A N tracing model to analyse N transformations in old grassland soil

A new ¹⁵N tracing model was developed to analyse nitrogen (N) transformations in old grassland soil. There was a need to develop a new model because existing models such as FLUAZ were not able to simulate the observed N dynamics. The new features of the model are: (a) simulation of heterotrophic nitrification, (b) simulation of dissimilatory nitrate (NO₃⁻) reduction to ammonium (NH₄⁺) (DNRA), (c) release of adsorbed or stored fertiliser N into the available mineral N pools and (d) immobilisation of NH₄⁺ and NO₃⁻ into two separate organic N pools with different re-mineralisation characteristics. The tracing model contains six N pools and nine simultaneous N transformations either at zero- or first-order kinetics. The model is set up in the modelling software ModelMaker which contains non-linear optimisation routines based on the Marquardt-Levenberg algorithm. The model is able to simulate data obtained from triple labelling studies where either the NH₄⁺, the NO₃⁻ or both pools were labelled with ¹⁵N. The flexible modelling environment allows the user to develop the model further.     

Effects of repeated fertilizer and cattle slurry applications over 38 years on N dynamics in a temperate grassland soil

The effects of repeated synthetic fertilizer or cattle slurry applications at annual rates of 50, 100 or 200 m³ ha⁻¹ yr⁻¹ over a 38 year period were investigated with respect to herbage yield, N uptake and gross soil N dynamics at a permanent grassland site. While synthetic fertilizer had a sustained and constant effect on herbage yield and N uptake, increasing cattle slurry application rates increased the herbage yield and N uptake linearly over the entire observation period. Cattle slurry applications, two and four times the recommended rate (50 m³ ha⁻¹ yr⁻¹, 170 kg N ha⁻¹), increased N uptake by 46 and 78%, respectively after 38 years. To explain the long-term effect, a ¹⁵N tracing study was carried out to identify the potential change in N dynamics under the various treatments. The analysis model evaluated process-specific rates, such as mineralization, from two organic-N pools, as well as nitrification from NH₄⁺ and organic-N oxidation. Total mineralization was similar in all treatments. However, while in an unfertilized control treatment more than 90% of NH₄⁺ production was related to mineralization of recalcitrant organic-N, a shift occurred toward a predominance of mineralization from labile organic-N in the cattle slurry treatments and this proportion increased with the increase in slurry application rate. Furthermore, the oxidation of recalcitrant organic-N shifted from a predominant NH₄⁺ production in the control treatment, toward a predominant NO₃⁻ production (heterotrophic nitrification) in the cattle slurry treatments. The concomitant increase in heterotrophic nitrification and NH₄⁺ oxidation with increasing cattle slurry application rate was mainly responsible for the increase in net NO₃⁻ production rate. Thus the increase in N uptake and herbage yield on the cattle slurry treatments could be related to NO₃⁻ rather than NH₄⁺ production. The ¹⁵N tracing study was successful in revealing process-specific changes in the N cycle in relationship to long-term repeated amendments.     

A method to assess whether ‘preferential use’ occurs after N ammonium addition; implication for the N isotope dilution technique

The robustness of the assumption of equilibrium between native and added N during ¹⁵N isotope dilution has recently been questioned by Watson et al. (Soil Biol Biochem 32 (2000) 2019-2030). We re-analyzed their raw data using equations that consider the added and native NH₄⁺ and NO₃⁻ pools as separate state variables. Gross mineralization rates and first-order rate constants for NH₄⁺ and NO₃⁻ consumption were obtained by combining analytical integration of the differential equations with a non-linear fitting procedure. The first-order rate constants for NH₄⁺ consumption and NO₃⁻ immobilization for the added NH₄⁺ and NO₃⁻ pool were used to estimate gross mineralization rates and first-order rate constants for nitrification of native NH₄⁺. The latter were 2-4 times lower than the first-order rate constants derived from the added N pool. This discrepancy between first-order rate constants for nitrification implies that one or more process rates estimated for the added N pools cannot be applied to the native N pools. Preferential use of the added N resulted in an overestimation of the gross mineralization by 1.5-2.5-fold, emphasizing the need for critical evaluation of the assumption of equilibrium before gross mineralization rates are calculated.     

