N2O and NO emissions from an Andisol field as influenced by pelleted poultry manure

Animal manures from intensive livestock operations can be pelleted to improve handlings and recyclings of embodied nutrients. The aim of this study was to evaluate the influence of pelleted poultry manure on N₂O and NO fluxes from an Andisol field. In autumn 2006 and summer 2007, poultry manure (PM), pelleted poultry manure (PP), and chemical fertilizer (CF) were applied at a rate of 120 kg N ha⁻¹ in each cultivation period to Komatsuna (Brassica rapa var. peruviridis). Nitrous oxide and NO fluxes were measured using an automated monitoring system. A soil incubation experiment was also conducted to determine the influence of intact and ground pelleted manure on N₂O, NO, and CO₂ production with a water-filled pore space (WFPS) of 30 or 50%. In the field measurements, N₂O emission rates from the organic fertilizer treatments were larger than that from the CF treatment, possibly because organic C stimulated denitrification. The highest N₂O flux was observed from the PP treatment after a rainfall following fertilization, and the cumulative emission rate (2.72 ± 0.22 kg N ha⁻¹ y⁻¹) was 3.9 and 7.1 times that from the PM and CF treatments, respectively. In contrast, NO emission rates were highest from the CF treatment. The NO/N₂O flux ratio indicated that nitrification was the dominant process for NO and N₂O production from the CF treatment. Cumulative N₂O emission rates from all treatments were generally higher during the wetter cultivation period (autumn 2006) than during the drier cultivation period (summer 2007). In contrast, NO emission rates were higher in the drier than in the wetter cultivation period. The incubation experiment results showed a synergistic effect of soil moisture and the pelleted manure form on N₂O emission rates. The intact pelleted manure with the 50% WFPS treatment produced the highest N₂O and CO₂ fluxes and resulted in the lowest soil NO₃⁻ content after the incubation. These results indicate that anaerobic conditions inside the pellets, caused by rainfall and heterotrophic microbial activities, led to denitrification, resulting in high N₂O fluxes. Controlling the timing of N application by avoiding wet conditions might be one mitigation option to reduce N₂O emission rates from the PP treatment in this study field.     

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

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

Microbial growth efficiencies across a soil moisture gradient assessed using C-acetic acid vapor and N-ammonia gas

One integrative measurement of microbial activity in soils is the efficiency by which microbes convert assimilated carbon (C) into biomass C. This efficiency, called the microbial growth efficiency (Y), is a key physiological characteristic that regulates soil carbon sequestration, nutrient immobilization, and greenhouse gas emissions. Changes in rainfall patterns and soil water content as the result of global climate change have the potential to influence microbial activity and lead to changes in Y and thus, nutrient cycling at the ecosystem level. Unfortunately, little information is available on how environmental variables such as soil moisture influence Y. We have developed a new method for injecting ¹³C-labeled carbon as acetic acid vapor into soil that will allow measurement of microbial growth efficiency (as Y_C) without increasing soil moisture content. We compare Y determined with this new approach with an alternate method where injected ¹⁵N-labeled ammonia gas is used to quantify microbial N immobilization, and microbial growth efficiency is calculated based on microbial C:N and respiration rate (as Y_N). We also include injections of a solution containing labeled ammonium and acetate in our experiment to compare the results of our vapor methods with more commonly employed liquid-based methods. The ¹³C-acetic acid vapor, which was supplied to soils with soil moisture content ranging from 0.05 to 0.21 g H₂O g⁻¹ soil, was readily assimilated and respired by microbes. Between 0.10 and 0.21 g H₂O g⁻¹ soil (−0.60 to −0.04 MPa), values of Y_C averaged 0.46, and were significantly lower than values of Y_N, with average values of 0.58. Over this range, soil moisture content had no significant effect on either Y_C or Y_N. However, at the lowest soil moisture content (0.05 g H₂O g⁻¹ soil; <−6.0 MPa), Y_C and Y_N diverged substantially, suggesting that in very dry soils, constraints on microbial growth cause differential uptake of C and N resources.     

