Denitrification Rate and Controlling Factors for Accumulated Nitrate in the Deep Subsoil of Intensive Farmlands: A Case Study in the North China Plain

Denitrification in subsoil(to a depth of 12 m) is an important mechanism to reduce nitrate(NO_3~-) leaching into groundwater.However, regulating mechanisms of subsoil denitrification, especially those in the deep subsoil beneath the crop root zone, have not been well documented. In this study, soil columns of 0–12 m depth were collected from intensively farmed fields in the North China Plain. The fields had received long-term nitrogen(N) fertilizer inputs at 0(N0), 200(N200) and 600(N600) kg N ha~(-1) year~(-1). Main soil properties related to denitrification, i.e., soil water content, NO_3~-, dissolved organic carbon(DOC), soil organic carbon(SOC),pH, denitrifying enzyme activity(DEA), and anaerobic denitrification rate(ADR), were determined. Statistical comparisons among the treatments were performed. The results showed that NO_3~- was more heavily accumulated in the entire soil profile of the N600 treatment, compared to the N0 and N200 treatments. The SOC, DOC, and ADR decreased with increasing soil depth in all treatments,whereas considerable DEA was observed throughout the subsoil. The long-term fertilizer rates affected ADR only in the upper 4 m soil layers. The ADRs in the N200 and N600 treatments were significantly correlated with DOC. Multiple regression analysis indicated that DOC rather than DEA was the key factor regulating denitrification beneath the root zone. Additional research is required to determine if carbon addition into subsoil can be a promising approach to enhance NO_3~- denitrification in the subsoil and consequently to mitigate groundwater NO_3~- contamination in the intensive farmlands.     

中国太湖流域水稻土中的反硝化细菌

This study attempted to determine the characteristics of the communities, the ecological factors, and the denitrifying enzyme activity for denitrifying bacteria found in the paddy soils of the Taihu Lake Basin, China. Samples of the six main soil types of the basin were taken from paddy fields with different fertilities. The total numbers of bacteria and denitrifying bacteria in the high fertility soils were much more than those in low fertility soils, and the number of denitrifying bacteria accounted for 49% to 80% of the total number of bacteria. The O2 content was an important ecological factor that affected denitrification. Of test the strains isolated from the paddy soils in the Taihu Lake Basin, some (e.g., Pseudomonas spp.) grew well under low oxygen partial pressure, while others (e.g., Bacillus spp.) had no strict predilection with O2 content. Another critical ecological factor was the nitrogen concentration. Three selected denitrifying bacteria grew better in aculture medium with 135 instead of 276mg L^-1 nitrogen. At the same time 67% of the test strains were able to reduce NO3^- to NO2^- and 56% had N2O reductase.     

Mitrification-Denitrification Loss of Added Nitrogen in Flooded RiceRhizosphere

Nitrification-denitrification losses of 15N-labelled nitrate and ammonium applied to the rhizos phere and nonrhizosphere of flooded rice were evaluated in 2 greenhouse rhizobox experiments.The loss of added N via denitrification was estimated directly by measuring the total fluxes of (N2O N2)12N,It was found that 67% and 51%-56% of 15N-nitrate added to rice rhizosphere were lost as (N2O N2)-15N in the 2 experiments,respectively,which were comparable to that added to norhizosphere soil(70%and 47%,respectively),implying that the denitrifying activity in rice rhizosphere was as high as that in nonrhizosphere soil.However,only trace amounts (0-0\3% of added N)were recovered as (N2O N2)-15N when 15N-ammonium was applied to either rhizosphere or nonrhizosphere,which seems to indicate that the nitrifying activity in the either rhizosphere of nonrhizosphere soils was quite low.The apparent denitrification calculated from 15N balance studies was 10%-47% higher than the total flux of (N2O N2)-15N.Reasons for the large differences can not be explained satisfactorily.Though the denitrifying activity in rhizosphere was high and comparable to that in nonrhizosphere soil.presumably due to the low nitrifying activity and /or the strong competition of N uptake against denitrification.the nitrification-denitrification taking place in rhizosphere could not be an important mechanism of loss of ammonium N in flooded rice-soil system.     

