Soil environmental conditions rather than denitrifier abundance and diversity drive potential denitrification after changes in land uses

Land‐use practices aiming at increasing agro‐ecosystem sustainability, e.g. no‐till systems and use of temporary grasslands, have been developed in cropping areas, but their environmental benefits could be counterbalanced by increased N₂O emissions produced, in particular during denitrification. Modelling denitrification in this context is thus of major importance. However, to what extent can changes in denitrification be predicted by representing the denitrifying community as a black box, i.e. without an adequate representation of the biological characteristics (abundance and composition) of this community, remains unclear. We analysed the effect of changes in land uses on denitrifiers for two different agricultural systems: (i) crop/grassland conversion and (ii) cessation/application of tillage. We surveyed potential denitrification (PD), the abundance and genetic structure of denitrifiers (nitrite reducers), and soil environmental conditions. N₂O emissions were also measured during periods of several days on control plots. Time‐integrated N₂O emissions and PD were well correlated among all control plots. Changes in PD were partly due to changes in denitrifier abundance but were not related to changes in the structure of the denitrifier community. Using multiple regression analysis, we showed that changes in PD were more related to changes in soil environmental conditions than in denitrifier abundance. Soil organic carbon explained 81% of the variance observed for PD at the crop/temporary grassland site, whereas soil organic carbon, water‐filled pore space and nitrate explained 92% of PD variance at the till/no‐till site, without any residual effect of denitrifier abundance. Soil environmental conditions influenced PD by modifying the specific activity of denitrifiers, and to a lesser extent by promoting a build‐up of denitrifiers. Our results show that an accurate simulation of carbon, oxygen and nitrate availability to denitrifiers is more important than an accurate simulation of denitrifier abundance and community structure to adequately understand and predict changes in PD in response to land‐use changes.     

The response of denitrifiers in a sandy loam soil affected by a long-term fertilization to organic carbon and nitrate

The response of denitrifiers to carbon in the form of glucose (Glc-C) and nitrate (NO ₃ ⁻ -N) amendments was studied in four differently fertilized plots of sandy-loam soil. Two basically different characteristics of denitrification activity were determined: (1) potential denitrification measured as nitrous oxide production during 1-d incubation in the presence of acetylene, and (2) denitrifying enzyme activity determined in soil slurries as a N₂O production in the presence of acetylene and chloramphenicol during 1 h of incubation. Potential denitrification was strongly influenced by both Glc-C and NO ₃ ⁻ -N amendments in their various combinations, but was also affected by the fertilization practice. The response of denitrifiers to Glc-C and NO ₃ ⁻ -N was generally lower in unfertilized and surprisingly also in highly fertilized soils than in organically and moderately fertilized soils. Denitrifying enzyme activity was stimulated by the fertilization and was, in contrast to potential denitrification, the highest in highly fertilized soil. The results indicate that although active denitrifiers were present in a highly fertilized soil, their ability to develop under optimal conditions was decreased (being similar to that of denitrifiers in unfertilized soil). This suggests long-term changes in soil microbial community in a highly fertilized soil, presumably connected to changes in soil chemistry caused by fertilization.     

Characterization of N<Subscript>2</Subscript>O emissions and associated microbial communities from the ant mounds in soils of a humid tropical rainforest