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.     

Ammonium immobilisation during chloroform fumigation

The assumption in using the chloroform fumigation technique for microbial biomass determination is that microbes are killed or at least inactivated by the treatment. Problems associated with transformations of the N released on or during fumigation have so far only been associated with the fumigation-incubation method. A laboratory and a field study were carried out to investigate the possible N transformations during biomass determination by the fumigation-extraction method. Labelled NH₄NO₃ (either the NO₃⁻, NH₄⁺ or both pools were ¹⁵N enriched) was applied to the soil and biomass determinations made at intervals subsequently. The size and enrichment of the ammonium (NH₄⁺), and nitrate (NO₃⁻) pools were determined before and after chloroform fumigation. The ¹⁵N enrichment of the NH₄⁺ pool after fumigation could only be explained if immobilisation of ammonium occurred at some time during the 24 h fumigation period. The extent of this immobilisation was calculated. In addition, there was evidence that nitrification occurred during the fumigation procedure at the start of the laboratory study and throughout the field study. The laboratory and field study differed mainly in the dynamics related to NO₃⁻ uptake and release. There was evidence for uptake of NO₃⁻ by the microbial biomass with and without utilization. We conclude that the ¹⁵N enrichment in the microbial biomass cannot be accurately determined when N transformations and release of non-utilized N occurs during fumigation. The possible immobilisation of mineral N during fumigation will affect the magnitude of the factor used to convert measured microbial biomass N to actual microbial biomass N in soil.     

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.     

Net and gross mineral N production rates at three levels of forest canopy retention: evidence that NH<Subscript>4</Subscript><Superscript>+</Superscript> and NO<Subscript>3</Subscript><Superscript>−</Superscript> dynamics are uncoupled

Alternative silvicultural systems were introduced in Coastal Western Hemlock forests of British Columbia, Canada, to reduce disturbance incurred by conventional clear-cutting and to maintain the forest influence on soil nutrient cycling. As we hypothesized, in situ pools and net mineralization of NH₄ ⁺ were lower under no and low disturbance (old-growth forest and shelterwood) compared to clear-cuts (high disturbance); in situ pools and net production of NO₃ ⁻ were very low across all treatments. Gross transformation rates of NH₄ ⁺ increased while those of NO₃ ⁻ decreased with increasing disturbance, suggesting that these processes were uncoupled. We conclude that shelterwood harvesting reduces the impact on forest floor NH₄ ⁺ cycling compared to clear-cutting, and that neither low nor high disturbance intensity results in substantial NO₃ ⁻ accumulation, as what occasionally occurs in other ecosystems. We hypothesize that the uncoupling of NH₄ ⁺ and NO₃ ⁻ dynamics may be due to the predominance of heterotrophic nitrification by lignin-degrading fungi that oxidize organic N rather than NH₄ ⁺–N, and whose activities are suppressed at high NH₄ ⁺ concentrations.     

Heterotrophic nitrification is the predominant NO<Stack><Subscript>3</Subscript><Superscript>−</Superscript></Stack> production mechanism in coniferous but not broad-leaf acid forest soil in subtropical China

A study was carried out to investigate the potential gross nitrogen (N) transformations in natural secondary coniferous and evergreen broad-leaf forest soils in subtropical China. The simultaneously occurring gross N transformations in soil were quantified by a ¹⁵N tracing study. The results showed that N dynamics were dominated by NH₄⁺ turnover in both soils. The total mineralization (from labile and recalcitrant organic N) in the broad-leaf forest was more than twice the rate in the coniferous forest soil. The total rate of mineral N production (NH₄⁺ + NO₃⁻) from the large recalcitrant organic N pool was similar in the two forest soils. However, appreciable NO₃⁻ production was only observed in the coniferous forest soil due to heterotrophic nitrification (i.e. direct oxidation of organic N to NO₃⁻), whereas nitrification in broad-leaf forest was little (or negligible). Thus, a distinct shift occurred from predominantly NH₄⁺ production in the broad-leaf forest soil to a balanced production of NH₄⁺ and NO₃⁻ in the coniferous forest soil. This may be a mechanism to ensure an adequate supply of available mineral N in the coniferous forest soil and most likely reflects differences in microbial community patterns (possibly saprophytic, fungal, activities in coniferous soils). We show for the first time that the high nitrification rate in these soils may be of heterotrophic rather than autotrophic nature. Furthermore, high NO₃⁻ production was only apparent in the coniferous but not in broad-leaf forest soil. This highlights the association of vegetation type with the size and the activity of the SOM pools that ultimately determines whether only NH₄⁺ or also a high NO₃⁻ turnover is present.     

