Evolution of nitrate reductase: molecular and structural variations on a common function

The biological transformation of nitrogen oxyanions is widespread in nature and gives rise to a robust biogeochemical cycle. The first step in nitrate reduction is carried out by the enzyme nitrate reductase (NR). Although NR always catalyzes the same chemical reaction (conversion of nitrate into nitrite), its location in the cell, structure, and function are organism-dependent. We use protein sequence data to determine phylogenetic relationships and to examine similarities in structure and function. Three distinct clades of NR are apparent: the eukaryotic assimilatory NR (Euk-NR) clade, the membrane-associated prokaryotic NR (Nar) clade, and a clade that includes both the periplasmic NR (Nap) and prokaryotic assimilatory NR (Nas). The high degree of sequence similarity and a phylogenetic distribution that follows taxonomic classification suggest a monophyletic origin for the Euk-NR early on in the evolution of eukaryotic cells. In contrast, sequence conservation, phylogenetic analysis, and physiology suggest that both Nar and Nap were acquired by horizontal gene transfer. Nap and Nas share a lesser degree of similarity, with Nap a subclade of Nas. Nap from strict anaerobic bacteria such as Desulfovibrio desulfuricans is ancestral to facultative species and may provide an evolutionary link between Nap and Nas. We observed conserved binding sites for molybdenum and pterin cofactors in all four proteins. In pathways involving Euk-NR, Nas, and Nar, for which ammonia is the end product, nitrite is reduced to ammonia by a siroheme nitrite reductase. Nap, however, is coupled to a pentaheme nitrite reductase. In denitrification, whether Nar or Nap is involved, nitrite is reduced to nitric oxide by either a cytochrome cd1 or a copper-containing nitrite reductase. This complexity underscores the importance of nitrate reduction as a key biological process.     

Subsoils: chemo-and biological denitrification, N2O and N2 emissions

Agricultural practices, soil characteristics and meteorological conditions are responsible for eventual nitrate accumulation in the subsoil. There is a lot of evidence that denitrification occurs in the subsoil and rates up to 60–70 kg ha^-1 yr^-1 might be possible. It has also been shown that in the presence of Fe²⁺ (formed through weathering of minerals) and an alkaline pH, nitrate can be chemically reduced. Another possible pathway of disappearance is through the formation of nitrite, which is unstable in acid conditions. With regard to the emission of N₂O and N₂, it can be stated that all conditions whereby the denitrification process becomes marginal are favourable for N₂O formation rather than for N₂. Because of its high solubility, however, an important amount of N₂O might be transported with drainage water.     

Seasonal N<Subscript>2</Subscript>O emissions respond differently to environmental and microbial factors after fertilization in wheat–maize agroecosystem

Biogeochemical processes regulating cropland soil nitrous oxide (N₂O) emissions are complex, and the controlling factors need to be better understood, especially for seasonal variation after fertilization. Seasonal patterns of N₂O emissions and abundances of archaeal ammonia monooxygenase (amoA), bacterial amoA, nitrate reductase (narG), nitrite reductase (nirS/nirK), and nitrous oxide reductase (nosZ) genes in long-term fertilized wheat–maize soils have been studied to understand the roles of microbes in N₂O emissions. The results showed that fertilization greatly stimulated N₂O emission with higher values in pig manure-treated soil (OM, 2.88 kg N ha^?1 year^?1) than in straw-returned (CRNPK, 0.79 kg N ha^?1 year^?1) and mineral fertilizer-treated (NPK, 0.90 kg N ha^?1 year^?1) soils. Most (52.2–88.9%) cumulative N₂O emissions occurred within 3 weeks after fertilization. Meanwhile, N₂O emissions within 3 weeks after fertilization showed a positive correlation with narG gene copy number and a negative correlation with soil NO₃^? contents. The abundances of narG and nosZ genes had larger direct effects (1.06) than ammonium oxidizers (0.42) on N₂O emissions according to partial least squares path modeling. Stepwise multiple regression also showed that log narG was a predictor variable for N₂O emissions. This study suggested that denitrification was the major process responsible for N₂O emissions within 3 weeks after fertilization. During the remaining period of crop growth, insufficient N substrate and low temperature became the primary limiting factors for N₂O emission according to the results of the regression models.     

