Tracing nitrogen transformations during spring development of winter wheat induced by 15N labeled cattle slurry applied with different techniques.

Slurry application is often associated with considerable nitrogen (N) losses: ammonia (NH3), nitrous oxide (N2O) and a mostly unknown contribution of dinitrogen (N2) emission, as well as N leaching. Thus, an outdoor lysimeter experiment with growing winter wheat in undisturbed soil cores was set up to follow the transformation of cattle slurry 15NH4+ and soil 15NO3- using a double labeling approach. Slurry treatments included the following application techniques: a trailing hose with/without acidification, and open slot injection with/without nitrification inhibitor. The fertilizer application rate was 67 kg N ha-1. In addition to NH3 emissions, N2O and N2 emissions were measured, as well as N contents and 15N enrichment of soil N pools and plant compartments. The major gaseous loss pathway was NH3 with up to 8 kg N ha-1 following trailing hose application, while slot injection significantly reduced NH3-N losses. Regardless of the application technique, N2O emissions were low (up to 0.1 kg N2O-N ha-1), while N2 emissions reached up to 3 kg N ha-1. No effect on N leaching from topsoil was found. 15N plant uptake was greater in slot injection than trailing hose treatments. An effect of the nitrification inhibitor was visible in the nitrate contents, but not in gaseous N losses or N leaching from topsoil. Impacts of the application techniques on individual soil N pools were small. The 15N recovery offered a chance to map the short-term effects and was highest in the soil Nt pool (32 % to 48 % of 15N applied) with a greater contribution of microbial N than mineral N at beginning of stem elongation. Indications for high N immobilization was derived from the applied N balance approach. In the present case, slot injection scored as the best application technology based on the highest NH3 reduction, while N2 and N2O emissions were not enhanced.

Sources of nitrous oxide emissions from hydroponic tomato cultivation: Evidence from stable isotope analyses.

Introduction: Hydroponic vegetable cultivation is characterized by high intensity and frequent nitrogen fertilizer application, which is related to greenhouse gas emissions, especially in the form of nitrous oxide (N2O). So far, there is little knowledge about the sources of N2O emissions from hydroponic systems, with the few studies indicating that denitrification could play a major role. Methods: Here, we use evidence from an experiment with tomato plants (Solanum lycopersicum) grown in a hydroponic greenhouse setup to further shed light into the process of N2O production based on the N2O isotopocule method and the 15N tracing approach. Gas samples from the headspace of rock wool substrate were collected prior to and after 15N labeling at two occasions using the closed chamber method and analyzed by gas chromatography and stable isotope ratio mass spectrometry. Results: The isotopocule analyses revealed that either heterotrophic bacterial denitrification (bD) or nitrifier denitrification (nD) was the major source of N2O emissions, when a typical nutrient solution with a low ammonium concentration (1-6 mg L-1) was applied. Furthermore, the isotopic shift in 15N site preference and in δ18O values indicated that approximately 80-90% of the N2O produced were already reduced to N2 by denitrifiers inside the rock wool substrate. Despite higher concentrations of ammonium present during the 15N labeling (30-60 mg L-1), results from the 15N tracing approach showed that N2O mainly originated from bD. Both, 15N label supplied in the form of ammonium and 15N label supplied in the form of nitrate, increased the 15N enrichment of N2O. This pointed to the contribution of other processes than bD. Nitrification activity was indicated by the conversion of small amounts of 15N-labeled ammonium into nitrate. Discussion/Conclusion: Comparing the results from N2O isotopocule analyses and the 15N tracing approach, likely a combination of bD, nD, and coupled nitrification and denitrification (cND) was responsible for the vast part of N2O emissions observed in this study. Overall, our findings help to better understand the processes underlying N2O and N2 emissions from hydroponic tomato cultivation, and thereby facilitate the development of targeted N2O mitigation measures.

Nitrite isotope characteristics and associated soil N transformations.

