Explaining the doubling of N2 O emissions under elevated CO2 in the Giessen FACE via in-field 15 N tracing.

Rising atmospheric CO2 concentrations are expected to increase nitrous oxide (N2 O) emissions from soils via changes in microbial nitrogen (N) transformations. Several studies have shown that N2 O emission increases under elevated atmospheric CO2 (eCO2 ), but the underlying processes are not yet fully understood. Here, we present results showing changes in soil N transformation dynamics from the Giessen Free Air CO2 Enrichment (GiFACE): a permanent grassland that has been exposed to eCO2 , +20% relative to ambient concentrations (aCO2 ), for 15 years. We applied in the field an ammonium-nitrate fertilizer solution, in which either ammonium ( NH4+ ) or nitrate ( NO3- ) was labelled with 15 N. The simultaneous gross N transformation rates were analysed with a 15 N tracing model and a solver method. The results confirmed that after 15 years of eCO2 the N2 O emissions under eCO2 were still more than twofold higher than under aCO2 . The tracing model results indicated that plant uptake of NH4+ did not differ between treatments, but uptake of NO3- was significantly reduced under eCO2 . However, the NH4+ and NO3- availability increased slightly under eCO2 . The N2 O isotopic signature indicated that under eCO2 the sources of the additional emissions, 8,407 µg N2 O-N/m2 during the first 58 days after labelling, were associated with NO3- reduction (+2.0%), NH4+ oxidation (+11.1%) and organic N oxidation (+86.9%). We presume that increased plant growth and root exudation under eCO2 provided an additional source of bioavailable supply of energy that triggered as a priming effect the stimulation of microbial soil organic matter (SOM) mineralization and fostered the activity of the bacterial nitrite reductase. The resulting increase in incomplete denitrification and therefore an increased N2 O:N2 emission ratio, explains the doubling of N2 O emissions. If this occurs over a wide area of grasslands in the future, this positive feedback reaction may significantly accelerate climate change.

Nitrous Oxide Fluxes, Soil Oxygen, and Denitrification Potential of Urine- and Non-Urine-Treated Soil under Different Irrigation Frequencies.

Despite increased use of irrigation to improve forage quality and quantity for grazing cattle ( Linnaeus), there is a lack of data that assess how irrigation practices influence nitrous oxide (NO) emissions from urine-affected soils. Irrigation effects on soil oxygen (O) availability, a primary controller of NO fluxes, is poorly understood. It was hypothesized that increased irrigation frequency would result in lower NO emissions by increasing soil moisture and decreasing soil O concentrations. This would favor more NO reduction to dinitrogen (N). We examined effects of high (3-d) versus low (6-d) irrigation frequency with and without bovine urine addition to pasture. Nitrous oxide fluxes were measured daily for 35 d. Soil O, temperature, and water content were continuously measured at multiple depths. Inorganic nitrogen, organic carbon, and soil pH were measured at 6-d intervals. Measurements of denitrification enzyme activity with and without acetylene inhibition were used to infer the NO/(NO + N) ratio. The NO/(NO + N) ratio was lower under high- compared with low-frequency irrigation, suggesting greater potential for NO reduction to N with more frequent irrigation. Although NO fluxes were increased by urine addition, they were not affected by irrigation frequency. Soil O decreased temporarily after urine deposition, but O dynamics did not explain NO dynamics. Relative soil gas diffusivity (/) was a better predictor of NO fluxes than O concentration. On a free-draining soil, increasing irrigation frequency while providing the same total water volume did not enhance NO emissions under ruminant urine patches in a grazed pasture.

Nitrous oxide emissions from in situ deposition of N-labeled ryegrass litter in a pasture soil.

