Nitrous oxide emission as affected by alternate anaerobic and aerobic conditions from soil suspensions enriched with ammonium sulfate

The effect of several anaerobic and aerobic cycles of varying duration on N₂O emission and labelled N loss was investigated in (¹⁵NH₄)₂SO₄ amended soil suspensions. No N₂O was evolved from the continuously-anaerobic treatment. The continuously-aerobic treatment produced approximately 0.8 μg N₂O-N g^?1 dry soil in 56 days. Alternate anaerobic-aerobic cycles increased the net N₂O evolution with 7.2 μg N₂O-N g^?1 dry soil produced in 56 days from the 7-day anaerobic, 7-day aerobic treatment. The net N₂O evolution increased further when the duration of the anaerobic and aerobic periods was increased from 7-7 days to 14-14 days (15.7μg N₂O-N g^?1 dry soil in 56 days), although the total ¹⁵N loss from the system was approximately the same for the two treatments. The results of this study show that N₂O evolution from soils is likely to be greater under fluctuating moisture conditions than under either continuously well-aerated conditions, or continuously excess-moisture conditions.     

Determination of kC and kNin situ for calibration of the chloroform fumigation-incubation method

¹⁴C-labelled glucose and ¹⁵N-labelled KNO₃ were added to soil and the microbial biomass during 42 days' incubation was estimated using the chloroform fumigation-incubation method (CFIM). By day 1, most of the glucose (1577 μgCg^?1 soil) was metabolized and 110 μg NO^?₃-Ng^?1 soil were immobilized. In situ values for the proportions of biomass C (k_C) and biomass N (k_N) mineralized during the 10 days after CHCl₃ fumigation were determined on the basis that the immobilized labelled C and N remaining in the soil at this time were present as living microbial cells and their associated metabolites. The tracer data indicated that biomass C could be calculated by applying a k_c value of 0.41 to the CO₂-C evolved from the fumigated sample without subtraction of an unfumigated “control”. Biomass N was estimated from the net NH₄^?-N accumulation during the fumigation-incubation. The problem of reimmobilization of NH⁺₄-N where organisms of wide C:N ratio occur was overcome by adjusting the value of k_N according to the ratio of CO₂-C evolved: net NH₄⁺-N accumulated during the fumigation-incubation (C_F:N_F).A C_F:N_F ratio of 6:1 resulted in a k_N of 0.30 whereas a ratio of 13:1 indicated a k_N of 0.20.     

Heterotrophic N2-fixation in arid soil crusts

The cryptogamic soil crusts of the Great Basin Artemisia, Ceratoides, and Atriplex plant communities contain a significant heterotrophic N₂-fixing microbial population in addition to the predominating filamentous cyanobacteria. The bacterial association with the cyanobacteria exhibits a phycosphere-like effect. Heterotrophically fixed N gains reached 17.5 μg N· g^?1 of soil (23.1% increase above the initial soil N content) and 45.9 μg N·g^?1 of soil (57.4% increase) after 3 and 5 weeks, respectively. (NH₄)₂SO₄ and native plant material amendments to soil resulted in a 41–100% reduction in N₂-fixation. The potential input of N to soil crusts may be reduced in the presence of shrub-produced allelochemic agents and by concurrent denitrification.     

Chloroform fumigation and the release of soil nitrogen: The effects of fumigation time and temperature

Fumigation with CHC1₃ (24 h, 25°C) increased the amount of NH₄-N and total N extracted by 0.5 M K₂SO₄ from two soils (one arable, one grassland). The amount of N released by CHC1₃ increased with the duration of fumigation up to 5 days, when it levelled off. Between about 10–34% of the total N released by CHC1₃ was in the form of NH₄-N, the proportion increasing with duration of exposure.When a grassland soil that had received a field application of ¹⁵N-labelled fertilizer 1 yr previously was fumigated, the N released by CHC1₃ was 4 times more heavily labelled than the soil N as a whole. Prolonging the exposure of this soil to CHC1₃ increased the amount of total N released, but hardly altered the proportion of labelled N in the CHC1₃-released N, suggesting that N is being released from a single soil fraction. The most likely soil fraction is the soil microbial biomass. It is suggested that CHC1₃ does not alter the K₂SO₄-extractability of soil-N fractions other than microbial N and that the extra N released by CHC1₃ and extracted by K₂SO₄ gives a direct measure of soil microbial biomass N.In contrast to fumigation done at lower temperatures, less total N was released by soil fumigated at 60°C, or above, than was released from unfumigated soil held at the same temperature. The greater release of N in the non-fumigated soils above 60°C could have been due to soil enzymic processes which were inhibited by CHC1₃ in the fumigated soil.     

