CH4 oxidation and emissions of CH4 and N2O from Lolium perenne swards under elevated atmospheric CO2

Emissions of N₂O and CH₄ and CH₄ oxidation rates were measured from Lolium perenne swards in a short-term study under ambient (36 Pa) and elevated (60 Pa) atmospheric CO₂ at the Free Air Carbon dioxide Enrichment experiment, Eschikon, Switzerland. Elevated pCO₂ increased (P<0.05) N₂O emissions from high N fertilised (11.2 g N m⁻²) swards by 69%, but had no significant effect on net emissions of CH₄. Application of ¹³C-CH₄ (11 μl l⁻¹; 11 at.% excess ¹³C) to closed chamber headspaces in microplots enabled determination of rates of ¹³C-CH₄ oxidation even when net CH₄ fluxes from main plots were positive. We found a significant interaction between fertiliser application rate and atmospheric pCO₂ on ¹³C-CH₄ oxidation rates that was attributed to differences in gross nitrification rates and C and N availability. CH₄ oxidation was slower and thought to be temporarily inhibited in the high N ambient pCO₂ sward. The most rapid CH₄ oxidation of 14.6 μg ¹³C-CH₄ m⁻² h⁻¹ was measured in the high fertilised elevated pCO₂ sward, and we concluded that either elevated pCO₂ had a stimulatory effect on CH₄ oxidation or inhibition of oxidation following fertiliser application was lowered under elevated pCO₂. Application of ¹⁴NH₄¹⁵NO₃ and ¹⁵NH₄¹⁵NO₃ (10 at.% excess ¹⁵N) to different replicates enabled determination of the respective contributions of nitrification and denitrification to N₂O emissions. Inhibition of CH₄ oxidation in the high fertilised ambient pCO₂ sward, due to competition between NH₃ and CH₄ for methane monooxygenase enzymes or toxic effects of NH₂OH or NO₂⁻ produced during nitrification, was hypothesised to increase gross nitrification (12.0 mg N kg dry soil⁻¹) and N₂O emissions during nitrification (327 mg ¹⁵N-N₂O m⁻² over 11 d). Our results indicate that increasing atmospheric concentrations of CO₂ may increase emissions of N₂O by denitrification, lower nitrification rates and either increase or decrease the ability of soil to act as a sink for atmospheric CH₄ depending on fertiliser management.     

Nitrogen fertiliser rate affects the frequency of nitrate-dissimilating Pseudomonas spp. in the rhizosphere of Lolium perenne grown under elevated pCO2 (Swiss FACE)

The effect of elevated pCO₂ (60 Pa) on the frequency of nitrate-dissimilating Pseudomonas (NDP) was investigated in the rhizosphere of fertilised Lolium perenne swards in the Swiss Free Air Carbon dioxide Enrichment (FACE) experiment. Numbers of cultivable root-associated Pseudomonas were greater under elevated (60 Pa) than under ambient (36 Pa) pCO₂ in both high and low N-fertilised swards. For both pCO₂ conditions, the NDP frequency decreased with closer root proximity to L. perenne roots in low fertilised swards. Anyway, in high N swards the NDP frequency was similar in root and soil fractions. Thus, N availability may be a major factor influencing NDP populations under elevated pCO₂, most likely due to increased competition for N between plant and nitrate-dissimilating bacteria.     

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.     

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.     

Nitrous oxide production by nitrification and denitrification in soil aggregates as affected by O2 concentration