Nitrite (NO2-) is a crucial compound in the N soil cycle. As an intermediate of nearly all N transformations, its isotopic signature may provide precious information on the active pathways and processes. NO2- analyses have already been applied in 15N tracing studies, increasing their interpretation perspectives. Natural abundance NO2- isotope studies in soils were so far not applied and this study aims at testing if such analyses are useful in tracing the soil N cycle. We conducted laboratory soil incubations with parallel natural abundance and 15N treatments, accompanied by isotopic analyses of soil N compounds (NO3-, NO2-, NH4+). The double 15N tracing method was used as a reference method for estimations of N transformation processes based on natural abundance nitrite dynamics. We obtained a very good agreement between the results from nitrite isotope model proposed here and the 15N tracing approach. Natural abundance nitrite isotope studies are a promising tool to our understanding of soil N cycling.

Nitrogen isotope analysis of aqueous ammonium and nitrate by membrane inlet isotope ratio mass spectrometry (MIRMS) at natural abundance levels.

RATIONALE: Existing methods for the measurement of the 15 N/14 N isotopic composition of ammonium and nitrate are either only suitable for labelled samples or require considerable sample preparation efforts (or both). Our goal was to modify an existing analytical approach to allow for natural abundance precision levels. METHODS: Published reaction protocols were used to convert ammonium into N2 by NaOBr and nitrate into N2 O by TiCl3 . A membrane inlet system was developed and coupled to an isotope ratio mass spectrometer to allow precise determination of the analytes. RESULTS: Concentrations of ≥35 µmol/L N for both ammonium or nitrate could be analysed for δ15 N values with precisions of better than 0.9 mUr. While ammonium analyses exhibited a small concentration dependency and an offset of 2.7 mUr at high ammonium concentrations irrespective of the standard isotopic composition, nitrate analysis showed no offset but a blank contribution visible at very low concentrations. CONCLUSIONS: The presented method is capable of fast measurement of δ15 N values in ammonium and nitrate from aqueous samples with reasonable accuracy at natural abundance levels. It will thus facilitate the application of isotopic methods to studies of nitrogen cycling in ecosystems.

Development and verification of a novel isotopic N2 O measurement technique for discrete static chamber samples using cavity ring-down spectroscopy.

RATIONALE: N2 O isotopomers are a useful tool to study soil N cycling processes. The reliability of such measurements requires a consistent set of international N2 O isotope reference materials to improve inter-laboratory and inter-instrument comparability and avoid reporting inaccurate results. All these are the more important given the role of N2 O in anthropogenic climate change and the pressing need to develop our understanding of soil N cycling and N2 O emission to mitigate such emissions. Cavity ring-down spectroscopy (CRDS) could potentially overcome resource requirements and technical challenges, making N2 O isotopomer measurements more feasible and less expensive than previous approaches (e.g., gas chromatography [GC] and isotope ratio mass spectrometry [IRMS]). METHODS: A combined laser spectrometer and small sample isotope module (CRDS & SSIM) method enabled N2 O concentration, δ15 Nbulk , δ15 Nα , δ15 Nß and site preference (SP) measurements of sample volumes <20 mL, such as static chamber samples. Sample dilution and isotopic mixing as well as N2 O concentration dependence were corrected numerically. A two-point calibration procedure normalised δ values to the international isotope-ratio scales. The CRDS & SSIM repeatability was determined using a reference gas (Ref Gas). CRDS & SSIM concentration measurements were compared with those obtained by GC, and the isotope ratio measurements from two different mass spectrometers were compared. RESULTS: The repeatability (mean ± 1σ; n = 10) of the CRDS & SSIM measurements of the Ref Gas was 710.64 ppb (± 8.64), 2.82‰ (± 0.91), 5.41‰ (± 2.00), 0.23‰ (± 0.22) and 5.18‰ (± 2.18) for N2 O concentration, δ15 Nbulk , δ15 Nα , δ15 Nß and SP, respectively. The CRDS & SSIM concentration measurements were strongly correlated with GC (r = 0.99), and they were more precise than those obtained using GC except when the N2 O concentrations exceeded the specified operating range. Normalising CRDS & SSIM δ values to the international isotope-ratio scales using isotopic N2 O standards (AK1 and Mix1) produced accurate results when the samples were bracketed within the range of the δ values of the standards. The CRDS & SSIM δ15 Nbulk and SP precision was approximately one order of magnitude less than the typical IRMS precision. CONCLUSIONS: CRDS & SSIM is a promising approach that enables N2 O concentrations and isotope ratios to be measured by CRDS for samples <20 mL. The CRDS & SSIM repeatability makes this approach suitable for N2 O "isotopomer mapping" to distinguish dominant source pathways, such as nitrification and denitrification, and requires less extensive lab resources than the traditionally used GC/IRMS. Current study limitations highlighted potential improvements for future users of this approach to consider, such as automation and physical removal of interfering trace gases before sample analysis.