Effect of ammonium-, nitrite- and nitrate nitrogen on anaerobic nitrogenase activity in soil

Sandy loam soil, with added glucose, was incubated anaerobically under N₂ and subjected to repeated 1-h C₂H₂ reduction assays. In the presence of 1% glucose the addition of 50 μg NH₄⁺ ?N/g or of 20 μg NO^?₃ N/g (untreated soil contained 1.2 μg NH⁺₄?N and 7.10 μg NO^?₃-N/g) caused at least some suppression of nitrogenase activity. Activity developed when the KCl-extractable soil inorganic nitrogen concentration dropped below 35 μg/g. In the presence of 0.1 or 0.05% glucose the addition of 5 μg NH⁺₄?N/g caused some suppression of nitrogenase activity. However, activity developed when the soil NH₄⁺-N concentration dropped below about 4 μg/g. With 0.1% glucose and 5 μg added NO^?₂ N/g, activity did not develop until the soil NO^?₂ -N concentration dropped to zero. Added NO^?₃ N was rapidly reduced and denitrified to NO^?₂- N, N₂O-N and NH⁺₄ N and furthermore caused some inhibition of CO₂ evolution. The data from NH₄^?-addition experiments are consistent with a nitrogenase repression/ derepression threshold of 4 and 35μg NH⁺₄-N/g at 0.05 and 1% glucose concentrations, respectively. The data from NO^?₂- and NO^?₃-addition experiments suggest a combination of repression and toxicity effects in the presence of added NO^?₃ N.     

A field study of gross rates of N mineralization and nitrification and their relationships to microbial biomass and enzyme activities in soils treated with dairy effluent and ammonium fertilizer

Abstract. Gross N mineralization and nitrification rates were measured in soils treated with dairy shed effluent (DSE) (i.e. effluent from the dairy milking shed, comprising dung, urine and water) or ammonium fertilizer (NH₄Cl) under field conditions, by injecting ¹⁵N-solution into intact soil cores. The relationships between gross mineralization rate, microbial biomass C and N and extracellular enzyme activities (protease, deaminase and urease) as affected by the application of DSE and NH₄Cl were also determined. During the first 16 days, gross mineralization rate in the DSE treated soil (4.3–6.1 μg N g^?1 soil day^?1) were significantly (P 14;< 14;0.05) higher than those in the NH₄Cl treated soil (2.6–3.4 μg N g^?1 soil day^?1). The higher mineralization rate was probably due to the presence of readily mineralizable organic substrates in the DSE, accompanied by stimulated microbial and extracellular enzyme activities. The stable organic N compounds in the DSE were slow to mineralize and contributed little to the mineral N pool during the period of the experiment. Nitrification rates during the first 16 days were higher in the NH₄Cl treated soil (1.7–1.2 μg N g^?1 soil day^?1) compared to the DSE treated soil (0.97–1.5 μg N g^?1 soil day^?1). Soil microbial biomass C and N and extracellular enzyme activities (protease, deaminase and urease) increased after the application of the DSE due to the organic substrates and nutrients applied, but declined with time, probably because of the exhaustion of the readily available substrates. The NH₄Cl application did not result in any significant increases in microbial biomass C, protease or urease activities due to the lack of carbonaceous materials in the ammonium fertilizer. However, it did increase microbial biomass N and deaminase activity. Significant positive correlations were found between gross N mineralization rate and soil microbial biomass, protease, deaminase and urease activities. Nitrification rate was significantly correlated to biomass N but not to the microbial biomass C or the enzyme activities. Stepwise regression analysis showed that the variations of gross N mineralization rate was best described by the microbial biomass C and N.     

