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.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

区分土壤中硝化与反硝化对N2O产生贡献的方法

土壤是产生N₂O的最主要来源之一。硝化和反硝化反应是产生N₂O的主要机理，由于硝化和反硝化微生物同时存在于土壤中，因而硝化和反硝化作用能同时产生N₂O。N₂O的来源可通过使用选择性抑制剂，杀菌剂以及加入的标记底物确定。通过对生成N₂O反应的每一步分析，主要从抑制反应发生的催化酶和细菌着手，总结了测量区分硝化、反硝化和DNRA反应对N₂O产生的贡献方法。并对¹⁵N标记底物法，乙炔抑制法和环境因子抑制法作了详细介绍。     

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.     

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.     

Gaseous nitrogen losses from fertilized soil during nitrification and denitrification using the acetylene blockage technique

Summary A sandy soil amended with different forms and amounts of fertilizer nitrogen (urea, ammonium sulphate and potassium nitrate) was investigated in model experiments for N₂O emission, which may be evolved during both oxidation of ammonia to nitrate and anaerobic respiration of nitrate. Since C₂H₂ inhibits both nitrification and the reduction of N₂O to N₂ during denitrification, the amount of N₂O evolved in the presence and absence of C₂H₂ represents the nitrogen released through nitrification and denitrification.Results show that amounts of N₂O-N lost from soils incubated anaerobically with 0.1% C₂H₂ and treated with potassium nitrate (23.1 µg N-NO ₃ ^– /g dry soil) exceeded those from soils incubated in the presence of 20% oxygen and treated with even larger amounts of nitrogen as urea and ammonium sulphate. This indicates that nitrogen losses by denitrification may potentially be higher than those occurring through nitrification.     

Nitrous oxide flux from soil amino acid mineralization

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

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.     

Nitrate leaching in soil: Tracing the NO3 sources with the help of stable N and O isotopes

Legumes increase the plant-available N pool in soil, but might also increase NO₃⁻ leaching to groundwater. To minimize NO₃⁻ leaching, N-release processes and the contribution of legumes to NO₃⁻ concentrations in soil must be known. Our objectives were (1) to quantify NO₃⁻-N export to >0.3 m soil depth from three legume monocultures (Medicago x varia Martyn, Onobrychis viciifolia Scop., Lathyrus pratensis L.) and from three bare ground plots. Furthermore, we (2) tested if it is possible to apply a mixing model for NO₃⁻ in soil solution based on its dual isotope signals, and (3) estimated the contribution of legume mineralization to NO₃⁻ concentrations in soil solution under field conditions. We collected rainfall and soil solution at 0.3 m soil depth during 1 year, and determined NO₃⁻ concentrations and δ¹⁵N and δ¹⁸O of NO₃⁻ for >11.5 mg NO₃⁻-N l⁻¹. We incubated soil samples to assess potential N release by mineralization and determined δ¹⁵N and δ¹⁸O signals of NO₃⁻ derived from mineralization of non-leguminous and leguminous organic matter.Mean annual N export to >0.3 m soil depth was highest in bare ground plots (9.7 g NO₃⁻-N m⁻²; the SD reflects the spatial variation) followed by Medicago x varia monoculture (6.0 g NO₃⁻-N m⁻²). The O. viciifolia and L. pratensis monocultures had a much lower mean annual N export (0.5 and 0.3 g NO₃⁻-N m⁻²). The averaged NO₃⁻-N leaching during 70 days was not significantly different between field estimates and incubation for the Medicago x varia Martyn monoculture.The δ¹⁵N and δ¹⁸O values in NO₃⁻ of rainfall (δ¹⁵N: 3.3±0.8‰; δ¹⁸O: 30.8±4.7‰), mineralization of non-leguminous SOM (9.3±0.9‰; 6.7±0.8‰), and mineralization of leguminous SOM (1.5±0.6‰; 5.1±0.9‰) were markedly different. Applying a linear mixing model based on these three sources to δ¹⁵N and δ¹⁸O values in NO₃⁻ of soil solution during winter 2003, we calculated 18-41% to originate from rainfall, 38-57% from mineralization of non-leguminous SOM, and 18-40% from mineralization of leguminous SOM.Our results demonstrate that (1) even under legumes NO₃⁻-N leaching was reduced compared to bare ground, (2) the application of a three-end-member mixing model for NO₃⁻ based on its dual isotope signals produced plausible results and suggests that under particular circumstances such models can be used to estimate the contributions of different NO₃⁻ sources in soil solution, and (3) in the 2nd year after establishment of legumes, they contributed approximately one-fourth to NO₃⁻-N loss.     