Biologically mediated release of endogenous N2O and NO2 gases in a hydrothermal, hypoxic subterranean environment.

The migration of geogenic gases in continental areas with geothermal activity and active faults is an important process releasing greenhouse gases (GHG) to the lower troposphere. In this respect, caves in hypogenic environments are natural laboratories to study the compositional evolution of deep-endogenous fluids through the Critical Zone. Vapour Cave (Alhama, Murcia, Spain) is a hypogenic cave formed by the upwelling of hydrothermal CO2-rich fluids. Anomalous concentrations of N2O and NO2 were registered in the cave's subterranean atmosphere, averaging ten and five times the typical atmospheric backgrounds, respectively. We characterised the thermal conditions, gaseous compositions, sediments, and microbial communities at different depths in the cave. We did so to understand the relation between N-cycling microbial groups and the production and transformation of nitrogenous gases, as well as their coupled evolution with CO2 and CH4 during their migration through the Critical Zone to the lower troposphere. Our results showed an evident vertical stratification of selected microbial groups (Archaea and Bacteria) depending on the environmental parameters, including O2, temperature, and GHG concentration. Both the N2O isotope ratios and the predicted ecological functions of bacterial and archaeal communities suggest that N2O and NO2 emissions mainly depend on the nitrification by ammonia-oxidising microorganisms. Denitrification and abiotic reactions of the reactive intermediates NH2OH, NO, and NO2- are also plausible according to the results of the phylogenetic analyses of the microbial communities. Nitrite-dependent anaerobic methane oxidation by denitrifying methanotrophs of the NC10 phylum was also identified as a post-genetic process during migration of this gas to the surface. To the best of our knowledge, our report provides, for the first time, evidence of a niche densely populated by Micrarchaeia, which represents more than 50% of the total archaeal abundance. This raises many questions on the metabolic behaviour of this and other archaeal phyla.

Quantifying N2O reduction to N2 during denitrification in soils via isotopic mapping approach: Model evaluation and uncertainty analysis.

The last step of denitrification, i.e. the reduction of N2O to N2, has been intensively studied in the laboratory to understand the denitrification process, predict nitrogen fertiliser losses, and to establish mitigation strategies for N2O. However, assessing N2 production via denitrification at large spatial scales is still not possible due to lack of reliable quantitative approaches. Here, we present a novel numerical "mapping approach" model using the δ15Nsp/δ18O slope that has been proposed to potentially be used to indirectly quantify N2O reduction to N2 at field or larger spatial scales. We evaluate the model using data obtained from seven independent soil incubation studies conducted under a He-O2 atmosphere. Furthermore, we analyse the contribution of different parameters to the uncertainty of the model. The model performance strongly differed between studies and incubation conditions. Re-evaluation of the previous data set demonstrated that using soils-specific instead of default endmember values could largely improve model performance. Since the uncertainty of modelled N2O reduction was relatively high, further improvements to estimate model parameters to obtain more precise estimations remain an on-going matter, e.g. by determination of soil-specific isotope fractionation factors and isotopocule endmember values of N2O production processes using controlled laboratory incubations. The applicability of the mapping approach model is promising with an increasing availability of real-time and field based analysis of N2O isotope signatures.