Conversion of forest to agricultural land affects the relative contribution of bacteria and fungi to nitrification in humid subtropical soils

Land-use and management practices can affect soil nitrification. However, nitrifying microorganisms responsible for specific nitrification process under different land-use soils remains unknown. Thus, we investigated the relative contribution of bacteria and fungi to specific soil nitrification in different land-use soils (coniferous forest, upland fields planted with corn and rice paddy) in humid subtropical region in China. ¹⁵N dilution technique in combination with selective biomass inhibitors and C₂H₂ inhibition method were used to estimate the relative contribution of bacteria and fungi to heterotrophic nitrification and autotrophic nitrification in the different land-use soils in humid subtropical region. The results showed that autotrophic nitrification was the predominant nitrification process in the two agricultural soils (upland and paddy), while the nitrate production was mainly from heterotrophic nitrification in the acid forest soil. In the upland soils, streptomycin reduced autotrophic nitrification by 94%, whereas cycloheximide had no effect on autotrophic nitrification, indicating that autotrophic nitrification was mainly driven by bacteria. However, the opposite was true in another agricultural soil (paddy), indicating that fungi contributed to the oxidation of NH₄⁺ to NO₃^?. In the acid forest soil, cycloheximide, but not streptomycin, inhibited heterotrophic nitrification, demonstrating that fungi controlled the heterotrophic nitrification. The conversion of forest to agricultural soils resulted in a shift from fungi-dominated heterotrophic nitrification to bacteria- or fungi-dominated autotrophic nitrification. Our results suggest that land-use and management practices, such as the application of N fertilizer and lime, the long-term waterflooding during rice growth, straw return after harvest, and cultivation could markedly influence the relative contribution of bacteria and fungi to specific soil nitrification processes.     

Nitrapyrin effectiveness in reducing nitrous oxide emissions decreases at low doses of urea in an Andosol

In the tropics,frequent nitrogen(N)fertilization of grazing areas can potentially increase nitrous oxide(N₂O)emissions.The application of nitrification inhibitors has been reported as an effective management practice for potentially reducing N loss from the soil-plant system and improving N use efficiency(NUE).The aim of this study was to determine the effect of the co-application of nitrapyrin(a nitrification inhibitor,NI)and urea in a tropical Andosol on the behavior of N and the emissions of N₂O from autotrophic and heterotrophic nitrification.A greenhouse experiment was performed using a soil(pH 5.9,organic matter content 78 g kg^-1,and N 5.6 g kg^-1)sown with Cynodon nlemfuensis at 60%water-filled pore space to quantify total N₂O emissions,N₂O derived from fertilizer,soil ammonium(NH₄⁺)and nitrate(NO₃^-),and NUE.The study included treatments that received deionized water only(control,NI).No significant differences were observed in soil NH₄⁺content between the UR and UR+NI treatments,probably because of soil mineralization and NO₃^-produced by heterotrophic nitrification,which is not effectively inhibited by nitrapyrin.After 56 d,N₂O emissions in UR(0.51±0.12 mg N₂O-N concluded that the soil organic N mineralization and heterotrophic nitrification are the main processes of NH₄⁺and NO₃^-production.Additionally,it was found that N₂O emissions were partially a consequence of the direct oxidation of the soil's organic N via heterotrophic nitrification coupled to denitrification.Finally,the results suggest that nitrapyrin would likely exert significant mitigation on N₂O emissions only if a substantial N surplus exists in soils with high organic matter content.     

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.     