Nitrous oxide emitted by soils can be produced either by denitrification in anoxic conditions or by nitrification in presence of O₂. The relative importance of the two processes, particularly under varied partial pressures of O₂, is not always known. This paper focuses on the influence of O₂ concentration on N₂O production by nitrification and denitrification in an arable Orthic Luvisol. Soil aggregates (2-3 mm size), water unsaturated, received 116 mg N kg⁻¹ as ammonium sulphate labelled with ¹⁵N and were incubated during 14 days at different O₂ partial pressures: 0, 0.35, 0.76, 1.5, 4.3 and 20.4 kPa. A ¹⁵N tracing technique was used to quantify nitrification and denitrification rates. ¹⁵N₂O and ¹⁵N₂ were measured. Oxygen pressure appeared to strongly influence both nitrification and denitrification rates and also N₂O emissions. Nitrification rates were reduced by a factor of 6-9 when O₂ decreased from 20.4 to 0.35 kPa. They were highly correlated with O₂ consumption rates. Denitrification mainly occurred in complete anoxic conditions. The proportion of N₂O emitted by denitrification was estimated by two independent methods: one based on ¹⁵N tracing using isotope composition of NH₄, NO₃ and N₂O, the other based on the measurement of the ¹⁵N₂O:¹⁵N₂ ratio. The two methods gave close results. The highest N₂O emissions were obtained under complete anoxic conditions and were due to denitrification. However, N₂O emissions almost as important were obtained at day 14 with 1.5 kPa O₂ pressure, and they were due to nitrification. Nitrification was the main source of N₂O at O₂ concentrations greater than 0.35 kPa. The amounts of N₂O-N emitted by nitrification were linearly related to the amounts of N nitrified, but the slope of the regression was highly dependent on O₂ concentration: it varied from 0.16 to 1.48% when O₂ concentration was reduced from 20.4 to 0.76 kPa. Emissions of N₂O by nitrification may then be quite significant if nitrification occurs at a reduced O₂ concentration.     

Denitrification enzyme activity and potential of subsoils under grazed grasslands assayed by membrane inlet mass spectrometer

The importance of subsoil denitrification on the fate of agriculturally derived nitrate (NO₃) leached to groundwater is crucial for budgeting N in an ecosystem and for identifying areas where the risk of excess NO₃ is reduced. However, the high atmospheric background of di-nitrogen (N₂) causes difficulties in assessing denitrification enzyme activity (DEA) and denitrification potential (DP) in soils directly. Here, we apply Membrane Inlet Mass Spectrometry (MIMS) technique to investigate indirectly DEA and DP in soils by measuring N₂/Ar ratio changes in headspace water over soil. Soils were collected from 0-10, 15-25 and 60-70 cm depths of a grazed ryegrass and grass-clover. The samples were amended with helium-flushed deionized water containing ranges of NO₃ and carbon (glucose-C) and were incubated for six hours in the dark at 21 °C. The peaks for N₂/Ar ratio, declined with increasing soil depth, indicating a reduced substrate requirements to initiate DEA en-masse (15-30 mg NO₃-N alone or with 60-120 mg glucose-C, kg⁻¹ soil). The dissolved N₂O concentrations were very small (0.004-0.269 μg N kg⁻¹ soil) but responded well to the added N and C, showing a reduction in DEA with soil depth. In three separate studies, only subsoils were incubated for 3 days at 12 °C with 20-30 mg NO₃-N ± 40-60 mg glucose-C, kg⁻¹ soil. Denitrification capacity (DC, NO₃ only treatment) was not statistically different to the control (no amendment) within a land use (0.03-0.05 vs. 0.07-0.22 mg N kg⁻¹ soil d⁻¹), the highest being in ryegrass subsoils receiving groundwater. The DP was significantly (P < 0.0001) higher in subsoils under ryegrass than under grass-clover (0.50-0.71 vs. 1.15 mg N kg⁻¹ soil d⁻¹). The rates of DP (NO₃ + glucose-C) increased significantly (P < 0.0001) in unsaturated and saturated subsoils (0.92 and 2.19 mg N kg⁻¹ soil d⁻¹, respectively) of grass-clover, due to the higher reductive state resulting from the 10 day pre-incubation. Available C accelerated denitrification in soils and superseded the temporary elevation in oxidative state due to NO₃ addition. The substrates load differences between the land uses regulated the degree of denitrification rates. Results suggest that both dissolved N₂O measured by gas chromatography and N₂/Ar ratio measured by MIMS to indirectly determine DEA, and the latter to quantify total DC/DP in soils can be used. However, interference of oxygen in the MIMS system should be considered if available C is added or is naturally elevated in soil or groundwater.     

