Soil Formate Regulates the Fungal Nitrous Oxide Emission Pathway

Fungal activity is a major driver in the global nitrogen cycle, and mounting evidence suggests that fungal denitrification activity contributes significantly to soil emissions of the greenhouse gas nitrous oxide (N₂O). The metabolic pathway and oxygen requirement for fungal denitrification are different from those for bacterial denitrification. We hypothesized that the soil N₂O emission from fungi is formate and O₂ dependent and that land use and landforms could influence the proportion of N₂O coming from fungi. Using substrate-induced respiration inhibition under anaerobic and aerobic conditions in combination with ¹⁵N gas analysis, we found that formate and hypoxia (versus anaerobiosis) were essential for the fungal reduction of ¹⁵N-labeled nitrate to ¹⁵N₂O. As much as 65% of soil-emitted N₂O was attributable to fungi; however, this was found only in soils from water-accumulating landforms. From these results, we hypothesize that plant root exudates could affect N₂O production from fungi via the proposed formate-dependent pathway.     

Implications of Competitive Inhibition in the Acetylene Reduction Assay for Dinitrogen Fixation

An earlier paper which analysed the implications of diffusionlimitation for acetylene reduction by intact legume noduleshas recently been criticized for ignoring competitive inhibitionof acetylene reduction by dinitrogen. Mathematical analysesof competitive inhibition show that the dependence of acetylenereduction rate on acetylene concentration under atmosphericN₂ is described adequately by an equation functionally equivalentto the Michaelis—Menten equation, despite competitiveinhibition. Therefore, no modification of the original analysisis required. However, competitive inhibition is shown to bea significant factor when experiments under atmospheric N₂ andunder argon are compared. Acetylene reduction, dinitrogen fixation, competitive inhibition, diffusion limitation, legume nodules     

Production of Nitrous Oxide by Ammonia-Oxidizing Chemoautotrophic Microorganisms in Soil

Gas chromatographic studies showed that nitrous oxide was produced in each instance when sterilized (autoclaved) soil was incubated after treatment with ammonium sulfate and inoculation with pure cultures of ammonia-oxidizing chemoautotrophic microorganisms (strains of Nitrosomonas, Nitrosospira, and Nitrosolobus). Production of N₂O in ammonium-treated sterilized soil inoculated with Nitrosomonas europaea increased with the concentration of ammonium and the moisture content of the soil and was completely inhibited by both nitrapyrin and acetylene. Similar effects of nitrapyrin, acetylene, ammonium concentration, and soil moisture content were observed in studies of factors affecting N₂O production in nonsterile soil treated with ammonium sulfate. These observations support the conclusion that, at least under some conditions, most of the N₂O evolved from soils treated with ammonium or ammonium-producing fertilizers is generated by chemoautotrophic nitrifying microorganisms during oxidation of ammonium to nitrite.     

Acetylene Reduction by Soil Cores of Maize and Sorghum in Brazil

Nitrogenase activity was measured by the C₂H₂ reduction method in large soil cores (29 cm in diameter by 20 cm in depth) of maize (Zea mays) and sorghum (Sorghum vulgare). The activity was compared to that obtained by a method in which the roots were removed from the soil and assayed for nitrogenase activity after an overnight preincubation in 1% O₂. In a total of six experiments and 28 soil cores, the nitrogenase activity of the cores was an average of 14 times less than the activity of roots removed from the same cores and preincubated. Nitrogenase activity in the cores was very low and extrapolated to an average nitrogen fixation rate of 2.8 g of N/hectare per day. It was shown that inadequate gas exchange was not a reason for the lower activity in the soil cores, and the core method gave satisfactory results for nitrogenase activity of soybeans (Glycine max) and Paspalum notatum.     

Inhibition of Methanogenesis in Marine Sediments by Acetylene and Ethylene: Validity of the Acetylene Reduction Assay for Anaerobic Microcosms

Methanogenesis was irreversibly inhibited in sediments by concentrations of acetylene employed in nitrogen fixation assays (1 to 20%, vol/vol). Ethylene, but not ethane, also stopped methane production, and the inhibition was reversed by gassing with hydrogen.     

Iron: The Forgotten Driver of Nitrous Oxide Production in Agricultural Soil

In response to rising interest over the years, many experiments and several models have been devised to understand emission of nitrous oxide (N₂O) from agricultural soils. Notably absent from almost all of this discussion is iron, even though its role in both chemical and biochemical reactions that generate N₂O was recognized well before research on N₂O emission began to accelerate. We revisited iron by exploring its importance alongside other soil properties commonly believed to control N₂O production in agricultural systems. A set of soils from California''s main agricultural regions was used to observe N₂O emission under conditions representative of typical field scenarios. Results of multivariate analysis showed that in five of the twelve different conditions studied, iron ranked higher than any other intrinsic soil property in explaining observed emissions across soils. Upcoming studies stand to gain valuable information by considering iron among the drivers of N₂O emission, expanding the current framework to include coupling between biotic and abiotic reactions.     

