂O flux rates. The wet topsoil conditions caused by irrigation favored the production of N₂O from NO₃^? fertilizers, but not from NH₄⁺ fertilizers, thus indicating that denitrification was the main process causing N₂O emissions. It is therefore argued that there is scope for reducing N₂O emission from irrigated cotton production; i.e. through the exclusive use of NH₄⁺ fertilizers. Advanced application and irrigation techniques such as subsurface fertilizer application, drip irrigation and fertigation may also minimize N₂O emission from this regionally dominant agro-ecosystem.     

Nitrous oxide emissions from a bermudagrass pasture: Interseeded winter rye and poultry litter

Adequate use of manure in grasslands may constitute an economical means of manure disposal and an abundant source of nutrients for plants; however, excessive nitrogen (N) additions to these soils could create new environmental risks such as increasing nitrous oxide (N₂O) emissions. These potentially adverse effects in grasslands may be mitigated by improved management practices. In pasture systems, the combined effects of poultry litter applications and interseeded rye (Secale cereale L.) on N₂O emissions are still not well established. This study was conducted to estimate the magnitude of soil surface N₂O fluxes as affected by interseeded winter rye forage, annually spring-applied composted turkey litter as well as by weather and soil parameters. Fluxes were measured by vented chambers during 2 yr in a bermudagrass (Cynodon dactylon [L.] Pers.) pasture in moderately well-drained Tonti gravelly silt loam (fine-loamy, active, mesic Typic Fragiudault) located in northwestern Arkansas, USA. During the 60 d following turkey litter applications, N₂O fluxes were frequently well correlated with soil nitrate (NO₃⁻; r: up to 0.82, P's < 0.05) implying substrate stimulation on soil N₂O production. Likewise, rainfall patterns strongly influenced N₂O fluxes. Large rainfalls of 91 and 32 mm occurred within 6 d prior to the maximum N₂O flux means (263 and 290 μg N m⁻² h⁻¹, respectively). Treatment effects on N₂O emissions were significant only in spring periods following manure addition, particularly in the second year of our study. In the spring of 2000, additions of composted turkey litter resulted in 1.5-fold increase in seasonal cumulative N₂O emissions (P = 0.04) which was directly associated to a numerically greater soil NO₃⁻. In the spring of 2001, soils planted to rye exhibited a pronounced significant effect on mitigating N₂O emissions (30 vs. 112 mg N m⁻²; P = 0.04). During the winter and early spring, rye growth also decreased quantities of both soil NO₃⁻ and water-filled pore space (WFPS) partly accounting for the lower N₂O emissions in these fields. These results suggest that because poultry litter additions increased and interseeded rye diminished N₂O emissions, the combined implementation of both management practices can produce environmental benefits while sustaining productivity in temperate pasture systems.     

Low sulfide concentrations affect nitrate transformations in freshwater and saline coastal retention pond sediments

Coastal areas in the southeastern USA are prone to hurricanes and strong storms that may cause salt-water influx to freshwater aquatic sediments. These changes in environmental conditions may impact sediment processes including nitrogen (N) cycling. The relative abilities of sediment microbial communities from two freshwater golf course retention ponds that drain into the adjacent wetlands, and two proximal saline wetland ponds, to remove nitrate (NO₃⁻) were compared to assess whether low concentrations of sulfide changed N-transformation processes. Microcosms were incubated with NO₃-N (300 μg g dw⁻¹) alone, and with NO₃-N and sulfide (H₂S) (100 and 200 μg g dw⁻¹). Nitrous oxide (N₂O), nitrite (NO₂⁻), NO₃⁻, ammonium (NH₄⁺), SO₄²⁻ and acid volatile sulfides were analyzed over time. The acetylene block technique was used to measure denitrification in sediment microcosms with no added H₂S. Denitrification was measured without acetylene (C₂H₂) addition in microcosms with added H₂S. With no added H₂S, denitrification was greater in the freshwater retention ponds than in the wetland ponds. Although low H₂S concentrations generally increased NO₃-N removal rates at all sites, lag periods were increased and denitrification was inhibited by low sulfide in the freshwater sediments, as evidenced by the greater concentrations of N₂O that accumulated compared to those in the wetland sediments. In addition to the inability of the freshwater sediments to convert N₂O to N₂ in the absence of C₂H₂, anomalously high transient NO₂-N concentrations accumulated in the retention pond samples. NH₄-N formation generally decreased due to H₂S addition at the freshwater sites; NH₄-N formation increased initially at the wetland sites, but was greater when no H₂S was added. Storm events that allow influx of SO₄²⁻-containing seawater into freshwater systems may change the dominant N species produced from nitrate reduction. Even low concentrations of sulfide produced incomplete denitrification and decreased formation of NH₄⁺ in these coastal freshwater sediments.     

