Microbial processes and the site of N2O production in a temperate grassland soil

To understand nitrous oxide (N₂O) emissions from terrestrial ecosystems it is necessary to understand the processes leading to N₂O production. Here, for the first time, results are presented which identify in situ the processes of N₂O production in a temperate grassland soil. A small portion of the nitrogen (N) applied in the summer to the grassland soil was rapidly transported below the main rooting zone (>20 cm) and resulted in large N₂O productions at depths of 20-50 cm. Preferential pathways must have been responsible for this movement because the soil conditions were not conducive to leaching by piston flow. The N₂O was entirely produced by nitrate (NO₃⁻) reduction which was surprising because the bulk soil was aerobic. Therefore, reduction processes can operate during times of the year when it is least expected and cause large N₂O concentrations deep in the soil profile.     

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.     

Dinitrogen emissions and the N2:N2O emission ratio of a Rendzic Leptosol as influenced by pH and forest thinning

Reduction of nitrous oxide (N₂O) to dinitrogen (N₂) by denitrification in soils is of outstanding ecological significance since it is the prevailing natural process converting reactive nitrogen back into inert molecular dinitrogen. Furthermore, the extent to which N₂O is reduced to N₂ via denitrification is a major regulating factor affecting the magnitude of N₂O emission from soils. However, due to methodological problems in the past, extremely little information is available on N₂ emission and the N₂:N₂O emission ratio for soils of terrestrial ecosystems. In this study, we simultaneously determined N₂ and N₂O emissions from intact soil cores taken from a mountainous beech forest ecosystem. The soil cores were taken from plots with distinct differences in microclimate (warm-dry versus cool-moist) and silvicultural treatment (untreated control versus heavy thinning). Due to different microclimates, the plots showed pronounced differences in pH values (range: 6.3–7.3). N₂O emission from the soil cores was generally very low (2.0 ± 0.5–6.3 ± 3.8 μg N m⁻² h⁻¹ at the warm-dry site and 7.1 ± 3.1–57.4 ± 28.5 μg N m⁻² h⁻¹ at the cool-moist site), thus confirming results from field measurements. However, N₂ emission exceeded N₂O emission by a factor of 21 ± 6–220 ± 122 at the investigated plots. This illustrates that the dominant end product of denitrification at our plots and under the given environmental conditions is N₂ rather than N₂O. N₂ emission showed a huge variability (range: 161 ± 64–1070 ± 499 μg N m⁻² h⁻¹), so that potential effects of microclimate or silvicultural treatment on N₂ emission could not be identified with certainty. However, there was a significant effect of microclimate on the magnitude of N₂O emission as well as on the mean N₂:N₂O emission ratio. N₂:N₂O emission ratios were higher and N₂O emissions were lower for soil cores taken from the plots with warm-dry microclimate as compared to soil cores taken from the cool-moist microclimate plots. We hypothesize that the increase in the N₂:N₂O emission ratio at the warm-dry site was due to higher N₂O reductase activity provoked by the higher soil pH value of this site. Overall, the results of this study show that the N₂:N₂O emission ratio is crucial for understanding the regulation of N₂O fluxes of the investigated soil and that reliable estimates of N₂ emissions are an indispensable prerequisite for accurately calculating total N gas budgets for the investigated ecosystem and very likely for many other terrestrial upland ecosystems as well.     

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.     

Emission of N2O, N2 and CO2 from soil fertilized with nitrate: effect of compaction, soil moisture and rewetting

Soil compaction and soil moisture are important factors influencing denitrification and N₂O emission from fertilized soils. We analyzed the combined effects of these factors on the emission of N₂O, N₂ and CO₂ from undisturbed soil cores fertilized with (150 kg N ha⁻¹) in a laboratory experiment. The soil cores were collected from differently compacted areas in a potato field, i.e. the ridges (ρ_D=1.03 g cm⁻³), the interrow area (ρ_D=1.24 g cm⁻³), and the tractor compacted interrow area (ρ_D=1.64 g cm⁻³), and adjusted to constant soil moisture levels between 40 and 98% water-filled pore space (WFPS).High N₂O emissions were a result of denitrification and occurred at a WFPS≥70% in all compaction treatments. N₂ production occurred only at the highest soil moisture level (≥90% WFPS) but it was considerably smaller than the N₂O-N emission in most cases. There was no soil moisture effect on CO₂ emission from the differently compacted soils with the exception of the highest soil moisture level (98% WFPS) of the tractor-compacted soil in which soil respiration was significantly reduced. The maximum N₂O emission rates from all treatments occurred after rewetting of dry soil. This rewetting effect increased with the amount of water added. The results show the importance of increased carbon availability and associated respiratory O₂ consumption induced by soil drying and rewetting for the emissions of N₂O.     