Use of the 15?N gas flux method to measure the source and level of N2O and N2 emissions from grazed grassland

Understanding the contribution of nitrification and denitrification to production of nitrous oxide (N₂O), a potent greenhouse gas, is important in devising effective mitigation strategies to reduce emissions. In this study the ¹⁵N gas flux method was used to investigate N₂O and N₂ emissions following an application of ¹⁵N labelled ammonium nitrate (0.71?mol?N?m^?2) to intensive grassland swards (grazed at 2.74 or 2.05 livestock units ha^?1 year^?1) at a site in Southern Ireland. The ¹⁵N labelled fertiliser (NO₃ moiety ¹⁵N labelled at 60 at. %) was applied to designated soil areas in the field, enclosed by static chambers, in June 2009, September 2009 and March 2010. Fluxes of N₂O and N₂ were determined over 12?days on each occasion. N₂O and N₂ emissions were significantly (P?<?0.001) lower in March 2010 than in June or September 2009. There was little difference between the two swards grazed at different stocking rates on N₂O or N₂ emissions. Mean cumulative N₂O emissions over 103?h were 212.9, 279.5 and 62.06?mg?m^?2 for June 2009, September 2009 and March 2010, respectively. Mean cumulative N₂ emissions for the three time periods were 818.8, 893.8 and 87?mg?m^?2, respectively. The N₂O mole fraction averaged 0.21 and 0.23 in June 2009 and September 2009, respectively, but increased to 0.41 in March 2010 which may have been due to changes in denitrifier community composition or due to N₂O reductase being sensitive to low soil temperatures. The results point to denitrification of nitrate as the major source of N₂O at this site which may have implications for choice of fertiliser in moist temperate climates.     

N2O and NO emissions from different N sources and under a range of soil water contents

Emissions of nitrous oxide (N₂O) and nitric oxide (NO) have been identified as one of the most important sources of atmospheric pollution from grasslands. Soils are major sources for the production of N₂O and NO, which are by-products or intermediate products of microbial nitrification and denitrification processes. Some studies have tried to evaluate the importance of denitrification or nitrification in the formation of N₂O or NO but there are few that have considered emissions of both gases as affected by a wide range of different factors. In this study, the importance of a number of factors (soil moisture, fertiliser type and temperature) was determined for N₂O and NO emissions. Nitrous oxide and NO evolution in time and the possibility of using the ratio NO:N₂O as an indicator for the processes involved were also explored. Dinitrogen (N₂) and ammonia (NH₃) emissions were estimated and a mass balance for N fluxes was performed. Nitrous oxide and NO were produced by nitrification and denitrification in soils fertilised with and by denitrification in soils fertilised with . Water content in the soil was the most important factor affecting N₂O and NO emissions. Our N₂O and NO data were fitted to quadratic (r=0.8) and negative exponential (r=0.7) equations, respectively. A long lag phase was observed for the N₂O emitted from soils fertilised with (denitrification), which was not observed for the soils fertilised with (nitrification) and was possibly due to a greater inhibiting effect of low temperatures on microbial activity controlling denitrification rather than on nitrification. The use of the NO:N₂O ratio as a possible indicator of denitrification or nitrification in the formation of N₂O and NO was discounted for soils fertilised with . The N mass balance indicated that about 50 kg N ha⁻¹ was immobilised by microorganisms and/or taken up by plant roots, and that most of the losses ocurred in wet soils (WFPS >60%) as N₂ and NH₃ losses (>55%).     

Nitrous oxide production from soil experiments: denitrification prevails over nitrification