Responses of soil organic matter and greenhouse gas fluxes to soil management and land use changes in a humid temperate region of southern Europe

We studied the effects of soil management and changes of land use on soils of three adjacent plots of cropland, pasture and oak (Quercus robur) forest. The pasture and the forest were established in part of the cropland, respectively, 20 and 40 yr before the study began. Soil organic matter (SOM) dynamics, water-filled pore space (WFPS), soil temperature, inorganic N and microbial C, as well as fluxes of CO₂, CH₄ and N₂O were measured in the plots over 25 months. The transformation of the cropland to mowed pasture slightly increased the soil organic and microbial C contents, whereas afforestation significantly increased these variables. The cropland and pasture soils showed low CH₄ uptake rates (<1 kg C ha⁻¹ yr⁻¹) and, coinciding with WFPS values >70%, episodes of CH₄ emission, which could be favoured by soil compaction. In the forest site, possibly because of the changes in soil structure and microbial activity, the soil always acted as a sink for CH₄ (4.7 kg C ha⁻¹ yr⁻¹). The N₂O releases at the cropland and pasture sites (2.7 and 4.8 kg N₂O-N ha⁻¹ yr⁻¹) were, respectively, 3 and 6 times higher than at the forest site (0.8 kg N₂O-N ha⁻¹ yr⁻¹). The highest N₂O emissions in the cultivated soils were related to fertilisation and slurry application, and always occurred when the WFPS >60%. These results show that the changes in soil properties as a consequence of the transformation of cropfield to intensive grassland do not imply substantial changes in SOM or in the dynamics of CH₄ and N₂O. On the contrary, afforestation resulted in increases in SOM content and CH₄ uptake, as well as decreases in N₂O emissions.     

Nitrous oxide emissions from boreal organic soil under different land-use

Nitrous oxide emissions were studied with a static chamber technique during 2 years from a drained organic soil in eastern Finland. After drainage, the soil was forested with birch (Betula pendula Roth) and 22 years later, part of the forest was felled and then used for cultivation of barley (Hordeum vulgare L.) and grass. The annual N₂O emissions from the cultivated soil (from 8.3 to 11.0 kg N₂O-N ha⁻¹ year⁻¹) were ca. twice the annual emission from the adjacent forest site (4.2 kg N₂O-N ha⁻¹ year⁻¹). The N₂O emissions from the soils without plants (kept bare by regular cutting or tilling) were also lower (from 6.5 to 7.1 kg N₂O-N ha⁻¹ year⁻¹) than those from the cultivated soil. There was a high seasonal variation in the fluxes with a maximum in spring and early summer. The N₂O fluxes during the winter period accounted for 15-60% of the total annual emissions. N₂O fluxes during the snow-free periods were related to the water table (WT) level, water-filled pore space, carbon mineralisation and the soil temperature. A linear regression model with CO₂ production, WT and soil temperature at the depth of 5 cm as independent variables explained 54% of the variation in the weekly mean N₂O fluxes during the snow-free periods. N₂O fluxes were associated with in situ net nitrification, which alone explained 58% of the variation in the mean N₂O fluxes during the snow-free period. The N₂O-N emissions were from 1.5 to 5% of net nitrification. The acetylene blockage technique indicated that most of the N₂O emitted in the snow-free period originated from denitrification.     

Denitrification,N2-fixation and fermentation during anaerobic incubation of soils amended with glucose and nitrate

Nitrate and glucose additions were investigated for their role in the C and N dynamics during anaerobic incubation of soil. A gas-flow soil core method was used, in which the net production of N₂, N₂O, NO, CO₂, and CH₄ under a He atmosphere could be monitored both accurately and frequently. In all experiments clayey silt loam soil samples were incubated for 9 days at 25 °C. Addition of nitrate (50 mg KNO₃-N kg^-1 soil) had no effect on total denitrification and CO₂ production rates, while the N₂O/N₂ ratio was affected considerably. The cumulative N₂O production exceeded the cumulative N₂ production for 6 days in the treatment with nitrate addition, compared to 1.2 days in the unamended treatment. Glucose addition stimulated the microbial activity considerably. The denitrification rates were limited by the growth rate of the denitrifying population. During denitrification no significant differences were observed between the treatments with 700 mg glucose-C kg^-1 and 4200 mg glucose-C kg^-1, both in combination with 50 mg KNO₃-N kg^-1. The N₂ production rates were remarkably low, until NO _inf3 ^sup- exhaustion caused rapid reduction of N₂O to N₂ at day 2. During the denitrification period 15–18 mg N kg^-1 was immobilised in the growing biomass. After NO _inf3 ^sup- shortage, a second microbial population, capable of N₂-fixation, became increasingly important. This change was clearly reflected in the CO₂ production rates. Net volatile fatty acid (VFA) production was monitored during the net N₂-fixation period with acetate as the dominant product. N₂-fixation faded out, probably due to N₂ shortage, followed by increased VFA production. In the high C treatment butyrate became the most important VFA, while in the low C treatment acetate and butyrate were produced at equal rates. During denitrification no VFA accumulation occurred; this does not prove, however, that denitrification and fermentation appeared sequentially. The experiments illustrate clearly the interactions of C-availability, microbial population and nitrate availability as influencing factors on denitrification and fermentation.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Chemical composition, or quality, of agroforestry residues influences N2O emissions after their addition to soil