During pasture grazing, freshly harvested herbage (litterfall) is dropped onto soils from the mouths of dairy cattle, potentially inducing nitrous oxide (NO) emissions. Although the Intergovernmental Panel on Climate Change (IPCC) recommends accounting for NO emissions from arable crop residues in national inventories, emissions from the litterfall of grazed pasture systems are not recognized. The objective of this study was to investigate the potential of litterfall to contribute to NO emissions in a field study located on a pasture site in Canterbury, New Zealand (43°38.50' S, 172°27.17' E). We applied N-labeled perennial ryegrass ( L.) to the surface of a pastoral soil (Temuka clay loam) and, for up to 139 d thereafter, quantified the contribution of herbage decomposition to NO production and soil N dynamics. Litterfall contributed to the N enrichment of soil NO-N and NO-N pools. After 49 d, N recovery as NO equated to 0.93% of the surface-applied litter N, with 38 to 75% of the cumulative NO flux occurring within 4 to 10 d of treatment application. Emissions of NO likely resulted from ammonification followed by a coupling of nitrification and denitrification during litter decomposition on the soil surface. The emission factor of the litter deposited in situ was 1.2 ± 0.2%, which is not substantially greater than the IPCC default emission factor value of 1% for crop residues. Further in situ studies using different pasture species and litterfall rates are required to understand the microbial processes responsible for litter-induced NO emissions.

Nitrate as an alternative electron acceptor destabilizes the mineral associated organic carbon in moisturized deep soil depths.

Numerous studies have investigated the effects of nitrogen (N) addition on soil organic carbon (SOC) decomposition. However, most studies have focused on the shallow top soils <0.2 m (surface soil), with a few studies also examining the deeper soil depths of 0.5-1.0 m (subsoil). Studies investigating the effects of N addition on SOC decomposition in soil >1.0 m deep (deep soil) are rare. Here, we investigated the effects and the underlying mechanisms of nitrate addition on SOC stability in soil depths deeper than 1.0 m. The results showed that nitrate addition promoted deep soil respiration if the stoichiometric mole ratio of nitrate to O2 exceeded the threshold of 6:1, at which nitrate can be used as an alternative acceptor to O2 for microbial respiration. In addition, the mole ratio of the produced CO2 to N2O was 2.57:1, which is close to the theoretical ratio of 2:1 expected when nitrate is used as an electron acceptor for microbial respiration. These results demonstrated that nitrate, as an alternative acceptor to O2, promoted microbial carbon decomposition in deep soil. Furthermore, our results showed that nitrate addition increased the abundance of SOC decomposers and the expressions of their functional genes, and concurrently decreased MAOC, and the ratio of MAOC/SOC decreased from 20% before incubation to 4% at the end of incubation. Thus, nitrate can destabilize the MAOC in deep soils by stimulating microbial utilization of MAOC. Our results imply a new mechanism on how above-ground anthropogenic N inputs affect MAOC stability in deep soil. Mitigation of nitrate leaching is expected to benefit the conservation of MAOC in deep soil depths.

Intensive cattle grazing affects pasture litter-fall: an unrecognized nitrous oxide source.

The rationale for this study came from observing grazing dairy cattle dropping freshly harvested plant material onto the soil surface, hereafter called litter-fall. The Intergovernmental Panel on Climate Change (IPCC) guidelines include NO emissions during pasture renewal but do not consider NO emissions that may result from litter-fall. The objectives of this study were to determine litter-fall rates and to assess indicative NO emission factors (EFs) for the dominant pasture species (perennial ryegrass [ L.] and white clover [ L.]). Herbage was vacuumed from intensively managed dairy pastures before and after 30 different grazing events when cows (84 cows ha) grazed for 24 h according to a rotational system; the interval between grazing events ranged from 21 to 30 d. A laboratory incubation study was performed to assess potential EF values for the pasture species at two soil moisture contents. Finely ground pasture material was incubated under controlled laboratory conditions with soil, and the NO emissions were measured until rates returned to control levels. On average, pre- and postgrazing dry matter yields per grazing event were 2516 ± 636 and 1167 ± 265 kg DM ha (±SD), respectively. Pregrazing litter was absent, whereas postgrazing fresh and senesced litter-fall rates were 53 ± 24 and 19 ± 18 kg DM ha, respectively. Annually, the rotational grazing system resulted in 12 grazing events where fresh litter-fall equaed to 16 kg N ha yr to the soil. Emission factors in the laboratory experiment indicated that the EF for perennial ryegrass and white clover ranged from 0.7 to 3.1%. If such EF values should also occur under field conditions, then we estimate that litter-fall induces an NO emission rate of 0.3 kg NO ha yr. Litter-fall as a source of NO in grazed pastures requires further assessment.

