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     

Nitrapyrin effectiveness in reducing nitrous oxide emissions decreases at low doses of urea in an Andosol

In the tropics,frequent nitrogen(N)fertilization of grazing areas can potentially increase nitrous oxide(N₂O)emissions.The application of nitrification inhibitors has been reported as an effective management practice for potentially reducing N loss from the soil-plant system and improving N use efficiency(NUE).The aim of this study was to determine the effect of the co-application of nitrapyrin(a nitrification inhibitor,NI)and urea in a tropical Andosol on the behavior of N and the emissions of N₂O from autotrophic and heterotrophic nitrification.A greenhouse experiment was performed using a soil(pH 5.9,organic matter content 78 g kg^-1,and N 5.6 g kg^-1)sown with Cynodon nlemfuensis at 60%water-filled pore space to quantify total N₂O emissions,N₂O derived from fertilizer,soil ammonium(NH₄⁺)and nitrate(NO₃^-),and NUE.The study included treatments that received deionized water only(control,NI).No significant differences were observed in soil NH₄⁺content between the UR and UR+NI treatments,probably because of soil mineralization and NO₃^-produced by heterotrophic nitrification,which is not effectively inhibited by nitrapyrin.After 56 d,N₂O emissions in UR(0.51±0.12 mg N₂O-N concluded that the soil organic N mineralization and heterotrophic nitrification are the main processes of NH₄⁺and NO₃^-production.Additionally,it was found that N₂O emissions were partially a consequence of the direct oxidation of the soil's organic N via heterotrophic nitrification coupled to denitrification.Finally,the results suggest that nitrapyrin would likely exert significant mitigation on N₂O emissions only if a substantial N surplus exists in soils with high organic matter content.     

Relationships between soil moisture content and nitrous oxide production during nitrification and denitrification

Summary The effect of soil water content [60%–100% water-holding capacity (WHC)] on N₂O production during autotrophic nitrification and denitrification in a loam soil was studied in a laboratory experiment by selectively inhibiting nitrification with a low C₂H₂ concentration (2.1 Pa). Nitrifiers usually produced more N₂O than denitrifiers. During an initial experimental period of 0–6 days the nitrifiers produced more N₂O than the denitrifiers by a factor ranging from 1.4 to 16.5, depending on the water content and length of incubation. The highest N₂O production rate by nitrifiers was observed at 90% WHC, when the soil had become partly anaerobic, as indicated by the high denitrification rate. At 100% WHC there were large gaseous losses from denitrification, while nitrification losses were smaller except for the first period of measurement, when there was still some O₂ remaining in the soil. The use of 10 kPa C₂H₂ to inhibit reduction of N₂O to N₂ stimulated the denitrification process during prolonged incubation over several days; thus the method is unsuitable for long-term studies.     

Contribution of Heterotrophic Nitrification to Nitrous Oxide Production in a Long-Term N-Fertilized Arable Black Soil

To understand the contribution of key microbial processes to nitrous oxide (N₂O) emission in intensively cultivated black soil, laboratory incubation were conducted at 70% water-holding capacity (WHC) and 25 °C, using different gases (air, oxygen, or argon) within the headspace of the incubation chambers to evaluate gas inhibition effects. Arable black soil was sampled from an experimental field that has received urea since October 1979. Nitrification contributed to 57% of total N₂O emission, of which as much as 67% resulted from heterotrophic nitrification. These data strongly suggest that high soil organic carbon concentrations and low pH values are more favorable to N₂O production through heterotrophic, rather than autotrophic, nitrification. Nitrous oxide produced by denitrification accounted for 28% of the total N₂O emission, and the nitrifier denitrification accounted for 15% of the N₂O emitted from the tested soil. These findings indicate that heterotrophic nitrification was the primary N₂O production process in the tested soil.     

