Biosensor Determination of the Microscale Distribution of Nitrate, Nitrate Assimilation, Nitrification, and Denitrification in a Diatom-Inhabited Freshwater Sediment

High-resolution NO₃⁻ profiles in freshwater sediment covered with benthic diatoms were obtained with a new microscale NO₃⁻ biosensor characterized by absence of interference from chemical species other than NO₂⁻ and N₂O. Analysis of the microprofiles obtained indicated no nitrification during darkness, high rates of nitrification and a tight coupling between nitrification and denitrification during illumination, and substantial rates of NO₃⁻ assimilation during illumination. Nitrification during darkness could be induced by purging the bulk water with O₂ gas, indicating that the stimulatory effect on nitrification by illumination was caused by algal production of O₂. NH₄⁺ addition did not stimulate nitrification during darkness when O₂ was restricted to the upper 1-mm layer, and there was thus a low nitrification potential in the permanently oxic top 1 mm of the sediment.     

Estimation of Nitrification and Denitrification from Microprofiles of Oxygen and Nitrate in Model Sediment Systems

The coupling between nitrification and denitrification and the regulation of these processes by oxygen were studied in freshwater sediment microcosms with O₂ and NO₃^- microsensors. Depth profiles of nitrification (indicated as NO₃^- production), denitrification (indicated as NO₃^- consumption), and O₂ consumption activities within the sediment were calculated from the measured concentration profiles. From the concentration profiles, it was furthermore possible to distinguish between the rate of denitrification based on the diffusional supply of NO₃^- from the overlying water and the rate based on NO₃^- supplied by benthic nitrification (D_w and D_n, respectively). An increase in O₂ concentration caused a deeper O₂ penetration while a decrease in D_w and an increase in D_n were observed. The relative importance for total denitrification of NO₃^- produced by nitrification thus increased compared with NO₃^- supplied from the water phase. The decrease in D_w at high oxygen was due to an increase in diffusion path for NO₃^- from the overlying water to the denitrifying layers in the anoxic sediment. At high O₂ concentrations, nitrifying activity was restricted to the lower part of the oxic zone where there was a continuous diffusional supply of NH₄⁺ from deeper mineralization processes, and the long diffusion path from the nitrification zone to the overlying water compared with the path to the denitrifying layers led to a stimulation in D_n.     

Denitrification, Dissimilatory Reduction of Nitrate to Ammonium, and Nitrification in a Bioturbated Estuarine Sediment as Measured with 15N and Microsensor Techniques

Nitrogen and oxygen transformations were studied in a bioturbated (reworked by animals) estuarine sediment (Norsminde Fjord, Denmark) by using a combination of ¹⁵N isotope (NO₃^-), specific inhibitor (C₂H₂), and microsensor (N₂O and O₂) techniques in a continuous-flow core system. The estuarine water was NO₃^- rich (125 to 600 μM), and NO₃^- was consistently taken up by the sediment on the four occasions studied. Total NO₃^- uptake (3.6 to 34.0 mmol of N m^-2 day^-1) corresponded closely to N₂ production (denitrification) during the experimental steady state, which indicated that dissimilatory, as well as assimilatory, NO₃^- reduction to NH₄⁺ was insignificant. When C₂H₂ was applied in the flow system, denitrification measured as N₂O production was often less (58 to 100%) than the NO₃^- uptake because of incomplete inhibition of N₂O reduction. The NO₃^- formed by nitrification and not immediately denitrified but released to the overlying water, uncoupled nitrification, was calculated both from ¹⁵NO₃^- dilution and from changes in NO₃^- uptake before and after C₂H₂ addition. These two approaches gave similar results, with rates ranging between 0 and 8.1 mmol of N m^-2 day^-1 on the four occasions. Attempts to measure total nitrification activity by the difference between NH₄⁺ fluxes before and after C₂H₂ addition failed because of non-steady-state NH₄⁺ fluxes. The vertical distribution of denitrification and oxygen consumption was studied by use of N₂O and O₂ microelectrodes. The N₂O profiles measured during the experimental steady state were often irregularly shaped, and the buildup of N₂O after C₂H₂ was added was much too fast to be described by a simple diffusion model. Only bioturbation by a dense population of infauna could explain these observations. This was corroborated by the relationship between diffusive and total fluxes, which showed that only 19 to 36 and 29 to 62% of the total O₂ uptake and denitrification, respectively, were due to diffusion-reaction processes at the regular sediment surface, excluding animal burrows.     