Emissions of N₂O were measured following addition of ¹⁵N-labelled (2.6-4.7 atom% excess ¹⁵N) agroforestry residues (Sesbania sesban, mixed Sesbania/Macroptilium atropurpureum, Crotalaria grahamiana and Calliandra calothyrsus) to a Kenyan oxisol at a rate of 100 mg N kg soil⁻¹ under controlled environment conditions. Emissions were increased following addition of residues, with 22.6 mg N m⁻² (124.4 mg N m⁻² kg biomass⁻¹; 1.1 mg ¹⁵N m⁻²; 1.03% of ¹⁵N applied) emitted as N₂O over 29 d after addition of both Sesbania and Macroptilium residues in the mixed treatment. Fluxes of N₂O were positively correlated with CO₂ fluxes, and N₂O emissions and available soil N were negatively correlated with residue lignin content (r=−0.49;P<0.05), polyphenol content (r=−0.94;P<0.05), protein binding capacity (r=−0.92;P<0.05) and with (lignin+polyphenol)-to-N ratio (r=−0.55;P<0.05). Lower emission (13.6 mg N m⁻² over 29 d; 94.5 mg N m⁻² kg biomass⁻¹; 0.6 mg ¹⁵N m⁻²; 0.29% of ¹⁵N applied) after addition of Calliandra residue was attributed to the high polyphenol content (7.4%) and high polyphenol protein binding capacity (383 μg BSA mg plant⁻¹) of this residue binding to plant protein and reducing its availability for microbial attack, despite the residue having a N content of 2.9%. Our results indicate that residue chemical composition, or quality, needs to be considered when proposing mitigation strategies to reduce N₂O emissions from systems relying on incorporation of plant biomass, e.g. improved-fallow agroforestry systems, and that this consideration should extend beyond the C-to-N ratio of the residue to include polyphenol content and their protein binding capacity.     

Shifts in rhizosphere microbial communities and enzyme activity of Poa alpina across an alpine chronosequence

This study quantifies the influence of Poa alpina on the soil microbial community in primary succession of alpine ecosystems, and whether these effects are controlled by the successional stage. Four successional sites representative of four stages of grassland development (initial, 4 years (non-vegetated); pioneer, 20 years; transition, 75 years; mature, 9500 years old) on the Rotmoos glacier foreland, Austria, were sampled. The size, composition and activity of the microbial community in the rhizosphere and bulk soil were characterized using the chloroform-fumigation extraction procedure, phospholipid fatty acid (PLFA) analysis and measurements of the enzymes β-glucosidase, β-xylosidase, N-acetyl-β-glucosaminidase, leucine aminopeptidase, acid phosphatase and sulfatase. The interplay between the host plant and the successional stage was quantified using principal component (PCA) and multidimensional scaling analyses. Correlation analyses were applied to evaluate the relationship between soil factors (C_org, N_t, C/N ratio, pH, ammonium, phosphorus, potassium) and microbial properties in the bulk soil. In the pioneer stage microbial colonization of the rhizosphere of P. alpina was dependent on the reservoir of microbial species in the bulk soil. As a consequence, the rhizosphere and bulk soil were similar in microbial biomass (ninhydrin-reactive nitrogen (NHR-N)), community composition (PLFA), and enzyme activity. In the transition and mature grassland stage, more benign soil conditions stimulated microbial growth (NHR-N, total amount of PLFA, bacterial PLFA, Gram-positive bacteria, Gram-negative bacteria), and microbial diversity (Shannon index H) in the rhizosphere either directly or indirectly through enhanced carbon allocation. In the same period, the rhizosphere microflora shifted from a G⁻ to a more G⁺, and from a fungal to a more bacteria-dominated community. Rhizosphere β-xylosidase, N-acetyl-β-glucosaminidase, and sulfatase activity peaked in the mature grassland soil, whereas rhizosphere leucine aminopeptidase, β-glucosidase, and phosphatase activity were highest in the transition stage, probably because of enhanced carbon and nutrient allocation into the rhizosphere due to better growth conditions. Soil organic matter appeared to be the most important driver of microbial colonization in the bulk soil. The decrease in soil pH and soil C/N ratio mediated the shifts in the soil microbial community composition (bacPLFA, bacPLFA/fungPLFA, G⁻, G⁺/G⁻). The activities of β-glucosidase, β-xylosidase and phosphatase were related to soil ammonium and phosphorus, indicating that higher decomposition rates enhanced the nutrient availability in the bulk soil. We conclude that the major determinants of the microflora vary along the successional gradient: in the pioneer stage the rhizosphere microflora was primarily determined by the harsh soil environment; under more favourable environmental conditions, however, the host plant selected for a specific microbial community that was related to the dynamic interplay between soil properties and carbon supply.     