Contributions of nitrification and denitrification to N2O emissions from soils at different water-filled pore space

A combination of stable isotope and acetylene (0.01% v/v) inhibition techniques were used for the first time to determine N₂O production during denitrification, autotrophic nitrification and heterotrophic nitrification in a fertilised (200 kg N ha^–1) silt loam soil at contrasting (20–70%) water-filled pore space (WFPS). ¹⁵N-N₂O emissions from ¹⁴NH₄¹⁵NO₃ replicates were attributed to denitrification and ¹⁵N-N₂O from ¹⁵NH₄¹⁵NO₃ minus that from ¹⁴NH₄¹⁵NO₃ replicates was attributed to nitrification and heterotrophic nitrification in the presence of acetylene, as there was no dissimilatory nitrate reduction to ammonium or immobilisation and remineralisation of ¹⁵N-NO₃^–. All of the N₂O emitted at 70% WFPS (31.6 mg N₂O-N m^–2 over 24 days; 1.12 g N₂O-N g dry soil^–1; 0.16% of N applied) was produced during denitrification, but at 35–60% WFPS nitrification was the main process producing N₂O, accounting for 81% of ¹⁵N-N₂O emitted at 60% WFPS, and 7.9 g ¹⁵N-N₂O m^–2 (0.28 ng ¹⁵N-N₂O g dry soil^–1) was estimated to be emitted over 7 days during heterotrophic nitrification in the 50% WFPS treatment and accounted for 20% of ¹⁵N-N₂O from this treatment. Denitrification was the predominant N₂O-producing process at 20% WFPS (2.6 g ¹⁵N-N₂O m^–2 over 7 days; 0.09 ng ¹⁵N-N₂O g dry soil^–1; 85% of ¹⁵N-N₂O from this treatment) and may have been due to the occurrence of aerobic denitrification at this WFPS. Our results demonstrate the usefulness of a combined stable isotope and acetylene approach to quantify N₂O emissions from different processes and to show that several processes may contribute to N₂O emission from agricultural soils depending on soil WFPS.     

Nitrogen transformation rates and N<Subscript>2</Subscript>O producing pathways in two pasture soils

Purpose

Better understanding of N transformations and the regulation of N₂O-related N transformation processes in pasture soil contributes significantly to N fertilizer management and development of targeted mitigation strategies.

Materials and methods

¹⁵N tracer technique combined with acetylene (C₂H₂) method was used to measure gross N transformation rates and to distinguish pathways of N₂O production in two Australian pasture soils. The soils were collected from Glenormiston (GN) and Terang (TR), Victoria, Australia, and incubated at a soil moisture content of 60% water-filled pore space (WFPS) and at temperature of 20 °C.

Results and discussion

Two tested pasture soils were characterized by high mineralization and immobilization turnover. The average gross N nitrification rate (n_tot) was 7.28 mg N kg^?1 day^?1 in TR soil () and 5.79 mg N kg^?1 day^?1 in GN soil. Heterotrophic nitrification rates (n_h), which accounting for 50.8 and 41.9% of n_tot, and 23.4 and 30.1% of N₂O emissions in GN and TR soils, respectively, played a role similar with autotrophic nitrification in total nitrification and N₂O emission. Denitrification rates in two pasture soils were as low as 0.003–0.004 mg N kg^?1 day^?1 under selected conditions but contributed more than 30% of N₂O emissions.

Conclusions

Results demonstrated that two tested pasture soils were characterized by fast N transformation rates of mineralization, immobilization, and nitrification. Heterotrophic nitrification could be an important NO₃^?–N production transformation process in studied pasture soils. Except for autotrophic nitrification, roles of heterotrophic nitrification and denitrification in N₂O emission in two pasture soils should be considered when developing mitigation strategies.

     

Heterotrophic nitrification is the predominant NO3 − production pathway in acid coniferous forest soil in subtropical China

To date, occurrence and stimulation of different nitrification pathways in acidic soils remains unclear. Laboratory incubation experiments, using the acetylene inhibition and ¹⁵N tracing methods, were conducted to study the relative importance of heterotrophic and autotrophic nitrification in two acid soils (arable (AR) and coniferous forest) in subtropical China, and to verify the reliability of the ¹⁵N tracing model. The gross rate of autotrophic nitrification was 2.28 mg?kg^?1?day^?1, while that of the heterotrophic nitrification (0.01 mg?kg^?1?day^?1) was negligible in the AR soil. On the contrary, the gross rate of autotrophic nitrification was very low (0.05 mg?kg^?1?day^?1) and the heterotrophic nitrification (0.98 mg?kg^?1?day^?1) was the predominant NO₃ ^? production pathway accounting for more than 95 % of the total nitrification in the coniferous forest soil. Our results showed that the ¹⁵N tracing model was reliable when used to study soil N transformation in acid subtropical soils.     

