Gross N transfer rates in field soils measured by 15N-pool dilution

Abstract

A micro-plot ¹⁵N-tracer experiment was established in three different soils of a long-term soil fertility field experiment. The nutrient-poor loam sand has been subjected to various treatments over the years and this has resulted in different organic C (0.35% – 0.86%), microbial biomass (38.3 – 100.0 µg C _mic g^?1 soil), clay and fine silt contents. Using the ¹⁵N-pool dilution technique, we assessed gross N-transfer rates in the field. Gross N mineralization rates varied strongly among the three plots and ranged between 0.4 and 4.2 µg N g^?1 soil d^?1. Gross nitrification rates were estimated to be between 0 and 2.1 µg N g^?1 soil d^?1. No correlation between gross N mineralization rates and the organic matter content of the soils was established. However, gross nitrate consumption rates increased with increasing soil C content. The ¹⁵N-pool dilution technique was successfully used to measure gross N transfer rates directly in the field.     

Cold storage and laboratory incubation of intact soil cores do not reflect in-situ nitrogen cycling rates of tropical forest soils

Measurements of N transformation rates in tropical forest soils are commonly conducted in the laboratory from disturbed or intact soil cores. On four sites with Andisol soils under old-growth forests of Panama and Ecuador, we compared N transformation rates measured from laboratory incubation (at soil temperatures of the sites) of intact soil cores after a period of cold storage (at 5 °C) with measurements conducted in situ. Laboratory measurements from stored soil cores showed lower gross N mineralization and NH₄⁺ consumption rates and higher gross nitrification and NO₃⁻ immobilization rates than the in-situ measurements. We conclude that cold storage and laboratory incubation change the soils to such an extent that N cycling rates do not reflect field conditions. The only reliable way to measure N transformation rates of tropical forest soils is in-situ incubation and mineral N extraction in the field.     

Potential importance of bacteria and fungi in nitrate assimilation in soil

Soil microorganisms can use a wide range of N compounds but are thought to prefer NH₄⁺. Nevertheless, ¹⁵N isotope dilution studies have shown that microbial immobilization of NO₃⁻ can be an important process in many soils, particularly relatively undisturbed soils. Our objective was to develop a method for measuring NO₃⁻ immobilization potential so that the relative contributions of bacteria and fungi could be determined. We modified and optimized a soil slurry method that included amendments of KNO₃, glucose, and methionine sulfoximine (an inhibitor of N assimilation) in the presence of two protein synthesis inhibitors: chloramphenicol, which inhibits bacteria, or cycloheximide, which inhibits fungi. By adding ¹⁵N-labeled KNO₃, we were able to measure gross rates of NO₃⁻ production (i.e., gross nitrification) and consumption (i.e., gross NO₃⁻ immobilization). We found that bacteria, not fungi, had the greatest potential for assimilating, or immobilizing, NO₃⁻ in these soils. This is consistent with their growth habit and distribution in the heterogeneous soil matrix.     

好氧条件下两种水稻土的氮素总转化速率比较

Although to date individual gross N transformations could be quantified by ~(15)N tracing method and models,studies are still limited in paddy soil.An incubation experiment was conducted using topsoil(0-20 cm) and subsoil(20-60 cm) of two paddy soils,alkaline and clay(AC) soil and neutral and silt loam(NSL) soil,to investigate gross N transformation rates.Soil samples were labeled with either ~(15)NH4_NO_3 or NH_4~(15)NO_3,and then incubated at 25 °C for 168 h at 60%water-holding capacity.The gross N mineralization(recalcitrant and labile organic N mineralization) rates in AC soil were 1.6 to 3.3 times higher than that in NSL soil,and the gross N nitrification(autotrophic and heterotrophic nitrification) rates in AC soil were 2.4 to 4.4 times higher than those in NSL soil.Although gross NO_3~- consumption(i.e.,NO_3~- immobilization and dissimilatory NO_3~- reduction to NH_4~+ rates increased with increasing gross nitrification rates,the measured net nitrification rate in AC soil was approximately 2.0 to 5.1 times higher than that in NSL soil.These showed that high NO_3~- production capacity of alkaline paddy soil should be a cause for concern because an accumulation of NO_3~- can increase the risk of NO_3~- loss through leaching and denitrification.     