Biochar incorporation into pasture soil suppresses in situ nitrous oxide emissions from ruminant urine patches.

Nitrous oxide (N2O) emissions from grazing animal excreta are estimated to be responsible for 1.5 Tg of the total 6.7 Tg of anthropogenic N2O emissions. This study was conducted to determine the in situ effect of incorporating biochar, into soil, on N2O emissions from bovine urine patches and associated pasture uptake of N. The effects of biochar rate (0-30 t ha(-1)), following soil incorporation, were investigated on ruminant urine-derived N2O fluxes, N uptake by pasture, and pasture yield. During an 86-d spring-summer period, where irrigation and rainfall occurred, the N2O fluxes from 15N labeled ruminant urine patches were reduced by >50%, after incorporating 30 t ha(-1) of biochar. Taking into account the N2O emissions from the control plots, 30 t ha(-1) ofbiochar reduced the N2O emission factor from urine by 70%. The atom% 15N enrichment of the N2O emitted was lower in the 30 t ha(-1) biochar treatment, indicating less urine-N contributed to the N2O flux. Soil NO3- -N concentrations were lower with increasing biochar rate during the first 30 d following urine deposition. No differences occurred, due to biochar addition, with respect to dry matter yields, herbage N content, or recovery of 15N applied in herbage. Incorporating biochar into the soil can significantly diminish ruminant urine-derived N2O emissions. Further work is required to determine the persistence of the observed effect and to fully understand the mechanism(s) of the observed reduction in N2O fluxes.

Biochar and the nitrogen cycle: introduction.

Nitrogen (N) is an essential nutrient, and research to date shows that biochar potentially has the ability to manipulate the rates of N cycling in soil systems by influencing nitrification rates and adsorption of ammonia and increasing NH4+ storage by enhancing cation exchange capacity in soils. Its influence on these processes may have further implications in terms of reducing gaseous N losses such as N2O and nitrate leaching. However, further detailed research is required to fully understand the transformation mechanisms and fate of N when associated with biochar treated soils. The three research papers that comprise this special collection of papers on biochar and the nitrogen cycle focus on biochar's diverse ability to influence N cycling processes. These papers show for the first time (i) how microbial nitrification communities and function differ with exposure to biochar, (ii) how the length of time the soil has been in contact with biochar influences N transformation and how this can vary with soil type, and (iii) how composting of organic materials with biochar can reduce N losses and enhance the nutrient status of the composted product. Considerable knowledge gaps still exist in terms of understanding the precise mechanisms through which biochar influences soil N transformations, and how biochar affects both plant and microbial N supply. The general direction that research on biochar should focus on with respect to the N cycle is the effect(s) that biochar has on N transformation in soils, both chemical and biological mechanisms, and the fate of N applied to biochar treated soils. This research needs to be performed at both field plot and microbial scales.

Application methods of tracers for N2O source determination lead to inhomogeneous distribution in field plots.