Tropical rainforest soils harbor a considerable diversity of soil fauna that contributes to emissions of N₂O. Despite their ecological dominance, there is limited information available about the contribution of epigeal ant mounds to N₂O emissions in these tropical soils. This study aimed to determine whether ant mounds contribute to local soil N emissions in the tropical humid rainforest. N₂O emission was determined in vitro from individual live ants, ant-processed mound soils, and surrounding reference soils for two trophically distinct and abundant ant species: the leaf-cutting Atta mexicana and omnivorous Solenopsis geminata. The abundance of total bacteria, nitrifiers (AOA and AOB), and denitrifiers (nirK, nirS, and nosZ) was estimated in these soils using quantitative PCR, and their respective mineral N contents determined. There was negligible N₂O emission detected from live ant individuals. However, the mound soils of both species emitted significantly greater (3-fold) amount of N₂O than their respective surrounding reference soils. This emission increased significantly up to 6-fold in the presence of acetylene, indicating that, in addition to N₂O, dinitrogen (N₂) is also produced from these mound soils at an equivalent rate (N₂O/N₂?=?0.57). Functional gene abundance (nitrifiers and denitrifiers) and mineral N pools (ammonium and nitrate) were significantly greater in mound soils than in their respective reference soils. Furthermore, in the light of the measured parameters and their correlation trends, nitrification and denitrification appeared to represent the major N₂O-producing microbial processes in ant mound soils. The ant mounds were estimated to contribute from 0.1 to 3.7% of the total N₂O emissions of tropical rainforest soils.     

Management of nitrogen fertilizer application, rather than functional gene abundance, governs nitrous oxide fluxes in hydroponics with rockwool

Aims

Nitrous oxide (N₂O) is a strong greenhouse effective gas (GHG); the primary human source of N₂O is agricultural activities. Excessive nitrogen (N) fertilization of agricultural land is now widely recognized as a major contributor. In soil, the microbial processes of nitrification and denitrification are the principal sources of N₂O. However, it remains poorly understood how conventional hydroponics influences GHG emission. Here, we compared GHG fluxes from soil and rockwool used for hydroponics under identical nutrient conditions.

Methods

Tomato plants (Solanum lycopersicum, momotaro) were grown in soil or by hydroponics using rockwool. In situ emissions of CH₄, CO₂, and N₂O, and the abundance of genes involved in nitrification and denitrification were measured during cultivation.

Results

Hydroponics with rockwool mitigated CO₂ emission by decreasing the microbial quantity in the rhizosphere. Dilution of the nutrient solution significantly decreased N₂O emission from rockwool. Although proliferation of nitrifiers or denitrifiers in the rhizosphere did not induce N₂O emission, reuse or long-term use of rockwool induced a 3.8-fold increase in N₂O emission.

Conclusions

Our data suggest that hydroponics has a lower environmental impact and that adequate fertilizer application, rather than bacterial control, governs N₂O fluxes in hydroponic cultivation using rockwool.     

N<Subscript>2</Subscript>O production by denitrification in an urban river: evidence from isotopes,functional genes,and dissolved organic matter

Rivers are important sources of N₂O emissions into the atmosphere. Nevertheless, N₂O production processes in rivers are not well identified. We measured concentrations and isotopic ratios of N₂O, NH₄ ⁺, NO₂ ^?, and NO₃ ^? in surface water to identify the microbial processes of N₂O production along the Tama River in Japan. We also measured the functional gene abundance of nitrifiers and denitrifiers (amoA-bacteria, nirK, nirS, nosZ clade I, nosZ clade II) together with concentrations of dissolved organic carbon (DOC) and fluorescence intensities of protein and humic components of dissolved organic matter (DOM) to support the elucidation of N₂O production processes. The observed nitrogen (δ¹⁵N) and oxygen (δ¹⁸O) of N₂O were within the expected isotopic range of N₂O produced by nitrate reduction, indicating that N₂O was dominantly produced by denitrification. The positive significant correlation between N₂O_Net concentration and nirK gene abundance implied that nitrifiers and denitrifiers are contributors to N₂O production. Fluorescence intensities of protein and humic components of DOM and concentrations of DOC did not show significant correlations with N₂O concentrations, which suggests that DOC and abundance of DOM components do not control dissolved N₂O. Measurement of isotope ratios of N₂O and its substrates was found to be a useful tool to obtain evidence of denitrification as the main source of N₂O production along the Tama River.     