Nitrous oxide is produced in soils and sediments essentially through the processes of nitrification and denitrification, although several rival processes could be competing. This study was undertaken in order to better understand the controlling factors of nitrification, denitrification and associated N₂O production as well as the contribution of these two processes to the average N₂O production by soils and sediments. With this aim, soil and sediment samples were taken in contrasting periods and different land use types, each time at different depths (upper and lower soil horizons). They were incubated in separate batches in specific conditions to promote denitrification and nitrification: (1) a complete anaerobic environment adding KNO₃ for the denitrification assay and (2) an aerobic environment (21 % O₂) with addition of NH₄Cl for the nitrification assay. Potentials of nitrification and denitrification were determined by the rates of nitrate either reduced (for denitrification) or produced (for nitrification). Overall, denitrification potential varied from 70 to 2,540 ng NO ₃ ^— -N g^?1 dry soil h^?1 and nitrification potential from 30 to 1,150 ng NO₃ ^?-N g^?1 dry soil h^?1. Nitrous oxide production by denitrification was significantly (P < 0.05) greater in topsoils (10–30 cm) than in subsoils (90–110 cm), ranging, respectively, from 26 to 250 ng N₂O-N g^?1 dry soil h^?1 versus 1.5 to 31 ng N₂O-N g^?1 dry soil h^?1, i.e., a mean 19.5 versus. 6.0 % of the NO₃ ^? denitrified for the upper and lower horizons, respectively. Considering the N₂O production in relation with the nitrate production (e.g., nitrification), no significant difference (P < 0.05) was found in the soil profile, which ranged from 0.03 to 6 ng N₂O-N g^?1 dry soil h^?1. This production accounts for 0.21 and 0.16 % of the mean of the NO₃ ^? produced for the top and subsoils, respectively. On the basis of the average production by both top- and subsoils, N₂O production by denitrification is clearly greater than by nitrification under the measurement conditions used in this study, from 30- to 100-fold higher. Such a high potential of N₂O emission must be taken into account when reducing nitrate contamination by increasing denitrification is planned as a curative measure, e.g. in rehabilitation/construction of wetlands.     

Nitrous oxide emission as affected by liming an acidic mineral soil used for arable agriculture

The effect of liming an acidic mineral soil (Dystric Nitosol from southern China), used for arable agriculture, on N₂O emission was studied in an incubation experiment. After the soil pH had been raised from pH 4.4 to 5.2, 6.7 and 8.1, soil samples were either amended with NH₄ ⁺ and incubated aerobically, favoring nitrification or, after application of NO₃ ^–, the incubation took place under anaerobic conditions, favoring denitrification. Gas sampling for N₂O determination and soil analyses were performed at regular intervals up to 13 days. Under nitrification conditions only small N₂O emission rates were observed (max. 6 g N kg^–1 d^–1) with significant differences between high and low pH values during the first 2 days of incubation. The nitrifying activity was low, even with high pH, and this, together with good aeration conditions, could partly explain the small N₂O evolution. During denitrification, however, cumulative N₂O emissions reached much higher values (1600 g N kg^–1 in comparison to 40 g N kg^–1 under nitrification conditions). N₂O emission during denitrification was significantly enhanced by increasing soil pH. Under alkaline conditions (pH 8.1) a large nitrite accumulation occurred, which was in line with the highest nitrate reductase activity determined in this treatment. The limited availability of organic carbon is probably the main reason for the absence of further reduction of NO₂ ^– to N₂O or N₂. At pH 6.7 the total N₂O emission was slightly higher than at pH 8.1, although the start of pronounced emissions was retarded and only small amounts of NO₂ ^– accumulated. Acid soil conditions caused either negligible (pH 4.4) or only small (pH 5.2) N₂O emissions. It can be concluded that these kinds of soil, used alternatively for production of upland crops or paddy rice, are prone to high N₂O emissions after flooding, particularly under neutral to alkaline conditions. In order to avoid major N₂O evolution and accumulation of nitrite, which can be leached into groundwater, the pH should not be raised to values above 5.5–6.     