Nitrification Inhibitor Reduces Nitrous Oxide Production from Different Soil Profiles of an Andosol Soil

A laboratory incubation was conducted to evaluate nitrous oxide (N₂O) production during nitrification and the effect of a nitrification inhibitor on N₂O production from different profiles in a Japanese orchard Andosol. Soils were collected from five profiles: A1, A2, Bw1, Bw2, and BC. The soils were treated with ammonium sulfate at the rate of 200 mg N kg^?1 with or without dicyandiamide (DCD) and incubated under aerobic conditions for 32 days. The net nitrification rate without DCD during the first 8 days was greater in the surface soils than in the subsurface soils. Accordingly, the surface soils showed a greater cumulative N₂O production than the subsurface soils. Application of DCD significantly reduced the nitrification rate and thus N₂O production from any depths of soils by 33.8 to 62.9%. Our study showed that substantial N₂O was produced from the subsurface soil, although the amount was less than from the surface soils.     

Isotopologue ratios of N2O emitted from microcosms with NH4 fertilized arable soils under conditions favoring nitrification

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 determine variability and control of the isotopic fingerprint of N₂O fluxes following NH₄⁺-fertilization and dominated by nitrification. We conducted a microcosm study with three arable soils fertilized with 0–140 mg NH₄⁺–N kg⁻¹. Fractions of N₂O derived from nitrification and denitrification were determined in parallel experiments using the ¹⁵N tracer and acetylene inhibition techniques or by comparison with unfertilized treatments. Soils were incubated for 3–10 days at low moisture (30–55% water-filled pore space) in order to establish conditions favoring nitrification. 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). N₂O originated mainly from nitrification (>80%) in all treatments and the proportion of NH₄⁺ nitrified that was lost as N₂O ranged between 0.07 and 0.45%. δ¹⁸O and SP of N₂O fluxes ranged from 15 to 28.4‰ and from 13.9 to 29.8‰, respectively. These ranges overlapped with isotopic signatures of N₂O from denitrification reported previously. There was a negative correlation between SP and δ¹⁸O which is opposite to reported trends in N₂O from denitrification. Variation of average ¹⁵N signatures of N₂O (δ¹⁵N^bulk) did not supply process information, apparently because a strong shift in precursor signatures masked process-specific effects on δ¹⁵N^bulk. Maximum SP of total N₂O fluxes and of nitrification fluxes was close to reported SP of N₂O from NH₄⁺ or NH₂OH conversion by autotrophic nitrifiers, suggesting that SP close to 30‰ is typical for autotrophic nitrification in soils following NH₄⁺-fertilization. The results suggest that the δ¹⁸O/SP fingerprint of N₂O might be used as a new indicator of the dominant source process of N₂O fluxes in soils.     

N2O emissions from different cropping systems and from aerated,nitrifying and denitrifying tanks of a municipal waste water treatment plant