Simulation Model of the Coupling between Nitrification and Denitrification in a Freshwater Sediment

A model was constructed to simulate the results of experiments which investigated nitrification and denitrification in the freshwater sediment of Lake Vilhelmsborg, Denmark (K. Jensen, N. P. Sloth, N. Risgaard-Petersen, S. Rysgaard, and N. P. Revsbech, Appl. Environ. Microbiol. 60:2094-2100, 1994). The model output faithfully represented the profiles of O₂ and NO₃^- and rates of nitrification, denitrification, and O₂ consumption as the O₂ concentration in the overlying water was increased from 10 to 600 μM. The model also accurately predicted the response, to increasing O₂ concentrations, of the integrated (micromoles per square meter per hour) rates of nitrification and denitrification. The simulated rates of denitrification of NO₃^- diffusing from the overlying water (D_w) and of NO₃^- generated by nitrification within the sediment (D_n) corresponded to the experimental rates as the O₂ concentration in the overlying water was altered. The predicted D_w and D_n rates, as NO₃^- concentration in the overlying water was changed, closely resembled those determined experimentally. The model was composed of 41 layers 0.1 mm thick, of which 3 represented the diffusive boundary layer in the water. Large first-order rate constants for nitrification and denitrification were required to completely oxidize all NH₄⁺ diffusing from the lower sediment layers and to remove much of the NO₃^- produced. In addition to the flux of NH₄⁺ from below, the model required a flux of an electron donor, possibly methane. Close coupling between nitrification and denitrification, achieved by allowing denitrification to tolerate some O₂ (~10 μM), was necessary to reproduce the real data. Spatial separation of the two processes (no toleration by denitrification of O₂) resulted in too high NO₃^- concentrations and too low rates of denitrification.     

1795–1995: Milestones in ROS research

Summary

Oxygen (O₂)-dependent and O₂-independent antimicrobial mechanisms are used by alveolar macrophages (AM) to maintain lung sterility, but these mechanisms are underdeveloped in neonatal AM. Nitric oxide (NO^.), a more recently described antimicrobial and immunomodulating molecule, has not been studied in neonatal AM. Lavaged AM from 3-day-old, 10-day-old, maternal and adult rats were treated with or without lipopolysaccharide (LPS) and/or interferon-γ (IFN-γ) and NO^. synthase activity was measured as its L-arginine metabolites: NO₂^?, NO₃^?, and citrulline. Superoxide anion (O₂^.-) production by suspended macrophages, initiated by either opsonized zymosan or phorbol, was used as a marker of O₂-dependent antimicrobial activity. Lysozyme content of AM was measured as a component of O₂-independent antimicrobial activity. Unstimulated 3-day-old macrophages generated >10-fold more NO₂^? + NO₃^? than did 10-day-old, maternal or adult AM. Twenty hours after LPS and IFN-γ stimulation, 3-day-old AM produced > 2 times more NO₂^? and NO₃^? than did the more mature macrophages. Basal and stimulated O₂^.- release was similar among 3-day-old, 10-day-old and adult AM, while lysozyme concentrations were > 4-fold higher in adult macrophages compared to AM from 3-day-old pups. Rather than having a role in NO^.-dependent antimicrobial activity, we propose that newborn AM have amplified NO^. production to modulate their own differentiation and replication after birth. The age-dependent differences in NO^. synthase expression by AM may lend insight into the regulation of this important enzyme.     

Selective Inhibition of Ammonium Oxidation and Nitrification-Linked N2O Formation by Methyl Fluoride and Dimethyl Ether