Concepts in modelling N2O emissions from land use

Modelling nitrous oxide (N₂O) emissions from soil is challenging because multiple biological processes are involved that each respond differently to various environmental and soil factors. Soil water content, organic carbon, temperature and pH are often used in models that predict N₂O emissions, yet for each of these factors there are concepts that are not fully understood. Though a ubiquitous measure of soil water for models, the application of functions based on water filled pore space across soils that vary in bulk density is not ideal. Diffusion of gases and solutes in soil are controlled by the volume fractions of air and water present. Across soils with different bulk densities, both of these terms vary at constant water filled pore space. Soil organic carbon influences N₂O emissions in two ways: as a source of energy for denitrifiers and also by driving biological oxygen demand and the creation of anaerobic zones in the soil. Soil temperature influences N₂O emissions through its effect on the activity of microorganisms and enzymes. A variety of temperature response functions have been proposed. The preferred response function should contain a temperature optimum that can be varied in response to climatic conditions to account for microbial adaptation. Soil pH can have direct and indirect influences on rates and product ratios of nitrification and denitrification. The concepts of pH optima and microbial adaptation need to be considered in modelling. Methodological issues such as microsite versus bulk soil measurements and apportioning N₂O fluxes to the various N transformation processes remain an impediment to characterising the influence of pH and other factors on N₂O emissions. Quantifying the response of N₂O emissions to individual factors using regression analysis requires all other factors to be controlled experimentally. Boundary line analysis provides a way of defining the response to a single input variable where other influencing variables are not controlled. Such analyses can aid in the definition of the shape and magnitude of response functions to be incorporated into process simulation models. Process/mechanistic simulation models offer a greater transferability than empirical models but careful consideration of temporal and spatial scale and the availability of data to run these models is critical in developing model structure.     

Relative Rates of Nitric Oxide and Nitrous Oxide Production by Nitrifiers, Denitrifiers, and Nitrate Respirers

Biogenic emissions of nitric and nitrous oxides have important impacts on the photochemistry and chemistry of the atmosphere. Although biogenic production appears to be the overwhelming source of N₂O, the magnitude of the biogenic emission of NO is very uncertain. In soils, possible sources of NO and N₂O include nitrification by autotrophic and heterotrophic nitrifiers, denitrification by nitrifiers and denitrifiers, nitrate respiration by fermenters, and chemodenitrification. The availability of oxygen determines to a large extent the relative activities of these various groups of organisms. To better understand this influence, we investigated the effect of the partial pressure of oxygen (pO₂) on the production of NO and N₂O by a wide variety of common soil nitrifying, denitrifying, and nitrate-respiring bacteria under laboratory conditions. The production of NO per cell was highest by autotrophic nitrifiers and was independent of pO₂ in the range tested (0.5 to 10%), whereas N₂O production was inversely proportional to pO₂. Nitrous oxide production was highest in the denitrifier Pseudomonas fluorescens, but only under anaerobic conditions. The molar ratio of NO/N₂O produced was usually greater than unity for nitrifiers and much less than unity for denitrifiers. Chemodenitrification was the major source of both the NO and N₂O produced by the nitrate respirer Serratia marcescens. Chemodenitrification was also a possible source of NO and N₂O in nitrifier cultures but only when high concentrations of nitrite had accumulated or were added to the medium. Although most of the denitrifiers produced NO and N₂O only under anaerobic conditions, chemostat cultures of Alcaligenes faecalis continued to emit these gases even when the cultures were sparged with air. Based upon these results, we predict that aerobic soils are primary sources of NO and that N₂O is produced only when there is sufficient soil moisture to provide the anaerobic microsites necessary for denitrification by either denitrifiers or nitrifiers.     

