Response of denitrifying communities to successive soil freeze–thaw cycles

The effect of soil freeze–thaw cycles on the denitrification potential was examined based on the C₂H₂ inhibition method. The gross N₂O production curve of the soil sample (incubation with C₂H₂) showed minor changes between the freeze–thaw treatment and the unfrozen control. However, kinetics analysis revealed that the initial production rate, an indicator of the population density of denitrifying communities, decreased (P = 0.043) and the specific growth rate constant, an indicator of the activity of denitrifying communities, increased (P = 0.039) as a result of the freeze–thaw cycles in five of six soil samples examined. The increase in the specific growth rate constant suggested the stimulation of the activity of denitrifying communities that survived after the freeze–thaw cycles and may explain the minor suppression on the gross N₂O production in spite of decreasing the population density of denitrifying communities that was suggested by the initial production rate. The net N₂O production curve of the soil sample (incubation without C₂H₂) showed a remarkable change in one out of six soil samples, and in that one soil sample, N₂O release to the atmosphere was largely stimulated (7.6 times) by the freeze–thaw cycles. However, the stimulation of the N₂O release by the freeze–thaw cycles was even observed in two other selected soil samples (4.6 and 1.8 times), suggesting that an imbalance in the N₂O-producing and N₂O-reducing activities of denitrifying communities might complementally explain the N₂O release stimulated by the freeze–thaw cycles.     

Nitrogen mineralization,N<Subscript>2</Subscript>O production and soil microbiological properties as affected by long-term applications of sewage sludge composts

A field study was conducted to investigate the long-term effect of surface application of sewage sludge composts vs chemical N fertilizer on total N, total C, soluble organic C, pH, EC, microbial biomass C and N, protease activity, deaminase activity, urease activity, gross and net rates of N mineralization and nitrification, CO₂ evolution, and N₂O production. Soil samples were taken from five depths (0–15, 15–20, 20–30, 30–40, and 40–50 cm) of a long-term experiment at the University of Tokyo, Japan. Three fields have been receiving sewage sludge composted with rice husk (RH), sawdust (SD), or mixed chemical fertilizer NPK (CF), applied at the rate of 240 kg N ha^–1 each in split applications in summer and autumn since 1978. Significantly higher amounts of total N and C and soluble organic C were found in the compost than in the CF treatments up to the 40-cm soil depth, indicating improved soil quality in the former. In the CF treatment, soil pH values were significantly lower and electrical conductivity values were significantly higher than those of compost-treated soils of up to 50 cm depth. Soil microbial biomass C and N, CO₂ evolution, protease, deaminase, and urease activities were significantly higher in the compost than in the CF treatments due to greater availability of organic substrates that stimulated microbial activity. Gross N mineralization rates determined by ¹⁵N dilution technique were eight and five times higher in the SD and RH treatments than in the CF treatment, respectively, probably due to high levels of microbial and enzyme activities. Net N mineralization rates were also significantly higher in the compost treatments and were negative in the CF treatment indicating immobilization. Net nitrification rates were higher in compost treatments and negative in the CF treatment. Nitrous oxide productions from compost treatments were higher than the CF treatment due to the greater availability of mineral N as a result of higher mineralization and nitrification rates and soluble organic C in the former. Most of the measured parameters were highest in the surface soil (0–15 cm) and were significantly higher in the SD treatment than in the RH treatment.     

Nitrate removal from drained and reflooded fen soils affected by soil N transformation processes and plant uptake