Fluxes of nitrous oxide and nitric oxide from experimental excreta patches in boreal agricultural soil

Nitric oxide (NO) and nitrous oxide (N₂O) emissions were measured from experimental dung and urine patches placed on boreal pasture soil during two growing seasons and one autumn period until soil freezing. N₂O emissions in situ were studied by a static chamber method. NO was measured with a dynamic chamber method using a NO analyser in situ. Mean emissions from the control plots were 47.6±4.5 μg N₂ON m⁻² h⁻¹ and 12.6±1.6 μg NON m⁻² h⁻¹. N₂O and NO emissions from urine plots (132±21.2 μg N₂ON m⁻² h⁻¹ and 51.9±7.6 μg NON m⁻² h⁻¹) were higher than those from dung plots (110.0±20.1 μg N₂ON m⁻² h⁻¹ and 14.7±2.1 μg NON m⁻² h⁻¹). There was a large temporal variation in N₂O and NO emissions. Maximum N₂O emissions were measured a few weeks after dung or urine application, whereas the maximum NO emissions were detected the following year. NO was responsible on average 14% (autumn) and 34% (summer) of total (NO+N₂O)N emissions from the pasture soil. NO emissions increased with increasing soil temperature and with decreasing soil moisture. N₂O emissions increased with increasing soil moisture, but did not correlate with soil temperature. Therefore we propose that N₂O and NO were produced mainly during different microbial processes, i.e., nitrification and denitrification, respectively. The results show that the overall conditions and mechanism especially for emissions of NO are still poorly understood but that there are differences in the mechanisms regulating N₂O and NO production.     

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.     

Contribution of nitrification and denitrification to N2O emissions from urine patches

Urine deposition by grazing livestock causes an immediate increase in nitrous oxide (N₂O) emissions, but the responsible mechanisms are not well understood. A nitrogen-15 (¹⁵N) labelling study was conducted in an organic grass-clover sward to examine the initial effect of urine on the rates and N₂O loss ratio of nitrification (i.e. moles of N₂O-N produced per moles of nitrate produced) and denitrification (i.e. moles of N₂O produced per moles of N₂O+N₂ produced). The effect of artificial urine (52.9 g N m⁻²) and ammonium solution (52.9 g N m⁻²) was examined in separate experiments at 45% and 35% water-filled pore space (WFPS), respectively, and in each experiment a water control was included. The N₂O loss derived from nitrification or denitrification was determined in the field immediately after application of ¹⁵N-labelled solutions. During the next 24 h, gross nitrification rates were measured in the field, whereas the denitrification rates were measured in soil cores in the laboratory. Compared with the water control, urine application increased the N₂O emission from 3.9 to 42.3 μg N₂O-N m⁻² h⁻¹, whereas application of ammonium increased the emission from 0.9 to 6.1 μg N₂O-N m⁻² h⁻¹. In the urine-affected soil, nitrification and denitrification contributed equally to the N₂O emission, and the increased N₂O loss resulted from a combination of higher rates and higher N₂O loss ratios of the processes. In the present study, an enhanced nitrification rate seemed to be the most important factor explaining the high initial N₂O emission from urine patches deposited on well-aerated soils.     

Nitrogen oxides emission from soils bearing a potato crop as influenced by fertilization with treated pig slurries and composts

Nitrous oxide, nitric oxide and denitrification losses from an irrigated soil amended with organic fertilizers with different soluble organic carbon fractions and ammonium contents were studied in a field study covering the growing season of potato (Solanum tuberosum). Untreated pig slurry (IPS) with and without the nitrification inhibitor dicyandiamide (DCD), digested thin fraction of pig slurry (DTP), composted solid fraction of pig slurry (CP) and composted municipal solid waste (MSW) mixed with urea were applied at a rate of 175 kg available N ha⁻¹, and emissions were compared with those from urea (U) and a control treatment without any added N fertilizer (Control). The cumulative denitrification losses correlated significantly with the soluble carbohydrates, dissolved N and total C added. Added dissolved organic C (DOC) and dissolved N affected the N₂O/N₂ ratio, and a lower ratio was observed for organic fertilizers than from urea or unfertilized controls. The proportion of N₂O produced from nitrification was higher from urea than from organic fertilizers. Accumulated N₂O losses during the crop season ranged from 3.69 to 7.31 kg N₂O-N ha⁻¹ for control and urea, respectively, whereas NO losses ranged from 0.005 to 0.24 kg NO-N ha⁻¹, respectively. Digested thin fraction of pig slurry compared to IPS mitigated the total N₂O emission by 48% and the denitrification rate by 33%, but did not influence NO emissions. Composted pig slurry compared to untreated pig slurry increased the N₂O emission by 40% and NO emission by 55%, but reduced the denitrification losses (34%). DCD partially inhibited nitrification rates and reduced N₂O and NO emissions from pig slurry by at least 83% and 77%, respectively. MSW+U, with a C:N ratio higher than that of the composted pig slurry, produced the largest denitrification losses (33.3 kg N ha⁻¹), although N₂O and NO emissions were lower than for the U and CP treatments.This work has shown that for an irrigated clay loam soil additions of treated organic fertilizers can mitigate the emissions of the atmospheric pollutants NO and N₂O in comparison with urea.     