Earthworms change the distribution and availability of phosphorous in organic substrates

In laboratory controlled soil microcosms, the distribution and availability of phosphorous (P) were determined in the surface-casts and the burrows-linings of the anecic earthworm L. terrestris and were compared with non-ingested soil. To simulate more realistic earthworm community conditions, a combination of L. terrestris plus the endogeic A. caliginosa was tested. For a 2-month period, the earthworms were given two organic food substrates: rye-grass littered onto the soil surface and sewage sludge mixed with soil. The following treatments were designed: (i) soil alone (S), (ii) soil and sewage sludge (SS), soil and rye-grass litter (SL), and (iv) soil, litter and sludge (SSL). Analyses were performed for P contents (total, available and organic), organic matter content (organic carbon, C_org and total nitrogen, N_tot) and the two acid and alkaline phosphatase activities (AcPA and AkPA). Earthworms enhanced AcPA and were also responsible for additional AkPA in soil. The two AcPA and AkPA increased not only in surface-casts but also in burrows-linings that paralleled with the decrease of organic P in SL and SSL treatments. The stimulation of AcPA began quickly and declined rapidly in casts (from 19 to 8 μmol phenol g⁻¹ dry wt h⁻¹, respectively at week 2 and 8 in the SL treatment) but it was initiated later and maintained at a high level for longer in burrows (more than 10 μmol phenol g⁻¹ dry wt h⁻¹ at week 8 in the SL treatment). Significant positive correlations were found between the AkPA activities and N_tot contents (r=0.95, p=0.001) and to a lesser extend with C_org contents (r=0.76, p=0.05) in casts from the SL treatment, while AcPA significantly correlated with N_tot (r=0.91, p=0.004) but not with C_org (r=0.72, p=0.06). P availability was always highest in casts. However, the available P contents decreased sharply over time in casts and were still low in burrow-linings, suggesting that a large part of inorganic P produced was rapidly immobilized for the microbial growth. Total P content was unchanged except in the SL treatment in which it increased in casts and burrows (ca. 725 μg g⁻¹, at week 4). Organic P was first the highest in casts and then decreased over time (from 168 at week 1 to 140 μg g⁻¹ at week 8 in the SL treatment). This study illustrates that earthworms facilitate P transfer downward increasing a P patchy distribution in the soil, and significantly change the biogeochemical status of P (availability, organic phosphorous pool, AcPA activities) in certain hot spots such as casts and burrow-linings.     

Dynamics of the nitrous oxide reducing community during adaptation to Zn stress in soil

Laboratory studies show that the nitrous oxide (N₂O) reduction rate in soil is strongly inhibited by trace metal contamination; however, this effect appears transient. Here we assess if this recovery is due to microbial adaptation associated with shifts in community composition. Soils were spiked with zinc chloride (0-5000 mg Zn kg⁻¹) in a factorial design with 3 application rates of organic matter (OM), i.e. 0, 2 and 4 g milled hay kg⁻¹, to accelerate growth and, potentially, adaptation rate. The soil treatments were incubated outdoors with free drainage during 1 year and periodically sampled. The potential N₂O reduction rate, measured in an anaerobic laboratory assay, was inhibited by Zn during the first 2 months after spiking with 50% inhibition at 500-1000 mg Zn kg⁻¹. After 6 months exposure, the N₂O reduction rate recovered to at least 80% of the rate in the control treatment in the series receiving OM up to the largest Zn dose, but strong inhibition remained in the series which did not receive OM. In this series recovery was only observed after 12 months exposure. Soil pore water Zn concentrations did not explain the recovery of the N₂O reduction rate in the control series suggesting that recovery is due to adaptation and not to reduced Zn bioavailability. The faster recovery in the series receiving OM was partially, but not fully related to the effects of OM on Zn bioavailability. The recovery at all Zn and OM treatments co-varied with a recovery of nosZ gene abundance from about 1 × 10⁷ copies g⁻¹ soil in the soil treatments with decreased activity to 5 × 10⁸ copies g⁻¹ soil in the other soil treatments. The nosZ gene DGGE profile of the soil microbial communities revealed minor changes in the nosZ containing community. This study strongly suggests that the transient effects of trace metal inhibition of N₂O reduction is due to the development of a Zn tolerant denitrifying community.     