Evidence that fungi can oxidize NH4+ to NO3− in a grassland soil

The contribution of bacteria and fungi to NH₄⁺ and organic N (N_org) oxidation was determined in a grassland soil (pH 6.3) by using the general bacterial inhibitor streptomycin or the fungal inhibitor cycloheximide in a laboratory incubation study at 20°C. Each inhibitor was applied at a rate of 3 mg g^?1 oven‐dry soil. The size and enrichment of the mineral N pools from differentially (NH₄¹⁵NO₃ and ¹⁵NH₄NO₃) and doubly labelled (¹⁵NH₄¹⁵NO₃) NH₄NO₃ were measured at 3, 6, 12, 24, 48, 72, 96 and 120 hours after N addition. Labelled N was applied to each treatment, to supply NH₄⁺‐N and NO₃^?‐N at 3.15 μmol N g^?1 oven‐dry soil. The N treatments were enriched to 60 atom % excess in ¹⁵N and acetate was added at 100 μmol C g^?1 oven‐dry soil, to provide a readily available carbon source. The oxidation rates of NH₄⁺ and N_org were analysed separately for each inhibitor treatment with a ¹⁵N tracing model. In the absence of inhibitors, the rates of NH₄⁺ oxidation and organic N oxidation were 0.0045 μmol N g^?1 hour^?1 and 0.0023 μmol N g^?1 hour^?1, respectively. Streptomycin had no effect on nitrification but cycloheximide inhibited the oxidation of NH₄⁺ by 89% and the oxidation of organic N by more than 30%. The current study provides evidence to suggest that nitrification in grassland soil is carried out by fungi and that they can simultaneously oxidize NH₄⁺ and organic N.     

Short-term bacterial growth,nutrient uptake,and ATP turnover in sterilized,inoculated and C-amended soil: The influence of N availability

Bacteria, Pseudomonas paucimobilis, were inoculated at two concentrations (6.56 × 10⁴ g^?1 and 6.56 × 10⁶g^?1) into sterilized soil amended with 700 μg glucose-C g^?1. Two levels of NH⁺₄-N (11.0μg g^?1 and 81.0 μg g^?1) were used. The subsequent development was followed for three days by measurement of several biological, chemical and physiological parameters.The amount of bacterial biomass-C (μg g^?1 soil) became twice as great in high as in low N treatments, and significantly decreased between 39.5 and 63.5 h for the high inoculum, high N level treatment due to decreasing cell size. By the end of the experiment the cumulative respired carbon was twice as great and more inorganic P was immobilized for high compared to low N treatments and all available NH⁺₄-N was taken up by the final sample time. Soil ATP concentrations were twice as large in high N treatments but the turnover times were twice as long compared to low N systems. The yield coefficient (Y), calculated from respiration and biomass-C values, equalled 0.61 while substrate was plentiful. Nitrogen limitation did not alter the efficiencey with which glucose was transformed into biomass, but rather controlled the total amount of glucose used and biomass produced.     

Symbiosis of Trifolium subterraneum with mycorrhizal fungi and Rhizobium trifolii as affected by ammonium sulphate and nitrification inhibitors

The symbioses between Trifolium subterraneum, mycorrhizal fungi and Rhizohium are affected by (NH₄)₂SO₄ and by the nitrification inhibitors 2-chloro-6 (trichloromethyl) pyridine (N-Serve) and 2-trichloromethyl pyridine (2TMP). At 50 μg · g^?1 soil N-Serve and 2TMP had toxic effects on plant growth, measured as leaf expansion, root length and dry weight. Lower concentrations of N-Serve also produced some toxic symptoms. The addition of (NH₄)₂SO₄ to the soil at 2 and 6 m-equiv NH⁺₄ per pot, resulted in reduced root length and nodulation. Shoot dry weight was reduced at 6 m-equiv NH⁺₄ per pot. In the presence of (NH₄)₂SO₄ the toxic effects of the nitrification inhibitors on plant growth were less.Both nitrification inhibitors reduced development of mycorrhizal entry-points and extent of root colonization (% infection). Percentage infection of the root system was also reduced by (NH₄)₂SO₄. Development of nodules on the lateral roots was increased in the presence of N-Serve at 5 and 15 μ · g^?1. This effect, however, was accompanied by a marked reduction in N₂ase activity. Smaller increases in nodulation were apparent with 2TMP and were associated with variable N₂ase activity.     

Immobilization and mineralization of nitrogen in soils incubated with pig slurry or ammonium sulphate

Nitrogen mineralization and immobilization were investigated in two soils incubated with ammonium sulphate or pig slurry over a range of temperatures and moisture contents. A reduction in the mineralization of soil organic N was observed in soils incubated with 100 μg NH₄⁺-Ng^?1 soil as ammonium sulphate at 30°C but not at lower temperatures. Addition of 100 μg NH₄⁺-N g^?1 soil as pig slurry resulted in a period of nett immobilization lasting up to 30 days at 5°C. Although the length of the immobilization phase was shorter at higher temperatures the total N immobilized was similar. The subsequent rate of mineralization in slurry-treated soils was not significantly greater (P = 0.05) than in untreated soils. There was no evidence of any subsequent increased mineralization arising from the immobilized N or slurry organic N for up to 175 days. The rate of immobilization was found to increase with increasing moisture content, though the period of nett immobilization was shorter, so that the amount of N immobilized was similar over a range of moisture contents from 10 to 40%. Approximately 40% of the NH₄⁺-N in the slurry was immobilized under the incubation conditions used.     