Source determination of N2O has often been performed using stable isotope incubation experiments. In situ experiments with isotopic tracers are an important next step. However, the challenge is to distribute the tracers in the field as homogeneously as possible. To examine this, a bromide solution was applied as a stand-in tracer using either a watering can, a sprayer, or syringes to a relatively dry (25% gravimetric moisture content) or wet (30%) silt loam. After 1 h, samples were taken from three soil depths (0-10 cm), and analyzed for their water content and bromide concentration. The application with syringes was unsuccessful due to blocked cannulas. Therefore, further laboratory experiments were conducted with side-port cannulas. Despite a larger calculated gravimetric soil moisture difference with watering can application, more Br- tracer was recovered in the sprayer treatment, probably due to faster transport of Br- through macropore flow in the wetter conditions caused by the watering can treatment. The losses of Br- (33% for the watering can, 28% for the sprayer treatment) are equivalent to potential losses of isotopic tracer solutions. For application of 60 at% 15NH4 +, this resulted in theoretical enrichments of 44-53 at% in the upper 2.5 cm and 7-48 at% in 5-10 cm. As there was hardly any NO3 - in the soil, extrapolations for 15NO3 - calculated enrichments were 57-59 at% in the upper 2.5 cm and 26-57 at% in 5-10 cm. Overall, no method, including the side-port cannulas, was able to achieve a homogeneous distribution of the tracer. Future search for optimal tracer application should therefore investigate methods that utilize capillary forces and avoid overhead pressure. We recommend working on rather dry soil when applying tracers, as tracer recovery was larger here. Furthermore, larger amounts of tracer lead to more uniform distributions. Further studies should also investigate the importance of plant surfaces.

Global Research Alliance N2 O chamber methodology guidelines: Introduction, with health and safety considerations.

Non-steady-state (NSS) chamber techniques have been used for decades to measure nitrous oxide (N2 O) fluxes from agricultural soils. These techniques are widely used because they are relatively inexpensive, easy to adopt, versatile, and adaptable to varying conditions. Much of our current understanding of the drivers of N2 O emissions is based on studies using NSS chambers. These chamber techniques require decisions regarding multiple methodological aspects (e.g., chamber materials and geometry, deployment, sample analysis, and data and statistical analysis), each of which may significantly affect the results. Variation in methodological details can lead to challenges in comparing results between studies and assessment of reliability and uncertainty. Therefore, the New Zealand Government, in support of the objectives of the Livestock Research Group of the Global Research Alliance on Agricultural Greenhouse Gases (GRA), funded two international projects to, first, develop standardized guidelines on the use of NSS chamber techniques and, second, refine them based on the most up to date knowledge and methods. This introductory paper summarizes a collection of papers that represent the revised guidelines. Each article summarizes existing knowledge and provides guidance and minimum requirements on chamber design, deployment, sample collection, storage and analysis, automated chambers, flux calculations, statistical analysis, emission factor estimation and data reporting, modeling, and "gap-filling" approaches. The minimum requirements are not meant to be highly prescriptive but instead provide researchers with clear direction on best practices and factors that need to be considered. Health and safety considerations of NSS chamber techniques are also provided with this introductory paper.

Effects of denitrification and transport on the isotopic composition of nitrate (δ18O, δ15N) in freshwater systems.

Nitrate isotopes (δ15N-NO3- and δ18O-NO3-) are a potentially powerful tool for tracking the biological removal of reactive nitrogen (N) as it is transported from land to sea. However, uncertainties about, 1) the variability of the strength of biological isotopic fractionation during anaerobic benthic NO3- reduction (the kinetic enrichment factor: εdenit), and, 2) how accurately these εdenit values are expressed in overlying aerobic surface waters (the effective enrichment factor: εeff), currently limit their use in freshwater systems. Here we used a combination of incubation experiments and numerical modelling to construct a simple framework for defining freshwater εdenit based on interactions between benthic denitrification and diffusive transport to surface waters. Under non-limited, anaerobic conditions the εdenit values produced in submerged soils (n = 3) and sediments (n = 4) with denitrification rates between 10 and 600 mg N m-2 d-1 ranged from -3‰ to -28‰. Critically, model results indicated that diffusive transport would homogenise this to an effective fractionation range of -6 ±â€¯4‰. Evidence for biological and hydrological variability of NO3- isotope fractionation means that values measured in aerobic surface water environments are most appropriately evaluated by a range of fractionation values, rather than commonly used single 'site specific' εdenit values.

