Spatial Variation in Denitrification and N2O Emission in Relation to Nitrate Removal Efficiency in a N-stressed Riparian Buffer Zone

Spatial variability in hydrological flowpaths and nitrate-removal processes complicates the overall assessment of riparian buffer zone functioning in terms of water quality improvement as well as enhancement of the greenhouse effect by N₂O emissions. In this study, we evaluated denitrification and nitrous oxide emission in winter and summer along two groundwater flowpaths in a nitrate-loaded forested riparian buffer zone and related the variability in these processes to controlling soil factors. Denitrification and emissions of N₂O were measured using flux chambers and incubation experiments. In winter, N₂O emissions were significantly higher (12.4 mg N m⁻² d⁻¹) along the flowpath with high nitrate removal compared with the flowpath with low nitrate removal (2.58 mg N m⁻² d⁻¹). In summer a reverse pattern was observed, with higher N₂O emissions (13.6 mg N m⁻² d⁻¹) from the flowpath with low nitrate-removal efficiencies. Distinct spatial patterns of denitrification and N₂O emission were observed along the high nitrate-removal transect compared to no clear pattern along the low nitrate-removal transect, where denitrification activity was very low. Results from this study indicate that spots with high nitrate-removal efficiency also contribute significantly to an increased N₂O emission from riparian zones. Furthermore, we conclude that high variability in N₂O:N₂ ratio and weak relationships with environmental conditions limit the value of this ratio as a proxy to evaluate the environmental consequences of riparian buffer zones.     

Influence of nutrient solution pH on N2O and N2 emissions from a soilless culture system

The influence of nutrient solution pH on the emission of N2O and N2 was investigated during cultivation of cucumbers in a closed-loop rockwool system. Between pH 4 and 7 these gaseous nitrogen losses increased from 1.6 to 21.1% of the N fertilizer input. This was equivalent to average flux rates of 0.06 and 0.85 kg nitrogen per hectare greenhouse area and day, respectively. The N2O/N2 ratio was inversely related to the total gaseous nitrogen losses. At neutral pH dinitrogen was the main emission product, whereas more acidic conditions favoured the emission of nitrous oxide. The pH effects were probably not indirectly affected by root respiration or exudation as much as by a direct inhibition of the activity of denitrifying microorganisms due to high H+ concentrations since similar results were obtained in unplanted nutrient solution systems with the addition of glucose as carbon source. Despite the low microbial denitrification activity under acidic conditions, nitrogen balance deficits of up to one-fifth of the N input still occurred. It is suggested these losses were predominantly caused by chemodenitrification.     

Nitrate limitation of N2O production and denitrification from tropical pasture and rain forest soils

Nitrous oxide production was measured in intact cores taken from active pasture and old-growth forest Inceptisols in the Atlantic Lowlands of Costa Rica. Following additions of aqueous KNO₃ or glucose, or the two combined amendments, the cores were incubated in the laboratory to determine if N₂O production rates were either N-limited or C-limited in the two land use types. Differences in rates of denitrification (N2₂O + N₂ production) among amended forest and pasture soils were determined by addition of 10% C₂H₂.The forest soils were relatively insensitive to all amendment additions, including the acetylene block. Forest N₂O production rates among the treatments did not differ from the controls, and were consistently lower than those of the pasture soils. With the addition of glucose plus nitrate to the forest soils, production of N₂O was three times greater than the controls, although this increase was not statistically significant. On the other hand, the pasture soils were definitely nitrogen-limited since N₂O production rates were increased substantially beyond controls by all the amendments which contained nitrate, despite the very low N level (5 mg N kg^–1 soil) relative to typical fertilizer applications. With respect to the nitrate plus glucose plus acetylene treatment, denitrification was high in the pasture soils; N₂O production in the presence of C₂H₂ was 150% of the rate of N₂O production measured in the absence of the acetylene block. The results are discussed in relation to the effects of agricultural land use practices and subsequent impacts of disturbance on N₂O release.     