Indications for enzymatic denitrification to N2O at low pH in an ammonia-oxidizing archaeon.

Nitrous oxide (N2O) is a key climate change gas and nitrifying microbes living in terrestrial ecosystems contribute significantly to its formation. Many soils are acidic and global change will cause acidification of aquatic and terrestrial ecosystems, but the effect of decreasing pH on N2O formation by nitrifiers is poorly understood. Here, we used isotope-ratio mass spectrometry to investigate the effect of acidification on production of N2O by pure cultures of two ammonia-oxidizing archaea (AOA; Nitrosocosmicus oleophilus and Nitrosotenuis chungbukensis) and an ammonia-oxidizing bacterium (AOB; Nitrosomonas europaea). For all three strains acidification led to increased emission of N2O. However, changes of 15N site preference (SP) values within the N2O molecule (as indicators of pathways for N2O formation), caused by decreasing pH, were highly different between the tested AOA and AOB. While acidification decreased the SP value in the AOB strain, SP values increased to a maximum value of 29‰ in N. oleophilus. In addition, 15N-nitrite tracer experiments showed that acidification boosted nitrite transformation into N2O in all strains, but the incorporation rate was different for each ammonia oxidizer. Unexpectedly, for N. oleophilus more than 50% of the N2O produced at pH 5.5 had both nitrogen atoms from nitrite and we demonstrated that under these conditions expression of a putative cytochrome P450 NO reductase is strongly upregulated. Collectively, our results indicate that N. oleophilus might be able to enzymatically denitrify nitrite to N2O at low pH.

Improvement of the 15 N gas flux method for in situ measurement of soil denitrification and its product stoichiometry.

RATIONALE: Field measurement of denitrification in agricultural ecosystems using the 15 N gas flux method has been limited by poor sensitivity because current isotope ratio mass spectrometry is not precise enough to detect low 15 N2 fluxes in the presence of a high atmospheric N2 background. For laboratory studies, detection limits are improved by incubating soils in closed systems and under N2 -depleted atmospheres. METHODS: We developed a new procedure to conduct the 15 N gas flux method suitable for field application using an artificially N2 -depleted atmosphere to improve the detection limit at the given precision of mass spectrometry. Laboratory experiments with and without 15 N-labelling and using different flushing strategies were conducted to develop a suitable field method. Subsequently, this method was tested in the field and results were compared with those obtained from the conventional 15 N gas flux method. RESULTS: Results of the two methods were in close agreement showing that the denitrification rates determined were not biased by the flushing procedure. Best sensitivity for N2 + N2 O fluxes was 10 ppb, which was 80-fold better than that of the reference method. Further improvement can be achieved by lowering the N2 background concentration below the values established in the present study. CONCLUSIONS: In view of this progress in sensitivity, the new method will be suitable to measure denitrification dynamics in the field beyond peak events.

Improved isotopic model based on 15 N tracing and Rayleigh-type isotope fractionation for simulating differential sources of N2 O emissions in a clay grassland soil.