Nitrous Oxide Emission Associated with Autotrophic Ammonium Oxidation in Acid Coniferous Forest Soil

Aerobic N₂O production was studied in nitrifying humus from urea-fertilized pine forest soil. Acetylene and nitrapyrin inhibited both NH₄⁺ oxidation and N₂O production, indicating that N₂O production was closely associated with autotrophic NH₄⁺ oxidation. N₂O production was enhanced by low soil pH; it was negligible above pH 4.7. When soil pH decreased from 4.7 to 4.1, the relative amount of N₂O-N produced from NH₄⁺-N oxidized increased exponentially to 20%. There was also some evidence that N₂O formation was stimulated by salts (potassium sulfate and sodium phosphates). The maximum rate of N₂O-N production was 0.17 μg of N₂O-N per g of soil per h. When humus was treated with NO₂⁻, N₂O evolved immediately, indicating chemical formation, but no N₂O was formed on the addition of NO₃⁻. The amount of N₂O-N evolved was 0.6 to 4.6% of NO₂⁻-N added. A high concentration of NO₂⁻ and low soil pH enhanced chemical production of N₂O. There was no accumulation of NO₂⁻ during nitrification. The calculations indicated that chemical formation of N₂O was not the main source of N₂O during NH₄⁺ oxidation. After the addition of inhibitors of NH₄⁺ oxidation the soils contained NO₃⁻, but no N₂O was produced. The results suggest that enhanced autotrophic NH₄⁺ oxidation is a potential source of N₂O in fertilized acid forest soil.     

Rapid Conversion of Added Nitrate to Nitrous Oxide and Dinitrogen in Northern Forest Soil

The aims of this study were to simulate wet deposition of atmospheric nitrate (NO₃^?) onto forest soils and trace its fate via conversion spatially and temporally into gaseous products nitrous oxide (N₂O) and dinitrogen (N₂). The most likely mechanism is microbial denitrification, but an intermediate product nitrite (NO₂^?) can fuel N₂O production via a chemical pathway involving reactions with iron and/or organic matter referred to as chemodenitrification. During summer months, we applied tracer amounts of ¹⁵N-labeled NO₃^? onto forest soils (pH ～ 4) at three sites in the White Mountain Region of New Hampshire, USA. We recovered ¹⁵N as N₂O in 210 of 504 measurements (42%) versus ¹⁵N as N₂ in 51 of 504 measurements (10%), suggesting partial microbial denitrification and/or chemodenitrification. When recovery occurred, the mean percent recovery of added ¹⁵N was just 1.1% as N₂O, although N₂ recovery was 33%. A site with old-growth trees had a larger percentage recovery as N₂ (48%), whereas a site that had burned 100 years ago had a small percentage recovery as N₂O (0.24%). The ¹⁵N composition of N₂O in ambient air, collected before addition of the label, was markedly enriched in ¹⁵N. Since flux measurements were made 2 h after the addition, the results suggest that denitrification enzymes and conditions for chemodenitrification are present throughout the summer months but account for small amounts of NO₃^? conversion into N₂O and N₂.     

乙炔抑制法在硝化与反硝化过程中的应用

硝化和反硝化作用在土壤氮素循环中扮演重要作用,由于硝化和反硝化作用一方面能够导致土壤中氮素的损失,另一方面能够产生温室气体-N2O,所以硝化和反硝化作用的研究备受关注.乙炔抑制法能同时测定硝化和反硝化作用,在硝化和反硝化作用中有着重要的应用.该文主要论述了乙炔抑制法的研究进展;以及对应用乙炔气体时存在的一些问题进行了综述.     