Determination of the fate of C labelled maize and wheat rhizodeposit-C in two agricultural soils in a greenhouse experiment under C-CO2-enriched atmosphere

A deeper understanding of the contribution of carbon (C) released by plant roots (rhizodeposition) to soil organic matter (SOM) can help to increase our knowledge of global C-cycling. These insights can eventually lead to sustainable management of SOM especially in agricultural systems. This study was conducted to determine the fate of ¹³C labelled rhizodeposit-C of maize and wheat plants. They were grown in a greenhouse in permeable nylon bags filled with upper soil material from two agricultural soils of the same location, but with different crop yields. The bags were placed into pots, which were also filled with soil surrounding the bags. Soil inside the bags was considered as rhizosphere soil, wheras the one outside the bags represented bulk soil. The contributions of rhizodeposits to water extractable organic carbon (WEOC), microbial biomass-C (MB-C), CO₂-C evolution, and total organic carbon (C_org) were investigated during a 7-week growing period. The WEOC, MB-C, CO₂-C, C_org contents and the respective δ¹³C values were determined regularly, and a newly developed method for determining δ¹³C values in soil extracts was applied.In both soils, regardless of crop yield potential, significant incorporation of rhizodeposition-derived C was observed in the MB-C, CO₂-C, and C_org pool, but not in the WEOC. The pattern of C incorporation into the different pools was the same for both soils with both plants, and rhizodeposit-derived C was recovered in the order MB-C<C_org<CO₂-C. This showed that rhizodeposits were mainly respired, but since C_org was the second largest pool of the overall balances, they were also stabilized in the soils at least in the short term. It is suggested that the increased SOM mineralization observed in this study (positive priming effects) was probably induced by C exchange processes between the soil matrix and soluble rhizodeposits. Moreover, soluble rhizodeposit-C was detected in MB-C and CO₂-C evolved outside the direct root zone, showing the availability of these C-components in the bulk soil.     

Microbial immobilization of ammonium and nitrate in relation to ammonification and nitrification rates in organic and conventional cropping systems

Agricultural systems that receive high or low organic matter (OM) inputs would be expected to differ in soil nitrogen (N) transformation rates and fates of ammonium (NH₄⁺) and nitrate (NO₃⁻). To compare NH₄⁺ availability, competition between nitrifiers and heterotrophic microorganisms for NH₄⁺, and microbial NO₃⁻ assimilation in an organic vs. a conventional irrigated cropping system in the California Central Valley, chemical and biological soil assays, ¹⁵N isotope pool dilution and ¹⁵N tracer techniques were used. Potentially mineralizable N (PMN) and hot minus cold KCl-extracted NH₄⁺ as indicators of soil N supplying capacity were measured five times during the tomato growing season. At mid-season, rates of gross ammonification and gross nitrification after rewetting dry soil were measured in microcosms. Microbial immobilization of NO₃⁻ and NH₄⁺ was estimated based on the uptake of ¹⁵N and gross consumption rates. Gross ammonification, PMN, and hot minus cold KCl-extracted NH₄⁺ were approximately twice as high in the organically than the conventionally managed soil. Net estimated microbial NO₃⁻ assimilation rates were between 32 and 35% of gross nitrification rates in the conventional and between 37 and 46% in the organic system. In both soils, microbes assimilated more NO₃⁻ than NH₄⁺. Heterotrophic microbes assimilated less NH₄⁺ than NO₃⁻ probably because NH₄⁺ concentrations were low and competition by nitrifiers was apparently strong. The high OM input organic system released NH₄⁺ in a gradual manner and, compared to the low OM input conventional system, supported a more active microbial biomass with greater N demand that was met mainly by NO₃⁻ immobilization.     

Mineralization of soil organic nitrogen solubilized by aqua ammonia fertilizer

Organic N solubilized by NH_3(aq) was extracted from ¹⁵N-labelled or unlabelled soil, concentrated and added to non-extracted soil, which was incubated under aerobic conditions at 27±1°C. Gross N mineralization, gross N immobilization, and nitrification in soils with or without addition of unlabelled soluble organic N were estimated by models based on the dilution of the NH ₄ ⁺ or NO _inf3 ^sup- pools, which were labelled with ¹⁵N at the beginning of incubation. Mineralization of labelled organic N was measured by the appearance of label in the mineral N pool. Although gross N mineralization and gross N immobilization were increased in two soils between day 0 and day 7 following addition of unlabelled organic N solubilized by NH_3(aq), there was no increase in net N mineralization. Solubilization of ¹⁵N-labelled organic N increased and the ¹⁵N enrichment of the soluble organic N decereased as the concentration of NH_3(aq) added increased. A constant proportion of approximately one-quarter of the labelled organic N added at different rates to non-extracted soil was recovered in the mineral N pool after an incubation period of 14 days, and the availability ratios calculated from net N mineralization data were 1.1:1 and 2.1:1 for 111 and 186 mg added organic-N kg^-1 soil, respectively, indicating that the mineralization of organic N was increased by solubilization.     