Mechanism leading to N<Subscript>2</Subscript>O production in wastewater treating biofilm systems

Wastewater treatment plants are known to be important point sources for nitrous oxide (N₂O) in the anthropogenic N cycle. Biofilm based treatment systems have gained increasing popularity in the treatment of wastewater, but the mechanisms and controls of N₂O formation are not fully understood. Here, we review functional groups of microorganism involved in nitrogen (N) transformations during wastewater treatment, with emphasis on potential mechanism of N₂O production in biofilms. Biofilms used in wastewater treatment typically harbour aerobic and anaerobic zones, mediating close interactions between different groups of N transforming organisms. Current models of mass transfer and biomass interactions in biofilms are discussed to illustrate the complex regulation of N₂O production. Ammonia oxidizing bacteria (AOB) are the prime source for N₂O in aerobic zones, while heterotrophic denitrifiers dominate N₂O production in anoxic zones. Nitrosative stress ensuing from accumulation of NO₂ ^? during partial nitrification or denitrification seems to be one of the most critical factors for enhanced N₂O formation. In AOB, N₂O production is coupled to nitrifier denitrification triggered by nitrosative stress, low O₂ tension or low pH. Chemical N₂O production from AOB intermediates (NH₂OH, HNO, NO) released during high NH₃ turnover seems to be limited to surface-near AOB clusters, since diffusive mass transport resistance for O₂ slows down NH₃ oxidation rates in deeper biofilm layers. The proportion of N₂O among gaseous intermediates (NO, N₂O, N₂) in heterotrophic denitrification increases when NO or nitrous acid (HNO₂) accumulates because of increasing NO₂ ^?, or when transient oxygen intrusion impairs complete denitrification. Limited electron donor availability due to mass transport limitation of organic substrates into anoxic biofilm zones is another important factor supporting high N₂O/N₂ ratios in heterotrophic denitrifiers. Biofilms accommodating Anammox bacteria release less N₂O, because Anammox bacteria have no known N₂O producing metabolism and reduce NO₂ ^? to N₂, thereby lowering nitrosative stress to AOB and heterotrophs.     

Competition for electrons favours N2O reduction in denitrifying Bradyrhizobium isolates

Bradyrhizobia are common members of soil microbiomes and known as N₂-fixing symbionts of economically important legumes. Many are also denitrifiers, which can act as sinks or sources for N₂O. Inoculation with compatible rhizobia is often needed for optimal N₂-fixation, but the choice of inoculant may have consequences for N₂O emission. Here, we determined the phylogeny and denitrification capacity of Bradyrhizobium strains, most of them isolated from peanut-nodules. Analyses of genomes and denitrification end-points showed that all were denitrifiers, but only ~1/3 could reduce N₂O. The N₂O-reducing isolates had strong preference for N₂O- over NO₃⁻-reduction. Such preference was also observed in a study of other bradyrhizobia and tentatively ascribed to competition between the electron pathways to Nap (periplasmic NO₃⁻ reductase) and Nos (N₂O reductase). Another possible explanation is lower abundance of Nap than Nos. Here, proteomics revealed that Nap was instead more abundant than Nos, supporting the hypothesis that the electron pathway to Nos outcompetes that to Nap. In contrast, Paracoccus denitrificans, which has membrane-bond NO₃⁻ reductase (Nar), reduced N₂O and NO₃⁻ simultaneously. We propose that the control at the metabolic level, favouring N₂O reduction over NO₃⁻ reduction, applies also to other denitrifiers carrying Nos and Nap but lacking Nar.     

Dinitrogen and nitrous oxide produced by denitrification and nitrification in soil with and without barley plants

Summary To examine the effect of barley roots on denitrification, a pot experiment was designed to compare N₂O production and denitrification in soils with and without barley plants. Denitrification, N₂O resulting from denitrification and nitrification, and respiration were estimated by incubating pots with soil with and without intact plants in plastic bags at high moisture levels. C₂H₂-inhibition of nitrous oxide reductase (partial pressure of 10 kPa C₂H₂) was used to determine total denitrification rates while incubations with ambient air and with C₂H₂ at partial pressures of 2.5–5 Pa were used to estimate the amounts of N₂O released from autotrophic nitrification and from denitrification processes. Other sources of N₂O were presumed to be negligible. Potential denitrification, nitrification and root biomass were measured in subsamples collected from four soil depths. A positive correlation was found between denitrification rates and root biomass. N₂ was the predominant denitrification product found close to roots; N₂O formed by non autotrophic nitrifiers, assumed to be denitrifiers originated in soil not affected by growing roots. Apparently, roots promote denitrification because they consumed oxygen, thereby increasing the anaerobic volume of the soil. The ratio of actual to potential denitrification rates increased over time, especially in the presence of roots.     