Knowledge about nitrate transformation processes and how they are affected by different plants is essential in order to reduce the loss of valuable N fertiliser as well as to prevent environmental pollution due to nitrate leaching or N₂O emission after fertilisation or the reflooding of degraded fens with nitrate-containing municipal sewage. Therefore four microcosm ¹⁵N tracer experiments were performed to evaluate the effect of common wetland plants (Phalaris arundinacea, Phragmites australis) combined with different soil moisture conditions (from dry to reflooded) on nitrate turnover processes. At the end of experiment, the total formation of gaseous N compounds was calculated using the ¹⁵N balance method. In two experiments (wet and reflooded soil conditions) the N₂O and N₂ emissions were also directly determined.Our results show that in degraded fen soils, which process mainly takes place—denitrification or transformation into organic N compounds—is determined by the soil moisture conditions. Under dry soil moisture conditions (water filled pore space: 31%) up to 80% of the ¹⁵N nitrate added was transformed into organic N compounds. This transformation process is not affected by plant growth. Under reflooded conditions (water filled pore space: 100%), the total gaseous N losses were highest (77-95% of the ¹⁵N-nitrate added) and the transformation into organic N compounds was very low (1.8% of ¹⁵N nitrate added). Under almost all soil conditions plant growth reduced the N losses by 20-25% of the ¹⁵N nitrate added due to plant uptake. The N₂ emissions exceeded the N₂O emissions by a factor of 10-20 in planted soil, and as much as 30 in unplanted soil. In the treatments planted with Phragmites australis, N₂O emission was about two times higher than in the corresponding unplanted treatment. 15% of the N₂O and N₂ formed was transported via the Phragmites shoots from the soil into the atmosphere. By contrast, Phalaris arundinacea did not affect N₂O emissions and no emission via the shoots was observed.     

Microbial biomass and C mineralization in agricultural soils as affected by atrazine addition

This study examines the effects of atrazine on both microbial biomass C and C mineralization dynamics in two contrasting agricultural soils (organic C, texture, and atrazine application history) located at Galicia (NW Spain). Atrazine was added to soils, a Humic Cambisol (H) and a Gleyic Cambisol (G), at a recommended agronomic dose and C mineralization (CO₂ evolved), and microbial biomass measurements were made in non-treated and atrazine-treated samples at different time intervals during a 12-week aerobic incubation. The cumulative curves of CO₂–C evolved over time fit the simple first-order kinetic model [Ct = Co (1 − e ^−kt )], whose kinetic parameters were quantified. Differences in these parameters were observed between the two soils studied; the G soil, with a higher content in organic matter and microbial biomass C and lower atrazine application history, exhibited higher values of the total C mineralization and the potentially mineralizable labile C pool than those for the H soil. The addition of atrazine modified the kinetic parameters and increased notably the C mineralized; by the end of the incubation the cumulative CO₂–C values were 33–41% higher than those in the corresponding non-added soils. In contrast, a variable effect or even no effect was observed on the soil microbial biomass following atrazine addition. The data clearly showed that atrazine application at normal agricultural rates may have important implications in the C cycling of these two contrasting acid soils.     

Microbiological characterization and nitrate reduction in subsurface soils

Summary Two borings (20 m depth) were performed in a sandy-clayey soil over a limestone bed and in a sandy soil with lumps of clay in some depths. Bacteria were found in the deeper soil layers of both profiles. The methods used to detect bacteria were those normally used for topsoil layers, plate counts of bacteria, ATP content, and direct microscopy. Measurements of CO₂ evolution showed that the potential for bacterial activity was present in all depths of the two profiles. However, the activity was strongly dependent on the presence of easily available organic C. An indication of the denitrification potential was obtained by measuring the N₂O evolution. Under aerobic incubation without the addition of glucose, N₂O was detected only in the topsoil. When glucose was added to the soil samples, N₂O was found at a low level in the deeper soil layers. Under anaerobic incubation, N₂O was detected in all deeper layers, and increased markedly when glucose was added to the soil samples.     