施用液态家畜废物的水稻田土壤中硝酸盐异化还原成铵过程的研究

Most studies on dissimilatory nitrate reduction to ammonium (DNRA) in paddy soils were conducted in the laboratory and in situ studies are in need for better understanding of the DNRA process.In this study,in situ incubations of soil DNRA using 15 N tracer were carried out in paddy fields under conventional water (CW) and low water (LW) managements to explore the potential of soil DNRA after liquid cattle waste (LCW) application and to investigate the impacts of soil redox potential (Eh) and labile carbon on DNRA.DNRA rates ranged from 3.06 to 10.40 mg N kg 1 dry soil d 1,which accounted for 8.55%-12.36% and 3.88%-25.44% of consumption of added NO 3-15 N when Eh at 5 cm soil depth ranged from 230 to 414 mV and 225 to 65 mV,respectively.DNRA rates showed no significant difference in paddy soils under two water managements although soil Eh and/or dissolved organic carbon (DOC) were more favorable for DNRA in the paddy soil under CW management 1 d before,or 5 and 7 d after LCW application.Soil DNRA rates were negatively correlated with soil Eh (P < 0.05,n=5) but positively correlated with soil DOC (P < 0.05,n=5) in the paddy soil under LW management,while no significant correlations were shown in the paddy soil under CW management.The potential of DNRA measured in situ was consistent with previous laboratory studies;and the controlling factors of DNRA in paddy soils might be different under different water managements,probably due to the presence of different microfloras of DNRA.     

中国亚热带地区土地利用和磷释放的数量关系

The increase of phosphorus concentration is a crucial factor causing the eutrophication of water body.while land use has an important impact on agricultural non-point sources(NPS) phosphorus discharge,Seven sites controlling the water in four sub-watersheds and the main exit of the Meicun Watershed of Xuancheng County,Anhui Provinec,were investigated by dynamic monitoring of stream water and nutrient discharge,integrating interpretation of areial image and GIS analysis to find out how the land use affects phosphorus loss with stream water in typical agriculture-forest watershed in subtropica China.These monitored sites are different in structure of typical agriculture-forest watershed in subtropical China.These monitored sitess are different in structure of land use,Phosphorus concentration of the stream water was analyzed every week and at the next day of rainfall,The velocity of flow was measured by kinemometer to calculate the runoff flux and phosphorus discharge.The results showed that the runoff flux and the discharges of dissolved phosphorus(DP),particle-associated phosphorus(PAP) and total phosphorus(TP) had significant exponential relationships with the area percentages of forest,pond and paddy field.There existed a significant exponential relationships with the area percentages of forest,pond and paddy field.There existed a significant linear relationship between the TP and PAP concentrations in stream water and the area percentages of forest,pond and paddy field,and the discharge of PAP was also significantly linearly correlated with the discharge of suspended soil particles.There was a logarithmic linear relationship between DP and PAP discharges,The study indicated that the adjustment of land used patterns and construction of ecologically sound aldnscape would be an important measure to reduce the runoff discharge of phosphorus,The results would be very useful in building the best management practices(BMPs) of agricultural watershed in subtropics.     

菜地土壤中氮肥的反硝化损失和N2O排放

A field experiment was conducted on Chinese cabbage （Brassica campestris L. ssp. pekinensis （Lour.） Olsson） in a Nanjing suburb in 2003. The experiment included 4 treatments in a randomized complete block design with 3 replicates： zero chemical fertilizer N （CK）; urea at rates of 300 kg N ha^-1 （U300） and 600 kg N ha^-1 （U600）, both as basal and two topdressings; and polymer-coated urea at a rate of 180 kg N ha^-1 （PCU180） as a basal application. The acetylene inhibition technique was used to measure denitrification （N2 ＋ N2O） from intact soil cores and N2O emissions in the absence of acetylene. Results showed that compared to （3K total denitrification losses were significantly greater （P ≤ 0.05） in the PCU180, U300, and U600 treatments,while N2O emissions in the U300 and U600 treatments were significantly higher （P ≤ 0.05） than （3K. In the U300 and U600 treatments peaks of denitrification and N2O emission were usually observed after N application. In the polymer-coated urea treatment （PCU180） during the period 20 to 40 days after transplanting, higher denitrification rates and N2O fluxes occurred. Compared with urea, polymer-coated urea did not show any effect on reducing denitrification losses and N2O emissions in terms of percentage of applied N. As temperature gradually decreased from transplanting to harvest, denitrification rates and N2O emissions tended to decrease. A significant （P ≤0.01） positive correlation occurred between denitrification （r = 0.872） or N2O emission （r = 0.781） flux densities and soil temperature in the CK treatment with a stable nitrate content during the whole growing season.     