Denitrification and N2O emission from urine-affected grassland soil

Denitrification and N₂O emission rates were measured following two applications of artificial urine (40 g urine-N m^–2) to a perennial rye-grass sward on sandy soil. To distinguish between N₂O emission from denitrification or nitrification, urine was also applied with a nitrification inhibitor (dicyandiamide, DCD). During a 14 day period following each application, the soil was frequently sampled, and incubated with and without acetylene to measure denitrification and N₂O emission rates, respectively.Urine application significantly increased denitrification and N₂O emission rates up to 14 days after application, with rates amounting to 0.9 and 0.6 g N m^–2 day^–1 (9 and 6 kg N ha^–1 day^–1), respectively. When DCD was added to the urine, N₂O emission rates were significantly lower from 3 to 7 days after urine application onwards. Denitrification was the main source of N₂O immediately following each urine application. 14 days after the first application, when soil water contents dropped to 15% (v/v) N₂O mainly derived from nitrification.Total denitrification losses during the 14 day periods were 7 g N m^–2, or 18% of the urine-N applied. Total N₂O emission losses were 6.5 and 3 g N m^–2, or 16% and 8% of the urine-N applied for the two periods. The minimum estimations of denitrification and N₂O emission losses from urine-affected soil were 45 to 55 kg N ha^–1 year^–1, and 20 to 50 kg N ha^–1 year^–1, respectively.     

Subsurface denitrification in a forest riparianzone: Interactions between hydrology and supplies ofnitrate and organic carbon

The influence of hydrology andpatterns of supply of electron donors and acceptors onsubsurface denitrification was studied in a forestriparian zone along the Boyne River in southernOntario that received high nitrogen inputs from a sandaquifer. Two hypotheses were tested: (1) subsurfacedenitrification is restricted to localized zones ofhigh activity; (2) denitrification zones occur atsites where groundwater flow paths transportNO₃ ^– to supplies of available organiccarbon. A plume of nitrate-rich groundwater withconcentrations of 10–30 mg N L^–1 flowed laterallyat depths of 1.5–5 m in sands beneath peat for ahorizontal distance of 100–140 m across the riparianzone to within 30–50 m of the river. In situ acetyleneinjections to piezometers revealed that significantdenitrification was restricted to a narrow zone ofsteep NO₃ ^– and N₂O decline at theplume margins. The location of these denitrificationsites in areas with steep gradients of groundwater DOCincrease supported hypothesis 2. Many of thesedenitrification hotspots occurred near interfacesbetween sands and either peats or buried river channeldeposits. Field experiments involving in situadditions of either glucose or NO₃ ^– topiezometers indicated that denitrification wasC-limited in a large subsurface area of the riparianzone, and became N-limited beyond the narrow zone ofNO₃ ^– consumption. These data suggest thatdenitrification may not effectively removeNO₃ ^– from groundwater transported at depththrough permeable riparian sediments unlessinteraction occurs with localized supplies of organicmatter.     

Denitrification and N2O emissions from a UK pasture soil following the early spring application of cattle slurry and mineral fertiliser