Mitigating N2O emissions from agricultural soils with fungivorous mites

Nitrous oxide (N₂O) is an important greenhouse gas and an ozone-depleting substance. Due to the long persistence of N₂O in the atmosphere, the mitigation of anthropogenic N₂O emissions, which are mainly derived from microbial N₂O-producing processes, including nitrification and denitrification by bacteria, archaea, and fungi, in agricultural soils, is urgently necessary. Members of mesofauna affect microbial processes by consuming microbial biomass in soil. However, how microbial consumption affects N₂O emissions is largely unknown. Here, we report the significant role of fungivorous mites, the major mesofaunal group in agricultural soils, in regulating N₂O production by fungi, and the results can be applied to the mitigation of N₂O emissions. We found that the application of coconut husks, which is the low-value part of coconut and is commonly employed as a soil conditioner in agriculture, to soil can supply a favorable habitat for fungivorous mites due to its porous structure and thereby increase the mite abundance in agricultural fields. Because mites rapidly consume fungal N₂O producers in soil, the increase in mite abundance substantially decreases the N₂O emissions from soil. Our findings might provide new insight into the mechanisms of soil N₂O emissions and broaden the options for the mitigation of N₂O emissions.Subject terms: Climate-change ecology, Climate-change ecology     

Importance of denitrifiers lacking the genes encoding the nitrous oxide reductase for N2O emissions from soil

Analyses of the complete genomes of sequenced denitrifying bacteria revealed that approximately 1/3 have a truncated denitrification pathway, lacking the nosZ gene encoding the nitrous oxide reductase. We investigated whether the number of denitrifiers lacking the genetic ability to synthesize the nitrous oxide reductase in soils is important for the proportion of N₂O emitted by denitrification. Serial dilutions of the denitrifying strain Agrobacterium tumefaciens C58 lacking the nosZ gene were inoculated into three different soils to modify the proportion of denitrifiers having the nitrous oxide reductase genes. The potential denitrification and N₂O emissions increased when the size of inoculated C58 population in the soils was in the same range as the indigenous nosZ community. However, in two of the three soils, the increase in potential denitrification in inoculated microcosms compared with the noninoculated microcosms was higher than the increase in N₂O emissions. This suggests that the indigenous denitrifier community was capable of acting as a sink for the N₂O produced by A. tumefaciens. The relative amount of N₂O emitted also increased in two soils with the number of inoculated C58 cells, establishing a direct causal link between the denitrifier community composition and potential N₂O emissions by manipulating the proportion of denitrifiers having the nosZ gene. However, the number of denitrifiers which do not possess a nitrous oxide reductase might not be as important for N₂O emissions in soils having a high N₂O uptake capacity compared with those with lower. In conclusion, we provide a proof of principle that the inability of some denitrifiers to synthesize the nitrous oxide reductase can influence the nature of the denitrification end products, indicating that the extent of the reduction of N₂O to N₂ by the denitrifying community can have a genetic basis.     

