Denitrification in the top and sub soil of grassland on peat soils

Denitrification is an important process in the nitrogen (N) balance of intensively managed grassland, especially on poorly drained peat soils. Aim of this study was to quantify the N loss through denitrification in the top and sub soil of grassland on peat soils. Sampling took place at 2 sites with both control (0 N) and N fertilised (+ N) treatments. Main difference between the sites was the ground water level. Denitrification was measured on a weekly basis for 2 years with a soil core incubation technique using acetylene (C₂H₂) inhibition. Soil cores were taken from the top soil (0–20 cm depth) and the sub soil (20–40 cm depth) and incubated in containers for 24 hours. The denitrification rate was calculated from the nitrous oxide production between 4 and 24 hours of incubation. Denitrification capacities of the soils and the soil layers were also determined.The top soil was the major layer for denitrification with losses ranging from 9 to 26 kg N ha^–1 yr^–1 from the O N treatment. Losses from the top soil of the + N treatment ranged from 13 to 49 kg N ha^–1 yr^–1. The sub soil contributed, on average, 20% of the total denitrification losses from the 0–40 layer. Losses from the 0–40 cm layer were 2 times higher on the + N treatment than on the O N treatment and totalled up to 70 kg N ha^–1 yr^–1. Significant correlation coefficients were found between denitrification activity on the one hand, and ground water level, water filled pore space and nitrate content on the other, in the top soil but not in the sub soil. The denitrification capacity experiment showed that the availability of easily decomposable organic carbon was an important limiting factor for the denitrification activity in the sub soil of these peat soils.     

Effect of substrate-dependent microbial ethylene production on plant growth

Various compounds have been identified as precursors/substrates for the synthesis of ethylene (C₂H₄) in soil. This study was designed to compare the efficiency of four substrates, namely L-methionine (L-MET), 2-keto-4-methylthiobutyric acid (KMBA), 1-aminocyclopropane-1-carboxylic acid (ACC), and calcium carbide (CaC₂), for ethylene biosynthesis in a sandy clay loam soil by gas chromatography. The classic “triple” response in etiolated pea seedling was employed as a bioassay to demonstrate the effect of substrate-dependent microbial production of ethylene on plant growth. Results revealed that an amendment with L-MET, KMBA, ACC (up to 0.10 g/kg soil) and CaC₂ (0.20 g/kg soil) significantly stimulated ethylene biosynthesis in soil. Overall, ACC proved to be the most effective substrate for ethylene production (1434 nmol/kg soil), followed by KMBA, L-MET, and CaC₂ in descending order. Results further revealed that ethylene accumulation in soil from these substrates caused a classic “triple” response in etiolated pea seedlings with different degrees of efficacy. A more obvious classic “triple” response was observed at 0.15, 0.10, and 0.20 g/kg soil of L-MET, KMBA/ACC, and CaC₂, respectively. Similarly, direct exposure of etiolated pea seedlings to commercial ethylene gas also modified the growth pattern in the same way. A significant direct correlation (r = 0.86 to 0.97) between substrate-derived C₂H₄ and the classic triple response in etiolated pea seedlings was observed. This study demonstrated that the presence of substrate(s) in soil may lead to increased ethylene concentration in the air of the soil, which may affect plant growth in a desired direction. Published in Russian in Mikrobiologiya, 2006, Vol. 75, No. 2, pp. 277–283. The text was submitted by the authors in English.     

Nitrogen mineralization and denitrification as influenced by crop residue particle size