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.     

Decomposition of C-labeled roots in a pasture soil exposed to 10 years of elevated CO2

The net flux of soil C is determined by the balance between soil C input and microbial decomposition, both of which might be altered under prolonged elevated atmospheric CO₂. In this study, we determined the effect of elevated CO₂ on decomposition of grass root material (Lolium perenne L.). ¹⁴C-labeled root material, produced under ambient (35 Pa pCO₂) or elevated CO₂ (70 Pa pCO₂) was incubated in soil for 64 days. The soils were taken from a pasture ecosystem which had been exposed to ambient (35 Pa pCO₂) or elevated CO₂ (60 Pa pCO₂) under FACE-conditions for 10 years and two fertilizer N rates: 140 and 560 kg N ha⁻¹ year⁻¹. In soil exposed to elevated CO₂, decomposition rates of root material grown at either ambient or elevated CO₂ were always lower than in the control soil exposed to ambient CO₂, demonstrating a change in microbial activity. In the soil that received the high rate of N fertilizer, decomposition of root material grown at elevated CO₂ decreased by approximately 17% after incubation for 64 days compared to root material grown at ambient CO₂. The amount of ¹⁴CO₂ respired per amount of ¹⁴C incorporated in the microbial biomass (q¹⁴CO₂) was significantly lower when roots were grown under high CO₂ compared to roots grown under low CO₂. We hypothesize that this decrease is the result of a shift in the microbial community, causing an increase in metabolic efficiency. Soils exposed to elevated CO₂ tended to respire more native SOC, both with and without the addition of the root material, probably resulting from a higher C supply to the soil during the 10 years of treatment with elevated CO₂. The results show the importance of using soils adapted to elevated CO₂ in studies of decomposition of roots grown under elevated CO₂. Our results further suggest that negative priming effects may obscure CO₂ data in incubation experiments with unlabeled substrates. From the results obtained, we conclude that a slower turnover of root material grown in an ‘elevated-CO₂ world’ may result in a limited net increase in C storage in ryegrass swards.     

Can a mixed stand of N2-fixing and non-fixing plants restrict N2O emissions with increasing CO2 concentration?

Initial effects of elevated atmospheric CO₂ concentration on N₂O fluxes and biomass production of timothy/red clover were studied in the laboratory. The experimental design consisted of two levels of atmospheric CO₂ (ca. 360 and 720 μmol CO₂ mol⁻¹) and two N fertilisation levels (5 and 10 g N m⁻²). There was a total of 36 mesocosms comprising sandy loam soil, which were equally distributed in four thermo-controlled greenhouses. In two of the greenhouses, the CO₂ concentration was kept at ambient concentration and in the other two at doubled concentration. Forage was harvested and the plants fertilised three times during the basic experiment, followed by harvest, a fertilisation with the double amount of nitrogen and rise of water level. Under elevated CO₂, harvestable and total aboveground dry biomass production of a mixed Trifolium/Phleum stand was increased at both N treatments compared to ambient CO₂. The N₂O flux rates under ambient CO₂ were significantly higher at both N treatments during the early growth of mixed Phleum/Trifolium mesocosms compared to the N₂O flux rate under elevated CO₂. However, when the conditions were favourable for denitrification at the end of the experiment, i.e. N availability and soil moisture were high enough, the elevated CO₂ concentration enhanced the N₂O efflux.     