CO2, CH4 and N2O fluxes of upland black spruce (Picea mariana) forest soils after forest fires of different intensity in interior Alaska

Abstract

Forest fires can change the greenhouse gase (GHG) flux of borea forest soils. We measured carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O) fluxes with different burn histories in black spruce (Picea mariana) stands in interior Alaska. The control forest (CF) burned in 1920; partially burned (PB) in 1999; and severely burned (SB1 and SB2) in 2004. The thickness of the organic layer was 22 ± 6 cm at CF, 28 ± 10 cm at PB, 12 ± 6 cm at SB1 and 4 ± 2 cm at SB2. The mean soil temperature during CO₂ flux measurement was 8.9 ± 3.1, 6.4 ± 2.1, 5.9 ± 3.4 and 5.0 ± 2.4°C at SB2, SB1, PB and CF, respectively, and differed significantly among the sites (P < 0.01). The mean CO₂ flux was highest at PB (128 ± 85 mg CO₂-C m^?2 h^?1) and lowest at SB1 (47 ± 19 mg CO₂-C m^?2 h^?1) (P < 0.01), and within each site it was positively correlated with soil temperature (P < 0.01). The CO₂ flux at SB2 was lower than that at CF when the soil temperature was high. We attributed the low CO₂ flux at SB1 and SB2 to low root respiration and organic matter decomposition rates due to the 2004 fire. The CH₄ uptake rate was highest at SB1 [–91 ± 21 μg CH₄-C m^?2 h^?1] (P < 0.01) and positively correlated with soil temperature (P < 0.01) but not soil moisture. The CH₄ uptake rate increased with increasing soil temperature because methanotroph activity increased. The N₂O flux was highest [3.6 ± 4.7 μg N₂O-N m^?2 h^?1] at PB (P < 0.01). Our findings suggest that the soil temperature and moisture are important factors of GHG dynamics in forest soils with different fire history.     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Nitrous oxide emissions from agricultural soils at low temperatures: a laboratory microcosm study

We studied in laboratory microcosms (intact soil cores) N₂O and CO₂ emissions from four different agricultural soil types (organic soil, clay, silt and loam) at low temperatures with or without freezing-thawing events. When the temperature of the frozen soil cores was increased stepwise from −8 °C the N₂O emissions began to increase at −0.5 °C, and peaked at −0.1 °C in the organic, clay and silt soils, and at +1.6 °C in the loam soils. However, a stepwise decrease in soil temperature from +15 °C also induced an increase in the N₂O emissions close to the 0 °C. These emissions peaked between −0.4 and +2.5 °C depending on the soil type and water content. However, the emission maxima were from 2 to 14.3% of those encountered in the experiments where frozen soils were thawed. Our results show that in addition to the well-documented thawing peak, soils also can have a maximum in their N₂O emission near 0 °C when soil temperature decrease. These emissions, however, are less than those emitted from thawing soils. The correlations between the N₂O and CO₂ emissions were weak. Our results suggest that N₂O is produced in soils down to a temperature of −6 °C.     

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.     

Nitrous oxide production and consumption in serially diluted soil suspensions as related to in situ N2O emission in submerged soils

A simple method for characterizing soil microbial community composition relevant to N₂O production and consumption was proposed. Ten-fold series soil dilution was prepared. Nitrate or N₂O was provided as the sole electron acceptor. Nitrous oxide concentration in the headspace gas across the serially diluted soil suspensions was measured against controls. Results showed that the patterns of N₂O production and consumption across the soil suspensions provided useful information on the microbial community composition relevant to N₂O production and consumption in these soils. An independent method, to that proposed here, was also employed to characterize denitrifier community compositions of the same soils. Data indicated that information on the soil microbial community composition characterized by both methods were compatible or mutually supporting and apparently related to in situ N₂O emissions. Soil samples from manure (applied with animal manure plus chemical fertilizer) plots had higher denitrification rates than the samples from normal fertilizer (applied with chemical fertilizer only) plots. It was concluded that functional characteristics of soil microbial communities relevant to N₂O production and consumption could be characterized at ecological levels and may potentially affect N₂O emissions.     