Initial recovery of soil carbon and nitrogen pools and dynamics following disturbance in jack pine forests: A comparison of wildfire and clearcut harvesting

Forests naturally maintained by stand-replacing wildfires are often managed with clearcut harvesting, yet we know little about how replacing wildfire with clearcutting affects soil processes and properties. We compared the initial recovery of carbon (C) and nitrogen (N) pools and dynamics following disturbance in jack pine (Pinus banksiana) stands in northern Lower Michigan, USA, by sampling soils (Oa+A horizons) from three “treatments”: 3-6-year-old harvest-regenerated stands, 3-6-year-old wildfire-regenerated stands and 40-55-year-old intact, mature stands (n=4 stands per treatment). We measured total C and N; microbial biomass and potentially mineralizable C and N; net nitrification; and gross rates of N mineralization and nitrification. Burned stands exhibited reduced soil N but not C, whereas clearcut and mature stands had similar quantities of soil organic matter. Both disturbance types reduced microbial biomass C compared to mature stands; however, microbial biomass N was reduced in burned stands but not in clearcut stands. The experimental C and N mineralization values were fit to a first-order rate equation to estimate potentially mineralizable pool size (C₀ and N₀) and rate parameters. Values for C₀ in burned and clearcut stands were approximately half that of the mature treatment, with no difference between disturbance types. In contrast, N₀ was lowest in the wildfire stands (170.2 μg N g⁻¹), intermediate in the clearcuts (215.4 μg N g⁻¹) and highest in the mature stands (244.6 μg N g⁻¹). The most pronounced difference between disturbance types was for net nitrification. These data were fit to a sigmoidal growth equation to estimate potential NO₃⁻ accumulation (Nit_max) and kinetic parameters. Values of Nit_max in clearcut soils exceeded that of wildfire and mature soils (149.2 vs. 83.5 vs. 96.5 μg NO₃⁻-N g⁻¹, respectively). Moreover, the clearcut treatment exhibited no lag period for net NO₃⁻ production, whereas the burned and mature treatments exhibited an approximate 8-week lag period before producing appreciable quantities of NO₃⁻. There were no differences between disturbances in gross rates of mineralization or nitrification; rather, lower NO₃⁻ immobilization rates in the clearcut soils, 0.20 μg NO₃⁻ g⁻¹ d⁻¹ compared to 0.65 in the burned soils, explained the difference in net nitrification. Because the mobility of NO₃⁻ and NH₄⁺ differs markedly in soil, our results suggest that differences in nitrification between wildfire and clearcutting could have important consequences for plant nutrition and leaching losses following disturbance.     

Nitrous oxide flux from soil amino acid mineralization

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

Elevated CO2 concentration and nitrogen fertilisation effects on N2O and CH4 fluxes and biomass production of Phleum pratense on farmed peat soil

The effects of elevated CO₂ supply on N₂O and CH₄ fluxes and biomass production of Phleum pratense were studied in a greenhouse experiment. Three sets of 12 farmed peat soil mesocosms (10 cm dia, 47 cm long) sown with P. pratense and equally distributed in four thermo-controlled greenhouses were fertilised with a commercial fertiliser in order to add 2, 6 or 10 g N m⁻². In two of the greenhouses, CO₂ concentration was kept at atmospheric concentration (360 μmol mol⁻¹) and in the other two at doubled concentration (720 μmol mol⁻¹). Soil temperature was kept at 15 °C and air temperature at 20 °C. Natural lighting was supported by artificial light and deionized water was used to regulate soil moisture. Forage was harvested and the plants fertilised three times during the basic experiment, followed by an extra fertilisations and harvests. At the end of the experiment CH₄ production and CH₄ oxidation potentials were determined; roots were collected and the biomass was determined. From the three first harvests the amount of total N in the aboveground biomass was determined. N₂O and CH₄ exchange was monitored using a closed chamber technique and a gas chromatograph. The highest N₂O fluxes (on average, 255 μg N₂O m⁻² h⁻¹ during period IV) occurred just after fertilisation at high water contents, and especially at the beginning of the growing season (on average, 490 μg N₂O m⁻² h⁻¹ during period I) when the competition of vegetation for N was low. CH₄ fluxes were negligible throughout the experiment, and for all treatments the production and oxidation potentials of CH₄ were inconsequential. Especially at the highest rates of fertilisation, the elevated supply of CO₂ increased above- and below-ground biomass production, but both at the highest and lowest rates of fertilisation, decreased the total amount of N in the aboveground dry biomass. N₂O fluxes tended to be higher under doubled CO₂ concentrations, indicating that increasing atmospheric CO₂ concentration may affect N and C dynamics in farmed peat soil.     