Total denitrification and nitrous oxide (N₂O) losses were measured from three contrasting dairy management systems representing good commercial practice (system 1), production maintained but with reduced N losses (system 2); and nitrate leaching less than 50 mg L^-1 but with reduced production (system 3). Measurements were made following mineral fertiliser application and from two plot experiments where four treatments were applied: control, NH₄NO₃ at 60 kg N ha^-1, cattle slurry applied to the surface (equivalent to 45 kg N ha^-1), and cattle slurry injected. Despite low soil temperatures (<6 °C) and low rainfall (<3 mm), total denitrification and N₂O losses peaked at 56 and 16 g N ha^-1 d^-1, respectively. Total denitrification losses decreased: system 1 system 2 > system 3, whereas N₂O losses decreased: system 2 > system 3 > system 1. Total denitrification losses tended to decrease with decreasing fertiliser application rate, whereas fertiliser application rate was not the sole determinant of the N₂O loss. The system 3 field was injected with cattle slurry for 2 yr, system 2 received some slurry by injection and system 1 received slurry to the surface. Thus, the amount, timing and method of previous cattle slurry application was important in determining the loss following subsequent fertiliser application. For the plot experiments, total denitrification and N₂O losses decreased in the order: slurry injected > mineral fertiliser > slurry applied to the surface > control for 5 days following application. However, 16 and 19 days after application, N₂O losses above the control were measured from plots that had received cattle slurry. It was inferred that the application of cattle slurry to the pasture soil stimulated greater N₂O production and increased losses over a longer time period compared with mineral fertiliser additions.     

Sources and sinks of nitrous oxide (N2O) in deep lakes

As reported from marine systems, we found that also in15 prealpine lakes N₂O concentrations werestrongly correlated with O₂ concentrations. Inoxic waters below the mixed surface layer, N₂Oconcentrations usually increased with decreasingO₂ concentrations. N₂O is produced in oxicepilimnia, in oxic hypolimnia and at oxic-anoxicboundaries, either in the water or at the sediment-waterinterface. It is consumed, however, incompletely anoxic layers. Anoxic water layers weretherefore N₂O undersaturated. All studied lakeswere sources for atmospheric N₂O, including thosewith anoxic, N₂O undersaturated hypolimnia.However, compared to agriculture, lakes seem not tocontribute significantly to atmospheric N₂Oemissions.     

Effect of ammonium and nitrate application on the NO and N2O emission out of different soils

The effect of nitrate and ammonium application (0, 50, 100 and 150 mg N kg^-1 soil) was studied in an incubation experiment. Four Belgian soils, selected for different soil characteristics, were used. The application of both nitrate and ammonium caused an increase of the NO and N₂O emission. The NO production from nitrate and ammonium was found to be of the same order of magnitude. At low pH the NO production was found to be highest from nitrate, at higher pH values the production was found to be higher from ammonium. This seems to be the result of the negative effect of low pH on nitrification.The ANOVA analysis was carried out to separate the effect of the form of nitrogen, quantily of N applied and soil characteristics. The total production of NO was found to depend for 97% on the soil characteristics and for 3% on the quantity of N added. The total N₂O production depended for 100% on the soil characteristics.Stepwise regression analysis showed that the total NO production was best predicted by a combination of the factors CaCO₃ content and NH₄ ⁺ concentration in the soil. Total N₂O production was best described by a combination of CaCO₃, water soluble carbon (WSC) and sand-content.The N₂O/NO ratio was found to be highly variable, indicating that their productions react differently to changes in conditions, or are partly independent.It may be concluded that to NO and N₂O from soils both nitrification and denitrification may be equally important, their relative importance depending on local conditions such as substrate availability, water content of the soil etc. However, the NO production seems to be more nitrification dependent than the N₂O production. ei]{gnE}{fnMerckx}{edSection editor}     

Seasonal variation in N2O emissions from urine patches: Effects of urine concentration, soil compaction and dung