Temporal and spatial dynamics of soil solution C and N concentrations during Lolium perenne L. sward establishment and the effects of elevated CO2 and N additions

There is now clear evidence for a prolonged increase in atmospheric CO₂ concentrations and enrichment of the biosphere with N. Understanding the fate of C in the plant-soil system under different CO₂ and N regimes is therefore of considerable importance in predicting the environmental effects of climate change and in predicting the sustainability of ecosystems. Swards of Lolium perenne were grown from seed in a Eutric Cambisol at either ambient (ca. 350 μmol mol⁻¹) or elevated (700 μmol mol⁻¹) atmospheric pCO₂ and subjected to two inorganic N fertilizer regimes (no added N and 70 kg N ha⁻¹ month⁻¹). After germination, soil solution concentrations of dissolved organic C (DOC), dissolved inorganic N (DIN), dissolved organic N (DON), phenolics and H⁺ were measured at five depths down the soil profile over 3 months. The exploration of soil layers down the soil profile by roots caused transient increases in soil solution DOC, DON and phenolic concentrations, which then subsequently returned to lower quasi-stable concentrations. In general, the addition of N tended to increase DOC and DON concentrations while exposure to elevated pCO₂ had the opposite effect. These treatment effects, however, gradually diminished over the duration of the experiment from the top of the soil profile downwards. The ambient pCO₂ plus added N regime was the only treatment to maintain a notable difference in soil solution solute concentration, relative to other treatments. This effect on soil solution chemistry appeared to be largely indirect resulting from increased plant growth and a decrease in soil moisture content. Our results show that although plant growth responses to elevated pCO₂ are critically dependent upon N availability, the organic chemistry of the soil solution is relatively insensitive to changes in plant growth once the plants have become established.     

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.     

Sources and rates of nitrous oxide emissions from grazed grassland after application of N-labelled mineral fertilizer and slurry

Nitrogen (N) fertilizer application and grazing are known to induce nitrous oxide (N₂O) emissions from grassland soils. In a field study, general information on rates of N₂O emission, the effect of cattle grazing and the type (mineral fertilizer, cattle slurry) and amount of N supply on the flux of N₂O from a sandy soil were investigated. N₂O emissions from permanent grassland managed as a mixed system (two cuts followed by two grazing cycles) were monitored over 11 months during 2001-2002 in northern Germany using the closed chamber method. The field experiment consisted of four regionally relevant fertilizer combinations, i.e. two mineral N application rates (0 and 100 kg N ha⁻¹ yr⁻¹) and two slurry levels (0 and 74 kg N ha⁻¹ yr⁻¹).Mean cumulative N₂O-N loss was 3.0 kg ha⁻¹ yr⁻¹, and the cumulative ¹⁵N-labelled N₂O emissions varied from 0.03% to 0.19% of the ¹⁵N applied. ¹⁵N labelling indicated that more N₂O was emitted from mineral N than from slurry treated plots, and in all treatments the soil N pool was always clearly the major source of N₂O. Regarding the total cumulative N₂O losses, differences among treatments were not significant, which was caused by: (i) a high variance in emissions during and after cattle grazing due to the random distribution of excrements and by (ii) high N₂ fixation of white clover in the 0 kg N ha⁻¹ treatments, which resulted in similar N status of all treatments. However before grazing started, treatments showed significant differences. After cattle grazing in summer, N₂O emission rates were higher than around the time of spring fertilizer application, or in winter. Grazing resulted in N₂O flux rates up to 489 μg N₂O-N m⁻² h⁻¹ and the grazing period contributed 31-57% to the cumulative N₂O emission. During freeze-thaw cycles in winter (December-February) N₂O emission rates of up to 147 μg N₂O-N m⁻² h⁻¹ were measured, which contributed up to 26% to the annual N₂O flux. The results suggest that N fertilizer application and grazing caused only short-term increases of N₂O flux rates whereas the major share of annual N₂O emission emitted from the soil N pool. The significantly increased N₂O fluxes during freeze-thaw cycles show the importance of emission events in winter which need to be covered by measurements for obtaining reliable estimates of annual N₂O emissions.     

