Carbon and oxygen controls on N2O and N2 production during nitrate reduction

Here we provide evidence that the form of carbon compound and O₂ concentration exert an inter-related regulation on the production and reduction of N₂O in soil. 6.7 mM d-glucose, 6.7 mM D-mannitol, 8 mM L-glutamic acid or 10 mM butyrate (all equivalent to 0.48 g C l⁻¹) were applied to slurries of a sandy loam soil. At the start of the experiment headspace O₂ concentrations were established at ∼2%, 10% and 21% O₂ v/v for each C treatment, and 2 mM K¹⁵NO₃ (25 atom % excess ¹⁵N) was applied, enabling quantification of ¹⁵N-N₂ production, ¹⁵N-(N₂O-to-N₂) ratios and DNRA. The form of C compound was most important in the initially oxic (21% O₂ v/v) soils, where addition of butyrate and glutamic acid resulted in greater N₂O production (0.61 and 0.3 μg N₂O-N g⁻¹ soil for butyrate and glutamic acid, respectively) than the addition of carbohydrates (glucose and mannitol). Although, there was no significant effect of C compound at low initial O₂ concentrations (∼2% O₂ v/v), production of ¹⁵N-N₂ was greatest where headspace O₂ concentrations were initially, or fallen to, ∼2% O₂ v/v, with greatest reduction of N₂O and lowering ¹⁵N-(N₂O-to-N₂) ratios (∼0-0.27). This may reflect that the effect of C is indirect through stimulation of heterotrophic respiration, lowering O₂ concentrations, providing sub-oxic conditions for dissimilatory nitrate reduction pathways. Addition of carbohydrates (glucose and mannitol) also resulted in greatest recovery of ¹⁵N in NH₄⁺ from applied ¹⁵N-NO₃⁻, indicative of the occurrence of DNRA, even in the slurries with initial 10% and 21% O₂ v/v concentrations. Our ¹⁵N approach has provided the first direct evidence for enhancement of N₂O reduction in the presence of carbohydrates and the dual regulation of C compound and O₂ concentration on N₂O production and reduction, which has implications for management of N₂O emissions through changing C inputs (exudates, rhizodeposition, residues) with plant species of differing C traits, or through plant breeding.     

The effect of moisture on nitrous oxide emissions from soil and the N<Subscript>2</Subscript>O/(N<Subscript>2</Subscript>O+N<Subscript>2</Subscript>) ratio under laboratory conditions

Nitrous oxide (N₂O) contributes to greenhouse effect; however, little information on the consequences of different moisture levels on N₂O/(N₂O+N₂) ratio is available. The aim of this work was to analyze the influence of different soil moisture values and thus of redox conditions on absolute and relative emissions of N₂O and N₂ at intact soil cores from a Vertic Argiudoll. For this reason, the effect of water-filled porosity space (WFPS) values of soil cores of 40, 80,100, and 120% (the last one with a 2-cm surface water layer) was investigated. The greatest N₂O emission occurred at 80% WFPS treatment where conditions were not reductive enough to allow the complete reduction to N₂. The N₂O/(N₂O+N₂) ratio was lowest (0–0.051) under 120% WFPS and increased with decreasing soil moisture content. N₂O/(N₂O+N₂) ratio values significantly correlated with soil Eh; redox conditions seemed to control the proportion of N gases emitted as N₂O. N₂O emissions did not correlate satisfactorily with N₂O/(N₂O+N₂) ratio values, whereas they were significantly explained by the amount of total N₂O+N₂ emissions.     