中国南方大棚蔬菜地氮平衡与损失

High rates of fertilizer nitrogen (N) are applied in greenhouse vegetable fields in southeastern China to maximize production;however,the N budgets of such intensive vegetable production remain to be explored.The goal of this study was to determine the annual N balance and loss in a greenhouse vegetable system of annual rotation of tomato,cucumber,and celery at five N (urea) application rates (0,348,522,696,and 870 kg N ha-1 year-1).Total N input to the 0-50 cm soil layer ranged from 531 to 1 053 kg ha-1,and N fertilizer was the main N source,accounting for 66%-83% of the total annual N input.In comparison,irrigation water,wet deposition,and seeds in total accounted for less than 1% of the total N input.The fertilizer N use efficiency was only 18% under the conventional application rate of 870 kg N ha-1 and decreased as the application rate increased from 522 to 870 kg N ha-1.Apparent N losses were 196-201 kg N ha-1,of which 71%-86% was lost by leaching at the application rates of 522-870 kg N ha-1.Thus,leaching was the primary N loss pathway at high N application rates and the amount of N leached was proportional to the N applied during the cucumber season.Moreover,dissolved organic N accounted for 10% of the leached N,whereas NH3 volatilization only contributed 0.1%-0.6% of the apparent N losses under the five N application rates in this greenhouse vegetable system.     

城郊结合区蔬菜种植地土壤性质特征

Human activity and urbanization result in urban-rural environmental gradients.Undersanding effect of the gradients on soil properties is necessary for management of the soils around urban areas.In this study,soil quality of some vegetable fields was characterized along an urban-rural gradient in shaoxing County,Zhejiang Province.Fifteen soilphysical and chemical properties were evaluated by using principal component analysis.Results showed that there was a great variation in the soil quality along the gradient.From rural to urban zones,soil organic matter,water-stable aggregates,cation exchangeable capacity(CEC),total N and P,and available K increased,whereas soil pH value decreased.In addition,Pb,Cu,Ni,Co,Zn and Cr in the soils tended to be accumulated toward the urban zone.Sequential chemical extraction showed that mobility of all the heavy metals in the soils tended toincrease from the rural to the urban zones.The variation of soil properties accounted for by the first principal component was significantly explained bythe difference in application rates of municipal wastes.     

中国东部合肥城市土壤氮素矿化特点

Nitrogen cycling has been poorly characterized in urban ecosystems. In this study, the in-situ buried bag incubation technique was used to quantify net rates of N mineralization and nitrification in soils of two urban sites, a street greening belt and a university campus, and a suburban site , a forest park, in Hefei, East China. The average concentration of extractable NO3- in the surface soil (0--10 cm) was significantly higher at the urban sites than the suburban park site, whereas extractable NH4+ concentration was significantly higher at the suburban park site than the urban sites. The forest park soil had greater potential N mineralization (148.1 μg N cm-3) than the soils from the campus (138.3 μg N cm-3) and street (99.8 μg N cm-3). It was estimated that the net mineralization rates varied between 1.63 and 2.69 μg N cm-3 d-1 and net nitrification rates between 0.82 and 1.02 μg N cm-3 d-1 at the suburban forest park site, but the rates varied from 1.27 to 2.41 μg N cm-3 d-1 and from 1.07 to 1.49 μg N cm-3 d-1, respectively, at the urban campus site. Both net mineralization and nitrification rates were lower during dry seasons. Results from regression analysis indicated that net N mineralization was significantly and positively correlated with soil moisture and soil C/N ratio, and was negatively correlated with soil pH. Relative nitrification was, however, significantly and negatively correlated with soil moisture and soil C/N ratio, and was positively correlated with soil pH. Mean relative nitrification was 0.763, indicating the dominance of nitrate cycling relative to ammonium cycling at the urban sites. The urban soils had the great potential for N losses compared to the suburban soils.     