Urine patches in pastures rank among the highest sources of the greenhouse gas nitrous oxide (N₂O) from animal production systems. Previous laboratory studies indicate that N₂O emissions for urine-N in pastures may increase with a factor five or eight in combination with soil compaction and dung, respectively. These combinations of urine, compaction and dung occur regularly in pastures, especially in so-called camping areas. The aims of this study were (i) to experimentally quantify the effect of compaction and dung on emission factors of N₂O from urine patches under field conditions; (ii) to detect any seasonal changes in emission from urine patches; and (iii) to quantify possible effects of urine concentration and -volume. A series of experiments on the effects of compaction, dung, urine-N concentration and urine volume was set up at a pasture on a sandy soil (typic Endoaquoll) in Wageningen, the Netherlands. Artificial urine was applied 8 times in the period August 2000–November 2001, and N₂O emissions were monitored for a minimum of 1 month after each application. The average emission factor for urine-only treatments was 1.55%. Over the whole period, only soil compaction had a clear significant effect, raising the average N₂O emissions from urine patches from 1.30% to 2.92% of the applied N. Dung had no consistent effect; although it increased the average emissions from 1.60% to 2.82%, this was clearly significant (P< 0.01) for only one application date and marginally significant (P=0.054) for the whole experiment. Both compaction and dung increased water-filled pore space (WFPS) of the topsoil for a more prolonged time than high urine volumes. No effect of amount of urine-N or urine volume on N₂O emissions relative to added N was detected for the whole experiment. There were clear differences between application dates, with highest emissions for urine-only treatments of 4.25% in October, 2000, and lowest of –0.11% in June, 2001. Emissions peaked at 60–70% WFPS, and decreased rapidly with both higher and lower WFPS. We conclude that compaction leads to a considerable increase in the N₂O emissions under field conditions, mainly through higher WFPS. Dung addition may have the same effect, although this was not consistent throughout our experiment. Seasonal variations seemed mainly driven by differences in WFPS. Based on this study, mitigation strategies should focus on minimizing the grazing period with wet conditions leading to WFPS > 50%, avoiding camping areas in pastures, and on avoiding grazing under moist soil conditions. Greenhouse gas budgets for grazing conditions should include the effects of soil compaction and dung to represent actual emissions.     

Loss of nitrogen in compacted grassland soil by simultaneous nitrification and denitrification

The soils of mid-Wales in grazed permanent pasture usually exhibit stagnogley features in the top 4–10 cm even though on sloping sites, they are freely drained. Nitrogen is often poorly recovered under these conditions. Our previous studies suggest that continuing loss of available N through concurrent nitrification and denitrification might provide an explanation for poor response to fertilizer N. The work described was designated to further test this proposition. When NH ₄ ⁺ –N was applied to the surface of intact cores, equilibrated at –5kPa matric potential, about 70% of NH ₄ ⁺ –N initially present was lost within 56 days of incubation. Study of different sections of the cores showed a rise in NO ₃ ^- level in the surface 0–2.5 cm soil layer but no significant changes below this depth. The imbalance between NO ₃ ^- accumulation and NH ₄ ⁺ disappearance during the study indicated a simultaneous nitrification and denitrification in the system. Furthermore, the denitrification potential of the soil was 3–4 times greater than nitrification potential so no major build-up of NO ₃ ^- would be expected when two processes occur simultaneously in micro-scale. When nitrification was inhibited by nitrapyrin, a substantial amount of NH ₄ ⁺ –N remained in the soil and persisted till the end of the incubation. The apparent recovery of applied N increased and of the total amount of N applied, 50% more was recovered relative to without nitrapyrin. It appears that addition of nitrapyrin inhibited nitrification, and consequently denitrification, by limiting the supply of NO ₃ ^- for denitrifying organisms. Emission of N₂O from the NH ₄ ⁺ amended soil cores further confirmed that loss of applied N was the result of both nitrification and denitrification, which occurred simultaneously in adjacent sites at shallow depths. This N loss could account for the poor response to fertilizer N often observed in pastoral agriculture in western areas of the UK.     