Methyl fluoride (CH₃F) and dimethyl ether (DME) inhibited nitrification in washed-cell suspensions of Nitrosomonas europaea and in a variety of oxygenated soils and sediments. Headspace additions of CH₃F (10% [vol/vol]) and DME (25% [vol/vol]) fully inhibited NO₂^- and N₂O production from NH₄⁺ in incubations of N. europaea, while lower concentrations of these gases resulted in partial inhibition. Oxidation of hydroxylamine (NH₂OH) by N. europaea and oxidation of NO₂^- by a Nitrobacter sp. were unaffected by CH₃F or DME. In nitrifying soils, CH₃F and DME inhibited N₂O production. In field experiments with surface flux chambers and intact cores, CH₃F reduced the release of N₂O from soils to the atmosphere by 20- to 30-fold. Inhibition by CH₃F also resulted in decreased NO₃^- + NO₂^- levels and increased NH₄⁺ levels in soils. CH₃F did not affect patterns of dissimilatory nitrate reduction to ammonia in cell suspensions of a nitrate-respiring bacterium, nor did it affect N₂O metabolism in denitrifying soils. CH₃F and DME will be useful in discriminating N₂O production via nitrification and denitrification when both processes occur and in decoupling these processes by blocking NO₂^- and NO₃^- production.     

Nitrification and Denitrification in Lake and Estuarine Sediments Measured by the 15N Dilution Technique and Isotope Pairing

The transformation of nitrogen compounds in lake and estuarine sediments incubated in the dark was analyzed in a continuous-flowthrough system. The inflowing water contained ¹⁵NO₃^-, and by determination of the isotopic composition of the N₂, NO₃^-, and NH₄⁺ pools in the outflowing water, it was possible to quantify the following reactions: total NO₃^- uptake, denitrification based on NO₃^- from the overlying water, nitrification, coupled nitrification-denitrification, and N mineralization. In sediment cores from both lake and estuarine environments, benthic microphytes assimilated NO₃^- and NH₄⁺ for a period of 25 to 60 h after darkening. Under steady-state conditions in the dark, denitrification of NO₃^- originating from the overlying water accounted for 91 to 171 μmol m^-2 h^-1 in the lake sediments and for 131 to 182 μmol m^-2 h^-1 in the estuarine sediments, corresponding to approximately 100% of the total NO₃^- uptake for both sediments. It seems that high NO₃^- uptake by benthic microphytes in the initial dark period may have been misinterpreted in earlier investigations as dissimilatory reduction to ammonium. The rates of coupled nitrification-denitrification within the sediments contributed to 10% of the total denitrification at steady state in the dark, and total nitrification was only twice as high as the coupled process.     

Role of Nitrate in Photosynthetic Electron Transport of Chlorella Vulgaris

Addition of nitrate to a suspension of NO₃ ^--depleted Chlorella vulgaris cells raised the O₂-evolving capacity of the organism by 60%. The rate of O₂-evolution under flash irradiation of the depleted cells was drastically reduced, which could be restored by addition of NO₃ ^-. The 2,5-dibromo-3-methyl-6-isopropyl-p-benzoquinone (DBMIB)-insensitive O₂-evolution, i.e., photosystem (PS) 2 activity of NO₃--depleted cells, showed a 75% stimulation by addition of NO₃ ^-. PS1-mediated electron transport was also stimulated (50%) by addition of NO₃ ^-. Fluorescence yields of the NO₃ ^--depleted cells were significantly reduced. A normal fluorescence response was restored by the addition of NO₃ ^-. The fluorescence yield of the NO₃ ^--depleted and DCMU-treated-cells increased significantly after addition of NO₃ ^- ions, indicating a further reduction of the primary acceptor of PS2 (Q). In addition, the low temperature fluorescence emission spectra showed that energy transfer to PS2 and PS1 was much higher when nitrate was present. Hence nitrate accelerates the light-induced charge transfer from the intact O₂-evolving system to the primary electron acceptor of PS2 and stimulates the PS1-mediated electron transport. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nitrification inhibitors

Summary The effects of furano compounds, furfural (furfuraldehyde) and furfuryl alcohol (5, 10, 20 and 30% of N applied) on nitrification of ammonium sulfate and urea N were studied in a sandy clay loam in laboratory. Both furfural and furfuryl alcohol significantly retarded the nitrification rates of both the fertilizers by inhibiting the conversion of NH₄ ⁺ to NO₂ ^- without affecting the oxidation of NO₂ ^- to NO₃ ^--N. 10, 20 and 30% concentrations of the compounds were effective up to 75 days with ammonium sulfate but more or less up to 45 days with urea. re]19760322     

Potential rates of nitrification and denitrification in an oligotrophic freshwater sediment system