Effects of C and N availability and soil-water potential interactions on N2O evolution and PLFA composition

Mitigation of agricultural N₂O emissions via management requires quantitative information about the regulation of the underlying processes. In this laboratory study, short-term evolution of N₂O from repacked soil was determined using an arable sandy loam soil adjusted to three water potentials (−15, −30 or −100 hPa) that were reached by adjustment of partly air-dried soil with nutrient solutions or water; a water retention curve of repacked soil had been determined prior to the incubation experiment. The amendments included a control treatment receiving water (CTL), and aqueous solutions of carbon in the form of glucose (C), ammonium sulfate (N), or both (CN). Rates of CO₂ and N₂O evolution were followed during 14 days. Soil inorganic N and phospholipid fatty acid (PLFA) composition were analyzed by the end of incubation. Across all nutrient treatments, the soil at the lower moisture content (−100 hPa) showed little or no N₂O evolution irrespective of nutrient treatment. Adding glucose alone reduced N₂O evolution relative to CTL. The addition of N alone had no effect on soil respiration, but significantly increased nitrate accumulation and N₂O evolution. The CN treatment resulted in higher respiration than with C amendment alone, but less N₂O evolution than with N alone, at least at −15 and −30 hPa. Whole-soil PLFA fingerprints at the end of incubation reflected the complex response of gaseous emissions. At −15 hPa growth of Gram negative bacteria, probably including denitrifiers, in the CN treatment was indicated by low cyclopropane-to-precursor ratios. At −100 hPa differentiation of branched-chain fatty acids was taken as evidence for an effect of C amendment on Gram positive bacteria. The highest potential for N₂O evolution was observed at the intermediate soil wetness level; the corresponding gas diffusivities indicated that this parameter may be a better predictor of N₂O emissions than water-filled pore space.     

Microbial activity and bacterial composition of H2-treated soils with net CO2 fixation

The effects of H₂ gas treatment of an agricultural soil cultivated previously with a mixture of clover (Trifolium pratense) and alfalfa (Medicago sativa) on CO₂ dynamics and microbial activity and composition were analyzed. The H₂ emission rate of 250 nmol H₂ g⁻¹ soil h⁻¹ was similar to the upper limit of estimated H₂ amounts emitted from N₂ fixing nodules into the surrounding soil ([Dong, Z., Layzell, D.B., 2001. H₂ oxidation, O₂ uptake and CO₂ fixation in hydrogen treated soil. Plant and Soil 229, 1-12.]). After 1 week of H₂ supply to soil samples simultaneously with H₂ uptake net CO₂ production declined continuously and this finally led to a net CO₂ fixation rate in the H₂-treated soil of 8 nmol CO₂ g⁻¹ soil h⁻¹. The time course of H₂ uptake and CO₂ fixation in the soils corresponded with an increase in microbial activity and biomass of the H₂-treated soil determined by microcalorimetric measurements, fluorescence in situ hybridization analysis (FISH) and DNA staining (DAPI). Shifts in the bacterial community structure caused by the supply of H₂ were recorded. While the H₂ treatment stimulated β-and γ-subclasses of Proteobacteria, it had no significant effect on α-Proteobacteria. In addition, FISH-detectable bacteria of the Cytophaga-Flavobacterium-Bacteroides phylum increased in numbers.     

Seasonal variation and fire effects on CH4, N2O and CO2 exchange in savanna soils of northern Australia