Identification and isolation of active N2O reducers in rice paddy soil

Dissolved N₂O is occasionally detected in surface and ground water in rice paddy fields, whereas little or no N₂O is emitted to the atmosphere above these fields. This indicates the occurrence of N₂O reduction in rice paddy fields; however, identity of the N₂O reducers is largely unknown. In this study, we employed both culture-dependent and culture-independent approaches to identify N₂O reducers in rice paddy soil. In a soil microcosm, N₂O and succinate were added as the electron acceptor and donor, respectively, for N₂O reduction. For the stable isotope probing (SIP) experiment, ¹³C-labeled succinate was used to identify succinate-assimilating microbes under N₂O-reducing conditions. DNA was extracted 24 h after incubation, and heavy and light DNA fractions were separated by density gradient ultracentrifugation. Denaturing gradient gel electrophoresis and clone library analysis targeting the 16S rRNA and the N₂O reductase gene were performed. For culture-dependent analysis, the microbes that elongated under N₂O-reducing conditions in the presence of cell-division inhibitors were individually captured by a micromanipulator and transferred to a low-nutrient medium. The N₂O-reducing ability of these strains was examined by gas chromatography/mass spectrometry. Results of the SIP analysis suggested that Burkholderiales and Rhodospirillales bacteria dominated the population under N₂O-reducing conditions, in contrast to the control sample (soil incubated with only ¹³C-succinate). Results of the single-cell isolation technique also indicated that the majority of the N₂O-reducing strains belonged to the genera Herbaspirillum (Burkholderiales) and Azospirillum (Rhodospirillales). In addition, Herbaspirillum strains reduced N₂O faster than Azospirillum strains. These results suggest that Herbaspirillum spp. may have an important role in N₂O reduction in rice paddy soils.     

Characterization of microbial NO production, N2O production and CH4 oxidation initiated by aeration of anoxic rice field soil

Intermittent drainage of rice fields isdiscussed as an option to mitigate emission ofCH₄, an important greenhouse gas. HoweverN₂O, a potentially more effective greenhouse gas,may be emitted during the aeration phase. Therefore,the metabolism of NO, N₂O, NH ,NO and NO and the kinetics ofCH₄ oxidation were measured after aeration ofmethanogenic rice field soil. Before aeration, thesoil contained NH in relatively highconcentrations (about 4 mM), while NO andNO were almost undetectable. Immediatelyafter aeration both NO and N₂O were produced withrates of about 15 pmol h^-1 gdw^-1 and 5 pmolh^-1 gdw^-1, respectively. Simultaneously,NH decreased while NO accumulated. Later on, NO was depletedwhile NO concentrations increased.Characteristic phases of nitrogen turnover wereassociated with the activities of ammonium oxidizers,nitrite oxidizers and denitrifiers. Oxidation ofNH and production of NO and N₂O wereinhibited by 10 Pa acetylene demonstrating thatnitrification was obligatory for the initiation ofnitrogen turnover and production of NO and N₂O.Ammonium oxidation was not limited by the availableNH and thus, concomittant production of NOand N₂O was not stimulated by addition ofNH . However, addition of NO stimulated production of NO and N₂O in bothanoxic and aerated rice soil slurries. In this case,10 Pa acetylene did not inhibit the production of NOand N₂O demonstrating that it was due todenitrification which was obviously limited by theavailability of NO . In the aerated soilslurries CH₄ was only oxidized if present atelevated concentrations >50 ppmv CH₄). Atatmospheric CH₄ concentrations (1.7 ppmv)CH₄ was not consumed, but was even slightly produced.CH₄ oxidation activity increased afterpreincubation at 20% CH₄, and then CH₄was also oxidized at atmospheric concentrations. CH₄oxidation kinetics exhibited sigmoid characteristicsat low CH₄ concentrations presumably because ofinhibition of CH₄ oxidation by NH .     