Transformations of nitrogen fertilizers in soil

The effects of ¹⁵N-labelled urea, (NH₄)₂SO₄ and KNO₃ on immobilization, mineralization, nitrification and ammonium fixation were examined under aerobic conditions in an acid tropical soil (pH 4.0) and in a neutral temperate soil (pH 6.8). Urea, (NH₄)₂SO₄ and KNO₃ slightly increased net mineralization of soil organic nitrogen in both soils. There was also an apparent Added Nitrogen Interaction (ANI) i.e. added labelled NH₄-N stood proxy for unlabelled NH₄-N that would otherwise have been immobilized. So far as immobilization and nitrification were concerned, urea and (NH₄)₂SO₄ behaved very similarly in each soil. Immobilization of NO₃-N was negligible in both soils. Some of the added labelled NH₄-N was rapidly fixed, more by the temperate soil than by the tropical soil. This labelled fixed NH₄-N decreased during incubation, in contrast to labelled organic N, which did not decline.     

Sulfur and nitrogen mineralization in soils compared using two incubation techniques

Net mineralization of sulfur and nitrogen was studied in three Canadian Prairie soils using two commonly used incubation methods. In the open system technique, where the soils were leached periodically II.3–11.8 μ g SO^2?₄ -S g^?1 soil was mineralized in 17 weeks. Little mineralization or a net immobilization of sulfur (from 1.4 to 1.3 μ g SO^2?₄-S g^?1 soil) was observed in a closed system where the soils were left undisturbed throughout incubation. Changes in the specific activity of ³⁵S-labelled soil solution sulfate during the closed incubation indicated that mineralization-immobilization processes were occurring simultaneously resulting in minimal net changes in CaCl₂-extractable SO_2?⁴ concentrations. The amounts of mineralized nitrogen (32.6–57.8 μg N g_?1 soil) were found to be independent of the incubation method employed.     

Chloroform fumigation and the release of soil nitrogen: A rapid direct extraction method to measure microbial biomass nitrogen in soil

A new “direct extraction” method for measuring soil microbial biomass nitrogen (biomass N) is described. The new method (fumigation-extraction) is based on CHC1₃ fumigation, followed by immediate extraction with 0.5 M K₂SO₄ and measurement of total N released by CHC1₃ in the soil extracts. The amounts of NH₄-N and total N extracted by K₂SO₄ immediately after fumigation increased with fumigation time up to 5 days. Total N released by CHC1₃ after 1 day fumigation (1 day CHC1₃-N) and after 5 days fumigation (5 day CHC1₃-N) were positively correlated with the flush of mineral N (F_N) in 37 soils that had been fumigated, the fumigant removed and the soils incubated for 10 days (fumigation-incubation). The regression equations were 1 day CHC1₃-N = (0.79 ± 0.022) F_N and 5 day CHC1₃-N = (1.01 ± 0.027) F_N, both regressions accounting for 92% of the variance in the data.In field soils previously treated with ¹⁵N-labelled fertilizer, the amounts of labelled N, measured after fumigation-extraction, were very similar to the amounts of labelled N mineralized during fumigation-incubation; both were about 4 times as heavily labelled as the soil N as a whole. These results suggest that fumigation-extraction and fumigation-incubation both measure the same fraction of the soil organic N (probably the cytoplasmic component of the soil microbial biomass) and that measurement of the total N released by CHC1₃ fumigation for 24 h provides a rapid method for measuring biomass N.     

The effect of inorganic nitrogen on the added nitrogen interaction of soils in incubation experiments

A laboratory incubation experiment was conducted to study the effect of indigenous inorganic N on the immobilization of applied N and on the occurrence of an added N interaction (ANI). Samples of six Mollisols from Illinois were incubated with ¹⁵N-labelled (NH₄)₂SO₄ (100 or 200 mg N kg^-1 soil), with or without the use of 0.01 M CaCl₂ to extract inorganic N (mainly NO _inf3 ^sup- ) before incubation. From 6 to 49% of the N applied was immobilized, higher percentages being obtained with unextracted soils than with the extracted soils and with the higher rate of N addition. Net mineralization of native N occurred in both the unextracted and extracted soils, but was more extensive in the unextracted soil and increased with the addition of N. The increases were accompanied by a positive ANI, which usually exceeded the amount of applied N immobilized and increased with the rate of addition. The ANI values observed with extracted soils were attributed to increased mineralization of native organic N.     