应用15N稀释法测定土壤氮素初级转化速率的一些关键技术

本文综合评述了应用~(15)N库稀释法测定土壤氮素初级转化速率的一些关键技术,即~(15)N标记土壤氮库的方法、~(15)N的加入量、丰度和标记物种类的选择,以及初始取样时间的确定。只有合理地运用这些关键技术,才能更准确地测定土壤氮素初级转化速率,进而更真实地表征土壤氮素的实际周转状况。     

Microbial C,N, and P relationships in moisture‐stressed soils of Potohar,Pakistan

In 11 rain‐fed arable soils of the Potohar plateau, Pakistan, the amounts of microbial‐biomass C (C_mic), biomass N (N_mic), and biomass P (P_mic) were analyzed in relation to the element‐specific total storage compartment, i.e., soil C_org, N_t, and P_t. The effects of climatic conditions and soil physico‐chemical properties on these relationships were highlighted with special respect to crop yield levels. Average contents of soil C_org, N_t, and P_t were 3.9, 0.32, and 0.61 mg (g soil)^–1, respectively. Less than 1% of P_t was extractable with 0.5 M NaHCO₃. Mean contents of C_mic, N_mic, and P_mic were 118.4, 12.0, and 3.9 µg (g soil)^–1. Values of C_mic, N_mic, P_mic, soil C_org, and N_t were all highly significantly interrelated. The mean crop yield level was closely connected with all soil organic matter– and microbial biomass–related properties, but showed also some influence by the amount of precipitation from September to June. Also the fraction of NaHCO₃‐extractable P was closely related to soil organic matter, soil microbial biomass, and crop yield level. This reveals the overwhelming importance of biological processes for P turnover in alkaline soils.     

Natural N abundances in a tallgrass prairie ecosystem exposed to 8-y of elevated atmospheric CO2

After 8-y of elevated CO₂, we previously detected greater amounts of total soil nitrogen, suggesting that rates of ecosystem N flux into or out of tallgrass prairie had been altered. Denitrification and associative N fixation rates are the two primary biological processes that are known to control N loss and accumulation in tallgrass prairie soil. Therefore, our objective was to assess the natural abundance of plant and soil ¹⁵N isotopes as a cumulative index of potential change in efflux or influx of N into and out of the tallgrass prairie after 8-y of exposure to elevated CO₂. Aboveground plant delta ¹⁵N values of Andropogon gerardii were close to zero and more positive as a result of elevated CO₂, but whole-soil values at the 5-30 cm depth were significantly reduced (6.8 vs 7.3; P<0.05) under elevated CO₂-chamber (EC) relative to ambient CO₂- chamber (AC). Total, aboveground plant biomass, root-in-growth, extractable N, microbial biomass N, and soil pools collectively exhibited a range of delta ¹⁵N values from −2.8 to 7.3. Measurements of surface soil ¹⁵N indicate that a change in N inputs and outputs has occurred as a result of elevated atmospheric CO₂. In addition to possible changes in denitrification and N₂ fixation, other sources of N such as the re-translocation of N to the surface from deeper soil layers are needed to explain how soil N accrues in surface soils as a consequence of elevated CO₂. Our results support the notion that C accrual may promote N accrual, possibly driven by high plant and microbial N demand amplified by soil N limitation.     

Effects of decreasing water potential on gross ammonification and nitrification in an acid coniferous forest soil