Net ecosystem carbon exchange for Bermuda grass growing in mesocosms as affected by irrigation frequency

Intensification of grazed grasslands following conversion from dryland to irrigated farming has the potential to alter ecosystem carbon (C) cycling and affect components of carbon dioxide (CO₂) exchange that could lead to either net accumulation or loss of soil C. While there are many studies on the effect of water availability on biomass production and soil C stocks, much less is known about the effect of the frequency of water inputs on the components of CO₂ exchange. We grew Bermuda grass (Cynodon dactylon L.) in mesocosms under irrigation frequencies of every day (I₁ treatment, 30 d), every two days (I₂ treatment, 12 d), every three days (I₃ treatment, 30 d), and every six days (I₆ treatment, 18 d, after I₂ treatment). Rates of CO₂ exchange for estimating net ecosystem CO₂ exchange (F_N), ecosystem respiration (R_E), and soil respiration (R_S) were measured, and gross C uptake by plants (F_G) and respiration from leaves (R_L) were calculated during two periods, 1–12 and 13–30 d, of the 30-d experiment. During the first 12 d, there were no significant differences in cumulative F_N (mean ±standard deviation, 61 ±30 g C m^-2, n = 4). During the subsequent 18 d, cumulative F_N decreased with decreasing irrigation frequency and increasing cumulative soil water deficit (W), with values of 70 ±22, 60 ±16, and 18 ±12 g C m^-2 for the I₁, I₃, and I₆ treatments, respectively. There were similar decreases in F_G, R_E, and R_L with increasing W, but differences in R_S were not significant. Use of the C₄ grass growing in a C₃-derived soil enabled partitioning of R_S into its autotrophic (R_A) and heterotrophic (R_H) components using a ¹³C natural abundance isotopic technique at the end of the experiment when differences in cumulative W between the treatments were the greatest. The values of R_H and its percentage contributions to R_S (43% ±8%, 42% ±8%, and 8% ±5% for the I₁, I₃, and I₆ treatments, respectively) suggested that R_H remained unaffected across a wide range of W and then decreased under extreme W. There were no significant differences in aboveground biomass between the treatments. Nitrous oxide (N₂O) emission was measured to determine if there was a trade-off effect between irrigation frequency and increasing W on net greenhouse gas emission, but no significant differences were found between the treatments. These findings suggest that over short periods in well-drained soil, irrigation frequency could be managed to manipulate soil water deficit in order to reduce net belowground respiratory C losses, particularly those from the microbial decomposition of soil organic matter, with no significant effect on biomass production and N₂O emission.     

N2O emissions and product ratios of nitrification and denitrification as affected by freezing and thawing

Agricultural soils contribute significantly to atmospheric nitrous oxide (N₂O). A considerable part of the annual N₂O emission may occur during the cold season, possibly supported by high product ratios in denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O-N/(NO₃⁻-N+NO₂⁻-N)) at low temperatures and/or in response to freeze-thaw perturbation. Water-soluble organic materials released from frost-sensitive catch crops and green manure may further increase winter emissions. We conducted short-term laboratory incubations under standardized moisture and oxygen (O₂) conditions, using nitrogen (N) tracers (¹⁵N) to determine process rates and sources of emitted N₂O after freeze-thaw treatment of soil or after addition of freeze-thaw extract from clover. Soil respiration and N₂O production was stimulated by freeze-thaw or addition of plant extract. The N₂O emission response was inversely related to O₂ concentration, indicating denitrification as the quantitatively prevailing process. Denitrification product ratios in the two studied soils (pH 4.5 and 7.0) remained largely unaltered by freeze-thaw or freeze-thaw-released plant material, refuting the hypothesis that high winter emissions are due to frost damage of N₂O reductase activity. Nitrification rates estimated by nitrate (NO₃⁻) pool enrichment were 1.5-1.8 μg NO₃-N g⁻¹ dw soil d⁻¹ in freeze-thaw-treated soil when incubated at O₂ concentrations above 2.3 vol% and one order of magnitude lower at 0.8 vol% O₂. Thus, the experiments captured a situation with severely O₂-limited nitrification. As expected, the O₂ stress at 0.8 vol% resulted in a high nitrification product ratio (0.3 g g⁻¹). Despite this high product ratio, only 4.4% of the measured N₂O accumulation originated from nitrification, reaffirming that denitrification was the main N₂O source at the various tested O₂ concentrations in freeze-thaw-affected soil. N₂O emission response to both freeze-thaw and plant extract addition appeared strongly linked to stimulation of carbon (C) respiration, suggesting that freeze-thaw-induced release of decomposable organic C was the major driving force for N₂O emissions in our soils, both by fuelling denitrifiers and by depleting O₂. The soluble C (applied as plant extract) necessary to induce a CO₂ and N₂O production rate comparable with that of freeze-thaw was 20-30 μg C g⁻¹ soil dw. This is in the range of estimates for over-winter soluble C loss from catch crops and green manure plots reported in the literature. Thus, freeze-thaw-released organic C from plants may play a significant role in freeze-thaw-related N₂O emissions.     