Nitrous oxide emissions, nitrate, water-soluble carbon and biological O₂ demand (BOD₅) were quantified in different cropping systems fertilized with varying amounts of nitrogen (clayey loam, October 1991 to May 1992), in an aerated tank (March 1993 to March 1994), and in the nitrification-denitrification unit (March to July 1994) of a municipal waste water treatment plant. In addition, the N₂O present in the soil body at different depths was determined (February to July 1994). N₂O was emitted by all cropping systems (mean releases 0.13–0.35 mg N₂O m^-2 h^-1), and all the units of the domestic waste water treatment plant (aerated tank 0–6.2 mg N₂O m^-2 h^-1, nitrification tank 0–204,3 mg N₂O m^-2, h^-1, denitrifying unit 0–2.2 mg N₂O m^-2 h^-1). During the N₂O-sampling periods estimated amounts of 0.9, 1.5, 2.4 and 1.4 kg N₂O–N ha^-1, respectively, were released by the cropping systems. The aerated, nitrifying and denitrifying tanks of the municipal waste water treatment plant released mean amounts of 9.1, 71.6 and 1.8 g N₂O–N m^-2, respectively, during the sampling periods.The N₂O emission were significantly positively correlated with nitrate concentrations in the field plots which received no N fertilizer and with the nitrogen content of the aerated sludge tank that received almost exclusively N in the form of NH ₄ ⁺ . Available carbon, in contrast, was significantly negatively correlated with the N₂O emitted in the soil fertilized with 80 kg N ha^-1 year. The significant negative correlation between the emitted N₂O and the carbon to nitrate ratio indicates that the lower the carbon to nitrate ratio the higher the amount of N₂O released. Increasing N₂O emissions seem to occur at electron donorto-acceptor ratios (C_H2_O or BOD₅-to-nitrate ratios) below 50 in the cropping systems and below 1200–1400 in the waste water treatment plant. The trapped N₂O in the soil body down to a depth of 90 cm demonstrates that agricultural production systems seem to contain a considerable pool of N₂O which may be reduced to N₂ on its way to the atmosphere, which may be transported to other environments or which may be released at sometime in the future.Dedicated to Professor J.C.G. Ottow on the occasion of his 60th birthday     

Denitrification and nitrogen immobilization as affected by organic matter and different forms of nitrogen added to an anaerobic water-sediment system

The effects of wheat straw and different forms of N on denitrification and N immobilization were studied in an anaerobic water-sediment system. The water-sediment system was supplemented with various combinations of wheat straw and ¹⁵N-labelled and unlabelled (NH₄)₂SO₄ or KNO₃, and incubated anaerobically at 30°C for 10 days. ¹⁵N-labelled and unlabelled NO _inf3 ^sup- , NO _inf2 ^sup- , NH _inf4 ^sup+ , and organic N were determined in the water-sediment system. The gases evolved (N₂, CO₂, N₂O, and CH₄) were analyzed by gas chromatography at regular intervals. Larger quantities of ¹⁵N₂–N and organic ¹⁵N were formed in wheat straw-amended systems than in non-amended systems. Trends in CO₂ production were similar to those of N₂–N evolution. The evolution of N₂O and CH₄ was negligible. Denitrification processes accounted for about 22 and 71% of the added ¹⁵NO _inf3 ^sup- –N in the absence and presence of wheat straw, respectively. The corresponding denitrification rates were 3.4 and 12.4 g ¹⁵Ng^-1 dry sediment day^-1. In systems amended with ¹⁵NO _inf3 ^sup- –N and ¹⁵NO _inf3 ^sup- +NH _inf4 ^sup+ –N without wheat straw, 1.82 and 1.58%, respectively, of the added ¹⁵NH _inf3 ^sup- –N was immobilized. The corresponding figures for the same systems supplemented with wheat straw were 5.08 and 4.10%, respectively. Immobilization of ¹⁵NO _inf4 ^sup+ –N was higher than that of ¹⁵NO _inf3 ^sup- –N. The presence of NO _inf3 ^sup- –N did not stimulate NH _inf4 ^sup+ –N immobilization.     

Enhanced denitrification in plots of N2-fixing faba beans compared to plots of a non-fixing legume and non-legumes

Denitrification rates were studied using the C₂H₂ inhibition technique in a 2-year field experiment within plots of nodulated and non-nodulated faba beans, ryegrass, and cabbage. Denitrification rates ranged from 14.40 to 0.02 ng N₂O–N g^–1 soil dry weight h^–1. Mean denitrification increased fourfold in plots of N₂–fixing Vicia faba compared to non-nodulated V. faba mutant F48, Lolium perenne, and Brassica oleracea. The results with and without C₂H₂ treatment indicate that in the field the major part of this enhanced denitrification led to the endproduct N₂ rather than to the ozone-degrading N₂O. Higher denitrification rates of plots with N₂–fixing plants in September seemed to be caused by an increase in soil NO _inf3 ^sup- of about 20 kg ha^–1 found between July and August. Soil NO _inf3 ^sup- and soil moisture explained 67% of the variation in denitrification rates of the different soil samples over the growing seasons in the 2 years. Soil moisture explained 44% of the variation for soil planted with N₂–fixing plants and 62% for soil planted with non-fixing plants. Positive exponential relationships were obtained between denitrification rates and soil nitrate (r=0.71) and soil moisture (r=0.82).     