Changes in the soil water regime, predicted as a consequence of global climate change, might influence the N cycle in temperate forest soils. We investigated the effect of decreasing soil water potentials on gross ammonification and nitrification in different soil horizons of a Norway spruce forest and tested the hypotheses that i) gross rates are more sensitive to desiccation in the Oa and EA horizon as compared to the uppermost Oi/Oe horizon and ii) that gross nitrification is more sensitive than gross ammonification. Soil samples were adjusted by air drying to water potentials from about field capacity to around −1.0 MPa, a range that is often observed under field conditions at our site. Gross rates were measured using the ¹⁵N pool dilution technique. To ensure that the addition of solute label to dry soils and the local rewetting does not affect the results by re-mineralization or preferential consumption of ¹⁵N, we compared different extraction and incubation times.T₀ times ranging from 10 to 300 min and incubation times of 48 h and 72 h did not influence the rates of gross ammonification and nitrification. Even small changes of water potential decreased gross ammonification and nitrification in the O horizon. In the EA horizon, gross nitrification was below detection limit and the response of the generally low rates of gross ammonification to decreasing water potentials was minor. In the Oi/Oe horizon gross ammonification and nitrification decreased from 37.5 to 18.3 mg N kg⁻¹ soil d⁻¹ and from 15.4 to 5.6 mg N kg⁻¹ soil d⁻¹ when the water potential decreased from field capacity to −0.8 MPa. In the Oa horizon gross ammonification decreased from 7.4 to 4.0 mg N kg⁻¹ soil d⁻¹ when the water potential reached −0.6 MPa. At such water potential nitrification almost ceased, while in the Oi/Oe horizon nitrification continued at a rather high level. Hence, only in the Oa horizon nitrification was more sensitive to desiccation than ammonification. Extended drought periods that might result from climate change will cause a reduction in gross N turnover rates in forest soils even at moderate levels of soil desiccation.     

Comparison of NH4 pool dilution techniques to measure gross N fluxes in a coarse textured soil

We compared gross N fluxes by ¹⁵N pool dilution in a coarse-textured agricultural soil when ¹⁵N was applied to the soil NH₄⁺ pool by either: (i) mixing a ¹⁵NH₄NO₃ solution into disturbed soil or (ii) injection of ¹⁵NH₃ gas into intact soil cores. The two techniques produced similar results for gross N mineralization rates indicating that NH₄⁺ production in soil was not altered by soil disturbance, method of application (gas vs. solution), or amount of N applied. This was not the case for immobilization rates, which were twofold higher when ¹⁵N label was applied to the soil NH₄⁺ pool with the mixing technique compared to the injection technique. This was attributed to the fact that more NH₄⁺ was applied with the mixing technique. Estimates of gross nitrification were accompanied by large error terms meaning differences between ¹⁵N labeling methods could not be accurately assessed for this process rate.     

Short-term competition between crop plants and soil microbes for inorganic N fertilizer

Agricultural systems that receive high amounts of inorganic nitrogen (N) fertilizer in the form of either ammonium (NH₄⁺), nitrate (NO₃⁻) or a combination thereof are expected to differ in soil N transformation rates and fates of NH₄⁺ and NO₃⁻. Using ¹⁵N tracer techniques this study examines how crop plants and soil microbes vary in their ability to take up and compete for fertilizer N on a short time scale (hours to days). Single plants of barley (Hordeum vulgare L. cv. Morex) were grown on two agricultural soils in microcosms which received either NH₄⁺, NO₃⁻ or NH₄NO₃. Within each fertilizer treatment traces of ¹⁵NH₄⁺ and ¹⁵NO₃⁻ were added separately. During 8 days of fertilization the fate of fertilizer ¹⁵N into plants, microbial biomass and inorganic soil N pools as well as changes in gross N transformation rates were investigated. One week after fertilization 45-80% of initially applied ¹⁵N was recovered in crop plants compared to only 1-10% in soil microbes, proving that plants were the strongest competitors for fertilizer N. In terms of N uptake soil microbes out-competed plants only during the first 4 h of N application independent of soil and fertilizer N form. Within one day microbial N uptake declined substantially, probably due to carbon limitation. In both soils, plants and soil microbes took up more NO₃⁻ than NH₄⁺ independent of initially applied N form. Surprisingly, no inhibitory effect of NH₄⁺ on the uptake and assimilation of nitrate in both, plants and microbes, was observed, probably because fast nitrification rates led to a swift depletion of the ammonium pool. Compared to plant and microbial NH₄⁺ uptake rates, gross nitrification rates were 3-75-fold higher, indicating that nitrifiers were the strongest competitors for NH₄⁺ in both soils. The rapid conversion of NH₄⁺ to NO₃⁻ and preferential use of NO₃⁻ by soil microbes suggest that in agricultural systems with high inorganic N fertilizer inputs the soil microbial community could adapt to high concentrations of NO₃⁻ and shift towards enhanced reliance on NO₃⁻ for their N supply.     