Tropical savanna ecosystems are a major contributor to global CO₂, CH₄ and N₂O greenhouse gas exchange. Savanna fire events represent large, discrete C emissions but the importance of ongoing soil-atmosphere gas exchange is less well understood. Seasonal rainfall and fire events are likely to impact upon savanna soil microbial processes involved in N₂O and CH₄ exchange. We measured soil CO₂, CH₄ and N₂O fluxes in savanna woodland (Eucalyptus tetrodonta/Eucalyptus miniata trees above sorghum grass) at Howard Springs, Australia over a 16 month period from October 2007 to January 2009 using manual chambers and a field-based gas chromatograph connected to automated chambers. The effect of fire on soil gas exchange was investigated through two controlled burns and protected unburnt areas. Fire is a frequent natural and management action in these savanna (every 1-2 years). There was no seasonal change and no fire effect upon soil N₂O exchange. Soil N₂O fluxes were very low, generally between −1.0 and 1.0 μg N m⁻² h⁻¹, and often below the minimum detection limit. There was an increase in soil NH₄⁺ in the months after the 2008 fire event, but no change in soil NO₃⁻. There was considerable nitrification in the early wet season but minimal nitrification at all other times.Savanna soil was generally a net CH₄ sink that equated to between −2.0 and −1.6 kg CH₄ ha⁻¹ y⁻¹ with no clear seasonal pattern in response to changing soil moisture conditions. Irrigation in the dry season significantly reduced soil gas diffusion and as a consequence soil CH₄ uptake. There were short periods of soil CH₄ emission, up to 20 μg C m⁻² h⁻¹, likely to have been caused by termite activity in, or beneath, automated chambers. Soil CO₂ fluxes showed a strong bimodal seasonal pattern, increasing fivefold from the dry into the wet season. Soil moisture showed a weak relationship with soil CH₄ fluxes, but a much stronger relationship with soil CO₂ fluxes, explaining up to 70% of the variation in unburnt treatments. Australian savanna soils are a small N₂O source, and possibly even a sink. Annual soil CH₄ flux measurements suggest that the 1.9 million km² of Australian savanna soils may provide a C sink of between −7.7 and −9.4 Tg CO₂-e per year. This sink estimate would offset potentially 10% of Australian transport related CO₂-e emissions. This CH₄ sink estimate does not include concurrent CH₄ emissions from termite mounds or ephemeral wetlands in Australian savannas.     

The production of N2O in Douglas fir litter as affected by anoxic conditions within litter particles and pores

The development of anoxic conditions in forest litter and the relation with nitrous oxide (N₂O) production and emission rates are not completely understood. Water content is an important factor in the regulation of N₂O production due to its effect on the development of anoxic conditions. A combination of simulation modeling and incubation experiments was used to study (1) O₂ concentrations in water and organic matter at various water saturation fractions of inter-particle pores in Douglas fir litter (F2 horizon), (2) the relationship between N₂O production and moisture content of litter and (3) to test whether diffusion constraints of nitrate (NO₃⁻) could have explained measured N₂O production rates within litter fragments. Model simulations showed that the occurrence of high N₂O production rates in samples with extremely high water contents coincided with the development of anoxic conditions in water-filled inter-particle pores. Measured N₂O production rates started to increase exponentially after 1-2 days in glucose-amended samples, during which substantial microbial growth was established. For these latter samples model simulations showed that the increase in O₂ consumption due to microbial growth lead to anoxic conditions in water-filled pores at locations which were far from the O₂ saturated air-filled pores. It was concluded that anoxic conditions in water-filled pores was the crucial factor for the development of high N₂O production rates. Diffusion limitation of NO₃⁻ and glucose were estimated to be negligible in the highly fragmented litter material used. The occurrence of diffusion limitation depended on litter particle size, the NO₃⁻ reduction potential and the NO₃⁻ concentration. Therefore, diffusion limitation together with N₂O production in litter cannot be neglected under field conditions with a low NO₃⁻ concentration or a high NO₃⁻ reduction potential.     

Seasonal changes in the spatial structures of N2O, CO2, and CH4 fluxes from Acacia mangium plantation soils in Indonesia

We evaluated the spatial structures of nitrous oxide (N₂O), carbon dioxide (CO₂), and methane (CH₄) fluxes in an Acacia mangium plantation stand in Sumatra, Indonesia, in drier (August) and wetter (March) seasons. A 60 × 100-m plot was established in an A. mangium plantation that included different topographical elements of the upper plateau, lower plateau, upper slope and foot slope. The plot was divided into 10 × 10-m grids and gas fluxes and soil properties were measured at 77 grid points at 10-m intervals within the plot. Spatial structures of the gas fluxes and soil properties were identified using geostatistical analyses. Averaged N₂O and CO₂ fluxes in the wetter season (1.85 mg N m⁻² d⁻¹ and 4.29 g C m⁻² d⁻¹, respectively) were significantly higher than those in the drier season (0.55 mg N m⁻² d⁻¹ and 2.73 g C m⁻² d⁻¹, respectively) and averaged CH₄ uptake rates in the drier season (−0.62 mg C m⁻² d⁻¹) were higher than those in the wetter season (−0.24 mg C m⁻² d⁻¹). These values of N₂O fluxes in A. mangium soils were higher than those reported for natural forest soils in Sumatra, while CO₂ and CH₄ fluxes were in the range of fluxes reported for natural forest soils. Seasonal differences in these gas fluxes appears to be controlled by soil water content and substrate availability due to differing precipitation and mineralization of litter between seasons. N₂O fluxes had strong spatial dependence with a range of about 18 m in both the drier and wetter seasons. Topography was associated with the N₂O fluxes in the wetter season with higher and lower fluxes on the foot slope and on the upper plateau, respectively, via controlling the anaerobic-aerobic conditions in the soils. In the drier season, however, we could not find obvious topographic influences on the spatial patterns of N₂O fluxes and they may have depended on litter amount distribution. CO₂ fluxes had no spatial dependence in both seasons, but the topographic influence was significant in the drier season with lowest fluxes on the foot slope, while there was no significant difference between topographic positions in the wetter season. The distributions of litter amount and soil organic matter were possibly associated with CO₂ fluxes through their effects on microbial activities and fine root distribution in this A. mangium plantation.     