Managing the crop residue particle size has the potential to affect N conservation in agricultural systems. We investigated the influence of barley (Hordeum vulgare) and pea (Pisum sativum) crop residue particle size on N mineralization and denitrification in two laboratory experiments. Experiment 1: ¹⁵N-labelled ground (3 mm) and cut (25 mm) barley residue, and microcrystalline cellulose+glucose were mixed into a sandy loam soil with additional inorganic N. Experiment 2: inorganic¹⁵ N and C₂H₂ were added to soils with barley and pea material after 3, 26, and 109 days for measuring gross N mineralization and denitrification.Net N immobilization over 60 days in Experiment 1 cumulated to 63 mg N kg^-1 soil (ground barley), 42 (cut barley), and 122 (cellulose+glucose). More N was seemingly net mineralized from ground barley (3.3 mg N kg^-1 soil) than from cut barley (2.7 mg N kg^-1 soil). Microbial biomass peaked at day 4 with the barley treatments and at day 14 with the cellulose+glucose whereafter the biomass leveled out at values 79 mg C kg^-1 (ground), 104 (cut), and 242 (cellulose+glucose) higher than for the control soil. Microbial growth yields were similar for the two barley treatments, ca. 60 mg C g^-1 substrate C added, which was lower than the 142 mg C g^-1 C added with cellulose+glucose. This suggests that the 75% (w/w) holocelluloses and sugars contained with the barley material remained physically protected despite grinding. In Experiment 2 gross mineralization on day 3 was 4.8 mg N kg^-1 d^-1 with ground pea, twice as much as for all other treatments. On day 26 the treatment with ground barley had the greatest gross N mineralization. In static cores ground barley denitrified 11-fold more than did cut barley, whereas denitrification was similar for the two pea treatments. In suspensions denitrification was similar for the two treatments both with barley and pea residue.We conclude that the higher microbial activity associated with the initial decomposition of ground plant material is due to a more intimate plant residue-soil contact. On the long term, grinding the plant residues has no significant effect on N dynamics.     

Distinguishing between Nitrification and Denitrification as Sources of Gaseous Nitrogen Production in Soil

The source of N₂O produced in soil is often uncertain because denitrification and nitrification can occur simultaneously in the same soil aggregate. A technique which exploits the differential sensitivity of these processes to C₂H₂ inhibition is proposed for distinguishing among gaseous N losses from soils. Denitrification N₂O was estimated from 24-h laboratory incubations in which nitrification was inhibited by 10-Pa C₂H₂. Nitrification N₂O was estimated from the difference between N₂O production under no C₂H₂ and that determined for denitrification. Denitrification N₂ was estimated from the difference between N₂O production under 10-kPa C₂H₂ and that under 10 Pa. Laboratory estimates of N₂O production were significantly correlated with in situ N₂O diffusion measurements made during a 10-month period in two forested watersheds. Nitrous oxide production from nitrification was most important on well-drained sites of a disturbed watershed where ambient NO₃⁻ was high. In contrast, denitrification N₂O was most important on poorly drained sites near the stream of the same watershed. Distinction between N₂O production from nitrification and denitrification was corroborated by correlations between denitrification N₂O and water-filled pore space and between nitrification N₂O and ambient NO₃⁻. This technique permits qualitative study of environmental parameters that regulate gaseous N losses via denitrification and nitrification.     

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

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

Responses of N fluxes and pools to elevated atmospheric CO2 in model forest ecosystems with acidic and calcareous soils

The objectives of this study were to estimate how soil type, elevated N deposition (0.7 vs. 7 g N m^–2y^–1) and tree species influence the potential effects of elevated CO₂ (370 vs. 570 mol CO₂ mol^–1) on N pools and fluxes in forest soils. Model spruce-beech forest ecosystems were established on a nutrient-rich calcareous sand and on a nutrient-poor acidic loam in large open-top chambers. In the fourth year of treatment, we measured N concentrations in the soil solution at different depths, estimated N accumulation by ion exchange resin (IER) bags, and quantified N export in drainage water, denitrification, and net N uptake by trees. Under elevated CO₂, concentrations of N in the soil solution were significantly reduced. In the nutrient-rich calcareous sand, CO₂ enrichment decreased N concentrations in the soil solution at all depths (–45 to –100%). In the nutrient-poor acidic loam, the negative CO₂ effect was restricted to the uppermost 5 cm of the soil. Increasing the N deposition stimulated the negative impact of CO₂ enrichment on soil solution N in the acidic loam at 5 cm depth from –20% at low N inputs to –70% at high N inputs. In the nutrient-rich calcareous sand, N additions did not influence the CO₂ effect on soil solution N. Accumulation of N by IER bags, which were installed under individual trees, was decreased at high CO₂ levels under spruce in both soil types. Under beech, this decrease occurred only in the calcareous sand. N accumulation by IER bags was negatively correlated with current-years foliage biomass, suggesting that the reduction of soil N availability indices was related to a CO₂-induced growth enhancement. However, the net N uptake by trees was not significantly increased by elevated CO₂. Thus, we suppose that the reduced N concentrations in the soil solution at elevated CO₂ concentrations were rather caused by an increased N immobilisation in the soil. Denitrification was not influenced by atmospheric CO₂ concentrations. CO₂ enrichment decreased nitrate leaching in drainage by 65%, which suggests that rising atmospheric CO₂ potentially increases the N retention capacity of forest ecosystems.     