Measurement of nitrous oxide and di-nitrogen emissions from agricultural soils

Nitrous oxide can be produced during nitrification, denitrification, dissimilatory reduction of NO ₃ ^- to NH ₄ ⁺ and chemo-denitrification. Since soils are a mosaic of aerobic and anaerobic zones, it is likely that multiple processes are contributing simultaneously to N₂O production in a soil profile. The N₂O produced by all processes may mix to form one pool before being reduced to N₂ by denitrification. Reliable methods are needed for measuring the fluxes of N₂O and N₂ simultaneously from agricultural soils. The C₂H₂ inhibition and ¹⁵N gas-flux methods are suitable for use in undisturbed soils in the field. The main disadvantage of C₂H₂ is that as well as blocking N₂O reductase, it also blocks nitrification and dissimilatory reduction of NO ₃ ^- to NH ₄ ⁺ . Potentially the ¹⁵ N gas-flux method can give reliable measurements of the fluxes of N₂O and N₂ when all N transformation processes proceed naturally. The analysis of ¹⁵N in N₂ and N₂O is now fully automated by continuous-flow isotope-ratio mass spectrometry for 12-ml gas samples contained in septum-capped vials. Depending on the methodology, the limit of detection ranges from 4 to 11 g N ha^-1day^-1 for N₂ and 4 to 15 g N ha^-1day^-1 for N₂O. By measuring the ¹⁵N content and distribution of ¹⁵N atoms in the N₂O molecules, information can also be obtained to help diagnose the sources of N₂O and the processes producing it. Only a limited number of field studies have been done using the ¹⁵N gas-flux method on agricultural soils. The measured flux rates and mole fractions of N₂O have been highly variable. In rain-fed agricultural soils, soil temperature and water-filled pore space change with the weather and so are difficult to modify. Soil organic C, NO ₃ ^- and pH should be amenable to more control. The effect of organic C depends on the degree of anaerobiosis generated as a result of its metabolism. If conditions for denitrification are not limiting, split applications of organic C will produce more N₂O than a single application because of the time lag in the synthesis of N₂O reductase. Increasing the NO ₃ ^- concentration above the K _m value for NO ₃ ^- reductase, or decreasing soil pH from 7 to 5, will have little effect on denitrification rate but will increase the mole fraction of N₂O. The effect of NO ₃ ^- concentration on the mole fraction of N₂O is enhanced at low pH. Manipulating the interaction between NO ₃ ^- supply and soil pH offers the best hope for minimising N₂O and N₂ fluxes.     

Microbial processes responsible for nitrous oxide production from acid soils in different land-use patterns in Pasirmayang,central Sumatra,Indonesia

The most probable number (MPN) method and dicyandiamide (DCD), an autotroph-specific inhibitor, were used to identify microbial processes that produced N₂O in acid soils. Nitrous oxide production was investigated in six sites in four different land-use patterns at Pasirmayang in Jambi province, in Sumatra, Indonesia, using soils collected in October 1999, March 2000 and June 2000. The six sites were primary forests (P1 and P2), loggedover forests (L2), a young rubber plantation established after forest had been logged and burned in March 1998 (L1), a plantation of Gmelina arborea established after logging and burning of forest in August 1996 (O), and a rubber plantation of a small landholder established in 1993 (R). Very small numbers of autotrophic ammonia oxidizers and autotrophic nitrite oxidizers were detected. Heterotrophic nitrifiers were detected at all sites, with the highest MPNs at L1 and L2. In incubation experiments, the emission of N₂O from soils increased at all six sites after adding citrate, a substrate for heterotrophic nitrifiers. No correlation appeared between denitrification activity and N₂O flux. We conclude that heterotrophic nitrifiers are the main microbial contributors to N₂O emission from acid soils in Pasirmayang, Jambi.     

Impacts of Water Table Management on N<Subscript>2</Subscript>O and N<Subscript>2</Subscript> from a Sandy Loam Soil in Southwestern Quebec,Canada

Nitrous oxide (N₂O) is primarily produced as intermediate in denitrification and, to a lesser extent, through nitrification processes. Nitrous oxide emission and, consequently, its atmospheric impacts depend on the extent to which N₂O is reduced to dinitrogen gas (N₂) by denitrifiers. Field experiments were conducted from 1998 through 2000 growing seasons at St. Emmanuel, Quebec, Canada, to investigate the combined impact of water table management (WTM) and N fertilization rate on the soil denitrification (N₂O + N₂) rate, rate of N₂O production, and the N₂O:N₂O + N₂ ratio. Water table treatments included subirrigation (SI) with a target water table depth of 0.6 m and free drainage (FD) with open drains. The tile drains (75 mm diameter) were laid at a 1.0 m depth from the soil surface. Nitrogen fertilizer was applied at two rates:120 and 200 kg N ha⁻¹ as ammonium nitrate (34-0-0). The N₂O + N₂ evolution rates were greater in SI (12.9 kg N ha⁻¹) than in FD (5.8 kg N ha⁻¹) plots. The percentages of N₂O relative to overall N₂O + N₂ evolution were 35 and 11% for 1998, 29 and 8% for 1999, and 37 and 20% for 2000, under FD and SI, respectively. The reduced N₂O production under SI was due to a greater reduction of N₂O to N₂. Results indicate that greater N₂O + N₂ evolution under shallow water tables are not necessarily accompanied by higher N₂O emissions.     