Mineralization of carbon and nitrogen from fresh and anaerobically stored sheep manure in soils of different texture

A sandy loam soil was mixed with three different amounts of quartz sand and incubated with (¹⁵NH₄)₂SO₄ (60 g N g^-1 soil) and fresh or anaerobically stored sheep manure (60 g g^-1 soil). The mineralization-immobilization of N and the mineralization of C were studied during 84 days of incubation at 20°C. After 7 days, the amount of unlabelled inorganic N in the manure-treated soils was 6–10 g N g^-1 soil higher than in soils amended with only (¹⁵NH₄)₂SO₄. However, due to immobilization of labelled inorganic N, the resulting net mineralization of N from manure was insignificant or slightly negative in the three soil-sand mixtures (100% soil+0% quartz sand; 50% soil+50% quartz sand; 25% soil+75% quartz sand). After 84 days, the cumulative CO₂ evolution and the net mineralization of N from the fresh manure were highest in the soil-sand mixutre with the lowest clay content (4% clay); 28% fo the manure C and 18% of the manure N were net mineralized. There was no significant difference between the soil-sand mixtures containing 8% and 16% clay, in which 24% of the manure C and -1% to 4% of the manure N were net mineralized. The higher net mineralization of N in the soil-sand mixture with the lowest clay content was probably caused by a higher remineralization of immobilized N in this soil-sand mixture. Anaerobic storage of the manure reduced the CO₂ evolution rates from the manure C in the three soil-sand mixtures during the initial weeks of decomposition. However, there was no effect of storage on net mineralization of N at the end of the incubation period. Hence, there was no apparent relationship between net mineralization of manure N and C.     

Added nitrogen interaction as affected by soil nitrogen pool size and fertilization – significance of displacement of fixed ammonium

Displacement of NH₄⁺ fixed in clay minerals by fertilizer ¹⁵NH₄⁺ is seen as one mechanism of apparent added nitrogen interactions (ANI), which may cause errors in ¹⁵N tracer studies. Pot and incubation experiments were carried out for a study of displacement of fixed NH₄⁺ by ¹⁵N‐labeled fertilizer (ammonium sulfate and urea). A typical ANI was observed when ¹⁵N‐labeled urea was applied to wheat grown on soils with different N reserves that resulted from their long‐term fertilization history: Plants took up more soil N when receiving fertilizer. Furthermore, an increased uptake of ¹⁵N‐labeled fertilizer, induced by increasing unlabeled soil nitrogen supply, was found. This ANI‐like effect was in the same order of magnitude as the observed ANI. All causes of apparent or real ANI can be excluded as explanation for this effect. Plant N uptake‐related processes beyond current concepts of ANI may be responsible. NH₄⁺ fixation of fertilizer ¹⁵NH₄⁺ in sterilized or non‐sterile, moist soil was immediate and strongly dependent on the rate of fertilizer added. But for the tested range of 20 to 160 mg ¹⁵NH₄⁺‐N kg^–1, the NH₄⁺ fixation rate was low, accounting for only up to 1.3 % of fertilizer N added. For sterilized soil, no re‐mobilization of fixed ¹⁵NH₄⁺ was observed, while in non‐sterile, biologically active soil, 50 % of the initially fixed ¹⁵NH₄⁺ was released up to day 35. Re‐mobilization of ¹⁵NH₄⁺ from the pool of fixed NH₄⁺ started after complete nitrification of all extractable NH₄⁺. Our results indicate that in most cases, experimental error from apparent ANI caused by displacement of fixed NH₄⁺ in clay is unlikely. In addition to the low percentage of only 1.3 % of applied ¹⁵N, present in the pool of fixed NH₄⁺ after 35 days, there were no indications for a real exchange (displacement) of fixed NH₄⁺ by ¹⁵N.     