Nitrogen turnover rates in a riparian fen determined by 15N dilution

Summary Gross rates of N mineralization, assimilation, nitrification, and NO _in3 ^sup- reduction were determined in soil from a wet riparian fen by 1-day incubations of soil cores and slurries with ¹⁵N-labelled substrates. N mineralization transformed 0.1% of the total organic N pool daily in the soil cores, of which 25% was oxidized through autotrophic nitrification and 53%–70% was incorporated into microorganisms. N mineralization and nitrification were markedly inhibited below 5 cm in soil depth. At least 80% of the NO _in3 ^sup- reduction in aerated cores occurred through dissimilatory processes. Dissimilatory reduction to NH _in4 ^sup+ (DNRA) occurred only below 5 cm in depth. The results show that NH _in4 ^sup+ oxidation was limited by available substrate and was itself a strong regulator of NO _in3 ^sup- -reducing activity. NO _in3 ^sup- reduction was significantly increased when the soil was suspended under anaerobiosis; adding glucose to the soil slurries increased NO _in3 ^sup- reduction by 2.4–3.7 times. Between 3% and 9% (net) of the added NO _in3 ^sup- was reduced through DNRA in the soil slurries. The highest percentage was observed in soil samples from deeper layers that were pre-incubated anaerobically.     

Effect of the inhibitors nitrapyrin and sodium chlorate on nitrification and N2O formation in an acid forest soil

An acid forest soil from beech forest gaps, which were either limed or unlimed, and the undisturbed forest was investigated for the type of nitrifying populations and the process of N₂O evolution. To see whether nitrifiers were of heterotrophic or autotrophic origin, the nitrification inhibitors nitrapyrin and sodium chlorate were applied to disturbed soil samples which underwent laboratory incubations. Nitrapyrin inhibits autotrophic nitrification. In different studies, sodium chlorate has been identified as an inhibitor either of autotrophic or of heterotrophic nitrification. In the samples investigated only nitrapyrin inhibited the autotrophic nitrification occurring in the limed soil. Sodium chlorate effectively inhibited heterotrophic nitrification. In the limed forest floor samples, where most autotrophic nitrification occured, sodium chlorate showed no inhibitory effect. In another laboratory incubation experiment, N₂O evolution from undisturbed soil columns, to which the above inhibitors were applied, was investigated. In those samples, in which nitrification had been reduced, neither inhibitor significantly reduced N₂O evolution. Thus it was concluded that the contribution of nitrification to N₂O losses is negligible, and that N₂O evolution arises from the activity of denitrifying organisms. Microbial biomass and respiration measurements showed that the inhibitors did not affect microflora negatively.     

Temporal and spatial patterns of denitrification enzyme activity and nitrous oxide fluxes in three adjacent vegetated riparian buffer zones

The aim of this study was to investigate temporal and spatial patterns of denitrification enzyme activity (DEA) and nitrous oxide (N₂O) fluxes in three adjacent riparian sites (mixed vegetation, forest and grass). The highest DEA was found in the surface (0–30 cm) soil and varied between 0.7±0.1 mg N kg^–1 day^–1 at 5°C and 5.9±0.4 mg N kg^–1 day^–1 at 15°C. There was no significant difference (P >0.05) between the DEA in the uppermost (0–30 cm and 60–90 cm) soil depths under different vegetation covers. In the two deepest (120–150 cm and 180–210 cm) soil depths the DEA varied between 0.0±0.0 mg N kg^–1 day^–1 at 5°C and 4.4±0.9 mg N kg^–1 day^–1 at 15°C and was clearly associated with the accumulation of buried organic carbon (OC). Two threshold values of OC were observed before DEA started to increase significantly, namely 5 and 25 g OC kg^–1 soil at 10–15°C and 5°C, respectively. In the three riparian sites N₂O fluxes varied between a net N₂O uptake of –0.6±0.4 mg N₂O-N m^–2 day^–1 and a net N₂O emission of 2.5±0.3 mg N₂O-N m^–2 day^–1. The observed N₂O emission did not lead to an important pollution swapping (from water pollution to greenhouse gas emission). Especially in the mixed vegetation and forest riparian site highest N₂O fluxes were observed upslope of the riparian site. The N₂O fluxes showed no clear temporal trend.     