Rates and controls of nitrous oxide and nitric oxide emissions following conversion of forest to pasture in Rondônia

Tropical soils are important sources of nitrous oxide (N₂O) and nitric oxide (NO) emissions from the Earths terrestrial ecosystems. Clearing of tropical rainforest for pasture has the potential to alter N₂O and NO emissions from soils by altering moisture, nitrogen supply or other factors that control N oxide production. In this review we report annual rates of N₂O and NO emissions from forest and pastures of different ages in the western Brazilian Amazon state of Rondônia and examine how forest clearing alters the major controls of N oxide production. Forests had annual N₂O emissions of 1.7 to 4.3 kg N ha^-1 y^-1 and annual NO emissions of 1.4 kg N ha^-1 y^-1. Young pastures of 1–3 years old had higher N₂O emissions than the original forest (3.1–5.1 kg N ha^-1 y^-1) but older pastures of 6 years or more had lower emissions (0.1 to 0.4 kg N ha^-1 y^-1). Both soil moisture and indices of soil N cycling were relatively poor predictors of N₂O, NO and combined N₂O + NO emissions. In forest, high N₂O emissions occurred at soil moistures above 30 water-filled pore space, while NO emissions occurred at all measured soil moistures (18–43). In pastures, low N availability led to low N₂O and NO emissions across the entire range of soil moistures. Based on these patterns and results of field fertilization experiments, we concluded that: (1) nitrification was the source of NO from forest soils, (2) denitrification was not a major source of N₂O production from forest soils or was not limited by NO- supply, (3) denitrification was a major source of N₂O production from pasture soils but only when NO₃^- was available, and (4) nitrification was not a major source of 3 NO production in pasture soils. Pulse wettings after prolonged dry periods increased N₂O and NO₃^- emissions for only short periods and not enough to appreciably affect annual emission rates. We project that Basin-wide, the effect of clearing for pasture in the future will be a small reduction in total N₂O emissions if the extensive pastures of the Amazon continue to be managed in a way similar to current practices. In the future, both N₂Oand NO fluxes could increase if uses of pastures change to include greater use of N fertilizers or N-fixing crops. Predicting the consequences of these changes for N oxide production will require an understanding of how the processes of nitrification and denitrification interact with soil type and regional moisture regimes to control N₂O and NO production from these new anthropogenic N sources.     

Denitrification in aqueous FeEDTA solutions

The biological reduction of nitric oxide (NO) in aqueous solutions of FeEDTA is an important key reaction within the BioDeNOx process, a combined physico‐chemical and biological technique for the removal of NO_x from industrial flue gasses. To explore the reduction of nitrogen oxide analogues, this study investigated the full denitrification pathway in aqueous FeEDTA solutions, ie the reduction of NO₃^?, NO₂^?, NO via N₂O to N₂ in this unusual medium. This was done in batch experiments at 30 °C with 25 mmol dm^?3 FeEDTA solutions (pH 7.2 ± 0.2). Also Ca²⁺ (2 and 10 mmol dm^?3) and Mg²⁺ (2 mmol dm^?3) were added in excess to prevent free, uncomplexed EDTA. Nitrate reduction in aqueous solutions of Fe(III)EDTA is accompanied by the biological reduction of Fe(III) to Fe(II), for which ethanol, methanol and also acetate are suitable electron donors. Fe(II)EDTA can serve as electron donor for the biological reduction of nitrate to nitrite, with the concomitant oxidation of Fe(II)EDTA to Fe(III)EDTA. Moreover, Fe(II)EDTA can also serve as electron donor for the chemical reduction of nitrite to NO, with the concomitant formation of the nitrosyl‐complex Fe(II)EDTA–NO. The reduction of NO in Fe(II)EDTA was found to be catalysed biologically and occurred about three times faster at 55 °C than NO reduction at 30 °C. This study showed that the nitrogen and iron cycles are strongly coupled and that FeEDTA has an electron‐mediating role during the subsequent reduction of nitrate, nitrite, nitric oxide and nitrous oxide to dinitrogen gas. Copyright © 2004 Society of Chemical Industry     