Gross N transformation rates and net N mineralisation rates related to the C and N contents of soil organic matter fractions in grassland soils of different age

We investigated the relationship between soil organic matter (SOM) content and N dynamics in three grassland soils (0-10 and 10-20 cm depth) of different age (6, 14 and 50 y-old) with sandy loam textures. To study the distribution of the total C and N content the SOM was fractionated into light, intermediate and heavy density fractions of particulate macro-organic matter (150-2000 μm) and the 50-150 μm and <50 μm size fractions. The potential gross N transformation rates (mineralisation, nitrification, NH₄⁺ and NO₃⁻ immobilization) were determined by means of short-term, fully mirrored ¹⁵N isotope dilution experiments (7-d incubations). The long-term potential net N mineralisation and gross N immobilization rates were measured in 70-d incubations. The total C and N contents mainly tended to increase in the 0-10 cm layer with increasing age of the grassland soils. Significant differences in total SOM storage were detected for the long-term (50 y-old) conversion from arable land to permanent grassland. The largest relative increase in C and N contents had occurred in the heavy density fraction of the macro-organic matter, followed by the 50-150 and <50 μm fractions. Our results suggest that the heavy density fraction of the macro-organic matter could serve as a good indicator of early SOM accumulation, induced by converting arable land to permanent grassland. Gross N mineralisation, nitrification, and (long-term) gross N immobilization rates tended to increase with increasing age of the grasslands, and showed strong, positive correlations with the total C and N contents. The calculated gross N mineralisation rates (7-d incubations) and net N mineralisation rates (70-d incubations) corresponded with a gross N mineralisation of 643, 982 and 1876 kg N ha⁻¹ y⁻¹, and a net N mineralisation of 195, 208 and 274 kg N ha⁻¹ y⁻¹ in the upper 20 cm of the 6, 14 and 50 y-old grassland soils, respectively. Linear regression analysis showed that 93% of the variability of the gross N mineralisation rates could be explained by variation in the total N contents, whereas total N contents together with the C-to-N ratios of the <50 μm fraction explained 84% of the variability of the net N mineralisation rates. The relationship between long-term net N mineralisation rates and gross N mineralisation rates could be fitted by means of a logarithmic equation (net m=0.24Ln(gross m)+0.23, R²=0.69, P<0.05), which reflects that the ratio of gross N immobilization-to-gross N mineralisation tended to increase with increasing SOM contents. Microbial demand for N tended to increase with increasing SOM content in the grassland soils, indicating that potential N retention in soils through microbial N immobilization tends to be limited by C availability.     

Chemical fixation of ammonia to soil organic matter after application of urea

Chemical fixation of NH₃ to soil organic matter was studied in two Swedish soils with different contents of organic matter: a clay soil with 2.3% C and an organic soil with 36.6% C. ¹⁵N‐labelled urea was applied at different rates to both sterilized and non‐sterilized soils. After 10 days, the soils were extracted and washed with K₂SO₄ and determined for total N and atom% ¹⁵N excess. Urea N was recovered as non‐extractable N in sterilized soil corresponding to 9.7% of supplied ^l5N‐labelled urea in the organic soil and 2.2% in the clay soil. Since no biological immobilization is thought to occur in the sterile soil, this non‐extractable N is suggested to be chemically fixed to soil organic matter. Owing to urea hydrolysis in the clay soil, pH increased from 6.3 to 9.3 and in the organic soil from 5.7 to 6.9 and 8.8, respectively, at the low and high urea supply.     

C and N turnover in structurally intact soils of different texture

The turnover of native and applied C and N in undisturbed soil samples of different texture but similar mineralogical composition, origin and cropping history was evaluated at −10 kPa water potential. Cores of structurally intact soil with 108, 224 and 337 g clay kg⁻¹ were horizontially sliced and ¹⁵N-labelled sheep faeces was placed between the two halves of the intact core. The cores together with unamended treatments were incubated in the dark at 20 °C and the evolution of CO₂-C determined continuously for 177 d. Inorganic and microbial biomass N and ¹⁵N were determined periodically. Net nitrification was less in soil amended with faeces compared with unamended soil. When adjusted for the NO₃-N present in soil before faeces was applied, net nitrification became negative indicating that NO₃-N had been immobilized or denitrified. The soil most rich in clay nitrified least N and ¹⁵N. The amounts of N retained in the microbial biomass in unamended soils increased with clay content. A maximum of 13% of the faeces ¹⁵N was recovered in the microbial biomass in the amended soils. CO₂-C evolution increased with clay content in amended and unamended soils. CO₂-C evolution from the most sandy soil was reduced due to a low content of potentially mineralizable native soil C whereas the rate constant of C mineralization rate peaked in this soil. When the pool of potentially mineralizable native soil C was assumed proportional to volumetric water content, the three soils contained similar proportions of potentially mineralizable native soil C but the rate constant of C mineralization remained highest in the soil with least clay. Thus although a similar availability of water in the three soils was ensured by their identical matric potential, the actual volume of water seemed to determine the proportion of total C that was potentially mineralizable. The proportion of mineralizable C in the faeces was similar in the three soils (70% of total C), again with a higher rate constant of C mineralization in the soil with least clay. It is hypothesized that the pool of potentially mineralizable C and C rate constants fluctuate with the soil water content.     