Microbial biomass and enzyme activities in coniferous forest soils as affected by lignite-derived deposition

A forest ecosystem study was conducted along a deposition gradient of air pollutants in old Scots pine stands located near the industrial belt around the city of Bitterfeld in northeast Germany from 1999 to 2000. In order to estimate the impact of different atmospheric deposition loads on microbial biomass and enzyme activities, samples were taken from the forest floor (L, F, and H horizon) and the mineral topsoil (0–10 cm). The emission-induced increases in ferromagnetic susceptibility, soil pH, concentrations of mobile (NH ₄NO ₃ extractable) Cr and Ni, effective cation exchange capacity, and base saturation in the humus layer along the 25-km long transect reflected that great portions of the past depositions were characterized by alkaline fly ash. Alkaline depositions significantly ( P <0.05) decreased the microbial biomass C and N contents, microbial biomass C-to-organic C ratios, and microbial respiration rates, but increased the metabolic quotient (qCO ₂) of the mineral topsoil and forest floor. Variations in microbial biomass and activity can mainly be predicted ( r ² =0.60) by the concentrations of Ca, Zn and Cd in these forest soils. The specific activities (activity kg ^-1 organic C) of l-asparaginase, l-glutaminase, arylsulfatase, and in part, acid phosphatase were significantly ( P <0.05) higher at forest sites receiving higher fly ash loads than those of the other sites, and thus followed the trend of the qCO ₂. In contrast, the specific activity of ß-glucosidase was significantly ( P <0.05) decreased at heavily affected sites compared to moderate and less affected sites, suggesting an inhibition of C mineralization in the forest floor of pine stands affected by predominantly alkaline emissions. A great portion ( r ²=0.91) of the variation in the specific enzyme activity data in forest soils in emission areas can be predicted from a linear combination of the variables total organic C and NH ₄Cl-extractable Ca, pH and effective cation exchange capacity.     

Microbial respiration and biomass (substrate-induced respiration) in soils of old-growth and regenerating forests on northern Vancouver Island,British Columbia

In studying the basal respiration, microbial biomass (substrate-induced respiration, SIR), and metabolic quotient (qCO₂) in western red cedar (Thuja plicata Donn ex D. Don)-western hemlock [(Tsuga heterophylla Raf.) Sarg.] ecosystems (old-growth forests, 3- and 10-year-old plantations) on northern Vancouver Island, British Columbia, Canada, we predicted that (1) soil basal respiration would be reduced by harvesting and burning, reflecting the reduction in microbial biomass and activities; (2) the microbial biomass would be reduced by harvesting and slash-burning, due to the excessive heat of the burning or due to reduced substrate availability; (3) microbial biomass in the plantations would tend to recover to the preharvesting levels with growth of the trees and increased substrate availability; and (4) microbial biomass measured by the SIR method would compare well with that measured by the fumigation-extraction (FE) method. Decaying litter layer (F), woody F (Fw) and humus layer (H) materials were sampled four times in the summer of 1992. The results obtained supported the four predictions. Microbial biomass was reduced in the harvested and slash-burned plots. Both SIR and FE methods provided equally good estimates of microbial biomass in the samples [SIR microbial C (mg g^-1)=0.227+0.458 FE microbial C (mg g^-1), r=0.63, P=0.0001] and proved suitable for microbial biomass measurements in this strongly acidic soil. Basal respiration was significantly greater in the old-growth forests than in the young plantations (P<0.05) in both F and H layers, but not in the Fw layer. For the 3- and 10-year-old plantations, there was no difference in basal respiration in F, Fw, and H layers. Basal respiration was related to changes in air temperature, precipitation, and the soil moisture contant at the time of sampling. The qCO₂ values were higher in the old-growth stands than in the plantations. Clear-cutting followed by prescribed burning did not increase soil microbial respiration, but CO₂ released from slash-burning and that contributed from other sources may be of concern to increasing atmospheric CO₂ concentrations.     