Limitations on the Analysis of Acetylene Reduction by Soybean at Low Levels of Acetylene

The analysis of acetylene reduction at low concentrations ofacetylene involves a number of assumptions and both technicaland kinetic complexities. The major difficulty in convertingacetylene reduction rates to apparent N₂ reduction rates isdetermining the K_m for acetylene in the presence of N₂. Thissubstrate competition is dominant over diffusion limitationeffects, but both introduce equivalent deviations in the observedK_m. Because N₂ is a non-linear partial competitive inhibitorof acetylene reduction, correction for its presence is difficult.Two further complications are introduced by the non-linear responseof nitrogenase to acetylene concentration even in the absenceof N₂, and changes in the apparent K_m of acetylene and K₁ ofN₂ as a function of other variables in the enzyme assay. Itis proposed that transient analysis may be used for measurementof diffusion coefficients and calculations of possible diffusionlimitations. It is demonstrated that one proposed model forestimating diffusion limitation (Denison et al., 1983, PlantPhysiology 73, 648–51) confounds substrate competitionwith diffusion limitation. Acetylene reduction, nitrogen fixation, diffusion limitation     

Nitrate Reduction in a Sulfate-Reducing Bacterium, Desulfovibrio desulfuricans, Isolated from Rice Paddy Soil: Sulfide Inhibition, Kinetics, and Regulation

From the second-highest dilution in a most-probable-number dilution series with lactate and sulfate as substrates and rice paddy soil as the inoculum, a strain of Desulfovibrio desulfuricans was isolated. In addition to reducing sulfate, sulfite, and thiosulfate, the strain also reduced nitrate to ammonia. The latter process was studied in detail, since the ability to reduce nitrate was strongly influenced by the presence of sulfide. Sulfide inhibited both growth on nitrate and nitrate reduction. A 70% inhibition of the nitrate reduction rate was obtained at 127 μM sulfide, and growth was inhibited by 50% at approximately 320 μM sulfide and was not detectable above 700 μM sulfide. In contrast, sulfate reduction was not affected at concentrations of up to 5 mM. After growth with sulfate, an induction period of 2 to 4 days was needed before nitrate reduction started. When nitrate and sulfate were present simultaneously, only sulfate was reduced, except when sulfate was present at very low concentrations (4 μM). At higher sulfate concentrations (500 μM), nitrate reduction was temporarily halted. The affinity for nitrate uptake was extremely high (K_m = 0.05 μM) compared with that for sulfate uptake (K_m = 5 μM). Thus, at low nitrate concentrations this bacterium is favored relative to denitrifiers (K_m = 1.8 to 13.7 μM) or other nitrate ammonifiers (e.g., Clostridium spp. [K_m = 500 μM]).     

Denitrification, Acetylene Reduction, and Methane Metabolism in Lake Sediment Exposed to Acetylene

Samples of sediment from Lake St. George, Ontario, Canada, were incubated in the laboratory under an initially aerobic gas phase and under anaerobic conditions. In the absence of added nitrate (NO₃⁻) there was O₂-dependent production of nitrous oxide (N₂O), which was inhibited by acetylene (C₂H₂) and by nitrapyrin, suggesting that coupled nitrification-denitrification was responsible. Denitrification of added NO₃⁻ was almost as rapid under an aerobic gas phase as under anaerobic conditions. The N₂O that accumulated persisted in the presence of 0.4 atm of C₂H₂, but was gradually reduced by some sediment samples at lower C₂H₂ concentrations. Low rates of C₂H₂ reduction were observed in the dark, were maximal at 0.2 atm of C₂H₂, and were decreased in the presence of O₂, NO₃⁻, or both. High rates of light-dependent C₂H₂ reduction occurred under anaerobic conditions. Predictably, methane (CH₄) production, which occurred only under anaerobiosis, was delayed by added NO₃⁻ and inhibited by C₂H₂. Consumption of added CH₄ occurred only under aerobic conditions and was inhibited by C₂H₂.     

Role of Microorganisms in Emission of Nitrous Oxide and Methane in Pulse Cultivated Soil Under Laboratory Incubation Condition

Soil from a pulse cultivated farmers land of Odisha, India, have been subjected to incubation studies for 40 consecutive days, to establish the impact of various nitrogenous fertilizers and water filled pore space (WFPS) on green house gas emission (N₂O & CH₄). C₂H₂ inhibition technique was followed to have a comprehensive understanding about the individual contribution of nitrifiers and denitrifiers towards the emission of N₂O. Nevertheless, low concentration of C₂H₂ (5 ml: flow rate 0.1 kg/cm²) is hypothesized to partially impede the metabolic pathways of denitrifying bacterial population, thus reducing the overall N₂O emission rate. Different soil parameters of the experimental soil such as moisture, total organic carbon, ammonium content and nitrate–nitrogen contents were measured at regular intervals. Application of external N-sources under different WFPS conditions revealed the diverse role played by the indigenous soil microorganism towards green house gas emission. Isolation of heterotrophic microorganisms (Pseudomonas) from the soil samples, further supported the fact that denitrification might be prevailing during specific conditions thus contributing to N₂O emission. Statistical analysis showed that WFPS was the most influential parameter affecting N₂O formation in soil in absence of an inhibitor like C₂H₂.