N2O and NO emissions from an Andisol field as influenced by pelleted poultry manure

Animal manures from intensive livestock operations can be pelleted to improve handlings and recyclings of embodied nutrients. The aim of this study was to evaluate the influence of pelleted poultry manure on N₂O and NO fluxes from an Andisol field. In autumn 2006 and summer 2007, poultry manure (PM), pelleted poultry manure (PP), and chemical fertilizer (CF) were applied at a rate of 120 kg N ha⁻¹ in each cultivation period to Komatsuna (Brassica rapa var. peruviridis). Nitrous oxide and NO fluxes were measured using an automated monitoring system. A soil incubation experiment was also conducted to determine the influence of intact and ground pelleted manure on N₂O, NO, and CO₂ production with a water-filled pore space (WFPS) of 30 or 50%. In the field measurements, N₂O emission rates from the organic fertilizer treatments were larger than that from the CF treatment, possibly because organic C stimulated denitrification. The highest N₂O flux was observed from the PP treatment after a rainfall following fertilization, and the cumulative emission rate (2.72 ± 0.22 kg N ha⁻¹ y⁻¹) was 3.9 and 7.1 times that from the PM and CF treatments, respectively. In contrast, NO emission rates were highest from the CF treatment. The NO/N₂O flux ratio indicated that nitrification was the dominant process for NO and N₂O production from the CF treatment. Cumulative N₂O emission rates from all treatments were generally higher during the wetter cultivation period (autumn 2006) than during the drier cultivation period (summer 2007). In contrast, NO emission rates were higher in the drier than in the wetter cultivation period. The incubation experiment results showed a synergistic effect of soil moisture and the pelleted manure form on N₂O emission rates. The intact pelleted manure with the 50% WFPS treatment produced the highest N₂O and CO₂ fluxes and resulted in the lowest soil NO₃⁻ content after the incubation. These results indicate that anaerobic conditions inside the pellets, caused by rainfall and heterotrophic microbial activities, led to denitrification, resulting in high N₂O fluxes. Controlling the timing of N application by avoiding wet conditions might be one mitigation option to reduce N₂O emission rates from the PP treatment in this study field.     

Spatial and seasonal variations of atmospheric N2O and CO2 fluxes from a subtropical mangrove swamp and their relationships with soil characteristics

Marine ecosystems are a known net source of greenhouse gases emissions but the atmospheric gas fluxes, particularly from the mangrove swamps occupying inter-tidal zones, are characterized poorly. Spatial and seasonal fluxes of nitrous oxide (N₂O) and carbon dioxide (CO₂) from soil in Mai Po mangrove swamp in Hong Kong, South China and their relationships with soil characteristics were investigated. The N₂O fluxes averaged from 32.1 to 533.7 μg m⁻² h⁻¹ and the CO₂ fluxes were between 10.6 and 1374.1 mg m⁻² h⁻¹. Both N₂O and CO₂ fluxes in this swamp showed large spatial and seasonal variations. The fluxes were higher at the landward site than the foreshore bare mudflat, and higher fluxes were recorded in warm, rather than cold, seasons. The landward site had the highest content of soil organic carbon (OC), total Kjeldahl nitrogen (TKN), nitrate (NO₃⁻–N) and total phosphorus (TP), while the bare mudflat had the highest ammonium nitrogen (NH₄⁺–N) concentration and soil denitrification potential activity. The N₂O flux was related, positively, to CO₂ flux. Soil NO₃⁻–N and TP increased N₂O flux, while soil OC and TP concentrations contributed to the CO₂ flux. The results indicated that the Mai Po mangrove swamp emitted significant amounts of greenhouse gases, and the N₂O emission was probably due to soil denitrifcation.     