亚热带松林中强酸性土壤自养和异养硝化作用的研究

The occurrence of nitrification in some acidic forest soils is still a subject of debate. Identification of main nitrification pathways in acidic forest soils is still largely unknown. Acidic yellow soil (Oxisol) samples were selected to test whether nitrification can occur or not in acidic subtropical pine forest ecosystems. Relative contributions of autotrophs and heterotrophs to nitrification were studied by adding selective nitrification inhibitor nitrapyrin. Soil NH₄⁺-N concentrations decreased, but NO₃^--N concentrations increased significantly for the no-nitrapyrin control during the first week of incubation, indicating that nitrification did occur in the acidic subtropical soil. The calculated net nitrification rate was 0.49 mg N kg^-1 d^-1 for the no-nitrapyrin control during the first week of incubation. Nitrapyrin amendment resulted in a significant reduction of NO₃^--N concentration. Autotrophic nitrification rate averaged 0.28 mg N kg^-1 d^-1 and the heterotrophic nitrification rate was 0.21 mg N kg^-1 d^-1 in the first week. Ammonia-oxidizing bacteria (AOB) abundance increased slightly during incubation, but nitrapyrin amendment significantly decreased AOB amoA gene copy numbers by about 80%. However, the ammonia-oxidizing archaea (AOA) abundance showed significant increases only in the last 2 weeks of incubation and it was also decreased by nitrapyrin amendment. Our results indicated that nitrification did occur in the present acidic subtropical pine forest soil, and autotrophic nitrification was the main nitrification pathway. Both AOA and AOB were the active biotic agents responsible for autotrophic nitrification in the acidic subtropical pine forest soil.     

Conversion of forest to agricultural land affects the relative contribution of bacteria and fungi to nitrification in humid subtropical soils

Land-use and management practices can affect soil nitrification. However, nitrifying microorganisms responsible for specific nitrification process under different land-use soils remains unknown. Thus, we investigated the relative contribution of bacteria and fungi to specific soil nitrification in different land-use soils (coniferous forest, upland fields planted with corn and rice paddy) in humid subtropical region in China. ¹⁵N dilution technique in combination with selective biomass inhibitors and C₂H₂ inhibition method were used to estimate the relative contribution of bacteria and fungi to heterotrophic nitrification and autotrophic nitrification in the different land-use soils in humid subtropical region. The results showed that autotrophic nitrification was the predominant nitrification process in the two agricultural soils (upland and paddy), while the nitrate production was mainly from heterotrophic nitrification in the acid forest soil. In the upland soils, streptomycin reduced autotrophic nitrification by 94%, whereas cycloheximide had no effect on autotrophic nitrification, indicating that autotrophic nitrification was mainly driven by bacteria. However, the opposite was true in another agricultural soil (paddy), indicating that fungi contributed to the oxidation of NH₄⁺ to NO₃^?. In the acid forest soil, cycloheximide, but not streptomycin, inhibited heterotrophic nitrification, demonstrating that fungi controlled the heterotrophic nitrification. The conversion of forest to agricultural soils resulted in a shift from fungi-dominated heterotrophic nitrification to bacteria- or fungi-dominated autotrophic nitrification. Our results suggest that land-use and management practices, such as the application of N fertilizer and lime, the long-term waterflooding during rice growth, straw return after harvest, and cultivation could markedly influence the relative contribution of bacteria and fungi to specific soil nitrification processes.     