不同土地利用方式土壤氨氧化微生物和反硝化微生物时空分布特征

研究不同土地利用方式下氮循环相关微生物在不同土壤剖面的分布,可为认识和理解土壤氮转化过程提供科学依据。土壤氨氧化微生物和反硝化微生物在调节氮肥利用率、硝态氮淋溶和氧化亚氮(N₂O)排放等方面有着重要作用。以北京郊区农田和林地两种土地利用方式为研究对象,分析土壤氨氧化潜势和亚硝酸盐氧化潜势在0—100 cm土壤剖面上的季节分布(春季和秋季),并通过实时荧光定量PCR方法表征土壤氨氧化和反硝化微生物的时空分布特征。结果表明,农田土壤氨氧化潜势、亚硝酸盐氧化潜势、氨氧化微生物和反硝化微生物丰度均显著高于林地土壤,且随土壤深度增加而显著降低。除氨氧化古菌amoA基因丰度在不同季节间无显著差异外,春季土壤氨氧化细菌(amoA基因)、反硝化微生物nirS、nirK和典型nosZ I基因的丰度均显著高于秋季。土壤有机质、总氮、NH~+₄-N、NO~-₃-N含量与氨氧化微生物和反硝化微生物的功能基因丰度显著相关。综上,不同土地利用方式下土壤氮循环相关微生物的丰度与土壤氮素的可利用性和转化过程紧密相关,研究结果对土壤氮素利用和养分管理提供...     

Denitrification in the vadose zone and in surficial groundwater of a sandy entisol with citrus production

A portion of nitrate (NO ₃ ⁻ ), a final breakdown product of nitrogen (N) fertilizers, applied to soils and/or that produced upon decomposition of organic residues in soils may leach into groundwater. Nitrate levels in water excess of 10 mg L⁻¹ (NO₃–N) are undesirable as per drinking water quality standards. Nitrate concentrations in surficial groundwater can vary substantially within an area of citrus grove which receives uniform N rate and irrigation management practice. Therefore, differences in localized conditions which can contribute to variations in gaseous loss of NO ₃ ⁻ in the vadose zone and in the surficial aquifer can affect differential concentrations of NO₃–N in the groundwater at different points of sampling. The denitrification capacity and potential in a shallow vadose zone soil and in surficial groundwater were studied in two large blocks of a citrus grove of ‘Valencia’ orange trees (Citrus sinensis (L.) Obs.) on Rough lemon rootstock ( Citrus jambhiri (L.)) under a uniform N rate and irrigation program. The NO₃–N concentration in the surficial groundwater sampled from four monitoring wells (MW) within each block varied from 5.5- to 6.6-fold. Soil samples were collected from 0 to 30, 30 to 90, or 90 to 150 cm depths, and from the soil/groundwater interface (SGWI). Groundwater samples from the monitoring wells (MW) were collected prior to purging (stagnant water) and after purging five well volumes. Without the addition of either C or N, the denitrification capacity ranged from 0.5 to 1.53, and from 0.0 to 2.25 mg N₂O–N kg⁻¹ soil at the surface soil and at the soil/groundwater interface, respectively. The denitrification potential increased by 100-fold with the addition of 200 mg kg⁻¹ each of N and C. The denitrification potential in the groundwater also followed a pattern similar to that for the soil samples. Denitrification potential in the soil or in the groundwater was greatest near the monitor well with shallow depth of vadose zone (MW3). Cumulative N₂O–N emission (denitrification capacity) from the SGWI soil samples and from stagnant water samples strongly correlated to microbial most probable number (MPN) counts (r² = 0.84 – 0.89), and dissolved organic C (DOC) (r² = 0.96 – 0.97). Denitrification capacity of the SGWI samples moderately correlated to water-filled pore space (WFPS) (r² = 0.52). However, extractable NO₃-N content of the SGWI soil samples poorly (negative) correlated to denitrification capacity (r² = 0.35). However, addition C, N or both to the soil or water samples resulted in significant increase in cumulative N₂O emission. This study demonstrated that variation in denitrification capacity, as a result of differences in denitrifier population, and the amount of readily available carbon source significantly (at 95% probability level) influenced the variation in NO₃–N concentrations in the surficial groundwater samples collected from different monitoring wells within an area with uniform N management. This revised version was published online in June 2006 with corrections to the Cover Date.     