同位素异位体溯源土壤N2O的方法、技术和展望

土壤是N₂O的重要排放来源之一。土壤中N₂O产生途径众多、受多种因素调控，深入分析土壤N₂O产生途径才能采取针对性的减排策略。稳定同位素技术已广泛用于研究土壤N₂O排放，N₂O同位素异位体法是近年来新兴的研究方法。该研究方法通过测定土壤N₂O的同位素组成(δ¹⁵NSPN₂O、δ¹⁸ON₂O和δ¹⁵N^bulkN₂O)分析N₂O排放贡献，因无需添加标记物、对土壤系统干扰小、成本低，适合在野外田间研究N₂O排放，是15N标记方法的有力补充。本文详细介绍了N₂O同位素异位体法的原理、质谱测定方法、定量分析方法、影响该方法的因素及其应用前景。     

Carbon and oxygen controls on N2O and N2 production during nitrate reduction

Here we provide evidence that the form of carbon compound and O₂ concentration exert an inter-related regulation on the production and reduction of N₂O in soil. 6.7 mM d-glucose, 6.7 mM D-mannitol, 8 mM L-glutamic acid or 10 mM butyrate (all equivalent to 0.48 g C l⁻¹) were applied to slurries of a sandy loam soil. At the start of the experiment headspace O₂ concentrations were established at ∼2%, 10% and 21% O₂ v/v for each C treatment, and 2 mM K¹⁵NO₃ (25 atom % excess ¹⁵N) was applied, enabling quantification of ¹⁵N-N₂ production, ¹⁵N-(N₂O-to-N₂) ratios and DNRA. The form of C compound was most important in the initially oxic (21% O₂ v/v) soils, where addition of butyrate and glutamic acid resulted in greater N₂O production (0.61 and 0.3 μg N₂O-N g⁻¹ soil for butyrate and glutamic acid, respectively) than the addition of carbohydrates (glucose and mannitol). Although, there was no significant effect of C compound at low initial O₂ concentrations (∼2% O₂ v/v), production of ¹⁵N-N₂ was greatest where headspace O₂ concentrations were initially, or fallen to, ∼2% O₂ v/v, with greatest reduction of N₂O and lowering ¹⁵N-(N₂O-to-N₂) ratios (∼0-0.27). This may reflect that the effect of C is indirect through stimulation of heterotrophic respiration, lowering O₂ concentrations, providing sub-oxic conditions for dissimilatory nitrate reduction pathways. Addition of carbohydrates (glucose and mannitol) also resulted in greatest recovery of ¹⁵N in NH₄⁺ from applied ¹⁵N-NO₃⁻, indicative of the occurrence of DNRA, even in the slurries with initial 10% and 21% O₂ v/v concentrations. Our ¹⁵N approach has provided the first direct evidence for enhancement of N₂O reduction in the presence of carbohydrates and the dual regulation of C compound and O₂ concentration on N₂O production and reduction, which has implications for management of N₂O emissions through changing C inputs (exudates, rhizodeposition, residues) with plant species of differing C traits, or through plant breeding.     

高氮和NO2-对中亚热带森林土壤N2O和NO产生的影响

利用15N同位素标记方法,研究在两种水分条件即60％和90％ WHC下,添加硝酸盐(NH4NO3,N 300 mg kg-1)和亚硝酸盐(NaNO2,N 1 mg kg-1)对中亚热带天然森林土壤N2O和NO产生过程及途径的影响.结果表明,在含水量为60％ WHC的情况下,高氮输入显著抑制了N2O和NO的产生(p＜0.01);但当含水量增为90％ WHC后,实验9h内抑制N2O产生,之后转为促进.所有未灭菌处理在添加NO2-后高氮抑制均立即解除并大量产生N2O和NO,与对照成显著差异(p＜0.01),在60％ WHC条件下,这种情况维持时间较短(21 h),但如果含水量高(90％ WHC)这种情况会持续很长时间(2周以上),说明水分有效性的提高和外源NO2-在高氮抑制解除中起到重要作用.本实验中N2O主要来源于土壤反硝化过程,而且加入未标记NO2-后导致杂合的N2O(14N15NO)分子在实验21 h内迅速增加,表明这种森林土壤的反硝化过程可能主要是通过真菌的“共脱氮”来实现,其贡献率可多达80％以上.Spearman秩相关分析表明未灭菌土壤NO的产生速率与N2O产生速率成显著正相关性(p＜0.05),土壤含水量越低二者相关性越高.灭菌土壤添加NO2-能较未灭菌土壤产生更多的NO,但却几乎不产生N2O,表明酸性土壤的化学反硝化对NO的贡献要大于N2O.     