NH3 Volatilization,N2O Emission and Microbial Biomass Turnover from 15N-Labeled Manure Under Laboratory Conditions

On irrigated agricultural soils from semi-arid and arid regions, ammonia (NH₃) volatilization and nitrous oxide (N₂O) emission can be a considerable source of N losses. This study was designed to test the capture of ¹⁵N loss as NH₃ and N₂O from previous and recent manure application using a sandy, calcareous soil from Oman amended one or two times with ¹⁵N labeled manure to elucidate microbial turnover processes under laboratory conditions. The system allowed to detect ¹⁵N enrichments in evolved N₂O-N and NH₃-N of up to 17% and 9%, respectively, and total N, K₂SO₄ extractable N and microbial N pools from previous and recent ¹⁵N labeled manure applications of up to 7%, 8%, and 15%. One time manured soil had higher cumulative N₂O-N emissions (141 µg kg^?1) than repeatedly manured soil with 43 µg kg^?1 of which only 22% derived from recent manure application indicating a priming effect.     

Effects of pentachlorophenol on asymbiotic nitrogen fixation in soil

The effect of 50, 100, 150, and 400 μg sodium pentachlorophenate (Na-PCP) per gram soil was studied in nonsterile soil incubated under aerobic and anaerobic conditions, and in sterilized soil inoculated withAzotobacter sp. isolated from the soil. N₂ fixation was determined by acetylene reduction. Pentachlorophenate at a concentration of 50 μg g^?1 had an inhibitory effect in nonsterile soil incubated aerobically while strong inhibition of dinitrogen fixation in nonsterile soil occurred in the presence of 100 μg g^?1 and above. The EC₅₀ values for the inhibition of nitrogenase activity in nonsterile soil incubated aerobically and anaerobically and in sterilized soil inoculated withAzotobacter sp. suspensions were 49.8±1.4 μg Na-PCP g^?1, 186.8±2.8 μg Na-PCP g^?1, and 660.8±29.3 μg Na-PCP g^?1, respectively.     

Chloride interference in total nitrogen analysis by the Kjeldahl method

Abstract

Very low recovery of NH₄+‐N was observed in total N determination of (NH₄)₂SO₄ in KC1 solutions by a semimicro Kjeldahl method using permanganate and reduced iron to recover NO₃‐ and NO₂‐, whereas complete recovery was obtained in analysis of NH₄+‐N in water, and of NO₃ ^?‐N or NO₂ ^?‐N in either water or KC1 solutions. The loss of NH₄ ⁺‐N observed with KC1 was attributed to the formation of NCl₃ upon reaction of NH₄ ⁺ with Cl₂ generated during oxidation of Cl^? by MnO₄ ^?. This difficulty is avoided by using K₂SO₄ instead of KC1 for extraction of inorganic N from soil. Complete recovery was obtained by adding ¹⁵N‐labeled NH₄+, NO₃‐, or NO₂‐ to 0.5 M K₂SO₄ soil extracts, and total ¹⁵N analyses of the labeled extracts were in good agreement with values calculated from the additions of ¹⁵N and the total N contents of the soil extracts.     

Effects of nitrogen dioxide on selected soil processes

Nitrogen dioxide gas was rapidly absorbed by soil. After a 15 min incubation at 25°C, soil at a moisture content of 16% absorbed 99% of the NO₂ introduced into the gas-phase volume of a closed system. The presence of microorganisms hatl no influence on the rate of absorption of the gas by soil. The absorption of NO₂ by sandy clay loam soil was not an oxygen- or temperature-dependent process nor did it depend upon the moisture content of the soil. These physical factors acquired significance only in determining the initial rate of absorption of the gas and the rate at which NO₂ diffused through the soil. Exposure of soil to NO₂ resulted in substantial increases in the levels of NO _inf2 ^sup? N in the soil. Chemical oxidation of the NO _inf2 ^sup? N resulted in an increase in NO _inf3 ^sup? N levels. During a 14-day incubation, NO _inf2 ^sup? N concentrations in sterile soil exposed to an atmosphere containing 100 μg ml^?1 of NO₂ decreased from 190 μg g^?1 of soil to 105 μg g^?1 with an accompanying increase in NO _inf3 ^sup? N from 2 μg g^? ₁ to 63 μg g^?1 of soil. Nitrogen dioxide severely inhibited the growth of both aerobic and anaerobic asymbiotic N₂-fixing bacteria in soil. After a 48 h incubation at 25°C, soil aggregates exposed to an atmosphere containing 100 μg ml^?1 of NO₂ contained 88% and 98% fewer aerobic and anaerobic N₂-fixing bacteria, respectively. C₂H₂-reduction measurements showed that nitrogenase synthesis and activity in artificial soil aggregates amended with 2% glucose were inhibited by 20% and 48%, respectively, when exposed to atmospheric concentrations of 35 and 3.5 μg ml^?1 of NO₂, respectively.