Microbial activities in forest soils exposed to chronic depositions from a lignite power plant

Atmospheric emissions of fly ash and SO₂ from lignite-fired power plants strongly affect large forest areas in Germany. The impact of different deposition loads on the microbial biomass and enzyme activities was studied at three forest sites (Picea abies (L.) Karst.) along an emission gradient of 3, 6, and 15 km downwind of a coal-fired power plant (sites Ia, II, and III, respectively), representing high, moderate and low emission rates. An additional site (site Ib) at a distance of 3 km from the power plant was chosen to study the influence of forest type on microbial parameters in coniferous forest soils under fly ash and SO₂ emissions. Soil microbial biomass C and N, CO₂ evolved and activities of l-asparaginase, l-glutaminase, β -glucosidase, acid phosphatase and arylsulfatase (expressed on dry soil and organic C basis) were determined in the forest floor (L, Of and Oh horizon) and mineral top soil (0-10 cm). The emission-induced increases in ferromagnetic susceptibility, soil pH, concentrations of mobile (NH₄NO₃ extractable) Cd, Cr, and Ni, effective cation exchange capacity and base saturation in the humus layer along the 15 km long transect significantly (P<0.05) reflected the effect of past depositions of alkaline fly ash. Soil microbial and biochemical parameters were significantly (P<0.05) affected by chronic fly ash depositions. The effect of forest type (i.e. comparison of sites Ia and Ib) on the studied parameters was generally dominated by the deposition effect. Alkaline depositions significantly (P<0.05) decreased the microbial biomass C and N, microbial biomass C-to-N ratios and microbial biomass C-to-organic C ratios. Microbial respiration, metabolic quotient (qCO₂) and the activities of l-asparaginase, l-glutaminase, β-glucosidase, acid phosphatase and arylsulfatase were increased by long-term depositions from the power plants. Acid phosphatase had the highest specific (enzyme activities expressed per unit organic C) activity values among the enzymes studied and arylsulfatase the lowest. The responses of the microbial biomass and soil respiration data to different atmospheric deposition loads were mainly controlled by the content of organic C and cation exchange capacity, while those of enzyme activities were governed by the soil pH and concentrations of mobile heavy metals. We concluded that chronic fly ash depositions decrease litter decomposition by influencing specific microbial and enzymatic processes in forest soils.     

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     

Microbial biomass and activity indices after organic substrate addition to a selenium-contaminated soil

High concentrations of Se in soil might have negative effects on microorganisms. For this reason, the effect of organic substrate addition (glucose + maize straw) on Se volatilisation in relation to changes in microbial biomass and activity indices was investigated using an artificially Se-contaminated soil. Microbial biomass N was reduced on average by more than 50% after substrate addition, but adenylate energy charge (AEC) and metabolic quotient qCO₂ were both increased. The Se content decreased by nearly 30% only with the addition of the organic substrate at 25°C. No significant Se loss occurred without substrate at 25°C or with substrate at 5°C. In the two treatments with substrate addition, the substrate-derived CO₂ evolution was about 30% lower with Se addition than without. In contrast, Se had no effect on any of the other soil microbial indices analysed, i.e. microbial biomass C, microbial biomass N, adenosine triphosphate (ATP), AEC, ATP-to-microbial biomass C, and qCO₂.     