The effect of grass maturing and root decay on N2O production in soil

The N₂O flux from the surface of grass-covered pots was only significant following grass maturing. Removal of the above-ground plant material resulted in an immediate and long-lasting increase in N₂O production in the soil. The results suggest that easily available organic matter from the roots stimulates the denitrification when the plants are damaged. Grass cutting might therefore result in a marked nitrogen loss through denitrification. The quantitative effect was equal in soil with and without succinate added. The size of the anaerobic zone around the roots is therefore sufficient to allow for denitrification activity mediated by increased organic matter availability because of plant cutting.     

Elevated temperature shifts soil N cycling from microbial immobilization to enhanced mineralization,nitrification and denitrification across global terrestrial ecosystems

We assessed the response of soil microbial nitrogen (N) cycling and associated functional genes to elevated temperature at the global scale. A meta‐analysis of 1,270 observations from 134 publications indicated that elevated temperature decreased soil microbial biomass N and increased N mineralization rates, both in the presence and absence of plants. These findings infer that elevated temperature drives microbially mediated N cycling processes from dominance by anabolic to catabolic reaction processes. Elevated temperature increased soil nitrification and denitrification rates, leading to an increase in N₂O emissions of up to 227%, whether plants were present or not. Rates of N mineralization, denitrification and N₂O emission demonstrated significant positive relationships with rates of CO₂ emissions under elevated temperatures, suggesting that microbial N cycling processes were associated with enhanced microbial carbon (C) metabolism due to soil warming. The response in the abundance of relevant genes to elevated temperature was not always consistent with changes in N cycling processes. While elevated temperature increased the abundances of the nirS gene with plants and nosZ genes without plants, there was no effect on the abundances of the ammonia‐oxidizing archaea amoA gene, ammonia‐oxidizing bacteria amoA and nirK genes. This study provides the first global‐scale assessment demonstrating that elevated temperature shifts N cycling from microbial immobilization to enhanced mineralization, nitrification and denitrification in terrestrial ecosystems. These findings infer that elevated temperatures have a profound impact on global N cycling processes with implications of a positive feedback to global climate and emphasize the close linkage between soil microbial C and N cycling.     

Effect of Soil pH Increase by Biochar on NO,N2O and N2 Production during Denitrification in Acid Soils

Biochar (BC) application to soil suppresses emission of nitrous- (N₂O) and nitric oxide (NO), but the mechanisms are unclear. One of the most prominent features of BC is its alkalizing effect in soils, which may affect denitrification and its product stoichiometry directly or indirectly. We conducted laboratory experiments with anoxic slurries of acid Acrisols from Indonesia and Zambia and two contrasting BCs produced locally from rice husk and cacao shell. Dose-dependent responses of denitrification and gaseous products (NO, N₂O and N₂) were assessed by high-resolution gas kinetics and related to the alkalizing effect of the BCs. To delineate the pH effect from other BC effects, we removed part of the alkalinity by leaching the BCs with water and acid prior to incubation. Uncharred cacao shell and sodium hydroxide (NaOH) were also included in the study. The untreated BCs suppressed N₂O and NO and increased N₂ production during denitrification, irrespective of the effect on denitrification rate. The extent of N₂O and NO suppression was dose-dependent and increased with the alkalizing effect of the two BC types, which was strongest for cacao shell BC. Acid leaching of BC, which decreased its alkalizing effect, reduced or eliminated the ability of BC to suppress N₂O and NO net production. Just like untreated BCs, NaOH reduced net production of N₂O and NO while increasing that of N₂. This confirms the importance of altered soil pH for denitrification product stoichiometry. Addition of uncharred cacao shell stimulated denitrification strongly due to availability of labile carbon but only minor effects on the product stoichiometry of denitrification were found, in accordance with its modest effect on soil pH. Our study indicates that stimulation of denitrification was mainly due to increases in labile carbon whereas change in product stoichiometry was mainly due to a change in soil pH.     