Potential rates of nitrification and denitrification were measured in an oligotrophic sediment system. Nitrification potential was estimated using the CO oxidation technique, and potential denitrification was measured by the acetylene blockage technique. The sediments demonstrated both nitrifying and denitrifying activity. E_h, O₂, and organic C profiles showed two distinct types of sediment. One type was low in organic C, had high O₂ and E_h, and had rates of denitrification 1,000 times lower than the other which had high organic C, low O₂, and low E_h. Potential nitrification and denitrification rates were negatively correlated with E_h. This suggests that environmental heterogeneity in denitrifier and nitrifier populations in oligotrophic sediment systems may be assessed using E_h before sampling protocols for nitrification or denitrification rates are established. There was no correlation between denitrification and nitrification rates or between either of these processes and NH₄ ⁺ or NO₃ ^– concentrations. The maximum rate of denitrification was 0.969 nmole N cm^–3 hour^–1, and the maximum rate of nitrification was 23.6 nmole cm^–3 hour^–1, suggesting nitrification does not limit denitrification in these oligotrophic sediments. Some sediment cores had mean concentrations of 6.0 mg O₂/liter and still showed both nitrification and denitrification activity.     

Effects of Methane Metabolism on Nitrification and Nitrous Oxide Production in Polluted Freshwater Sediment

We report the effect of CH₄ and of CH₄ oxidation on nitrification in freshwater sediment from Hamilton Harbour, Ontario, Canada, a highly polluted ecosystem. Aerobic slurry experiments showed a high potential for aerobic N₂O production in some sites. It was suppressed by C₂H₂, correlated to NO₃^- production, and stimulated by NH₄⁺ concentration, supporting the hypothesis of a nitrification-dependent source for this N₂O production. Diluted sediment slurries supplemented with CH₄ (1 to 24 μM) showed earlier and enhanced nitrification and N₂O production compared with unsupplemented slurries (≤1 μM CH₄). This suggests that nitrification by methanotrophs may be significant in freshwater sediment under certain conditions. Suppression of nitrification was observed at CH₄ concentrations of 84 μM and greater, possibly through competition for O₂ between methanotrophs and NH₄⁺ -oxidizing bacteria and/or competition for mineral N between these two groups of organisms. In Hamilton Harbour sediment, the very high CH₄ concentrations (1.02 to 6.83 mM) which exist would probably suppress nitrification and favor NH₄⁺ accumulation in the pore water. Indeed, NH₄⁺ concentrations in Hamilton Harbour sediment are higher than those found in other lakes. We conclude that the impact of CH₄ metabolism on N cycling processes in freshwater ecosystems should be given more attention.     

Denitrification and oxygen respiration in biofilms studied with a microsensor for nitrous oxide and oxygen

Depth distributions of O₂ respiration and denitrification activity were studied in 1- to 2-mm thick biofilms from nutrient-rich Danish streams. Acetylene was added to block the reduction of N₂O, and micro-profiles of O₂ and N₂O in the biofilm were measured simultaneously with a polarographic microsensor. The specific activities of the two respiratory processes were calculated from the microprofiles using a one-dimensional diffusion-reaction model. Denitrification only occurred in layers where O₂ was absent or present at low concentrations (of a fewM). Introduction of O₂ into deeper layers inhibited denitrification, but the process started immediately after anoxic conditions were reestablished. Denitrification activity was present at greater depth in the biofilm when the NO₃ ^– concentration in the overlying water was elevated, and the deepest occurrence of denitrification was apparently determined by the depth penetration of NO₃ ^–. The denitrification rate within each specific layer was not affected by an increase in NO₃ ^– concentration, and the half-saturation concentration (K_m) for NO₃ ^– therefore considered to be low (<25M). Addition of 0.2% yeast extract stimulated denitrification only in the uppermost 0.2 mm of the denitrification zone indicating a very efficient utilization of the dissolved organic matter within the upper layers of the biofilm.     