Acetylene Metabolism and Stimulation of Denitrification in an Agricultural Soil

The effects of C₂H₂ metabolism on N₂O production were examined in soil slurries. Enrichment of C₂H₂ consumption activity occurred only in aerobic incubations. Rapid disappearance of subsequent C₂H₂ additions, stimulation of CO₂ production, and most-probable-number enumerations of C₂H₂ utilizers indicated enrichment of the population responsible. During C₂H₂ consumption in slurries incubated statically under air, maximal rates of N₂O evolution were 19 times higher than those in anaerobic incubations. After 20 days of enrichment with C₂H₂, the production of N₂O by slurries supplemented with C₂H₂ and nitrate was 10 times higher than that in the unenriched controls. A Nocardia- or Arthrobacter-like bacterium was isolated that grew on C₂H₂ but did not denitrify. The behavior of soil inoculated with this bacterium became similar to that of C₂H₂-enriched soil incubated aerobically. Ethanol, acetate, and acetaldehyde were identified in enrichment experiments, and denitrification in soil slurries was stimulated by addition of the supernatant from a pure culture grown on mineral medium with C₂H₂. These results indicate that denitrification can be stimulated by the actions of an aerobic, nondenitrifying C₂H₂-metabolizing population. Utilization of intermediate metabolites by denitrifiers and enhanced O₂ consumption are two possible mechanisms for this stimulation.     

Patterns of denitrification loss from grazed grassland: Effects of N fertilizer inputs at different sites

Factors influencing the seasonal and daily variation in denitrification rates in grazed swards were examined at 5 experimental sites in England with wide ranging environmental/geographic conditions. There was a wide range of fertilizer inputs at each site. Rates of denitrification were estimated by a coring and field incubation technique using acetylene to inhibit the reduction of N₂O to N₂. Major features of the detailed results from two of the sites were: (i) the large ranges in rates of loss, (ii) the relatively low contributions to total annual loss during autumn and winter, (iii) the apparent association of high rates of loss with fertilizer additions made when the soil was wet or immediately preceding a rainfall event, and (iv) significant losses from soil at 10–30 cm in the profile. Multiple quadratic regression analysis of the effects of soil NO₃ ^--N, soil temperature and water was used to explain variability in rates of loss. When separate regressions were fitted within each site × year × season × fertilizer level subset, 51% of the variation in loss was explained on a poorly drained fine loam/silt but only 38% on a freely drained loam.     

Denitrification in Low pH Spodosols and Peats Determined with the Acetylene Inhibition Method

Potential denitrification rates were determined for predominantly acid (pH ≥ 3.6) horizons of forestal, miry, and agricultural soils from 22 locations in southern Finland. The acetylene inhibition method was used with nitrate-amended water-logged soils incubated in an N₂ atmosphere containing 2.5 or 5% C₂H₂. Complete inhibition of the reduction of N₂O to N₂ was observed in 99.3% of the samples. The denitrification rates varied from 0.12 to 53.8 μg of N·cm^-3·day^-1. Correlation between denitrification rate and soil pH was highly significant: r = 0.84 on a volume basis, and r = 0.44 on a weight basis. Vegetation type and amount of soil organic matter had a minor or no effect, respectively. In spodosolized soils the rates were significantly higher for B horizons than for A horizons. These results show that denitrification can occur in acid soils.     

River and lake sediments as sources of infective Frankia (Alnus)

Exudation of carboxylic anions and protons by plant roots plays an important role in mobilizing soil P under P-deficiency conditions. The objective of this work was to quantify short-term (6 h) carboxylate and H⁺ exudation by tomato roots in response to P concentration (0, 0.1, 0.5 and 1.0 mt M P) in nutrient solution (C_p). The exudation rate of tri- and dicarboxylates decreased exponentially with increasing C_p, from 0.3 to 0.03 mol plant^–1 6h^–1. At low C_p the predominant exudates were fumarate, citrate and succinate, while at C_p=0.5 and 1.0 mt M the prevalent anions were succinate and citrate. The solution pH declined sharply as C_p was lowered from 0.1 (pH=4.2) to 0 mt M P (pH=3.7).     