Measurement and modeling of nitrous and nitric oxide emissions from a tea field in subtropical central China

Tea fields represent an important source of nitrous oxide (N₂O) and nitric oxide (NO) emissions due to high nitrogen (N) fertilizer applications and very low soil pH. To investigate the temporal characteristics of N₂O and NO emissions, daily emissions were measured over 2½ years period using static closed-chamber/gas chromatograph and chemiluminescent measurement system in a tea field of subtropical central China. Our results revealed that N₂O and NO fluxes showed similar temporal trends, which were generally driven by temporal variations in soil temperature and soil moisture content and were also affected by fertilization events. The measured average annual N₂O and NO emissions were 10.9 and 3.3 kg N ha^?1 year^?1, respectively, highlighting the high N₂O and NO emissions from tea fields. To improve our understanding of N-cycling processes in tea ecosystems, we developed a new nitrogenous gas emission module for the water and nitrogen management model (WNMM, V2) that simulated daily N₂O and NO fluxes, in which the NO was simulated as being emitted from both nitrification and nitrite chemical decomposition. The results demonstrated that the WNMM captured the general temporal dynamics of N₂O (NSE = 0.40; R² = 0.52, RMSE = 0.03 kg N ha^?1 day^?1, P < 0.001) and NO (NSE = 0.41; R² = 0.44, RMSE = 0.01 kg N ha^?1 day^?1, P < 0.001) emissions. According to the simulation, denitrification was identified as the dominant process contributing 76.5% of the total N₂O emissions, while nitrification and nitrite chemical decomposition accounted for 52.3 and 47.7% of the total NO emissions, respectively.     

Catalytic vapour phase epoxidation of propene with nitrous oxide as an oxidant II. Development of a kinetic model

The vapour phase epoxidation of propene with nitrous oxide (N₂O) was experimentally investigated in a fixed bed reactor using a CsO_x/FeO_y/SiO₂ catalyst over a broad range of residence times. The influence of feed composition on the conversion and product distribution was determined for the reactants propene, propylene oxide (PO), propionaldehyde (PA), and N₂O. The experimental results were used to derive a formal kinetic model to describe the reactions in a network deduced in the first part of this publication [Thömmes, T., Zürcher, S., Wix, A., Reitzmann, A., Kraushaar-Czarnetzki, B., 2007. Catalytic vapour phase epoxidation of propene with nitrous oxide as an oxidant: I. Reaction network and product distribution. Applied Catalysis A 318, 160-169]. Self-inhibition of the propene conversion was observed, and an inhibition of the PO conversion through PO isomerisation products. The N₂O concentration has almost no effect on the conversion of propene and PO, but the PA conversion is accelerated significantly through N₂O. Propene related and N₂O related PO selectivities display opposed dependencies from reactant concentration. The modelling results indicate that coke is predominantly formed from oxygenated products. It is the key issue for future developments to prevent the fast consecutive conversion of PO.     

Measurements of N2O and NO emissions during tomato cultivation using a flow-through chamber system in a glasshouse

Nitrous oxide (N₂O) and nitric oxide (NO) fluxes resulting from long-term tomato cultivation in a glasshouse were continuously determined using the flow-through chamber method over the course of three cultivation periods. Gas concentrations were measured using an nondispersive infrared (gas filter correlation/infra-red) analyzer and a chemiluminescence-based analyzer, respectively. Following a basal application of fertilizer, daily N₂O and NO emission rates increased, with peaks lasting from 40 to 140 days. Short-term fluctuations in daily N₂O and NO emissions were affected by differences in nitrogen application, soil water, and soil temperature. Diurnal changes in N₂O and NO fluxes during the period of peak emissions depended primarily on soil temperature. Following the application of a top dressing (N as urea or calcium nitrate) in the irrigation water, the N₂O and NO fluxes increased immediately, with a very short period of peak emissions (1–5 h) after urea application. The duration of the peak period in daily accumulated N₂O and NO emissions following application of the top dressing ranged from 3 to 10 days.     