Mechanisms and kinetics of nitric and nitrous oxide production during nitrification in agricultural soil

Laboratory experiments were conducted with three California agricultural soils to examine substrate and process controls over temporal variability of NO and N₂O production during nitrification, and to quantify the kinetics of HNO₂‐mediated chemical reactions. Gross NO production rates were highly correlated (r² = 0.93–0.97) with calculated concentrations of HNO₂, which were shown to originate from autotrophic microbial oxidation of NH₄ ⁺ to NO₂ ^? Production of NO was not correlated with NH₄ ⁺ or NO₃^–, or with the overall nitrification rate. Distinct periods of high NO₂^– accumulation occurred below critical pH values in each soil, apparently due to inhibition of microbial NO₂^– oxidation. Data suggest that even during periods of relatively low NO₂^– accumulation and rapid overall nitrification, HNO₂‐mediated reactions may have been the primary source of NO. Rate coefficients (k_PNO) relating NO production to HNO₂ concentrations were determined for sterile (λ‐irradiated) soils, and were similar to k_PNO values in 2 of 3 nonsterile soils undergoing nitrification. Production of N₂O was correlated with HNO₂ (r² = 0.88–0.99) in sterile soils, and with NO₂^– and NO₃^– (R² = 0.72–0.91) in nonsterile soils. Experiments using ¹⁵N confirmed that dissimilatory NO₃^– reduction contributed to N₂O production even under primarily aerobic conditions. Sterile kPNO and kPN2O values were correlated (r² = 0.90 and 0.82) with soil organic matter content. Overall, the results demonstrate that both steps of the nitrification sequence, together with abiotic reactions involving NO₂^–/HNO₂ need to be considered in developing improved models of NO and N₂O emissions from soils.     

Identification of Heterotrophic Nitrification in a Sierran Forest Soil

A potential for heterotrophic nitrification was identified in soil from a mature conifer forest and from a clear-cut site. Potential rates of NO₂⁻ production were determined separately from those of NO₃⁻ by using acetylene to block autotrophic NH₄⁺ oxidation and chlorate to block NO₂⁻ oxidation to NO₃⁻ in soil slurries. Rates of NO₂⁻ production were similar in soil from the forest and the clear-cut site and were strongly inhibited by acetylene. The rate of NO₃⁻ production was much greater than that of NO₂⁻ production, and NO₃⁻ production was not significantly affected by acetylene or chlorate. Nitrate production was partially inhibited by cycloheximide, but was not significantly reduced by streptomycin. Neither the addition of ammonium nor the addition of peptone stimulated NO₃⁻ production. ¹⁵N labeling of the NH₄⁺ pool demonstrated that NO₃⁻ was not coming from NH₄⁺. The potential for heterotrophic nitrification in these forest soils was greater than that for autotrophic nitrification.     

Nitrous oxide in brackish Lake Nakaumi, Japan II: the role of nitrification and denitrification in N2O accumulation

In order to understand the role of nitrification and denitrification in the accumulation of nitrous oxide (N₂O) in the hypolimnetic water of brackish Lake Nakaumi, the effects of dissolved oxygen (DO) concentration on these activities were investigated by incubation experiments. N₂O was produced during the oxidation of NH₄ ⁺ to NO₂ ⁻ in nitrification and during the reduction of NO₃ ⁻ to N₂ in denitrification. N₂O-producing activity by nitrification (N₂O_N) increased markedly with decreasing concentrations of DO. Low DO (10%–30% saturation) induced high N₂O_N. In contrast to nitrification, N₂O-producing activity by denitrification (N₂O_D) decreased with decreasing concentrations of DO. Little N₂O was accumulated during denitrification under low-level conditions of DO (10%–30%), because of further reduction of N₂O to N₂. It can therefore be assumed that N₂O produced as the by-product of nitrification is concurrently reduced to N₂ by denitrification under low-DO conditions. This would result in no substantial accumulation of N₂O during active nitrification in the hypolimnetic water of Lake Nakaumi. Received: July 6, 2001 / Accepted: December 10, 2001     

The effect of nitrate and nitrite on oxygen evolution and carbon-dioxide assimilation and the reduction of nitrate and nitrite by intact chloroplasts