Nitrous oxide flux from soil amino acid mineralization

Nitrous oxide (N₂O) is a greenhouse gas produced during microbial transformation of soil N that has been implicated in global climate warming. Nitrous oxide efflux from N fertilized soils has been modeled using NO₃⁻ content with a limited success, but predicting N₂O production in non-fertilized soils has proven to be much more complex. The present study investigates the contribution of soil amino acid (AA) mineralization to N₂O flux from semi-arid soils. In laboratory incubations (−34 kPa moisture potential), soil mineralization of eleven AAs (100 μg AA-N g⁻¹ soil) promoted a wide range in the production of N₂O (156.0±79.3 ng N₂O-N g⁻¹ soil) during 12 d incubations. Comparison of the δ¹³C content (‰) of the individual AAs and the δ¹³C signature of the respired AA-CO₂-C determined that, with the exception of TYR, all of the AAs were completely mineralized during incubations, allowing for the calculation of a N₂O-N conversion rate from each AA. Next, soils from three different semi-arid vegetation ecosystems with a wide range in total N content were incubated and monitored for CO₂ and N₂O efflux. A model utilizing CO₂ respired from the three soils as a measure of organic matter C mineralization, a preincubation soil AA composition of each soil, and the N₂O-N conversion rate from the AA incubations effectively predicted the range of N₂O production by all three soils. Nitrous oxide flux did not correspond to factors shown to influence anaerobic denitrification, including soil NO₃⁻ contents, soil moisture, oxygen consumption, and CO₂ respiration, suggesting that nitrification and aerobic nitrifier denitrification could be contributing to N₂O production in these soils. Results indicate that quantification of AA mineralization may be useful for predicting N₂O production in soils.     

N2O emissions and product ratios of nitrification and denitrification as affected by freezing and thawing

Agricultural soils contribute significantly to atmospheric nitrous oxide (N₂O). A considerable part of the annual N₂O emission may occur during the cold season, possibly supported by high product ratios in denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O-N/(NO₃⁻-N+NO₂⁻-N)) at low temperatures and/or in response to freeze-thaw perturbation. Water-soluble organic materials released from frost-sensitive catch crops and green manure may further increase winter emissions. We conducted short-term laboratory incubations under standardized moisture and oxygen (O₂) conditions, using nitrogen (N) tracers (¹⁵N) to determine process rates and sources of emitted N₂O after freeze-thaw treatment of soil or after addition of freeze-thaw extract from clover. Soil respiration and N₂O production was stimulated by freeze-thaw or addition of plant extract. The N₂O emission response was inversely related to O₂ concentration, indicating denitrification as the quantitatively prevailing process. Denitrification product ratios in the two studied soils (pH 4.5 and 7.0) remained largely unaltered by freeze-thaw or freeze-thaw-released plant material, refuting the hypothesis that high winter emissions are due to frost damage of N₂O reductase activity. Nitrification rates estimated by nitrate (NO₃⁻) pool enrichment were 1.5-1.8 μg NO₃-N g⁻¹ dw soil d⁻¹ in freeze-thaw-treated soil when incubated at O₂ concentrations above 2.3 vol% and one order of magnitude lower at 0.8 vol% O₂. Thus, the experiments captured a situation with severely O₂-limited nitrification. As expected, the O₂ stress at 0.8 vol% resulted in a high nitrification product ratio (0.3 g g⁻¹). Despite this high product ratio, only 4.4% of the measured N₂O accumulation originated from nitrification, reaffirming that denitrification was the main N₂O source at the various tested O₂ concentrations in freeze-thaw-affected soil. N₂O emission response to both freeze-thaw and plant extract addition appeared strongly linked to stimulation of carbon (C) respiration, suggesting that freeze-thaw-induced release of decomposable organic C was the major driving force for N₂O emissions in our soils, both by fuelling denitrifiers and by depleting O₂. The soluble C (applied as plant extract) necessary to induce a CO₂ and N₂O production rate comparable with that of freeze-thaw was 20-30 μg C g⁻¹ soil dw. This is in the range of estimates for over-winter soluble C loss from catch crops and green manure plots reported in the literature. Thus, freeze-thaw-released organic C from plants may play a significant role in freeze-thaw-related N₂O emissions.     