RATIONALE: Isotopic signatures of N2 O can help distinguish between two sources (fertiliser N or endogenous soil N) of N2 O emissions. The contribution of each source to N2 O emissions after N-application is difficult to determine. Here, isotopologue signatures of emitted N2 O are used in an improved isotopic model based on Rayleigh-type equations. METHODS: The effects of a partial (33% of surface area, treatment 1c) or total (100% of surface area, treatment 3c) dispersal of N and C on gaseous emissions from denitrification were measured in a laboratory incubation system (DENIS) allowing simultaneous measurements of NO, N2 O, N2 and CO2 over a 12-day incubation period. To determine the source of N2 O emissions those results were combined with both the isotope ratio mass spectrometry analysis of the isotopocules of emitted N2 O and those from the 15 N-tracing technique. RESULTS: The spatial dispersal of N and C significantly affected the quantity, but not the timing, of gas fluxes. Cumulative emissions are larger for treatment 3c than treatment 1c. The 15 N-enrichment analysis shows that initially ~70% of the emitted N2 O derived from the applied amendment followed by a constant decrease. The decrease in contribution of the fertiliser N-pool after an initial increase is sooner and larger for treatment 1c. The Rayleigh-type model applied to N2 O isotopocules data (δ15 Nbulk -N2 O values) shows poor agreement with the measurements for the original one-pool model for treatment 1c; the two-pool models gives better results when using a third-order polynomial equation. In contrast, in treatment 3c little difference is observed between the two modelling approaches. CONCLUSIONS: The importance of N2 O emissions from different N-pools in soil for the interpretation of N2 O isotopocules data was demonstrated using a Rayleigh-type model. Earlier statements concerning exponential increase in native soil nitrate pool activity highlighted in previous studies should be replaced with a polynomial increase with dependency on both N-pool sizes.

Nitrous oxide effluxes from plants as a potentially important source to the atmosphere.

The global budget for nitrous oxide (N2 O), an important greenhouse gas and probably dominant ozone-depleting substance emitted in the 21st century, is far from being fully understood. Cycling of N2 O in terrestrial ecosystems has traditionally exclusively focused on gas exchange between the soil surface (nitrification-denitrification processes) and the atmosphere. Terrestrial vegetation has not been considered in the global budget so far, even though plants are known to release N2 O. Here, we report the N2 O emission rates of 32 plant species from 22 different families measured under controlled laboratory conditions. Furthermore, the first isotopocule values (δ15 N, δ18 O and δ15 Nsp ) of N2 O emitted from plants were determined. A robust relationship established between N2 O emission and CO2 respiration rates, which did not alter significantly over a broad range of changing environmental conditions, was used to quantify plant-derived emissions on an ecosystem scale. Stable isotope measurements (δ15 N, δ18 O and δ15 Nsp ) of N2 O emitted by plants clearly show that the dual isotopocule fingerprint of plant-derived N2 O differs from that of currently known microbial or chemical processes. Our work suggests that vegetation is a natural source of N2 O in the environment with a large fraction released by a hitherto unrecognized process.

NO Reduction to N2O Improves Nitrate 15N Abundance Analysis by Membrane Inlet Quadrupole Mass Spectrometry.

Earlier an automated sample preparation unit for inorganic nitrogen (SPIN) coupled to a membrane inlet quadrupole mass spectrometer (MIMS) was developed for automated and sensitive determination of the 15N abundances and concentrations of nitrate, nitrite and ammonium of aqueous solutions without any sample preparation. Here we describe an alternative analytical protocol to convert NO3- to N2O instead of NO before measurement. This is advantageous because NO strongly interacts with surfaces, requires long purge times, and still shows considerable carryover between samples, all of which is avoided when N2O is used as analyte. The sensitivity of the measurement of NO3- as N2O is comparable to the earlier measurements with NO as analyte.

Preliminary assessment of stable nitrogen and oxygen isotopic composition of USGS51 and USGS52 nitrous oxide reference gases and perspectives on calibration needs.