Transfer of N fixed by a legume tree to the associated grass in a tropical silvopastoral system

Below-ground transfer of nitrogen (N) fixed by legume trees to associated non-N₂-fixing crops has received little attention in agroforestry, although the importance of below-ground interactions is shown in other ecosystems. We used ¹⁵N natural abundance to estimate N transfer from the legume tree Gliricidia sepium (Jacq.) Kunth ex Walp. to C₄ grass Dichanthium aristatum (Poir.) C.E. Hubb. in a silvopastoral system, where N was recycled exclusively by below-ground processes and N₂ fixation by G. sepium was the sole N input to the system. Finding a suitable reference plant, a grass without contact with tree roots or litter, was problematic because tree roots invaded adjacent grass monocrop plots and soil isotopic signature in soil below distant grass monocrops differed significantly from the agroforestry plots. Thus, we used grass cultivated under greenhouse conditions in pots filled with agroforestry soil as the reference. A model of soil ¹⁵N fractionation during N mineralization was developed for testing the reliability of that estimate. Experimental and theoretical results indicated that 9 months after greenhouse transplanting, the percentage of fixed N in the grass decreased from 35% to <1%, due to N export in cut grass and dilution of fixed N with N taken up from the soil. The effect of soil ¹⁵N fractionation on the estimate of the reference value was negligible. This indicates that potted grass is a suitable reference N transfer studies using ¹⁵N natural abundance. About one third of N in field-grown grass was of atmospheric origin in agroforestry plots and in adjacent D. aristatum grassland invaded by G. sepium roots. The concentration of fixed N was correlated with fine root density of G. sepium but not with soil isotopic signature. This suggests a direct N transfer from trees to grass, e.g. via root exudates or common mycorrhizal networks.     

Contrasting response of two forest soils to nitrogen input: rapidly altered NO and N2O emissions and nirK abundance

Denitrification represents one of the main microbial processes producing the primary and secondary greenhouse gases nitrous oxide (N₂O) and nitric oxide (NO) in soils. It is well established that abiotic factors like the soil water content and the availability of nitrogen (N) are key parameters determining the activity of denitrifiers in soils. However, soils differing regarding their characteristics such as the content of C_org, the soil texture or the pH value may respond in specific manners to equivalent changes in soil moisture and N input. Thus, short-term incubation experiments were performed to test and compare the capacity of two contrasting Austrian forest soils to respond to mineral N application at increased soil water contents. Soils from the pristine Rothwald forest (rich in C_org) and the more acidic Schottenwald forest (poor in C_org) were amended with either NH ₄ ⁺ -N or NO ₃ ^? -N and were incubated at 40% and 70% water-filled pore space for 4 days. Changes in mineral N pools, nitrite reductase activity and NO and N₂O emission rates were measured, and the abundance and structural community composition of the functional group involved in nitrite reduction were analysed via quantitative real-time polymerase chain reaction and terminal restriction fragment length polymorphism analysis of the nirK gene. Rapid and distinct activity responses to increased soil moisture and altered mineral nitrogen availability were observed in two contrasting forest soils. In both soils, nitrogen oxide emission rates were stimulated by N inputs and, depending on the soil moisture status, either NO or N₂O emission was prevailing. However, different N cycling processes appeared to predominate in either soil under equivalent treatment. Nitrogen oxide emissions peaked following NO ₃ ^? application in Schottenwald soils but were the highest after NH ₄ ⁺ application in Rothwald soils. Denitrifying (nirK) communities differed significantly in Rothwald and Schottenwald soils; however, changes in the community structure were marginal during the short-term incubation. Abundances of nirK genes remained unaffected by N application in either soil. The soil water content affected nirK gene abundances only in Rothwald soil, indicating a distinct reaction of nitrite reducing communities in the two soils.