Soils,a sink for N2O? A review

Soils are the main sources of the greenhouse gas nitrous oxide (N₂O). The N₂O emission at the soil surface is the result of production and consumption processes. So far, research has concentrated on net N₂O production. However, in the literature, there are numerous reports of net negative fluxes of N₂O, (i.e. fluxes from the atmosphere to the soil). Such fluxes are frequent and substantial and cannot simply be dismissed as experimental noise.
Net N₂O consumption has been measured under various conditions from the tropics to temperate areas, in natural and agricultural systems. Low mineral N and large moisture contents have sometimes been found to favour N₂O consumption. This fits in with denitrification as the responsible process, reducing N₂O to N₂. However, it has also been reported that nitrifiers consume N₂O in nitrifier denitrification. A contribution of various processes could explain the wide range of conditions found to allow N₂O consumption, ranging from low to high temperatures, wet to dry soils, and fertilized to unfertilized plots. Generally, conditions interfering with N₂O diffusion in the soil seem to enhance N₂O consumption. However, the factors regulating N₂O consumption are not yet well understood and merit further study.
Frequent literature reports of net N₂O consumption suggest that a soil sink could help account for the current imbalance in estimated global budgets of N₂O. Therefore, a systematic investigation into N₂O consumption is necessary. This should concentrate on the organisms, reactions, and environmental factors involved.     

Symbiotic relationships between soil fungi and plants reduce N2O emissions from soil

N₂O is a potent greenhouse gas involved in the destruction of the protective ozone layer in the stratosphere and contributing to global warming. The ecological processes regulating its emissions from soil are still poorly understood. Here, we show that the presence of arbuscular mycorrhizal fungi (AMF), a dominant group of soil fungi, which form symbiotic associations with the majority of land plants and which influence a range of important ecosystem functions, can induce a reduction in N₂O emissions from soil. To test for a functional relationship between AMF and N₂O emissions, we manipulated the abundance of AMF in two independent greenhouse experiments using two different approaches (sterilized and re-inoculated soil and non-mycorrhizal tomato mutants) and two different soils. N₂O emissions were increased by 42 and 33% in microcosms with reduced AMF abundance compared to microcosms with a well-established AMF community, suggesting that AMF regulate N₂O emissions. This could partly be explained by increased N immobilization into microbial or plant biomass, reduced concentrations of mineral soil N as a substrate for N₂O emission and altered water relations. Moreover, the abundance of key genes responsible for N₂O production (nirK) was negatively and for N₂O consumption (nosZ) positively correlated to AMF abundance, indicating that the regulation of N₂O emissions is transmitted by AMF-induced changes in the soil microbial community. Our results suggest that the disruption of the AMF symbiosis through intensification of agricultural practices may further contribute to increased N₂O emissions.     

Tree species, root decomposition and subsurface denitrification potential in riparian wetlands

Patches of organic matter have been found to be important `hotspots' of denitrification in both surface and subsurface soils, but the factors controlling the formation and maintenance of these patches are not well established. We compared the concentration of patches of organic matter and root biomass in the subsurface (saturated zone) beneath poorly drained riparian wetland soils at four sites in Rhode Island, USA - two dominated by red maple (Acer rubrum) and two dominated by white pine (Pinus strobus). Denitrification enzyme activity (DEA) and carbon (C) content of patch material were compared between sites and between patches with different visual characteristics. Root decomposition was measured in an 8-week ex-situ incubation experiment that compared the effects of water content, root species, and soil matrix origin on CO₂ evolution. We observed significantly greater concentrations of patches at 55 cm at one red maple site than all other sites. DEA and percent C in patches was generally higher in patches than matrix soil and did not vary between sites or by patch type. White pine roots decomposed at a faster rate than red maple roots under unsaturated conditions. Our results suggest that faster root decomposition could result in lower concentrations of patches of organic material in subsurface soils at sites dominated by white pine. Tree species composition and root decomposition may play a significant role in the formation of patches and the creation and maintenance of groundwater denitrification hotspots in the subsurface of riparian wetlands. Abbreviations: DEA – denitrification enzyme activity; DOC – dissolved organic carbon; PD – poorly drained; RM-1 – red maple-1 site; RM-2 – red maple-2 site; WP-1 – white pine-1 site; WP-2 – white pine-2 site.     