Comparison of field and laboratory measurement of denitrification and N2O production in the saturated zone of hydromorphic soils

We evaluated a new method to measure in situ denitrification under field conditions in a number of water-saturated subsoils that had a broad range of biogeochemical properties. A test solution containing ¹⁵NO₃⁻ and/or C₂H₂ was introduced to the subsoil and the subsequent production of dissolved denitrification products was measured to quantify denitrification activity. The method showed a clear production of denitrification products over time. Results were compared to laboratory-based measurements from the same soil incubated as anaerobic slurries with added ¹⁵NO₃⁻. Rates of denitrification with the in situ and the laboratory methods ranged from 1-2800 and 1-1700 μg N kg⁻¹ d⁻¹, respectively. Generally the methods gave good agreement and we consider both to be valid. However, there were some significant deviations, which we attribute to spatial heterogeneity and laboratory effects. Because the laboratory method is so much easier to perform, we suggest it should be the preferred method for large-scale studies of denitrification from the soil types we investigated. However, the two methods showed poor agreement in determining the proportion of N₂O in the total denitrification output. This was because this proportion is subject to delicate and complex control. We conclude that neither method was suitable for quantifying N₂O emission from the denitrification measurements.     

Nitrogen fertilizers promote denitrification

A laboratory study was conducted to compare the effects of different N fertilizers on emission of N₂ and N₂O during denitrification of NO₃ ^– in waterlogged soil. Field-moist samples of Drummer silty clay loam soil (fine-silty, mixed, mesic Typic Haplaquoll) were incubated under aerobic conditions for 0, 2, 4, 7, 14, 21, or 42 days with or without addition of unlabelled (NH₄)₂SO₄, urea, NH₄H₂PO₄, (NH₄)₂HPO₄, NH₄NO₃ (200 or 1000 mg N kg^–1 soil), or liquid anhydrous NH₃ (1000 mg N kg^–1 soil). The incubated soil samples were then treated with ¹⁵N-labelled KNO₃ (250 mg N kg^–1 soil, 73.7 atom% ¹⁵N), and incubation was carried out under waterlogged conditions for 5 days, followed by collection of atmospheric samples for ¹⁵N analyses to determine labelled N₂ and N₂O. Compared to samples incubated without addition of unlabelled N, all of the fertilizers promoted denitrification of ¹⁵NO₃ ^–. Emission of labelled N₂ and N₂O decreased in the order: Anhydrous NH₃>urea<$>\gg<$> (NH₄)₂HPO₄>(NH₄)₂SO₄≃NH₄NO₃≃NH₄H₂PO₄. The highest emissions observed with anhydrous NH₃ or urea coincided with the presence of NO₂ ^–, and ¹⁵N analyses indicated that these emissions originated from NO₂ ^– rather than NO₃ ^–. Emissions of labelled N₂ and N₂O were significantly correlated with fertilizer effects on soil pH and water-soluble organic C. Received: 17 January 1996     

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.     

Chemical composition, or quality, of agroforestry residues influences N2O emissions after their addition to soil

Emissions of N₂O were measured following addition of ¹⁵N-labelled (2.6-4.7 atom% excess ¹⁵N) agroforestry residues (Sesbania sesban, mixed Sesbania/Macroptilium atropurpureum, Crotalaria grahamiana and Calliandra calothyrsus) to a Kenyan oxisol at a rate of 100 mg N kg soil⁻¹ under controlled environment conditions. Emissions were increased following addition of residues, with 22.6 mg N m⁻² (124.4 mg N m⁻² kg biomass⁻¹; 1.1 mg ¹⁵N m⁻²; 1.03% of ¹⁵N applied) emitted as N₂O over 29 d after addition of both Sesbania and Macroptilium residues in the mixed treatment. Fluxes of N₂O were positively correlated with CO₂ fluxes, and N₂O emissions and available soil N were negatively correlated with residue lignin content (r=−0.49;P<0.05), polyphenol content (r=−0.94;P<0.05), protein binding capacity (r=−0.92;P<0.05) and with (lignin+polyphenol)-to-N ratio (r=−0.55;P<0.05). Lower emission (13.6 mg N m⁻² over 29 d; 94.5 mg N m⁻² kg biomass⁻¹; 0.6 mg ¹⁵N m⁻²; 0.29% of ¹⁵N applied) after addition of Calliandra residue was attributed to the high polyphenol content (7.4%) and high polyphenol protein binding capacity (383 μg BSA mg plant⁻¹) of this residue binding to plant protein and reducing its availability for microbial attack, despite the residue having a N content of 2.9%. Our results indicate that residue chemical composition, or quality, needs to be considered when proposing mitigation strategies to reduce N₂O emissions from systems relying on incorporation of plant biomass, e.g. improved-fallow agroforestry systems, and that this consideration should extend beyond the C-to-N ratio of the residue to include polyphenol content and their protein binding capacity.