Denitrification in San Francisco Bay Intertidal Sediments

The acetylene block technique was employed to study denitrification in intertidal estuarine sediments. Addition of nitrate to sediment slurries stimulated denitrification. During the dry season, sediment-slurry denitrification rates displayed Michaelis-Menten kinetics, and ambient NO₃⁻ + NO₂⁻ concentrations (≤26 μM) were below the apparent K_m (50 μM) for nitrate. During the rainy season, when ambient NO₃⁻ + NO₂⁻ concentrations were higher (37 to 89 μM), an accurate estimate of the K_m could not be obtained. Endogenous denitrification activity was confined to the upper 3 cm of the sediment column. However, the addition of nitrate to deeper sediments demonstrated immediate N₂O production, and potential activity existed at all depths sampled (the deepest was 15 cm). Loss of N₂O in the presence of C₂H₂ was sometimes observed during these short-term sediment incubations. Experiments with sediment slurries and washed cell suspensions of a marine pseudomonad confirmed that this N₂O loss was caused by incomplete blockage of N₂O reductase by C₂H₂ at low nitrate concentrations. Areal estimates of denitrification (in the absence of added nitrate) ranged from 0.8 to 1.2 μmol of N₂ m⁻² h⁻¹ (for undisturbed sediments) to 17 to 280 μmol of N₂ m⁻² h⁻¹ (for shaken sediment slurries).     

Production of N2O in soil during decomposition of dead yeast cells with different spatial distributions

Production and sources of N₂O were determined in soil columns amended with autoclaved yeast cells either mixed into or added as 0.5 cm³ lumps to the soil in combination with no or 200 g NO₃ ^--N g^-1. At four occasions over a two-week study period, subsets of cores were measured for N₂O production during 4-hour incubations under atmospheres of ambient air, 10 Pa of C₂H₂, and N₂, respectively. Denitrification enzyme activity (DEA) was assessed in subsamples of cores that had been incubated continuously under air.Autoclaved yeast provided a C-source readily available for denitrifying bacteria in the soil. Nitrous oxide production was negligible in unamended columns whereas accumulated N₂O losses in the presence of yeast material were substantial, varying between 15 to 49 ng N₂O-N g^-1 h^-1. Mixing yeast into the soil caused the highest production of N₂O followed by the yeast lump and no yeast treatments. Incubation in the presence of 10 Pa C₂H₂ indicated that denitrification was the sole source of N₂O, in accordance with an increase in DEA. Nitrous oxide production and DEA peaked after 4–7 days of incubation, and both were unaffected by additional NO₃ ^-. Two-to four-fold responses to anaerobiosis and accumulation of NO₃ ^- and NH₄ ⁺ in proximity of the lumps indicated that N₂O production here was limited by relatively low C-availability. In contrast, 10- to 12-fold responses to anaerobiosis and no accumulation of inorganic N suggested a higher C-availability where yeast was mixed into the soil.     

Denitrification rate determined by nitrate disappearance is higher than determined by nitrous oxide production with acetylene blockage

A mixed beech and spruce forest soil was incubated under potential denitrification assay (PDA) condition with 10% acetylene (C₂H₂) in the headspace of soil slurry bottles. Nitrous oxide (N₂O) concentration in the headspace, as well as nitrate, nitrite and ammonium concentrations in the soil slurries were monitored during the incubation. Results show that nitrate disappearance rate was higher than N₂O production rate with C₂H₂ blockage during the incubation. Sum of nitrate, nitrite, and N₂O with C₂H₂ blockage could not recover the original soil nitrate content, showing an N imbalance in such a closed incubation system. Changes in nitrite and ammonium concentration during the incubation could not account for the observed faster nitrate disappearance rate and the N imbalance. Non-determined nitric oxide (NO) and N₂ production could be the major cause, and the associated mechanisms could vary for different treatments. Commonly applied PDA measurement likely underestimates the nitrate removal capacity of a system. Incubation time and organic matter/nitrate ratio are the most critical factors to consider using C₂H₂ inhibition technique to quantify denitrification. By comparing the treatments with and without an antibiotic, the results suggest that microbial N uptake probably played a minor role in N balance, and other denitrifying enzymes but nitrate reductase could be substantially synthesized during the incubation.     