RATIONALE: Despite a long history and growing interest in isotopic analyses of N2 O, there is a lack of isotopically characterized N2 O isotopic reference materials (standards) to enable normalization and reporting of isotope-delta values. Here we report the isotopic characterization of two pure N2 O gas reference materials, USGS51 and USGS52, which are now available for laboratory calibration (https://isotopes.usgs.gov/lab/referencematerials.html). METHODS: A total of 400 sealed borosilicate glass tubes of each N2 O reference gas were prepared from a single gas filling of a high vacuum line. We demonstrated isotopic homogeneity via dual-inlet isotope-ratio mass spectrometry. Isotopic analyses of these reference materials were obtained from eight laboratories to evaluate interlaboratory variation and provide preliminary isotopic characterization of their δ15 N, δ18 O, δ15 Nα , δ15 Nß and site preference (SP ) values. RESULTS: The isotopic homogeneity of both USGS51 and USGS52 was demonstrated by one-sigma standard deviations associated with the determinations of their δ15 N, δ18 O, δ15 Nα , δ15 Nß and SP values of 0.12 mUr or better. The one-sigma standard deviations of SP measurements of USGS51 and USGS52 reported by eight laboratories participating in the interlaboratory comparison were 1.27 and 1.78 mUr, respectively. CONCLUSIONS: The agreement of isotope-delta values obtained in the interlaboratory comparison was not sufficient to provide reliable accurate isotope measurement values for USGS51 and USGS52. We propose that provisional values for the isotopic composition of USGS51 and USGS52 determined at the Tokyo Institute of Technology can be adopted for normalizing and reporting sample data until further refinements are achieved through additional calibration efforts.

Estimating N2 O processes during grassland renewal and grassland conversion to maize cropping using N2 O isotopocules.

RATIONALE: Enhanced nitrous oxide (N2 O) emissions can occur following grassland break-up for renewal or conversion to maize cropping, but knowledge about N2 O production pathways and N2 O reduction to N2 is very limited. A promising tool to address this is the combination of mass spectrometric analysis of N2 O isotopocules and an enhanced approach for data interpretation. METHODS: The isotopocule mapping approach was applied to field data using a δ15 NspN2O and δ18 ON2O map to simultaneously determine N2 O production pathways contribution and N2 O reduction for the first time. Based on the isotopic composition of N2 O produced and literature values for specific N2 O pathways, it was possible to distinguish: (i) heterotrophic bacterial denitrification and/or nitrifier denitrification and (ii) nitrification and/or fungal denitrification and the contribution of N2 O reduction. RESULTS: The isotopic composition of soil-emitted N2 O largely resembled the known end-member values for bacterial denitrification. The isotopocule mapping approach indicated different effects of N2 O reduction on the isotopic composition of soil-emitted N2 O for the two soils under study. Differing N2 O production pathways in different seasons were not observed, but management events and soil conditions had a significant impact on pathway contribution and N2 O reduction. N2 O reduction data were compared with a parallel 15 N-labelling experiment. CONCLUSIONS: The field application of the isotopocule mapping approach opens up new prospects for studying N2 O production and consumption of N2 O in soil simultaneously based on mass spectrometric analysis of natural abundance N2 O. However, further studies are needed in order to properly validate the isotopocule mapping approach.

Stability of buried carbon in deep-ploughed forest and cropland soils - implications for carbon stocks.

Accumulation of soil organic carbon (SOC) may play a key role in climate change mitigation and adaptation. In particular, subsoil provides a great potential for additional SOC storage due to the assumed higher stability of subsoil SOC. The fastest way in which SOC reaches the subsoil is via burial, e.g. via erosion or deep ploughing. We assessed the effect of active SOC burial through deep ploughing on long-term SOC stocks and stability in forest and cropland subsoil. After 25-48 years, deep-ploughed subsoil contained significantly more SOC than reference subsoils, in both forest soil (+48%) and cropland (+67%). However, total SOC stocks down to 100 cm in deep-ploughed soil were greater than in reference soil only in cropland, and not in forests. This was explained by slower SOC accumulation in topsoil of deep-ploughed forest soils. Buried SOC was on average 32% more stable than reference SOC, as revealed by long-term incubation. Moreover, buried subsoil SOC had higher apparent radiocarbon ages indicating that it is largely isolated from exchange with atmospheric CO2. We concluded that deep ploughing increased subsoil SOC storage and that the higher subsoil SOC stability is not only a result of selective preservation of more stable SOC fractions.