Denitrification Activity of a Remarkably Diverse Fen Denitrifier Community in Finnish Lapland Is N-Oxide Limited

Peatlands cover more than 30% of the Finnish land area and impact N₂O fluxes. Denitrifiers release N₂O as an intermediate or end product. In situ N₂O emissions of a near pH neutral pristine fen soil in Finnish Lapland were marginal during gas chamber measurements. However, nitrate and ammonium fertilization significantly stimulated in situ N₂O emissions. Stimulation with nitrate was stronger than with ammonium. N₂O was produced and subsequently consumed in gas chambers. In unsupplemented anoxic microcosms, fen soil produced N₂O only when acetylene was added to block nitrous oxide reductase, suggesting complete denitrification. Nitrate and nitrite stimulated denitrification in fen soil, and maximal reaction velocities (v_max) of nitrate or nitrite dependent denitrification where 18 and 52 nmol N₂O h^-1 g_DW ^-1, respectively. N₂O was below 30% of total produced N gases in fen soil when concentrations of nitrate and nitrite were <500 μM. v_max for N₂O consumption was up to 36 nmol N₂O h^-1 g_DW ^-1. Denitrifier diversity was assessed by analyses of narG, nirK/nirS, and nosZ (encoding nitrate-, nitrite-, and nitrous oxide reductases, respectively) by barcoded amplicon pyrosequencing. Analyses of ~14,000 quality filtered sequences indicated up to 25 species-level operational taxonomic units (OTUs), and up to 359 OTUs at 97% sequence similarity, suggesting diverse denitrifiers. Phylogenetic analyses revealed clusters distantly related to publicly available sequences, suggesting hitherto unknown denitrifiers. Representatives of species-level OTUs were affiliated with sequences of unknown soil bacteria and Actinobacterial, Alpha-, Beta-, Gamma-, and Delta-Proteobacterial sequences. Comparison of the 4 gene markers at 97% similarity indicated a higher diversity of narG than for the other gene markers based on Shannon indices and observed number of OTUs. The collective data indicate (i) a high denitrification and N₂O consumption potential, and (ii) a highly diverse, nitrate limited denitrifier community associated with potential N₂O fluxes in a pH-neutral fen soil.     

Global change could amplify fire effects on soil greenhouse gas emissions

Background

Little is known about the combined impacts of global environmental changes and ecological disturbances on ecosystem functioning, even though such combined impacts might play critical roles in shaping ecosystem processes that can in turn feed back to climate change, such as soil emissions of greenhouse gases.

Methodology/Principal Findings

We took advantage of an accidental, low-severity wildfire that burned part of a long-term global change experiment to investigate the interactive effects of a fire disturbance and increases in CO₂ concentration, precipitation and nitrogen supply on soil nitrous oxide (N₂O) emissions in a grassland ecosystem. We examined the responses of soil N₂O emissions, as well as the responses of the two main microbial processes contributing to soil N₂O production – nitrification and denitrification – and of their main drivers. We show that the fire disturbance greatly increased soil N₂O emissions over a three-year period, and that elevated CO₂ and enhanced nitrogen supply amplified fire effects on soil N₂O emissions: emissions increased by a factor of two with fire alone and by a factor of six under the combined influence of fire, elevated CO₂ and nitrogen. We also provide evidence that this response was caused by increased microbial denitrification, resulting from increased soil moisture and soil carbon and nitrogen availability in the burned and fertilized plots.

Conclusions/Significance

Our results indicate that the combined effects of fire and global environmental changes can exceed their effects in isolation, thereby creating unexpected feedbacks to soil greenhouse gas emissions. These findings highlight the need to further explore the impacts of ecological disturbances on ecosystem functioning in the context of global change if we wish to be able to model future soil greenhouse gas emissions with greater confidence.