太湖地区水稻最适宜施氮量研究

To determine the optimal amount of nitrogen(N) fertilizer for achieving a sustainable rice production at the Taihu Lake region of China,two-year on-farm field experiments were performed at four sites using various N application rates.The results showed that 22%-30% of the applied N was recovered in crop and 7%-31% in soils at the rates of 100-350 kg N ha 1.Nitrogen losses increased with N application rates,from 44% of the applied fertilizer N at the rate of 100 kg N ha 1 to 69% of the N applied at 350 kg N ha 1.Ammonia volatilization and apparent denitrification were the main pathways of N losses.The N application rate of 300 kg N ha 1,which is commonly used by local farmers in the study region,was found to lead to a significant reduction in economic and environmental efficiency.Considering the cost for mitigating environmental pollution and the maximum net economic income,an application rate of 100-150 kg N ha 1 would be recommended.This recommended N application rate could greatly reduce N loss from 199 kg N ha 1 occurring at the N application rate of 300 kg N ha 1 to 80-110 kg N ha 1,with the rice grain yield still reaching 7 300-8 300 kg DW ha 1 in the meantime.     

Sediment denitrification in waterways in a rice-paddy-dominated watershed in eastern China

Purpose

Rice-paddy-dominated watersheds in eastern China are intensively cultivated, and lands with two crops receive as much as 550–600 kg?ha^–1?year^–1 of nitrogen (N), mainly through the addition of N-based fertilizers. However, stream N concentrations have been found to be relatively low. Waterways in the watersheds are assumed to be effective “sinks” for N, minimizing its downstream movement. We directly measured net sediment denitrification rates in three types of waterways (ponds, streams/rivers, and a reservoir) and determined the key factors that control net sediment denitrification. Such information is essential for evaluating the impact of the agricultural N cycle on the quality of surface water.

Materials and methods

The pond–stream–reservoir continuum was sampled every 2 months at nine sites in an agricultural watershed between November 2010 and December 2011. Net sediment N₂ fluxes/net sediment denitrification rates were determined by membrane inlet mass spectrometry and the N₂/Ar technique. A suite of parameters known to influence denitrification were also measured.

Results and discussion

Net denitrification rates ranged between 28.2?±?18.2 and 674.3?±?314.5 μmol N₂–N?m^–2?h^–1 for the streams, 23.7?±?23.9 and 121.2?±?38.7 μmol N₂–N?m^–2?h^–1 for the ponds, and 41.8?±?17.7 and 239.3?±?49.8 μmol N₂–N?m^–2?h^–1 for the reservoir. The mean net denitrification rate of the stream sites (173.2?±?248.4 μmol N₂–N?m^–2?h^–1) was significantly higher (p?<?0.001) than that of the pond sites (48.3?±?44.5 μmol N₂–N?m^–2?h^–1), and the three types of waterways all had significantly higher (p?<?0.01) mean net denitrification rates in summer than in other seasons. Linear regression and linear mixed effect model analysis showed that nitrate (NO₃ ^?–N) concentration in surface water was the primary controlling factor for net sediment denitrification, followed by water temperature. Using monitoring data on NO₃ ^?–N concentrations and temperature of the surface water of waterways and an established linear mixed effect model, total N removed through net sediment denitrification in the pond–stream–reservoir continuum was estimated at 46.8?±?24.0 t?year^–1 from July 2007 to June 2009, which was comparable with earlier estimates based on the mass balance method (34.3?±?12.7 t?year^–1), and accounted for 83.4 % of the total aquatic N. However, the total aquatic N was only 4.4 % of the total N input to the watershed, and thus most of the surplus N in the watershed was likely to be either denitrified or stored in soil.