Fluxes and production pathways of nitrous oxide in different types of tropical forest soils in Thailand

Nitrous oxide (N₂O) emissions from the soil surface of five different forest types in Thailand were measured using the closed chamber method. Soil samples were also taken to study the N₂O production pathways. The monthly average emissions (±SD, n?=?12) of N₂O from dry evergreen forest (DEF), hill evergreen forest (HEF), moist evergreen forest (MEF), mixed deciduous forest (MDF) and acacia reforestation (ARF) were 13.0?±?8.2, 5.7?±?7.1, 1.2?±?12.1, 7.3?±?8.5 and 16.7?±?9.2?µg N m^?2 h^?1, respectively. Large seasonal variations in fluxes were observed. Emission was relatively higher during the wet season than during the dry season, indicating that soil moisture and denitrification were probably the main controlling factors. Net N₂O uptake was also observed occasionally. Laboratory studies were conducted to further investigate the influence of moisture and the N₂O production pathways. Production rates at 30% water holding capacity (WHC) were 3.9?±?0.2, 0.5?±?0.06 and 0.87?±?0.01?ng N₂O-nitrogen (N) g-dw^?1day^?1 in DEF, HEF and MEF respectively. At 60% WHC, N₂O production rates in DEF, HEF and MEF soils increased by factors of 68, 9 and 502, respectively. Denitrification was found to be the main N₂O production pathway in these soils except in MEF.     

Denitrification potential in a grassland subsoil: effect of carbon substrates

We examined the potential of a subsoil to denitrify nitrate under optimal anaerobic conditions in a laboratory-based incubation when supplied with a range of C substrates of increasing recalcitrance. Both topsoil and its associated subsoil were supplied with nitrate and either glucose, starch or cellulose. Microbial respiration and the evolution of N₂O and N₂ were measured. The subsoil supported low amounts of microbial activity and responded only to the glucose treatment; with less than one-fifth of the N₂O production measured in the top soil. Overall, our findings demonstrated that the denitrification potential of this particular subsoil is relatively low and that only simple carbohydrates could be utilised readily by the resident microorganisms.     

Elevated CO2 differentially alters belowground plant and soil microbial community structure in reed canary grass-invaded experimental wetlands

Several recent studies have indicated that an enriched atmosphere of carbon dioxide (CO₂) could exacerbate the intensity of plant invasions within natural ecosystems, but little is known of how rising CO₂ impacts the belowground characteristics of these invaded systems. In this study, we examined the effects of elevated CO₂ and nitrogen (N) inputs on plant and soil microbial community characteristics of plant communities invaded by reed canary grass, Phalaris arundinacea L. We grew the invasive grass under two levels of invasion: the invader was either dominant (high invasion) at >90% plant cover or sub-dominant (low invasion) at <50% plant cover. Experimental wetland communities were grown for four months in greenhouses that received either 600 or 365 μl l⁻¹ (ambient) CO₂. Within each of three replicate rooms per CO₂ treatment, the plant communities were grown under high (30 mg l⁻¹) or low (5 mg l⁻¹) N. In contrast to what is often predicted under N limitation, we found that elevated CO₂ increased native graminoid biomass at low N, but not at high N. The aboveground biomass of reed canary grass did not respond to elevated CO₂, despite it being a fast-growing C3 species. Although elevated CO₂ had no impact on the plant biomass of heavily invaded communities, the relative abundance of several soil microbial indicators increased. In contrast, the moderately invaded plant communities displayed increased total root biomass under elevated CO_2, while little impact occurred on the relative abundance of soil microbial indicators. Principal components analysis indicated that overall soil microbial community structure was distinct by CO₂ level for the varying N and invasion treatments. This study demonstrates that even when elevated CO₂ does not have visible effects on aboveground plant biomass, it can have large impacts belowground.