Contribution of nitrification and denitrification to N2O production in peat, clay and loamy sand soils under different soil moisture conditions

Agricultural soils are a significant source of nitrous oxide (N₂O). Since mitigation of greenhouse gas emissions is needed in all sectors of society, it is important to identify the processes producing N₂O and the factors affecting the production rates in agricultural soils. This study aimed to elucidate the N₂O production in peat, clay and loamy sand at four different soil moisture conditions (40, 60, 80 and 100% Water Filled Pore Space). The ace­tylene inhibition technique was used to evaluate the contribution of nitrification to N₂O production. Nitrous oxide production responded markedly to soil moisture in all three soils. The highest N₂O production, measured at the wettest soils (100% WFPS), was up to four orders of magnitude higher than that at the dry soils (40% WFPS). In dry conditions N₂O production decreased in the order of peat > clay > loamy sand, while in wet conditions the highest N₂O production was measured in loamy sand, then in peat, and the lowest in clay soils. Nitrification was the dominant N₂O producing process in all the soils at 60% WFPS. In the sandy soil 70% of the total N₂O production originated from nitrification, while in the peat soil most of the total N₂O production originated from denitrification. Data on processes producing N₂O in agricultural soils are needed to develop process-based models that could reduce the uncertainty of the emission estimates in greenhouse gas inventories.     

Effects of type and amount of applied nitrogen fertilizer on nitrous oxide fluxes from intensively managed grassland

Five field experiments and one greenhouse experiment were carried out to assess the effects of nitrogen (N) fertilizer type and the amount of applied N fertilizer on nitrous oxide (N₂O) emission from grassland. During cold and dry conditions in early spring, emission of N₂O from both ammonium (NH ₄ ⁺ ) and nitrate (NO ₃ ^– ) containing fertilizers applied to a clay soil were relatively small, i.e. less than 0.1% of the N applied. Emission of N₂O and total denitrification losses from NO ₃ ^– containing fertilizers were large after application to a poorly drained sand soil during a wet spring. A total of 5–12% and 8–14% of the applied N was lost as N₂O and via denitrification, respectively. Emissions of N₂O and total denitrification losses from NH ₄ ⁺ fertilizers and cattle slurry were less than 2% of the N applied. Addition of the nitrification inhibitor dicyandiamide (DCD) reduced N₂O fluxes from ammonium sulphate (AS). However, the effect of DCD to reduce total N₂O emission from AS was much smaller than the effect of using NH ₄ ⁺ fertilizer instead of NO ₃ ^– fertilizer, during wet conditions. The greenhouse study showed that a high groundwater level favors production of N₂O from NO ₃ ^– fertilizers but not from NH ₄ ⁺ fertilizers. Inereasing calcium ammonium nitrate (CAN) application increased the emitted N₂O on grassland from 0.6% of the fertilizer application rate for a dressing of 50 kg N ha^–1 to 3.1% for a dressing of 300 kg N ha^–1. In another experiment, N₂O emission increased proportionally with increasing N rate. The results indicate that there is scope for reducing N₂O emission from grasslands by choosing the N fertilizer type depending on the soil moisture status. Avoiding excessive N application rates may also minimize N₂O emission from intensively managed grasslands.     

Effects of fertiliser type and the presence or absence of plants on nitrous oxide emissions from irrigated soils

Nitrous oxide (N₂O) emissions and denitrification losses from an irrigated sandy loam soil amended with composted municipal solid waste (MSW), sheep manure (SM), surface applied pig slurry (SPS), incorporated pig slurry (IPS) or urea (U) were studied under Mediterranean conditions. We quantified emissions, in both the presence and absence of maize and N₂O production, via denitrification and nitrification pathways using varying concentrations of acetylene. Discounting the N₂O lost in the Control, the percentages of N₂O lost in relation to the total N applied were greater for urea (1.80%) than for MSW (0.50%), SM (0.46%), SPS (1.02%) or IPS (1.27%). In general, plots treated with organic fertilisers emitted higher amounts of N₂O when under maize than bare soil plots. On the other hand, greater denitrification losses were also recorded for plots in the absence of plants (between 9.7 and 29.3 kg N₂O-N ha⁻¹) than for areas with plants (between 7.1 and 24.1 kg N₂O-N ha⁻¹). The proportion of N₂O produced via denitrification was greater from fertiliser treatments than for the controls and also greater without plants (between 66 and 91 % of the N₂O emitted) than with plants (between 48 and 81%).