Conclusions

High doses of N in a rice-paddy-dominated watershed did not lead to high stream N concentrations due to limited input of N into waterways and the high efficiency of waterways in removing N through denitrification.     

Nitrogen dynamics in different types of pasture in the Austrian Alps

 Soil N dynamics were compared in Alpine pastures on two mountains. N-pool sizes and N fluxes were measured relative to N losses via leaching and denitrification in summer. On each mountain, four types of pasture were studied: (1) forest pastures, (2) recently developed pastures formed by forest clearance ("new pastures"), (3) older established pastures, and (4) pastures planted with clover. At both study sites (Scheuchegg and Teufelstein) we obtained similar results. Compared with forest pasture soils, open pasture soils were found to have greater microbial biomass and faster mineralisation potentials, but net field mineralisation rates were slower. In the forest pastures, highest N losses via denitrification were found. Higher potential leaching of NO₃ ^–, estimated by accumulation of NO₃ ^– on ion-exchange resins, in the forest pasture soils suggests lower N uptake by microbes and herbaceous plants compared with open pastures. N₂O-production rates of the forest pasture soils at the Scheuchegg site (11.54 μg N₂O-N m^–2 h^–1) were of similar magnitude to those reported for spruce forests without pastures, but at Teufelstein (53.75 μg N₂O-N m^–2 h^–1) they were higher. However, if forest pastures are not overgrazed, no elevated N loss through N₂O production and leaching of NO₃ ^– is expected. Denitrification rates in the open pastures (0.83–7.50 μg N₂O-N m^–2 h^–1) were low compared with reports on lowland pastures. In soils of the new pastures, rates of microbial N processes were similar to those in the established pastures, indicating a high capacity of soils to restore their internal N cycle after forest clearance. Received: 19 August 1999     

Nitrate removal, denitrification and nitrous oxide production in the riparian zone of an ephemeral stream

Riparian zones are important features of the landscape that can buffer waterways from non-point sources of nitrogen pollution. Studies of perennial streams have identified denitrification as one of the dominant mechanisms by which this can occur. This study aimed to assess nitrate removal within the riparian zone of an ephemeral stream and characterise the processes responsible, particularly denitrification, using both in-situ and laboratory techniques. To quantify rates of groundwater nitrate removal and denitrification in-situ, nitrate was added to two separate injection-capture well networks in a perched riparian aquifer of a low order ephemeral stream in South East Queensland, Australia. Both networks also received bromide as a conservative tracer and one received acetylene to inhibit the last step of denitrification. An average of 77 ± 2% and 98 ± 1% of the added nitrate was removed within a distance of 40 cm from the injection wells (networks with acetylene and without, respectively). Based on rates of N₂O production in the network with added acetylene, denitrification was not a major mechanism of nitrate loss, accounting for only 3% of removal. Reduction of nitrate to ammonium was also not a major pathway in either network, contributing <4%. Relatively high concentrations of oxygen in the aquifer following recent filling by stream water may have reduced the importance of these two anaerobic pathways. Alternatively, denitrification may have been underestimated using the in-situ acetylene block technique. In the laboratory, soils taken from two depths at each well network were incubated with four nitrate-N treatments (ranging from ambient concentration to an addition of 15 mg N l⁻¹), with and without added acetylene. Potential rates of denitrification, N₂O production and N₂O:N₂ ratios increased with nitrate additions, particularly in shallow soils. Potential rates of denitrification observed in the laboratory were equivalent in magnitude to nitrate removal measured in the field (mean 0.26 ± 0.12 mg N kg of dry soil⁻¹ d⁻¹), but were two orders of magnitude greater than denitrification measured in the field with added acetylene. The relative importance of assimilatory vs. dissimilatory processes of nitrate removal depends on environmental conditions in the aquifer, particularly hydrology and its effects on dissolved oxygen concentrations. Depending on seasonal conditions, aquifers of ephemeral streams like the study site are likely to fluctuate between oxic and anoxic conditions; nevertheless they may still function as effective buffers. While denitrification to N₂ is a desirable outcome from a management perspective, assimilation into biomass can provide a rapid sink for nitrate, thus helping to reduce short-term delivery of nitrate downstream. Longer-term studies are needed to determine the overall effectiveness of riparian buffers associated with ephemeral streams in mitigating nitrate loads reaching downstream ecosystems.     