Dynamics of carbon and nitrogen in an extreme alkaline saline soil: A review

The soil of the former lake Texcoco is an ‘extreme’ alkaline saline soil with pH > 10 and electrolytic conductivity (EC) > 150 dS m⁻¹. These conditions have created a unique environment. Application of wastewater sludge to Texcoco soil showed that large amounts of NH₄⁺ were immobilized, NO₃⁻ was reduced aerobically, NO₂⁻ was formed and the mineralization of the organic material in the sludge was inhibited. A series of experiments were initiated to study the processes that inhibited the decomposition of organic material and affected the dynamics of mineral N. The large EC and pH inhibited the decomposition of easily decomposable organic material such as glucose and maize, although cellulolytic activity was observed in soil with pH 9.8 and EC 32.7 dS m⁻¹. The high soil pH favoured NH₃ volatilization of approximately 50 mg N kg⁻¹ soil within a day and a similar amount could be fixed on the soil matrix due to the dispersed minerals and their volcanic origin. Soil microorganisms immobilized large amounts of NH₄⁺ within a day when glucose was added to soil in excess of what was required for metabolic activity. Removal of NO₃⁻ from soil amended with glucose was not inhibited by 100% O₂ and NH₄⁺ indicating that the contribution of denitrification and assimilatory reduction to the reduction of NO₃⁻ was minimal while the formation of NO₂⁻ was not inhibited by 0.1% acetylene, known to inhibit nitrification. Additionally, the reduction of NO₃⁻ in the glucose-amended alkaline saline Texcoco soil was followed by an increase in the amount of NH₄⁺, which could not be due to denitrification. It was concluded that the reduction of NO₃⁻ and the formation of NO₂⁻ and NH₄⁺ in the glucose-amended soil was a result of aerobic NO₃⁻ reduction. A phylogenetic analysis of the archaeal community in the soil of the former lake Texcoco showed that some of the clones identified were capable of reducing NO₃⁻ aerobically to NO₂⁻ when glucose was added. A study of the diversity of the bacterial dissimilatory and respiratory nitrate-reducing communities indicated that bacteria could have contributed to the process.     

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.     

Rapid shift from denitrification to nitrification in soil after biogas residue application as indicated by nitrous oxide isotopomers

Nitrous oxide (N₂O) is one of the major greenhouse gases emitted from soils, where it is mainly produced by nitrification and denitrification. It is well known that rates of N₂O release from soils are mainly determined by the availability of substrates and oxygen, but N₂O source apportioning, highly needed to advance N₂O mitigation strategies, still remains challenging. In this study, using an automated soil incubation system, the N₂O site preference, i.e. the intramolecular ¹⁵N distribution, was analyzed to evaluate the progression in N₂O source processes following organic soil amendment. Biogas fermentation residue (BGR; originating from food waste fermentation) was applied to repacked grassland soil cores and compared to ammonium sulfate (AS) application, both at rates equivalent to 160 kg NH₄⁺-N ha⁻¹, and to unamended soil (control). The soil cores were incubated in a helium-oxygen atmosphere with 20 kPa O₂ for 43 days at 80% water-filled pore space. 43-day cumulative N₂O emissions were highest with BGR treated soil accounting for about 1.68 kg N₂O-N ha⁻¹ while application of AS caused much lower fluxes of c. 0.23 kg N₂O-N ha⁻¹. Also, after BGR application, carbon dioxide (CO₂) fluxes showed a pronounced initial peak with steep decline until day 21 whereas with ammonium addition they remained at the background level. N₂O dual isotope and isotopomer analysis of gas samples collected from BGR treated soil indicated bacterial denitrification to be the main N₂O generating process during the first three weeks when high CO₂ fluxes signified high carbon availability. In contrast, in the second half after all added labile carbon substrates had been consumed, nitrification, i.e. the generation of N₂O via oxidation of hydroxylamine, gained in importance reaching roughly the same N₂O production rate compared to bacterial denitrification as indicated by N₂O SP. Overall in this study, bacterial denitrification seemed to be the main N₂O forming process after application of biogas residues and fluxes were mainly driven by available organic carbon.     