Isotopologue fractionation during microbial reduction of N2O within soil mesocosms as a function of water-filled pore space

Isotopologue analyses of N₂O within soil mesocosm experiments were used to evaluate the influence of N₂O reduction on isotope fractionation. We investigated fractionation during N₂O reduction at 60%, 80% and 100% water-filled pore space (WFPS) and found net isotope effects (NIE) for δ¹⁵N of 4.2–7.8‰, δ¹⁸O of 12.5–19.1‰, δ¹⁵N^α of 6.4–9.7‰ and δ¹⁵N^β of 2.0–5.9‰. Consequently, N₂O reduction has a marked affect on isotopologue values and the importance of this process in flux chamber studies should not be ignored. With the exception of SP (the difference between the δ¹⁵N of the central, α, and terminal, β, atoms) inverse relationships between the NIE, reaction rate and reaction rate constant and WFPS were observed. Isotopic discrimination in SP during N₂O reduction was small and the average NIE for the treatments varied between 2.9‰ and 4.5‰. A strong correlation was evident between δ¹⁸O vs. δ¹⁵N and δ¹⁸O vs. δ¹⁵N^α during reduction with slopes of 2.6 and 1.9, respectively, which contrasts from a slope of <1 commonly observed for mixing between soil-derived and atmospheric N₂O in flux chambers.     

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.     

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.     

Elevated CO2 stimulates N2O emissions in permanent grassland

To evaluate climate forcing under increasing atmospheric CO₂ concentrations, feedback effects on greenhouse gases such as nitrous oxide (N₂O) with a high global warming potential should be taken into account. This requires long-term N₂O flux measurements because responses to elevated CO₂ may vary throughout annual courses. Here, we present an almost 9 year long continuous N₂O flux data set from a free air carbon dioxide enrichment (FACE) study on an old, N-limited temperate grassland. Prior to the FACE start, N₂O emissions were not different between plots that were later under ambient (A) and elevated (E) CO₂ treatments, respectively. However, over the entire experimental period (May 1998–December 2006), N₂O emissions more than doubled under elevated CO₂ (0.90 vs. 2.07 kg N₂O-N ha⁻¹ y⁻¹ under A and E, respectively). The strongest stimulation occurred during vegetative growth periods in the summer when soil mineral N concentrations were low. This was surprising because based on literature we had expected the highest stimulation of N₂O emissions due to elevated CO₂ when mineral N concentrations were above background values (e.g. shortly after N application in spring). N₂O emissions under elevated CO₂ were moderately stimulated during late autumn–winter, including freeze–thaw cycles which occurred in the 8th winter of the experiment. Averaged over the entire experiment, the additional N₂O emissions caused by elevated CO₂ equaled 4738 kg CO₂-equivalents ha⁻¹, corresponding to more than half a ton (546 kg) of CO₂ ha⁻¹ which has to be sequestered annually to balance the CO₂-induced N₂O emissions. Without a concomitant increase in C sequestration under rising atmospheric CO₂ concentrations, temperate grasslands may be converted into greenhouse gas sources by a positive feedback on N₂O emissions. Our results underline the need to include continuous N₂O flux measurements in ecosystem-scale CO₂ enrichment experiments.     

Compaction effects on CO2 and N2O production during drying and rewetting of soil

The effects of compaction on soil porosity and soil water relations are likely to influence substrate availability and microbial activity under fluctuating soil moisture conditions. We conducted a short laboratory incubation to investigate the effects of soil compaction on substrate availability and biogenic gas (CO₂ and N₂O) production during the drying and rewetting of a fine-loamy soil. Prior to initiating the drying and wetting treatments, CO₂ production (−10 kPa soil water content) from uncompacted soil was 2.3 times that of compacted soil and corresponded with higher concentrations of microbial biomass C (MBC) and dissolved organic C (DOC). In contrast, N₂O production was 67 times higher in compacted than uncompacted soil at field capacity. Soil aeration rather than substrate availability (e.g. NO₃⁻ and DOC) appeared to be the most important factor affecting N₂O production during this phase. The drying of compacted soil resulted in an initial increase in CO₂ production and a nearly two-fold higher average rate of C mineralization at maximum dryness (owing to a higher water-filled pore space [WFPS]) compared to uncompacted soil. During the drying phase, N₂O production was markedly reduced (by 93-96%) in both soils, though total N₂O production remained slightly higher in compacted than uncompacted soil. The increase in CO₂ production during the first 24 h following rewetting of dry soil was about 2.5 times higher in uncompacted soil and corresponded with a much greater release of DOC than in compacted soil. MBC appeared to be the source of the DOC released from uncompacted soil but not from compacted soil. The production of N₂O during the first 24 h following rewetting of dry soil was nearly 20 times higher in compacted than uncompacted soil. Our results suggest that N₂O production from compacted soil was primarily the result of denitrification, which was limited by substrates (especially NO₃⁻) made available during drying and rewetting and occurred rapidly after the onset of anoxic conditions during the rewetting phase. In contrast, N₂O production from uncompacted soil appeared to be primarily the product of nitrification that was largely associated with an accumulation of NO₃⁻ following rewetting of dry soil. Irrespective of compaction, the response to drying and rewetting was greater for N₂O production than for CO₂ production.     