Compared effects of long-term pig slurry applications and mineral fertilization on soil denitrification and its end products (N2O, N2)

The long-term (9 years) effect of pig slurry applications vs mineral fertilization on denitrifying activity, N₂O production and soil organic carbon (C) (extractable C, microbial biomass C and total organic C) was compared at three soil depths of adjacent plots. The denitrifying activities were measured on undisturbed soil cores and on sieved soil samples with acetylene method to estimate denitrification rates under field or potential conditions. Pig slurry applications had a moderate impact on the C pools. Total organic C was increased by +6.5% and microbial biomass C by ≥25%. The potential denitrifying activity on soil suspension was stimulated (×1.8, P<0.05) 12 days after the last slurry application. This stimulation was still apparent, but not significant, 10 months later and, according to both methods of denitrifying activity measurement (r ²=0.916, P<0.01 on sieved soil; r ²=0.845, P<0.001 on soil cores), was associated with an increase in microbial biomass C above a threshold of about 105 mg kg⁻¹. The effect of pig slurry on denitrification and N₂O reduction rates was detected on the surface layer (0–20 cm) only. However, no pig slurry effect could be detected on soil cores at field conditions or after NO₃ ⁻ enrichments at 20°C. Although the potential denitrifying activity in sieved soil samples was stimulated, the N₂O production was lower (P<0.03) in the plot fertilized with pig slurry, indicating a lower N₂O/(N₂O + N₂) ratio of the released gases. The pig-slurry-fertilized plot also showed a higher N₂O reduction activity, which is coherent with the lower N₂O production in anaerobiosis.     

Contribution of nitrification and denitrification to N2O emissions from urine patches

Urine deposition by grazing livestock causes an immediate increase in nitrous oxide (N₂O) emissions, but the responsible mechanisms are not well understood. A nitrogen-15 (¹⁵N) labelling study was conducted in an organic grass-clover sward to examine the initial effect of urine on the rates and N₂O loss ratio of nitrification (i.e. moles of N₂O-N produced per moles of nitrate produced) and denitrification (i.e. moles of N₂O produced per moles of N₂O+N₂ produced). The effect of artificial urine (52.9 g N m⁻²) and ammonium solution (52.9 g N m⁻²) was examined in separate experiments at 45% and 35% water-filled pore space (WFPS), respectively, and in each experiment a water control was included. The N₂O loss derived from nitrification or denitrification was determined in the field immediately after application of ¹⁵N-labelled solutions. During the next 24 h, gross nitrification rates were measured in the field, whereas the denitrification rates were measured in soil cores in the laboratory. Compared with the water control, urine application increased the N₂O emission from 3.9 to 42.3 μg N₂O-N m⁻² h⁻¹, whereas application of ammonium increased the emission from 0.9 to 6.1 μg N₂O-N m⁻² h⁻¹. In the urine-affected soil, nitrification and denitrification contributed equally to the N₂O emission, and the increased N₂O loss resulted from a combination of higher rates and higher N₂O loss ratios of the processes. In the present study, an enhanced nitrification rate seemed to be the most important factor explaining the high initial N₂O emission from urine patches deposited on well-aerated soils.     