Nitrification activity in submerged soils and its relation to denitrification loss

Summary Nitrification activity (formation of NO ₂ ^– + NO ₃ ^– per unit soil weight) was measured in the surface layer of 15 presubmerged soils incubated in petri dishes under flooded but aerobic conditions. soils with pH above 5 nitrified quickly, whereas soils with pH below this level did not nitrify or nitrified slowly. The pH values between 7 and 8.5 were optimal for nitrification. Organic-matter levels in the 15 soils of our study did not influence their nitrification activities. In a follow-up greenhouse pot study, after a period of 3 weeks, ¹⁵N-balance measurements showed that the loss of N through apparent denitrification did not follow the nitrification patterns of the soils observed in the petri dishes. Apparent denitrification accounted for 16.8% and 18.9% loss of ¹⁵N from a soil with insignificant nitrification activity and a soil with high nitrification activity, respectively. These results, thus, indicate a lack of correspondence between the nitrification activities of soil and the denitrification loss of N when the former was measured in the dark and the latter was estimated in the light. Soils that nitrified in the darkness of the incubator did not nitrify in the daylight in the greenhouse.     

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.     

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

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

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

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

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.     

农业土壤中的氧化亚氮排放: 为减排综述时空变化

This short review deals with soils as an important source of the greenhouse gas N2O. The production and consumption of N2O in soils mainly involve biotic processes: the anaerobic process of denitrification and the aerobic process of nitrification. The factors that significantly influence agricultural N2O emissions mainly concern the agricultural practices (N application rate, crop type, fertilizer type) and soil conditions (soil moisture, soil organic C content, soil pH and texture). Large variability of N2O fluxes is known to occur both at different spatial and temporal scales. Currently new techniques could help to improve the capture of the spatial variability. Continuous measurement systems with automatic chambers could also help to capture temporal variability and consequently to improve quantification of N2O emissions by soils. Some attempts for mitigating soil N2O emissions, either by modifying agricultural practices or by managing soil microbial functioning taking into account the origin of the soil N2O emission variability, are reviewed.     

土壤温度、水分和NH4+-N浓度对土壤硝化反应速度及N2O排放的影响

硝化反应是土壤、特别是干旱半干旱地区农业土壤N2O产生的重要途径之一。但是,目前环境条件对硝化反应中N2O排放的影响研究较少,而在国内外通用的几个模型中均用固定比例估算硝化反应过程中N2O的排放。本文通过砂壤土培养试验,研究了土壤温度、水分和NH4+-N浓度对硝化反应速度及硝化反应中N2O排放的影响,并用数学模型定量表示了各因素对硝化反应的作用,用最小二乘法最优拟合求得该土壤的最大硝化反应速度及N2O最大排放比例。结果表明,随着温度升高,硝化反应速度呈指数增长;水分含量由20%充水孔隙度(WFPS)增加到40%WFPS时,反应速度增加,水分含量增加到60%WFPS时反应速度略有降低;NH4+-N浓度增加对硝化反应速度起抑制作用。用米氏方程描述该土壤的硝化反应过程,其最大硝化反应速度为6.67mg·kg?1·d?1。硝化反应中N2O排放比例随温度升高而降低;随NH4+-N浓度增加而略有增加;20%和40%WFPS水分含量时,硝化反应中N2O排放比例为0.43%～1.50%,最小二乘法求得的最大比例为3.03%,60%WFPS时可能由于反硝化作用,N2O排放比例急剧增加,还需进一步研究水分对硝化反应中N2O排放的影响。