The mitigation potential of hippuric acid on N2O emissions from urine patches: An in situ determination of its effect

Previous laboratory studies have demonstrated that hippuric acid, a ruminant urine constituent, can mitigate nitrous oxide (N₂O) emissions from simulated urine patches. Hippuric acid has the potential to be a N₂O mitigation tool because animal diets can be manipulated to adjust its concentration in the urine. This study was conducted to determine if the effect observed in previous laboratory studies would also occur in situ under field conditions. In our field study, plots were treated with unadulterated bovine urine (56 mM hippuric acid), the same bovine urine amended with either benzoic acid (34 mM), dicyandiamide (DCD) or varying rates of hippuric acid (up to 90 mM). Soil inorganic-N, N₂O fluxes, and plant N responses were monitored over a 78 d period. Effects on microbial communities were monitored by determining the size and structure of nitrite oxidizer (nxrA) and nitrite reducer (nirS) bacterial populations using real-time PCR and denaturing gradient gel electrophoresis (DGGE), respectively. Decreases in N₂O emissions, with increasing hippuric and benzoic acid concentrations, were only seen on Day two of the trial. With the exception of the DCD treatment (0.60% of N applied) the amended urine treatments did not significantly affect emissions of N₂O as a percentage of N applied (1.28-1.65%). Soil inorganic-N and plant response were not affected by urinary amendment, except in the DCD treatment where nitrification inhibition occurred. Nitrite oxidizer community structures shifted and increased approximately 5.4-fold in size over 48 d in response to urine, although no specific response to elevated hippuric acid or benzoic acid was observed. No treatment effects were observed on community structure of the nitrite reducing bacteria but averaged over time the highest rate of hippuric acid significantly decreased nirS gene copy numbers g⁻¹ soil. We concluded that under the conditions of this field study, increasing hippuric or benzoic acid concentrations in bovine urine had no effect on N₂O mitigation in situ. We argue that the discrepancy with previous laboratory studies may be related to differences in soil pH, microbial communities and the presence of vegetation. Further research is needed to determine the potential for hippuric acid as a tool to mitigate N₂O emissions, and its effect(s) on resident N cycling microorganisms.     

Changes in the fungal-to-bacterial respiratory ratio and microbial biomass in agriculturally managed soils under free-air CO2 enrichment (FACE) - A six-year survey of a field study

In soil ecology, microbial parameters have been identified as sensitive indicators of changes in the soil environment. The Braunschweig FACE project provided the opportunity to study the effects of elevated CO₂ (550 μmol mol⁻¹) as compared to ambient CO₂ (370 μmol mol⁻¹) on total microbial biomass (C_mic), C_mic-to-C_org ratio and the fungal-to-bacterial respiratory ratio together with total C_org, N_t, C:N ratio and pH over a six-year period. Field management followed a typical crop rotation system of this region with either a crop-related full nitrogen supply (N100) or 50% reduced N supply (N50). The soil microbial parameters responded to the elevated CO₂ treatment in varying intensities and time spans. The fungal-to-bacterial respiratory ratio was the most sensitive parameter in responding to an elevated CO₂ treatment with highly significant differences to ambient CO₂-treated control plots in the third year of CO₂ fumigation. After six years bacterial respiratory activity had increased in ascending order to 34% in FACE-treated plots (N50 and N100) as compared to control plots. Soil microbial biomass (C_mic) responded more slowly to the FACE treatment with highly significant increases of >12% after the fourth year of CO₂ fumigation. The C_mic-to-C_org ratio responded very late in the last two years of the CO₂ treatment with a significant increase of >7.0% only in the N100 variant. Total C_org and N_t were slightly but significantly increased under FACE around 10.0% with ascending tendency over time starting with the second year of CO₂ treatment. No significant FACE effects could be recorded for the C:N ratio or pH.These results suggest that under FACE treatment changes in the soil microbial community will occur. In our study the fungal-to-bacterial respiratory ratio was superior to total C_mic as microbial bioindicators in reflecting changes in the soil organic matter composition.