Carbon-iron coupling reduced N2O emissions via promoting the conversion to N2 in paddy soils

The influence of redox reactions involving carbon-iron coupling (organic carbon and iron oxides) on nitrous oxide (N₂O) production in paddy soils remains poorly understood. In this study, two microcosm experiments were conducted to investigate the effects of carbon-iron coupling on N₂O emissions, and the underlying mechanisms were verified using quantitative denitrification functional genes (nirS, nirK, nosZI and nosZII) and high-throughput sequencing. The results showed that ferrihydrite (iron) significantly promoted N₂O-N emissions (p < 0.05) after adding ammonium nitrogen, while glucose (carbon) significantly inhibited N₂O-N emissions (p < 0.05). Carbon-iron coupling significantly decreased N₂O-N emissions (p < 0.05) but did not affect soil total nitrogen loss and increased nitrogen (N₂) emissions. After adding high concentrations of acetylene (10% C₂H₂), the N₂O-N emissions from carbon-iron coupling treatment increased significantly from 6.4 to 11.9 mg N kg⁻¹ (p < 0.05), which confirmed that the carbon-iron coupling reduced the N₂O emissions by promoting the conversion of N₂O to N₂. The mechanisms behind carbon-iron coupling promoting complete denitrification and reducing N₂O emissions were attributed to glucose promoting iron reduction and carbon-iron coupling enhancing the abundance of nosZI (42.7%) and nosZII (16.6%).     

Nitrous oxide dynamics during denitrification along a hydrological gradient of subtropical grasslands

Nitrous oxide (N₂O) dynamics during denitrification, including N₂O production and reduction, particularly as related to soil depth, are poorly understood. The objective of this study was to investigate the rates of N₂O production and reduction processes at various soil depths along a hydrological gradient in grazed subtropical grasslands. A batch incubation study was conducted on soils collected along a hydrological gradient representing isolated wetland (Center), transient edge (Edge) and pasture upland (Upland) in south-central Florida. Significantly different N₂O production and reduction rates between hydrological zones were observed for surface soils (0–10 cm) under ambient conditions, with average N₂O production rates of 0.368, 0.178 and 0.003 N₂O-N kg⁻¹ dry soil h⁻¹ for Center, Edge and Upland, respectively, and average N₂O reduction rates of 0.063, 0.132 and 0.002 N₂O-N kg⁻¹ dry soil h⁻¹. Nitrous oxide production and reduction in subsurface soils maintained low rates and showed small variations between depths and hydrological zones. Our results suggest that N₂O dynamics were affected by depth, mainly through labile organic carbon (C) and microbial biomass C, being influenced by hydrological zone primarily through soil NO₃^- content. The spatial distribution of N₂O fluxes from denitrification along the hydrological gradient is likely attributed to the differences in N₂O production and reduction in surface soils.     

Production of NO and N2O in the presence and absence of C2H2 by soil slurries and batch cultures of denitrifying bacteria

We evaluated the potential of the C₂H₂-catalyzed NO oxidation reaction to influence N₂O production during denitrification. We measured the total amount of free NO and N₂O produced by slurries of sandy loam soil and by batch cultures of denitrifying bacteria under both anaerobic and low O₂ conditions. The maximum amount of free NO released by anaerobic Nos⁺ (able to reduce N₂O to N₂) and Nos⁻ (unable to reduce N₂O to N₂) batch cultures of Cytophaga johnsonae strains catalyzing denitrification of nitrate was 17-79 nmol NO per bottle. In all cases the maximum headspace concentrations of NO-N measured in anaerobic cultures in the absence of C₂H₂ was less than 0.21% of those of N₂O-N measured in the presence of 10 kPa C₂H₂. Peak NO production was delayed when between 2.0 and 4.5% O₂ was present. Less NO accumulated in cultures in the presence of both O₂ and C₂H₂, and the maximum amount of NO-N measured in the absence of C₂H₂ was less than 0.13% of the total amount of N₂O-N measured in the presence of C₂H₂. For an agricultural sandy loam soil, the maximum concentrations of free NO released from slurries were 598-897 ng NO-N g⁻¹ of dry soil in the absence of C₂H₂ and 118-260 ng NO-N g⁻¹ of dry soil in the presence of 10 kPa C₂H₂. The maximum concentration of NO-N released in the absence of C₂H₂ was between 0.32 and 8.1% of the maximum concentration of N₂O-N accumulated in the presence of 10 kPa C₂H₂. We conclude that scavenging of NO by the C₂H₂-catalyzed NO oxidation reaction in the presence of trace amounts of O₂ does not cause a serious underestimate of long-term measurements of active denitrification in anaerobic soils containing adequate carbon and nitrate sources.     