Denitrification and total N losses from an irrigated sandy-clay loam under maize–wheat cropping system

Denitrification and total N losses were quantified from an irrigated field cropped to maize and wheat, each receiving urea at 100 kg N ha^-1. During the maize growing season (60 days), the denitrification loss measured directly by acetylene inhibition-soil cover method amounted 2.72 kg N ha^-1 whereas total N loss measured by ¹⁵N balance was 39 kg ha^-1. Most (87%) of the denitrification loss under maize occurred during the first two irrigation cycles. During the wheat growing season (150 days), the denitrification loss directly measured by acetylene inhibition-soil cover and acetylene inhibition-soil core methods was 1.14 and 3.39 kg N ha^-1, respectively in contrast to 33 kg N ha^-1 loss measured by ¹⁵N balance. Most (70-88%) of the denitrification loss under wheat occurred during the first three irrigation cycles. Soil moisture and NO ₃ ^- -N were the major factors limiting denitrification under both crops. Higher N losses measured by ¹⁵N balance than C₂H₂ inhibition method were perhaps due to underestimation of denitrification by C₂H₂ inhibition method and losses other than denitrification, most probably NH₃ volatilization.     

Phytotoxicity and distribution of copper in tropical soil amended with sewage sludge and copper sulfate

Abstract

A greenhouse experiment was conducted to evaluate phytotoxicity and distribution of Cu in a tropical soil amended with sewage sludge (Sw) and copper sulfate (CuSO₄.5H₂O). Samples of a clay soil from the State of Paraná, Brazil were collected at depth of 0–20; 20–40 and 40–60 cm, and brought to the laboratory to be properly accommodated in experimental units (PVC tubes). The Cu treatments were performed by the application of Sw (10 t ha-¹) amended with Cu (SB-T), and by CuSO₄. H₂O (WB-T). Lettuce plants were cultivated in the amended soil in order to predict the toxicity of the Cu. The experiment was conducted for 70 days, and then the lettuce plants and soil samples were collected for analysis. A sequential method was used to separate soil Cu into following fractions: exchangeable, amorphous iron oxide bound, crystalline iron oxide bound, organic matter bound and residual bound. The experimental results showed that Fe, Zn, K, P, Cu and organic matter amounts of the soil increased with the treatment SB-T. The toxic phyto-available Cu content in the soil for the lettuce plants was 80.00 mg kg^-1. A percolation study showed that the Cu contents were larger for the first 20 cm of depth, indicating that the metal was not transported down the soil profile. The Cu content of different fractions declined in an order residual > amorphous iron oxide > crystalline iron oxide > organic matter > exchangeable, regardless of treatment performed. Additionally, the Cu contents added from treatments were determined mainly in amorphous iron oxide fraction.     

Role of Microorganisms in Emission of Nitrous Oxide and Methane in Pulse Cultivated Soil Under Laboratory Incubation Condition

Soil from a pulse cultivated farmers land of Odisha, India, have been subjected to incubation studies for 40 consecutive days, to establish the impact of various nitrogenous fertilizers and water filled pore space (WFPS) on green house gas emission (N₂O & CH₄). C₂H₂ inhibition technique was followed to have a comprehensive understanding about the individual contribution of nitrifiers and denitrifiers towards the emission of N₂O. Nevertheless, low concentration of C₂H₂ (5 ml: flow rate 0.1 kg/cm²) is hypothesized to partially impede the metabolic pathways of denitrifying bacterial population, thus reducing the overall N₂O emission rate. Different soil parameters of the experimental soil such as moisture, total organic carbon, ammonium content and nitrate–nitrogen contents were measured at regular intervals. Application of external N-sources under different WFPS conditions revealed the diverse role played by the indigenous soil microorganism towards green house gas emission. Isolation of heterotrophic microorganisms (Pseudomonas) from the soil samples, further supported the fact that denitrification might be prevailing during specific conditions thus contributing to N₂O emission. Statistical analysis showed that WFPS was the most influential parameter affecting N₂O formation in soil in absence of an inhibitor like C₂H₂.     