Effect of elevated CO2 on soil N dynamics in a temperate grassland soil

The response of terrestrial ecosystems to elevated atmospheric CO₂ is related to the availability of other nutrients and in particular to nitrogen (N). Here we present results on soil N transformation dynamics from a N-limited temperate grassland that had been under Free Air CO₂ Enrichment (FACE) for six years. A ¹⁵N labelling laboratory study (i.e. in absence of plant N uptake) was carried out to identify the effect of elevated CO₂ on gross soil N transformations. The simultaneous gross N transformation rates in the soil were analyzed with a ¹⁵N tracing model which considered mineralization of two soil organic matter (SOM) pools, included nitrification from NH₄⁺ and from organic-N to NO₃⁻ and analysed the rate of dissimilatory NO₃⁻ reduction to NH₄⁺ (DNRA). Results indicate that the mineralization of labile organic-N became more important under elevated CO₂. At the same time the gross rate of NH₄⁺ immobilization increased by 20%, while NH₄⁺ oxidation to NO₃⁻ was reduced by 25% under elevated CO₂. The NO₃⁻ dynamics under elevated CO₂ were characterized by a 52% increase in NO₃⁻ immobilization and a 141% increase in the DNRA rate, while NO₃⁻ production via heterotrophic nitrification was reduced to almost zero. The increased turnover of the NH₄⁺ pool, combined with the increased DNRA rate provided an indication that the available N in the grassland soil may gradually shift towards NH₄⁺ under elevated CO₂. The advantage of such a shift is that NH₄⁺ is less prone to N losses, which may increase the N retention and N use efficiency in the grassland ecosystem under elevated CO₂.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

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.     

Pathways and dynamics of NO3 and NH4 applied in a mountain Picea abies forest and in a nearby meadow in central Switzerland

To evaluate the pathways and dynamics of inorganic nitrogen (N) deposition in previously N-limited ecosystems, field additions of ¹⁵N tracers were conducted in two mountain ecosystems, a forest dominated by Norway spruce (Picea abies) and a nearby meadow, at the Alptal research site in central Switzerland. This site is moderately impacted by N from agricultural and combustion sources, with a bulk atmospheric deposition of 12 kg N ha⁻¹ y⁻¹ equally divided between NH₄⁺ and NO₃⁻. Pulses of ¹⁵NH₄⁺ and ¹⁵NO₃⁻ were applied separately as tracers on plots of 2.25 m². Several ecosystem pools were sampled at short to longer-term intervals (from a few hours to 1 year), above and belowground biomass (excluding trees), litter layer, soil LF horizon (approx. 5-0 cm), A horizon (approx. 0-5 cm) and gleyic B horizon (5-20 cm). Furthermore, extractable inorganic N, and microbial N pools were analysed in the LF and A horizons. Tracer recovery patterns were quite similar in both ecosystems, with most of the tracer retained in the soil pool. At the short-term (up to 1 week), up to 16% of both tracers remained extractable or entered the microbial biomass. However, up to 30% of the added ¹⁵NO₃⁻ was immobilised just after 1 h, and probably chemically bound to soil organic matter. 16% of the NH₄⁺ tracer was also immobilised within hours, but it is not clear how much was bound to soil organic matter or fixed between layers of illite-type clay. While the extractable and microbial pools lost ¹⁵N over time, a long-term increase in ¹⁵N was measured in the roots. Otherwise, differences in recovery a few hours after labelling and 1 year later were surprisingly small. Overall, more NO₃⁻ tracer than NH₄⁺ tracer was recovered in the soil. This was due to a strong aboveground uptake of the deposited NH₄⁺ by the ground vegetation, especially by mosses.     

Temperature sensitivity of mineral N transformation rates, and heterotrophic nitrification: possible factors controlling the post-disturbance mineral N flush in forest floors