Diversity of denitrifying microflora and ability to reduce N2O in two soils

 The ozone-depleting gas N₂O is an intermediate in denitrification, the biological reduction of NO₃ ^– to the gaseous products N₂O and N₂ gas. The molar ratio of N₂O produced (N₂O/N₂O+N₂) varies temporally and spatially, and in some soils N₂O may be the dominant end product of denitrification. The fraction of NO₃ ^–-N emitted as N₂O may be due at least in part to the abundance and activity of denitrifying bacteria which possess N₂O reductase. In this study, we enumerated NO₃ ^–-reducing and denitrifying bacteria, and compared and contrasted collections of denitrifying bacteria isolated from two agricultural soils, one (Auxonne, soil A) with N₂O as the dominant product of denitrification, the other (Chalons, soil C) with N₂ gas as the dominant product. Isolates were tested for the ability to reduce N₂O, and the presence of the N₂O reductase (nosZ)-like gene was evaluated by polymerase chain reaction (PCR) using specific primers coupled with DNA hybridization using a specific probe. The diversity and phylogenetic relationships of members of the collections were established by PCR/restriction fragment length polymorphism of 16s rDNA. The two soils had similar numbers of bacteria which used NO₃ ^– as a terminal electron acceptor anaerobically. However, the soil A had many more denitrifiers which reduced NO₃ ^– to gaseous products (N₂O or N₂) than did soil C. Collections of 258 and 281 bacteria able to grow anaerobically in the presence of NO₃ ^– were isolated from soil A and soil C, respectively. These two collections contained 66 and 12 denitrifying isolates, respectively, the others reducing NO₃ ^– only as far as NO₂ ^–. The presence of nosZ sequences was generally a poor predictor of N₂O reducing ability: there was agreement between the occurrence of nosZ sequences and the N₂O reducing ability for only 42% of the isolates; 35% of the isolates (found exclusively in soil A) without detectable nosZ sequences reduced N₂O whereas 21% of the isolates carrying nosZ sequences did not reduce this gas under our assay conditions. Twenty-eight different 16S rDNA restriction patterns (using two restriction endonucleases) were distinguished among the 78 denitrifying isolates. Two types of patterns appeared to be common to both soils. Twenty-three and three types of patterns were found exclusively among bacteria isolated from soils A and C, respectively. The specific composition of denitrifying communities appeared to be different between the two soils studied. This may partly explain the differences in the behaviour of the soils concerning N₂O reduction during denitrification. Received: 31 October 1997     

The Effect of N Deposition on Nitrous Oxide and Nitric Oxide Emissions from Temperate Forest Soils

Long-term and short-term N deposition effects on N₂O and NO emissions from forest soils were compared. Long-term NH₃ deposition (> 20 years) from a poultry farm to a downwind woodland (decreasing from 73 to 18 kg N ha^-1 y^-1, 30 to 110 m downwind of the farm) resulted in the re-emission of 6% and 14% of NH₃-N deposited as N₂O-N and NO-N, respectively. However, when in short-term (2-3 years) field experiments the atmospheric N deposition to mature conifer plantations was raised by fumigation with NH₃ to 15 kg N ha^-1 y^-1 or by acid mist to 48 and 96 kg N ha^-1 y^-1 the N deposited was immobilised. In the acid mist experiment more than 2 years of acid mist (48 and 96 kg N ha-1 y^-1) were required to significantly increase N₂O emissions from -0.3 μg N₂O-N m^-2 h^-1 (control) to 0.5 and 5.7 μg N₂O-N m^-2 h^-1, respectively. This suggests, that N deposition simulation studies in soil ecosystems, which have previously not been exposed to high rates of N (by deposition or fertilisation), need to be long-term. Also, measurements of N₂O and/or NO may be a non-destructive, quick indicator of the N status of the soil.