Temporal changes in N2O-losses from two arable soils

N₂O-loss rates from two soils were measured over a continuous observation period of 2 years. The two soils, differing in texture (sandy loam and silty loam), are frequently used for intensive crop production. Rates were estimated using a closed soil cover box technique. N₂O-losses obtained were scrutinised with physical, chemical and microbiological properties of the soils as well as with climatic data.Large temporal changes in N₂O-emission rates were found. The data were approximately log-normal distributed. In spring maximal values of 20 g N₂O-N ha^-1 d^-1 were observed. According to this observation, two situations associated with high flux rates could be distinguished; 1. N₂O- production by soil at spring thaw and 2. N₂O-production within one week after N-fertilizer application. For both soils equal N₂O-losses were found, which are adequate to 1 kg N₂O-N ha^-1 per year. From this data was calculated that N₂O-losses ranged from 0.8–1.5% of the applied fertilizer N.     

Effects of urine on soil microbial biomass,methanogenesis, nitrification and denitrification in grassland soils

Urine was added under controlled conditions to intact turfs taken from long-term permanent pasture on clay loam and sandy loam soils in South West England. Methane exchanges were small (<+/−0.03 μg CH₄ m^-2 min^-1) and overall absorption equalled or exceeded emission in both soils. On the clay loam, wetting with water or urine increased soil microbial biomass C and N contents by about 20% but there was no specific effect of urine. Urine, however, caused an increase in soil respiration of >50% and the average increase was greater for cow's urine (30.8 mg CO₂ m^-2 min^-1) than for an artificial urine (20.1 mg CO₂ m^-2 min^-1). Emissions of nitric and nitrous oxides following urine application were substantial (on average 0.36 μg NO-N and 29 μg N₂O-N m^-2 min^-1) but short lived (<40 days). The high levels of ammonium found in the urine treated soils (>200 mg NH₄ ⁺-N kg^-1) were nitrified to nitrate over a period of 42 days. Qualitative changes in the soil microbial biomass were evidently not related to biomass size. Relationships between trace gas emissions and soil processes are discussed. ei]Section editor: R Merckx     

Fluxes of N2O,CH4 and CO2 on afforested boreal agricultural soils

After drainage of natural boreal peatlands, the decomposition of organic matter increases and peat soil may turn into a net source of CO₂ and N₂O, whereas CH₄ emission is known to decrease. Afforestation is a potential mitigation strategy to reduce greenhouse gas emission from organic agricultural soils. A static chamber technique was used to evaluate the fluxes of CH₄, N₂O and CO₂ from three boreal organic agricultural soils in western Finland, afforested 1, 6 or 23 years before this study. The mean emissions of CH₄ and N₂O during the growing seasons did not correlate with the age of the tree stand. All sites were sources of N₂O. The highest daily N₂O emission during the growing season, measured in the oldest site, was as high as 29 mg N₂O m^–2d^–1. In general, organic agricultural soils are sinks for methane. Here, the oldest site acted as a small sink for methane, whereas the two youngest afforested organic soils were sources for methane with maximum emission rates (up to 154 mg m^–2d^–1) similar to those reported for minerogenous natural peatlands. Soil respiration rates decreased with the age of the forest. The high soil respiration in the younger sites, probably resulted from the high biomass production of herbs, could create soil anaerobiosis and increase methane production. Our results show that afforestation of agricultural peat soils does not abruptly terminate the N₂O emissions during the first two decades, and afforestation can even enhance methane emission for a few years. The carbon accumulation in the developing tree stand can partly compensate the carbon loss from soil.     

The contribution of Rhizobium meliloti to soil denitrification

The ability of Rhizobium meliloti cells to denitrify in soils under several conditions was tested. All the strains tested were able to remove large amounts of N-NO₃ ^- from soils. Both water filled pore space above 36% and temperatures above 20°C greatly increased nitrogen losses. However, even with optimal conditions for denitrification and the highest rhizobial populations found in agricultural soils, the contribution of Rhizobium to the total denitrification was virtually negligible as compared to other soil microorganisms.To whom correspondence should be addressed