A major forest disturbance such as clearcutting may bring on a flush of mineral N in organic forest floor horizons, but the magnitude of this flush can vary markedly from one ecosystem to another. For example, it was previously established that clearcutting in a high elevation Engelmann spruce-subalpine fir (ESSF) ecosystem results in significantly higher NH₄⁺ and NO₃⁻ concentrations, whereas clearcutting in an old-growth coastal western hemlock (CWH) ecosystem has little effect on mineral N dynamics. We hypothesized that the higher mineral N flush observed in the ESSF ecosystem is due to a greater temperature sensitivity of mineral N transformation rates, and to a lower proportion of heterotrophic nitrifiers, compared to the CWH ecosystem. To test these two hypotheses, we sampled forest floors several times over the growing season from clearcut and old-growth plots in both ecosystems, and measured gross mineral N transformation rates at field temperatures and at 10 °C above field temperatures, as well as with and without acetylene to inhibit autotrophic nitrifiers. Gross NH₄⁺ transformations rates ranged between 20 and 120 μg N (g forest floor)⁻¹ day⁻¹ at the ESSF site, and between 15 and 40 μg N (g forest floor)⁻¹ day⁻¹ at the CWH site. Higher temperature increased gross NH₄⁺ transformation rates in forest floor samples at both sites, but the average Q₁₀ value was higher at the ESSF site (3.15) than at the CWH site (1.25). Temperature sensitivity at the ESSF site was greater in clearcut plots (Q₁₀=4.31) than in old-growth plots (Q₁₀=1.98). Gross NO₃⁻ transformation rates ranged between 10 and 32 μg N (g forest floor)⁻¹ day⁻¹ at the ESSF site, and between 10 and 24 μg N (g forest floor)⁻¹ day⁻¹ at the CWH site, but there were no significant effects of temperature or clearcutting on gross NO₃⁻ transformation rates at either site. Likewise, there were no significant differences in the proportion of heterotrophic nitrifiers between sites. Overall, our results support the view that the temperature sensitivity of microbial processes may explain the magnitude of the NH₄⁺ flush in some coniferous ecosystems, but we lack the evidence relating the magnitude of the NO₃⁻ flush to the proportion of heterotrophic nitrifiers.     

Controls on nitrification in a water-limited ecosystem: experimental inhibition of ammonia-oxidising bacteria in the Patagonian steppe

We studied controls on nitrification in an undisturbed water-limited ecosystem by inhibiting autotrophic nitrifying bacteria in soils with varying levels of vegetative cover. The activity of nitrifying bacteria was disrupted using nitrapyrin, 2-chloro-6-(trichloromethyl)-pyridine, under field conditions in three microenvironments (underneath shrubs, next to grasses and in bare soil). Ammonia-oxidising bacteria were detected by PCR analysis of DNA in soils. The inhibition of nitrification changed the concentrations of NO₃⁻ and NH₄⁺ in the soil, while the microenvironment was most important in determining the response of bacteria to the inhibitor. Nitrapyrin application resulted in a significant (p<0.05) reduction in soil NO₃⁻ concentration (39%) and a significant increase (p<0.001) in soil NH₄⁺ concentration (41%). Untreated bare-soil microenvironments had the lowest concentrations of NH₄⁺ (1.57 μg/g of dry soil) and NO₃⁻ (0.49 μg/g of dry soil) when compared to the other microenvironments, and showed the highest impacts of nitrification inhibition. For example, NH₄⁺ concentrations increased 288% and NO₃⁻ concentrations decreased 60% in inhibited bare-soil microenvironments. In contrast, untreated microenvironments underneath shrubs had the highest levels of NH₄⁺ (10.01 μg/g of dry soil) and NO₃⁻ (0.69 μg/g of dry soil), but showed no significant effects of inhibition of nitrification on soil nitrogen concentrations.     

Soil nitrogen conservation mechanisms in a pristine south Chilean Nothofagus forest ecosystem

A ¹⁵N tracing study was carried out to identify microbial and abiotic nitrogen (N) transformations in a south Chilean Nothofagus betuloides forest soil which is characterized by low N inputs and absence of human disturbance. Gross N transformation rates were quantified with a ¹⁵N tracing model in combination with a Markov chain Monte Carlo sampling algorithm for parameter estimation. The ¹⁵N tracing model included five different N pools (ammonium (NH₄⁺), nitrate (NO₃⁻), labile (N_lab) and recalcitrant (N_rec) soil organic matter and adsorbed NH₄⁺), and ten gross N transformation rates. The N dynamics in the N. betuloides ecosystem are characterized by low net but high gross mineralization rates. Mineralization in this soil was dominated by turnover of N_lab, while immobilization of NH₄⁺ predominantly entered the N_rec pool. A fast exchange between the NH₄⁺ and the adsorbed NH₄⁺ pool was observed, possibly via physical adsorption on and release from clay lattices, providing an effective buffer for NH₄⁺. Moreover, high NH₄⁺ immobilization rates into the N_rec pool ensure a sustained ecosystem productivity. Nitrate, the most mobile form of N in the system, is characterized by a slow turnover and was produced in roughly equal amounts from NH₄⁺ oxidation and organic N oxidation. More than 86% of the NO₃⁻ produced was immediately consumed again. This study showed for the first time that dissimilatory nitrate reduction to ammonium (DNRA) was almost exclusively (>99%) responsible for NO₃⁻ consumption. DNRA rather than NO₃⁻ immobilization ensures that NO₃⁻ is transformed into another available N form. DNRA may therefore be a widespread N retention mechanism in ecosystems that are N-limited and receive high rainfalls.     