Comparing Denitrification Estimates for a Texas Estuary by Using Acetylene Inhibition and Membrane Inlet Mass Spectrometry

Characterizing denitrification rates in aquatic ecosystems is essential to understanding how systems may respond to increased nutrient loading. Thus, it is important to ensure the precision and accuracy of the methods employed for measuring denitrification rates. The acetylene (C₂H₂) inhibition method is a simple technique for estimating denitrification. However, potential problems, such as inhibition of nitrification and incomplete inhibition of nitrous oxide reduction, may influence rate estimates. Recently, membrane inlet mass spectrometry (MIMS) has been used to measure denitrification in aquatic systems. Comparable results were obtained with MIMS and C₂H₂ inhibition methods when chloramphenicol was added to C₂H₂ inhibition assay mixtures to inhibit new synthesis of denitrifying enzymes. Dissolved-oxygen profiles indicated that surface layers of sediment cores subjected to the MIMS flowthrough incubation remained oxic whereas cores incubated using the C₂H₂ inhibition methods did not. Analysis of the microbial assemblages before and after incubations indicated significant changes in the sediment surface populations during the long flowthrough incubation for MIMS analysis but not during the shorter incubation used for the C₂H₂ inhibition method. However, bacterial community changes were also small in MIMS cores at the oxygen transition zone where denitrification occurs. The C₂H₂ inhibition method with chloramphenicol addition, conducted over short incubation intervals, provides a cost-effective method for estimating denitrification, and rate estimates are comparable to those obtained by the MIMS method.     

The effect of nitrate and nitrous oxide on hydrogen and methane accumulation in anaerobically-incubated soils

Summary The effect of KNO₃ and N₂O on the accumulation of CH₄, H₂ and denitrification products in two North Dakota soils during anaerobic incubation at 30°C was studied by means of gas chromatography. KNO₃ and N₂O (500 ppm N) reduced the rate of accumulation of CH₄ by a Tetonka soil regardless of whether the soil was in an air-dried condition or had been pre-incubated and actively producing CH₄ prior to the treatment application. Both KNO₃ and N₂O completely suppressed H₂ accumulation by the remoistened air-dried soil; no H₂ either in the presence or absence of added KNO₃ or N₂O was accumulated by the pre-incubated Tetonka soil subsequent to the treatment application. KNO₃ (250 ppm N) reduced the rate of accumulation of CH₄ by a Cavour loam during anaerobic incubation. No H₂ was accumulated by this soil during anaerobic incubation. At equivalent K⁺ concentrations, KNO₃ suppressed CH₄ accumulation by the Tetonka and Cavour soils to a greater extent than did KCl.     

Ammonium, nitrate, and total nitrogen in the soil water of feedlot and field soil profiles

A level feedlot, located in an area consisting of Wann silt loam changing with depth to sand, appears to contribute no more NO₃^- nitrogen, NH₄⁺ nitrogen, and total nitrogen to the shallow water table beneath it than an adjacent cropped field. Soil water samples collected at 46, 76, and 107 cm beneath the feedlot surface generally showed NO₃^- nitrogen concentrations of less than 1 μg/ml. During the summer months, soil water NO₃^- nitrogen increased at the 15-cm depth, indicating that nitrification took place at the feedlot surface. However, the low soil water NO₃^- nitrogen values below 15 cm indicate that denitrification takes place beneath the surface.     

Adaptation of Denitrifying Populations to Low Soil pH

Natural denitrification rates and activities of denitrifying enzymes were measured in an agricultural soil which had a 20-year past history of low pH (pH ca. 4) due to fertilization with acid-generating ammonium salts. The soil adjacent to this site had been limed and had a pH of ca. 6.0. Natural denitrification rates of these areas were of similar magnitude: 158 ng of N g⁻¹ of soil day⁻¹ for the acid soil and 390 ng of N g⁻¹ of soil day⁻¹ at the neutral site. Estimates of in situ denitrifying enzyme activity were higher in the neutral soil, but substantial enzyme activity was also detected in the acid soil. Rates of nitrous oxide reduction were very low, even when NO₃⁻ and NO₂⁻ were undetectable, and were ca. 400 times lower than the rates of N₂O production from NO₃⁻. Denitrification rates measured in slurries of the acid and neutral soil showed distinctly different pH optima (pH 3.9 and pH 6.3) which were near the pH values of the two soils. This suggests that an acid-tolerant denitrifying population had been selected during the 20-year period of low pH.