Interactive effects of ammonium application rates and temperature on nitrous oxide emission from tropical agricultural soil

Emissions of nitrous oxide (N₂O), a potent greenhouse gas, from agricultural soil have been recognized to be affected by nitrogen (N) application and temperature. Most of the previous studies were carried out to determine effects of temperature on N₂O emissions at a fixed N application rate or those of N application rates at a specific temperature. Knowledge about the effects of different ammonium (NH₄⁺) application rates and temperatures on N₂O emissions from tropical agricultural soil and their interactions is limited. Five grams of air-dried sandy loam soil, collected in Central Vietnam, were adjusted to 0, 400, 800 and 1200 mg NH₄-N kg^–1 soil (abbreviated as 0 N, 400 N, 800 N and 1200 N, respectively) at 60% water holding capacity were aerobically incubated at 20°C, 25°C, 30°C or 35°C for 28 days. Mineral N contents and N₂O emission rates were determined on days 1, 3, 5, 7, 14, 21 and 28. Cumulative N₂O emissions for 28 days increased with increasing NH₄⁺ application rates from 0 to 800 mg N kg^–1 and then declined to 1200 mg N kg^–1. Cumulative N₂O emissions increased in the order of 35°C, 20°C, 30°C and 25°C. This lowest emission at 35°C occurred because N₂O production was derived only from autotrophic nitrification while other N₂O production processes, e.g., nitrifier denitrification and coupled nitrification-denitrification occurred at lower temperatures. More specifically, cumulative N₂O emissions peaked at 800 N and 25°C, and the lowest emissions occurred at 1200 N and 35°C. In conclusion, N₂O emissions were not exponentially correlated with NH₄⁺ application rates or temperatures. Higher NH₄⁺ application rates at higher temperatures suppressed N₂O emissions.     

The effect of mixing organic biological waste materials and high-N crop residues on the short-time N2O emission from horticultural soil in model experiments

Manipulating the N release from high-N crop residues by simultaneous mixing of these residues with organic biological waste (OBW) materials seems to be a possible method to reduce NO₃^– leaching. The aim of this study was to examine whether the incorporation of OBW materials together with a high-N crop residue (celery) had also an effect on N₂O emission from horticultural soil under short-term and optimised laboratory conditions. A sandy loam soil and celery residues were mixed with different OBW materials and brought into PVC tubes at 80% water-filled pore space and 15°C. Every 2.5 h, a gas sample was taken and analysed by gas chromatography for its N₂O concentration. The soil amended with only celery residues had a cumulative N₂O emission of 9.6 mg N kg^–1 soil in 50 h. When the celery residues were mixed with an OBW material, the N₂O emission was each time lower than the emission from the celery-only treatment (between 3.8 and 5.9 mg N kg^–1 soil during maximum 77 h), except with paper sludge (17.2 mg N kg^–1 soil in 100 h). The higher N₂O emission from the paper sludge treatment was probably due to its unusually low C:N ratio. Straw, green waste compost 1 (GWC1) and 2 (GWC2), saw dust, and tannic acid reduced the N₂O emission of the celery treatment by 40 to 60%. Although the N₂O reduction potential can be expected to be lower and with differing dynamics under field conditions, this study indicates that apart from reducing NO₃^– leaching, OBW application may at the same time reduce N₂O emissions after incorporation of high-N crop residues.     

Processes responsible for the nitrous oxide emission from a Costa Rican Andosol under a coffee agroforestry plantation

We used the inhibitor acetylene (C₂H₂) at partial pressures of 10 Pa and 10 kPa to inhibit autotrophic nitrification and the reduction of nitrous oxide (N₂O) to N₂, respectively. Soils (Andosol) from a Coffea arabica plantation shaded by Inga densiflora in Costa Rica were adjusted to 39, 58, 76 and 87% water-filled pore space (WFPS) and incubated for 6 days in the absence or presence of C₂H₂. Soil respiration, nitrification rates and N₂O emissions by both processes were measured in relation to soil moisture conditions. At all WFPS studied, rates of N₂O and N₂ productions were small (4.8; 14.7; 23 and 239.6 ng N–N₂O g⁻¹ d.w. d⁻¹ at 39, 58, 76 and 87% WFPS, respectively), and despite a low soil pH (4.7), N₂O was mainly produced by nitrification, which was responsible for 85, 91, 84 and 87% of the total N₂O emissions at 39, 58, 76 and 87% WFPS, respectively. At the three smaller values of WFPS, a linear relationship was established between WFPS, soil respiration, nitrification and N₂O released by nitrification; no N₂ was produced by denitrification. At more anaerobic conditions achieved by a WFPS of 87%, a large rate of N₂O production was measured during nitrification, and N₂ production accounted for 84% of the gaseous N fluxes caused by denitrification.