Emissions and spatial variability of N2O, N2 and nitrous oxide mole fraction at the field scale, revealed with N isotopic techniques

The accurate measurement of nitrous oxide (N₂O) and dinitrogen (N₂) during the denitrification process in soils is a challenge which will help to estimate the contribution of soil N₂O emissions to global warming. Oxygen concentration, nitrate concentration and carbon availability are generally the main factors that control soil denitrification rate and the amount of N₂O or N₂ emitted. The aim of this paper is to present a database of the N₂O mole fraction measured at the field scale, and to test hypotheses concerning its regulation. A ¹⁵N-nitrate tracer solution was added to 36 undisturbed soil cores on a 20 m×20 m cultivated field plot. Fluxes of CO₂, N₂O and N₂ from the soil surface were monitored for 24 h. Soil moisture, bulk density, carbon, nitrogen and mineral nitrogen concentration were also measured to investigate possible spatial relationships between their variations and those of N₂O, N₂ and nitrous oxide mole fraction. Under high water content, nitrous oxide and N₂ emissions were highly variable with variation coefficients of 70-140%. N₂O emission rates were about twice as high as those of N₂, with a total denitrification rate ranging from 269 to 3843 g N ha⁻¹ d⁻¹. After 24 h of incubation, the values of nitrous oxide mole fraction ranged from 0.15 to 0.94 and no significant decline during incubation time was observed. Spatial variability of N₂O, N₂ and nitrous oxide mole fraction was high and no spatial dependence was observed at the scale of the experimental plot. Only tenuous relationships between gaseous nitrogen emissions and soil properties (mainly nitrate concentration and moisture content) were found. Meanwhile, a positive correlation was observed between N₂ and CO₂ emissions. This result supports the hypothesis that an increase in soil available organic carbon leads to N₂ emissions as the end product of denitrification.     

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.     

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.     

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

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

生物炭施用下中国农田土壤N2O排放的Meta分析

为明确施加生物炭对中国农田土壤N_2O排放的影响和主要控制因素,以公开发表的试验数据为研究对象,采用Meta-analysis法定量分析了施加生物炭条件下,气候、土壤性质、田间管理方式、生物炭性质与施加量对土壤N_2O排放的影响,并对各影响因素进行通径分析。结果表明,当年降雨量≥600 mm时,生物炭显著降低土壤N_2O排放量(P0.05),且随年降雨量的增加而增强;当年日照时数大于1 000 h时,生物炭对土壤N_2O的减排效果随年日照时数的增加而减弱。当土壤p H≥6.5时,生物炭对土壤N_2O的减排效果随土壤p H的增加呈先增后减趋势;在壤土中施加生物炭对N_2O的减排效果显著(P0.05),而砂土和黏土不显著(P0.05)。生物炭对覆膜土壤N_2O的减排效果优于不覆膜土壤;生物炭对土壤N_2O的减排效果随施氮肥量增加而减弱,而随生物炭比表面积的增加而增强。当生物炭C/N处于30~500时,生物炭施用下土壤N_2O排放量显著降低(P0.05);当生物炭施加量处于20~160 t×hm-2时,生物炭对土壤N_2O的减排效果随施加量增加而增强。生物炭对土壤N_2O减排的影响存在显著的区域性特征,对华南、华东、华中和东北地区影响显著(P0.05),而对西北地区不显著(P0.05);施氮肥量、生物炭施加量、年均温和年降雨量是影响生物炭减排效果的最主要因素,这些因素的相互作用共同影响生物炭对土壤N_2O的减排效果。该研究可为生物炭在我国农区的推广应用和农田N_2O减排提供参考。     

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.     

Nitrification controls N2O production rates in a frozen boreal forest soil

The winter season has been identified as a significant contributor to N₂O emissions from boreal soils, but our understanding of the processes regulating these emissions is fragmentary. We investigated potential N-sources and pathways involved in N₂O formation in a frozen boreal forest soil by labeling soil samples with ¹⁵N-containing substrates, and measured rates of ¹⁵N₂O/¹⁵N₂ formation under both oxic and anoxic conditions. Our results showed that all N₂O produced in the frozen samples originate from denitrification, but the rate-limiting factor is NO₃⁻ availability, which is largely governed by nitrification. This suggests that N₂O formation in frozen boreal soils may be sustained for a prolonged period of time, but is governed by a delicate balance of the O₂ regime.