Measuring 15N Abundance and Concentration of Aqueous Nitrate, Nitrite, and Ammonium by Membrane Inlet Quadrupole Mass Spectrometry.

An automated sample preparation unit for inorganic nitrogen (SPIN) coupled to a membrane inlet quadrupole mass spectrometer (MIMS) was developed for automated and sensitive determination of the 15N abundances and concentrations of nitrate, nitrite, and ammonium in aqueous solutions without any sample preparation. The minimum N concentration for an accurate determination of the 15N abundance is 7 µmol/L for nitrite and nitrate, with a relative standard deviation (RSD) of repeated measurements of <1%, and 70 µmol/L with an RSD < 0.4% in the case of ammonium. The SPIN-MIMS system provides a wide dynamic range (up to 3500 µmol/L) for all three N species for both isotope abundance and concentration measurements. The comparison of parallel measurements of 15N-labeled NH4+ and NO3- from soil extracts with the denitrifier method and the SPIN-MIMS system shows a good agreement between both methods.

Use of oxygen isotopes to differentiate between nitrous oxide produced by fungi or bacteria during denitrification.

RATIONALE: Fungal denitrifiers can contribute substantially to N2 O emissions from arable soil and show a distinct site preference for N2 O (SP(N2 O)). This study sought to identify another process-specific isotopic tool to improve precise identification of N2 O of fungal origin by mass spectrometric analysis of the N2 O produced. METHODS: Three pure bacterial and three fungal species were incubated under denitrifying conditions in treatments with natural abundance and stable isotope labelling to analyse the N2 O produced. Combining different applications of isotope ratio mass spectrometry enabled us to estimate the oxygen (O) exchange accelerated by denitrifying enzymes and the ongoing microbial pathway in parallel. This experimental set-up allowed the determination of δ18 O(N2 O) values and isotopic fractionation of O, as well as SP(N2 O) values, as a perspective to differentiate between microbial denitrifiers. RESULTS: Oxygen exchange during N2 O production was lower for bacteria than for fungi, differed between species, and depended also on incubation time. Apparent O isotopic fractionation during denitrification was in a similar range for bacteria and fungi, but application of the fractionation model indicated that different enzymes in bacteria and fungi were responsible for O exchange. This difference was associated with different isotopic fractionation for bacteria and fungi. CONCLUSIONS: δ18 O(N2 O) values depend on isotopic fractionation and isotopic fractionation may differ between processes and organism groups. By comparing SP(N2 O) values, O exchange and the isotopic signature of precursors, we propose here a novel tool for differentiating between different sources of N2 O.

Gas entrapment and microbial N2O reduction reduce N2O emissions from a biochar-amended sandy clay loam soil.

Nitrous oxide (N2O) is a potent greenhouse gas that is produced during microbial nitrogen transformation processes such as nitrification and denitrification. Soils represent the largest sources of N2O emissions with nitrogen fertilizer application being the main driver of rising atmospheric N2O concentrations. Soil biochar amendment has been proposed as a promising tool to mitigate N2O emissions from soils. However, the underlying processes that cause N2O emission suppression in biochar-amended soils are still poorly understood. We set up microcosm experiments with fertilized, wet soil in which we used 15N tracing techniques and quantitative polymerase chain reaction (qPCR) to investigate the impact of biochar on mineral and gaseous nitrogen dynamics and denitrification-specific functional marker gene abundance and expression. In accordance with previous studies our results showed that biochar addition can lead to a significant decrease in N2O emissions. Furthermore, we determined significantly higher quantities of soil-entrapped N2O and N2 in biochar microcosms and a biochar-induced increase in typical and atypical nosZ transcript copy numbers. Our findings suggest that biochar-induced N2O emission mitigation is based on the entrapment of N2O in water-saturated pores of the soil matrix and concurrent stimulation of microbial N2O reduction resulting in an overall decrease of the N2O/(N2O + N2) ratio.