Effect of reforestation on turnover of 15N-labelled nitrate and ammonium in relation to changes in soil microflora

The effects of incubation time, vegetation type (represented by a pine plantation, a protected and a periodically burnt eucalypt forest), lime and finely ground pine needles on the transformation of (¹⁵NH₄)₂SO₄ and K¹⁵NO₃ were studied in incubation experiments with a sandy lateritic podzolic soil from south-east Queensland. Microorganisms were counted so as to relate N transformations to particular groups of microorganisms.The heterotrophic miroflora utilized NH⁺₄ as a source of N in preference to NO^?₃, and autotrophic nitrifiers seemed to be weak competitors for NH⁺₄. Lime caused a slight loss of NO^?₃ and this was accompanied by an increase in the population of denitrifying bacteria.Lime promoted immobilization of NH⁺₄ by heterotrophic bacteria and subsequent mineralization by nitrifying bacteria, but when pine needles were also added the nitrifiers were suppressed and immobilization by heterotrophic bacteria dominated. Pine needles alone stimulated fungi to immobilize NH⁺₄.While reforestation with exotic pines caused a loss of total-N there was evidence of increased turnover, i.e. more rapid immobilization and nitrification, in pine plantation soils. Prescribed burning also promoted nitrification while reducing total-N.     

The N2O product ratio of nitrification and its dependence on long-term changes in soil pH

The contribution of nitrification to the emission of nitrous oxide (N₂O) from soils may be large, but its regulation is not well understood. The soil pH appears to play a central role for controlling N₂O emissions from soil, partly by affecting the N₂O product ratios of both denitrification (N₂O/(N₂+N₂O)) and nitrification (N₂O/(NO₂⁻+NO₃⁻). Mechanisms responsible for apparently high N₂O product ratios of nitrification in acid soils are uncertain. We have investigated the pH regulation of the N₂O product ratio of nitrification in a series of experiments with slurries of soils from long-term liming experiments, spanning a pH range from 4.1 to 7.8. ¹⁵N labelled nitrate (NO₃⁻) was added to assess nitrification rates by pool dilution and to distinguish between N₂O from NO₃⁻ reduction and NH₃ oxidation. Sterilized soil slurries were used to determine the rates of chemodenitrification (i.e. the production of nitric oxide (NO) and N₂O from the chemical decomposition of nitrite (NO₂⁻)) as a function of NO₂⁻ concentrations. Additions of NO₂⁻ to aerobic soil slurries (with ¹⁵N labelled NO₃⁻ added) were used to assess its potential for inducing denitrification at aerobic conditions. For soils with pH?5, we found that the N₂O product ratios for nitrification were low (0.2-0.9‰) and comparable to values found in pure cultures of ammonia-oxidizing bacteria. In mineral soils we found only a minor increase in the N₂O product ratio with increasing soil pH, but the effect was so weak that it justifies a constant N₂O product ratio of nitrification for N₂O emission models. For the soils with pH 4.1 and 4.2, the apparent N₂O product ratio of nitrification was 2 orders of magnitude higher than above pH 5 (76‰ and 14‰). This could partly be accounted for by the rates of chemodenitrification of NO₂⁻. We further found convincing evidence for NO₂⁻-induction of aerobic denitrification in acid soils. The study underlines the role of NO₂⁻, both for regulating denitrification and for the apparent nitrifier-derived N₂O emission.