Emissions of CO2, CH4 and N2O from Southern European peatlands

Peatlands play an important role in emissions of the greenhouse gases CO₂, CH₄ and N₂O, which are produced during mineralization of the peat organic matter. To examine the influence of soil type (fen, bog soil) and environmental factors (temperature, groundwater level), emission of CO₂, CH₄ and N₂O and soil temperature and groundwater level were measured weekly or biweekly in loco over a one-year period at four sites located in Ljubljana Marsh, Slovenia using the static chamber technique. The study involved two fen and two bog soils differing in organic carbon and nitrogen content, pH, bulk density, water holding capacity and groundwater level. The lowest CO₂ fluxes occurred during the winter, fluxes of N₂O were highest during summer and early spring (February, March) and fluxes of CH₄ were highest during autumn. The temporal variation in CO₂ fluxes could be explained by seasonal temperature variations, whereas CH₄ and N₂O fluxes could be correlated to groundwater level and soil carbon content. The experimental sites were net sources of measured greenhouse gases except for the drained bog site, which was a net sink of CH₄. The mean fluxes of CO₂ ranged between 139 mg m⁻² h⁻¹ in the undrained bog and 206 mg m⁻² h⁻¹ in the drained fen; mean fluxes of CH₄ were between −0.04 mg m⁻² h⁻¹ in the drained bog and 0.05 mg m⁻² h⁻¹ in the drained fen; and mean fluxes of N₂O were between 0.43 mg m⁻² h⁻¹ in the drained fen and 1.03 mg m⁻² h⁻¹ in the drained bog. These results indicate that the examined peatlands emit similar amounts of CO₂ and CH₄ to peatlands in Central and Northern Europe and significantly higher amounts of N₂O.     

Do earthworms increase N2O emissions in ploughed grassland?

Earthworm activity has been reported to lead to increased production of the greenhouse gas nitrous oxide (N₂O). This is due to emissions from worms themselves, their casts and drilosphere, as well as to general changes in soil structure. However, it remains to be determined how important this effect is on N₂O fluxes from agricultural systems under realistic conditions in terms of earthworm density, soil moisture, tillage activity and residue loads. We quantified the effect of earthworm presence on N₂O emissions from a pasture after simulated ploughing of the sod (‘grassland renovation’) for different soil moisture contents during a 62-day mesocosm study. Sod (with associated soil) and topsoil were separately collected from a loamy Typic Fluvaquent. Treatments included low (L), medium (M) and high (H) moisture content, in combination with: only soil (S); soil+incorporated sod (SG); soil+incorporated sod+the anecic earthworm Aporrectodea longa (SGE). Nitrous oxide and carbon dioxide (CO₂) fluxes were measured for 62 d. At the end of the incubation period, we determined N₂O production under water-saturated conditions, potential denitrification and potential mineralization of the soil after removing the earthworms. Cumulative N₂O and CO₂ fluxes over 62 d from incorporated sod were highest for treatment HSGE (973 μg N₂O-N and 302 mg CO₂-C kg⁻¹ soil) and lowest for LSG (64 μg N₂O-N and 188 mg CO₂-C kg⁻¹ soil). Both cumulative fluxes were significantly different for soil moisture (p<0.001), but not for earthworm presence. However, we observed highly significant earthworm effects on N₂O fluxes that reversed over time for the H treatments. During the first phase (day 3-day 12), earthworm presence increased N₂O emissions with approximately 30%. After a transitional phase, earthworm presence resulted in consistently lower (approximately 50%) emissions from day 44 onwards. Emissions from earthworms themselves were negligible compared to overall soil fluxes. After 62 d, original soil moisture significantly affected potential denitrification, with highest fluxes from the L treatments, and no significant earthworm effect. We conclude that after grassland ploughing, anecic earthworm presence may ultimately lead to lower N₂O emissions after an initial phase of elevated emissions. However, the earthworm effect was both determined and exceeded by soil moisture conditions. The observed effects of earthworm activity on N₂O emissions were due to the effect of earthworms on soil structure rather than to emissions from the worms themselves.     