Influence of copper on expression of nirS, norB and nosZ and the transcription and activity of NIR, NOR and N2 OR in the denitrifying soil bacteria Pseudomonas stutzeri.

Reduction of the potent greenhouse gas nitrous oxide (N(2)O) occurs in soil environments by the action of denitrifying bacteria possessing nitrous oxide reductase (N(2)OR), a dimeric copper (Cu)-dependent enzyme producing environmentally benign dinitrogen (N(2)). We examined the effects of increasing Cu concentrations on the transcription and activity of nitrite reductase (NIR), nitric oxide reductase (NOR) and N2 OR in Pseudomonas stutzeri grown anaerobically in solution over a 10-day period. Gas samples were taken on a daily basis and after 6 days, bacterial RNA was recovered to determine the expression of nirS, norB and nosZ encoding NIR, NOR and N(2)OR respectively. Results revealed that 0.05 mM Cu caused maximum conversion of N(2)O to N(2) via bacterial reduction of N(2)O. As soluble Cu generally makes up less than 0.001% of total soil Cu, extrapolation of 0.05 mg l(-l) soluble Cu would require soils to have a total concentration of Cu in the range of, 150-200 µg g(-1) to maximize the proportion of N(2)O reduced to N(2). Given that many intensively farmed agricultural soils are deficient in Cu in terms of plant nutrition, providing a sufficient concentration of biologically accessible Cu could provide a potentially useful microbial-based strategy of reducing agricultural N(2)O emissions.

The effect of lignite on nitrogen mobility in a low-fertility soil amended with biosolids and urea.

Lignite has been proposed as a soil amendment that reduces nitrate (NO3(-)) leaching from soil. Our objective was to determine the effect of lignite on nitrogen (N) fluxes from soil amended with biosolids or urea. The effect of lignite on plant yield and elemental composition was also determined. Batch sorption and column leaching experiments were followed by a lysimeter trial where a low fertility soil was amended with biosolids (400 kg N/ha equivalent) and urea (200 kg N/ha equivalent). Treatments were replicated three times, with and without lignite addition (20 t/ha equivalent). Lignite did not reduce NO3(-) leaching from soils amended with either biosolids or urea. While lignite decreased NO3(-) leaching from an unamended soil, the magnitude of this effect was not significant in an agricultural context. Furthermore, lignite increased cumulative N2O production from soils receiving urea by 90%. Lignite lessened the beneficial growth effects of adding biosolids or urea to soil. Further work could investigate whether coating urea granules with lignite may produce meaningful environmental benefits.

Dairy farm effluent effects on urine patch nitrous oxide and carbon dioxide emissions.

Dairy farm effluent (DFE) comprises animal feces, urine, and wash-down water collected at the milking shed. This is collected daily during the milking season and sprayed onto grazed dairy pastures. Urine patches in grazed pastures make a significant contribution to anthropogenic N(2)O emissions. The DFE could potentially mitigate N(2)O emissions by influencing the N(2)O to dinitrogen (N(2)) ratio, since it contains water-soluble carbon (WSC). Alternatively, DFE may enhance N(2)O emissions from urine patches. The application of DFE may also provide a substrate for the production of CO(2) in pasture soils. The effects of DFE on the CO(2) and N(2)O emissions from urine patches are unknown. Thus a laboratory experiment was performed where repeated DFE applications were made to repacked soil cores. Dairy farm effluent was applied at 0, 7, or 14 d after urine deposition. The urine was applied once on Day 0. Urine contained (15)N-enriched urea. Measurements of N(2)O, N(2), and carbon dioxide (CO(2)) fluxes, soil pH, and soil inorganic N concentrations were made. After 43 d the DFE had not mitigated N(2)O fluxes from urine patches. A small increase in the N(2)O flux occurred from the urine-treated soils where DFE was applied 1 wk after urine deposition. The amount of WSC applied in the DFE proved to be insignificant compared with the amount of soil C released as CO(2) following urine application. The priming of soil C in urine patches has implications for the understanding of soil C processes in grazed pasture ecosystems and the budgeting of C within these ecosystems.