Summary Intact chloroplasts isolated from spinach reduced NO₃ ^- and NO₂ ^- in the light without the addition of either co-factors or added enzymes. The maximum rate observed, however, for the reduction of NO₃ ^- was approximately 3Moles hr^-1 mg^-1 (chlorophyll) and for NO₂ ^- 6 Moles hr^-1 mg^-1 (chlorophyll). These rates were consistent with the enzyme content of whole chloroplasts, but much lower than those found in whole leaf extracts.The addition of both NO₃ ^- and NO₂ ^- in low concentrations resulted in transient increases in both O₂ evolution and CO₂ fixation. The increases in oxygen evolution were not consistent in amount and bore no relation to the amount of substrate reduced. Similar transients were observed in a number of experiments when NaCl or NH₄Cl were added.The addition of NO₂ ^- at concentrations of 10^-4 M and above resulted in marked inhibition of both O₂ evolution and CO₂ fixation. NO₂ ^- appears to inhibit by blocking the reduction of NADP. NO₃ ^- at similar concentrations had no such effect.An increase in the soluble amino nitrogen content of the chloroplasts was observed when NO₃ ^- or NO₂ ^- was reduced. There was, however, no increase in the incorporation of ¹⁴C from ¹⁴CO₂ into amino acids under these conditions. Even with the addition of ammonia the amount of ¹⁴C incorporated into the amino acids was not changed from less than 5% of the total ¹⁴C fixed. We conclude that while intact chloroplasts do have the ability to reduce both NO₃ ^- and NO₂ ^- at low rates, they do not synthesize appreciable amounts of amino acid directly, and this fact must be considered when formulating any pathways for nitrogen metabolism during photosynthesis.Supported in part by the National Research Council of Canada.     

Oxygen limitation of N2 fixation in stem-girdled and nitrate-treated soybean

The effects of increasing rhizosphere pO₂on nitrogenase activity and nodule resistance to O₂diffusion were investigated in soybean plants [Glycine max (L.) Merr. cv. Harosoy 63] in which nitrogenase (EC 1.7.99.2) activities were inhibited by (a) removal of the phloem tissue at the base of the stem (stem girdling), (b) exposure of roots to 10 mM NO₃over 5 days (NO₃-treated), or (c) partial inactivation of nitrogenase activity by an exposure of nodulated roots to 100 kPa O₂(O₂-inhibitcd). In control plants and in plants which had been treated with 100 kPa O₂, increasing rhizosphere O₂concentrations in 10 kPa increments from 20 to 70 kPa did not alter the steady-state nitrogenase activity. In contrast, in plants in which nitrogenase activities were depressed by stem girdling or by exposure to NO₃, increasing rhizosphere pO₂resulted in a recovery of 57 or 67%, respectively, of the initial, depressed rates of nitrogenase activity. This suggests that the nitrogenase activity of stem-girdled and NO₃-treated soybeans was O₂-limited. For each treatment, theoretical resistance values for O₂diffusion into nodules were estimated from measured rates of CO₂exchange, assuming a respiratory quotient of 1.1 and 0 kPa of O₂in the infected cells. At an external partial pressure of 20 kPa O₂, the stem-girdled and NO₃^--treated plants displayed resistance values which were 4 to 8.6 times higher than those in the nodules of the control plants. In control and O₂-inhibited plants, increases in pO₂from 20 to 70 kPa in 10 kPa increments resulted in a 2.5- to 3.9-fold increase in diffusion resistance to O₂, and had little effect on either respiration or nitrogenase activity. In contrast, in stem-girdled and NO₃^--treated plants, increases in external pO₂had little effect on diffusion resistance to O₂, but resulted in a 2.3- to 3.2-fold increase in nodule respiration and nitrogenase activity. These results are consistent with stem-girdling and NO₃^--inhibition treatments limiting phloem supply to nodules causing an increase in diffusion resistance to O₂at 20 kPa and an apparent insensitivity of diffusion resistance to increases in external pO₂.     

晋西北不同年限小叶锦鸡儿灌丛土壤氮矿化和硝化作用

利用PVC管顶盖埋管法研究了晋西北黄土高原区小叶锦鸡儿人工灌丛不同定植年限(5,10,20,30,40a)土壤氮矿化与硝化速率的动态和净矿化与硝化总量。结果表明,⑴小叶锦鸡儿灌丛土壤无机氮主要以NO_-~3-N形式存在,不同生长年限相同月份的土壤硝态氮(NO-3-N)含量分别是铵态氮(NH+4-N)含量的1.5—15.4倍;⑵土壤氮素硝化速率和矿化速率随生长年限延长而加快,30年生时达到高峰,数值达40.2,44.1 mg m~(-2)d~(-1)。从季节性变化看,7—8月份是硝化速率和矿化速率快速增长期,30年生小叶锦鸡儿灌丛土壤硝化速率和矿化速率分别达到86.9,93.1 mg m~(-2)d~(-1),显著高于其它生长年限(P0.05);(3)土壤氮素硝化与矿化总量同样随小叶锦鸡儿生长年限延长而增加,30年生时达到最高,与5年生相比,分别增加了3.7和3.1倍。(4)5—10月份小叶锦鸡儿生长期内,各年限土壤全氮量的2.3%被矿化成无机氮,其中87%最终被转化成NO-3-N形式存在于土体中。     