Land use effects on gross nitrogen mineralization, nitrification, and N2O emissions in ephemeral wetlands

Stable ¹⁵N isotope dilution and tracer techniques were used in cultivated (C) and uncultivated (U) ephemeral wetlands in central Saskatchewan, Canada to: (1) quantify gross mineralization and nitrification rates and (2) estimate the relative proportion of N₂O emissions from these wetlands that could be attributed to denitrification versus nitrification-related processes. In-field incubation experiments were repeated in early May, mid-June and late July. Mean gross mineralization and nitrification rates (10.3 and 3.1 mg kg⁻¹ d⁻¹, respectively) did not differ between C and U wetlands on any given date. Despite these similarities, the mean NH₄⁺ pool size in the U wetlands (17.2 mg kg⁻¹) was two to three times that of the C wetlands (6.7 mg kg⁻¹) whereas the mean NO₃⁻ pool size in U wetlands (2.2 mg kg⁻¹) was less than half that of C wetlands (5.8 mg kg⁻¹). Mean N₂O emissions from the C wetlands decreased from 112.8 to 17.0 ng N₂O m² s⁻¹ from May to July, whereas mean U-wetland N₂O emissions ranged only from 31.8 to 51.1 ng N₂O m² s⁻¹ over the same period. This trend is correlated to water-filled pore space in C wetlands, demonstrating a soil moisture influence on emissions. Denitrification is generally considered the dominant emitter of N₂O under anaerobic conditions, but in the C wetlands, only 49% of the May emissions could be directly attributed to denitrification, decreasing to 29% in July. In contrast, more than 75% of the N₂O emissions from the U wetlands arose from denitrification of the soil NO₃⁻ pool throughout the season. These land use differences in emission sources and rates should be taken into consideration when planning management strategies for greenhouse gas mitigation.     

The dynamics of N2O near the groundwater table and the transfer of N2O into the unsaturated zone: A case study from a sandy aquifer in Germany

The research area was the Fuhrberger Feld aquifer (FFA) in northern Germany. It is situated about 30km northeast of the city of Hannover and covers about 300km². Six multilevel sampling wells along a representative strip under predominantly arable land along a groundwater flow-line were sampled from the groundwater table down to a depth of 10m below the soil surface. We measured N₂O, CO₂, NO₃⁻, SO₄²⁻, DOC, pH, redox potentials and O₂ concentrations.N₂O accumulated at four out of six wells close to the groundwater table. About 20% of N₂O that occurred between the groundwater table and 7–8m below it resided in the top 0.4m. An exchange zone for N₂O at the interface between the saturated and the unsaturated zone extended 0.55 ± 0.22m below the groundwater table and acted as a source and sink for N₂O. N₂O below the exchange zone cannot be transferred from the groundwater to the atmosphere. The upward fluxes from the exchange zone into the unsaturated zone at the six wells ranged between 0.0009 and 0.3kg N₂O ha^{− 1} year^{− 1}. The yearly downward fluxes into the exchange zones had about the same order of magnitude as the upward fluxes. The upward and downward fluxes of N₂O at the (fluctuating) water table did cancel out each other, but this does not yet imply, that the N₂O fluxes at the soil surface also cancel out each other.N₂O–N:NO₃–N ratios were highly variable ranging from 0.0002 to 0.0417.A multiple regression for the monthly N₂O amounts in the exchange zone could explain 66% of the yearly variation. The significant variables were NO₃⁻, CO₂, pH, and O₂. Therefore, a combination of the land use (NO₃⁻), the geochemical boundary conditions (pH) and the type of denitrification reaction (O₂ and CO₂ indicate the importance of a heterotrophic denitrification process) governed the N₂O dynamics in the surface groundwater of the FFA and its transfer into the unsaturated zone.