Confirmation of co-denitrification in grazed grassland.

Pasture-based livestock systems are often associated with losses of reactive forms of nitrogen (N) to the environment. Research has focused on losses to air and water due to the health, economic and environmental impacts of reactive N. Di-nitrogen (N2) emissions are still poorly characterized, both in terms of the processes involved and their magnitude, due to financial and methodological constraints. Relatively few studies have focused on quantifying N2 losses in vivo and fewer still have examined the relative contribution of the different N2 emission processes, particularly in grazed pastures. We used a combination of a high (15)N isotopic enrichment of applied N with a high precision of determination of (15)N isotopic enrichment by isotope-ratio mass spectrometry to measure N2 emissions in the field. We report that 55.8 g N m(-2) (95%, CI 38 to 77 g m(-2)) was emitted as N2 by the process of co-denitrification in pastoral soils over 123 days following urine deposition (100 g N m(-2)), compared to only 1.1 g N m(-2) (0.4 to 2.8 g m(-2)) from denitrification. This study provides strong evidence for co-denitrification as a major N2 production pathway, which has significant implications for understanding the N budgets of pastoral ecosystems.

Effect of nitrogen and waterlogging on denitrifier gene abundance, community structure and activity in the rhizosphere of wheat.

Microbial denitrification plays a key role in determining the availability of soil nitrogen (N) to plants. However, factors influencing the structure and function of denitrifier communities in the rhizosphere remain unclear. Waterlogging can result in root anoxia and increased denitrification, leading to significant N loss from soil and potential nitrous oxide (N(2)O) emissions. This study investigated denitrifier gene abundance, community structure and activity in the rhizosphere of wheat in response to anoxia and N limitation. Denitrifier community structure in the rhizosphere differed from that in bulk soil, and denitrifier gene copy numbers (nirS, nirK, nosZ) and potential denitrification activity were greater in the rhizosphere. Anoxia and N limitation, and in particular a combination of both, reduced the magnitude of this effect on gene abundance (in particular nirS) and activity, with N limitation having greater impact than waterlogging in rhizosphere soil, in contrast to bulk soil where the impact of waterlogging was greater. Increased N supply to anoxic plants improved plant health and increased rhizosphere soil pH, which resulted in enhanced reduction of N(2)O. Both anoxia and N limitation significantly influenced the structure and function of denitrifier communities in the rhizosphere, with reduced root-derived carbon postulated to play an important role.

Biogeochemistry and community ecology in a spring-fed urban river following a major earthquake.

In February 2011 a MW 6.3 earthquake in Christchurch, New Zealand inundated urban waterways with sediment from liquefaction and triggered sewage spills. The impacts of, and recovery from, this natural disaster on the stream biogeochemistry and biology were assessed over six months along a longitudinal impact gradient in an urban river. The impact of liquefaction was masked by earthquake triggered sewage spills (~20,000 m(3) day(-1) entering the river for one month). Within 10 days of the earthquake dissolved oxygen in the lowest reaches was <1 mg l(-1), in-stream denitrification accelerated (attenuating 40-80% of sewage nitrogen), microbial biofilm communities changed, and several benthic invertebrate taxa disappeared. Following sewage system repairs, the river recovered in a reverse cascade, and within six months there were no differences in water chemistry, nutrient cycling, or benthic communities between severely and minimally impacted reaches. This study highlights the importance of assessing environmental impact following urban natural disasters.