Nitrite-driven nitrous oxide production under aerobic soil conditions: kinetics and biochemical controls

Nitrite (NO₂⁻) can accumulate during nitrification in soil following fertilizer application. While the role of NO₂⁻ as a substrate regulating nitrous oxide (N₂O) production is recognized, kinetic data are not available that allow for estimating N₂O production or soil‐to‐atmosphere fluxes as a function of NO₂⁻ levels under aerobic conditions. The current study investigated these kinetics as influenced by soil physical and biochemical factors in soils from cultivated and uncultivated fields in Minnesota, USA. A linear response of N₂O production rate () to NO₂⁻ was observed at concentrations below 60 μg N g⁻¹ soil in both nonsterile and sterilized soils. Rate coefficients (K_p) relating to NO₂⁻ varied over two orders of magnitude and were correlated with pH, total nitrogen, and soluble and total carbon (C). Total C explained 84% of the variance in K_p across all samples. Abiotic processes accounted for 31–75% of total N₂O production. Biological reduction of NO₂⁻ was enhanced as oxygen (O₂) levels were decreased from above ambient to 5%, consistent with nitrifier denitrification. In contrast, nitrate (NO₃⁻)‐reduction, and the reduction of N₂O itself, were only stimulated at O₂ levels below 5%. Greater temperature sensitivity was observed for biological compared with chemical N₂O production. Steady‐state model simulations predict that NO₂⁻ levels often found after fertilizer applications have the potential to generate substantial N₂O fluxes even at ambient O₂. This potential derives in part from the production of N₂O under conditions not favorable for N₂O reduction, in contrast to N₂O generated from NO₃⁻ reduction. These results have implications with regard to improved management to minimize agricultural N₂O emissions and improved emissions assessments.     

Effects of Mg(OH)2-fertilization on nutrient cycling in a heavily damaged Norway spruce ecosystem (NE Bavaria/FRG)

Main objective of this study was to test the effects of Mg(OH)₂-fertilization in a Norway spruce ecosystem showing severe symptoms of Mg-deficiency.The site is characterized by high atmospheric inputs with deposition rates of 1.25 kg H, 42 kg S, and 32 kg N per ha and year. The typic Dystrochrept derived from granite is acidified down to greater depths. The pH-values in soil solution of the organic surface layer and the upper mineral soil are around 3.5. Concentrations of Al, SO₄ ^2-, and especially NO₃ ^- and DOC are very high. The element balance indicates a significant influence of N-inputs and processes of N-turnover on the chemical status of the soil and probably on tree nutrition. Nitrification in the upper mineral soil leads to a transformation of a major part of NH₄ ⁺ into NO₃ ^-, which is quantitatively leached, resulting in an ecosystem-internal H⁺-production of 1.8 keq ha^-1yr^-1. NO₃ ^- and SO₄ ^2- govern the seepage output from the ecosystem.Mg(OH)₂ fertilization resulted in manifold increased Mg²⁺ concentrations in soil solution down to 70 cm soil depth and to a significant increase of pH down to 25 cm mineral soil depth. Nitrate concentrations were elevated after fertilization, but decreased within 15 months below the level of the control plot. As a mean over the whole experimental period, N-output was not increased by fertilization. Despite an elevated internal proton production due to nitrification, acid buffering in the soil was clearly increased, but enhanced Al-mobilization was not observed. Mg/Al- and Ca/H-ratios in soil solution indicate much more favourable conditions for fine root growth. Fertilization also increased the amount of exchangeable Mg down to 40cm mineral soil depth. Mg contents in current-year needles increased after three vegetation periods. Thirty months after application, only 10% and 4% of the fertilized Mg had left the organic surface layer and the mineral soil with seepage water output, respectively.