Cutting regime determines allocation of fixed nitrogen in white clover

Leguminous leys are important sources of nitrogen (N), especially in forage-based animal production and organic cropping. Models for estimating total N₂ fixation of leys—including below-ground plant-derived N (BGN)—are based on grazed or harvested leys. However, green manure leys can have different proportions of above-ground plant-derived N (AGN) and BGN when subjected to different cutting regimes. To investigate the effects of cutting on N distribution in white clover, a pot experiment was carried out using ¹⁵N techniques to determine N₂ fixation, N rhizodeposition and root C and N content of cut and uncut white clover (Trifolium repens L. cv. Ramona) plants. Percentage N derived from air (%Ndfa) was lower in uncut (63%) than in cut (72%) plants, but total Ndfa was not significantly affected by cutting. The higher reliance on N₂ fixation in cut plants was thus counterbalanced by lower biomass and total N content. With BGN taken into account, total plant-derived N increased by approximately 50% compared with AGN only. Cutting did not affect the proportion of BGN to standing shoot biomass N after regrowth, but decreased the proportion of BGN to total shoot biomass production during the entire growth period. Thus, estimates of N fixation in green manure leys should consider management practices such as cutting regime, as this can result in differences in above- and below-ground proportions of plant-derived N.     

Effects of drought on isolates of <Emphasis Type="Italic">Bradyrhizobium elkanii</Emphasis> cultured from <Emphasis Type="Italic">Albizia adianthifolia</Emphasis> seedlings of different provenances

Albizia adianthifolia (Schumach) W. F. Wright, a N-fixing legume tree, has a wide distribution in Africa, in Ghana occurring in high rainfall forests and in seasonally droughted forests, and is associated in the Ghanaian forest zone with dry, infertile sites. We hypothesised that A. adianthifolia hosted different rhizobial strains in different forest types, and that these different strains would show different growth responses to moisture stress and different motility and mortality in droughted soil. Three isolates, extracted from seedlings of A. adianthifolia growing in three forest types differing in seasonal drought, were identified as Bradyrhizobium elkanii and exposed to varying levels of osmotic stress. Growth responses varied between the three strains, one of which displayed clear signs of drought tolerance. A novel approach using soil leaching columns was used to test the effects of soil pore water (in terms of neck diameter) on both the survival and movement of wet and dry forest rhizobial isolates through soil columns. The responses of the isolates were significantly different, the pore neck diameter, marginally insignificant and the drought treatment insignificant. Thus the dry forest isolate survived better in all treatments, and showed less response to the treatments, than the isolate from the wet forest. The results offer preliminary evidence that Bradyrhizobium elkanii strains from A. adianthifolia in Ghana have evolved in response to local differences in seasonal water availability. These differences could assist the selection of A. adianthifolia provenances for agroforestry or land rehabilitation.     

Evaluation of nitrogen fixation by different strains of theAzolla-Anabaena symbiosis in the presence of a high level of ammonium

Summary The proportion of N derived from N₂ fixation for 99 strains ofAzolla spp. (comprising all known species) in the presence of ammonium (40 mg/1) was assessed using a¹⁵N-dilution technique. The percentage of N derived from air varied from 29.5% to 79.9%. Although the N concentration ofAzolla spp. was not correlated with fertilizer N, it correlated fairly well with N₂ fixation. Regression analysis suggests that the N yield ofAzolla spp. is more dependent on N₂ fixation than on ammonium assimilation. The high correlation between N yield and isotopically determined, fixed N₂ indicates that the N yield could be used as a parameter in the selection ofAzolla spp. strains that are capable of maintaining high N₂ fixation in the presence of a high level of ammonium.     

Response of field-grown bean (Phaseolus vulgaris L.) to Rhizobium inoculation and nitrogen fertilization in two Cerrados soils

 Most soils sown with field beans (Phaseolus vulgaris L.) contain indigenous rhizobia which might interfere with the establishment of inoculated strains. As a consequence, the benefits of bean inoculation are usually questioned, and the use of N fertilizer is gradually becoming a common practice. The present study had the objective of evaluating the effectiveness of inoculation and N fertilization in field soil with (site 1) and without (site 2) a previous bean-cropping history. At site 1, which had a rhizobial population of 7×10² cells g^–1 soil, inoculation had no effect on nodulation or yield, whereas at site 2 (<10 cells g^–1 soil) inoculation increased nodulation, nodule occupancy by the inoculated strain and grain yield. N fertilizer decreased nodulation at both sites, but increased grain yield at site 1 but not at site 2, indicating that the response to inoculation and N fertilization depends on the cropping history. When bean was cultivated for the first time, indigenous populations of rhizobia were low and high yields were accomplished solely with seed inoculation, with no further response to N fertilizer. In contrast, previous cultivation of bean increases soil rhizobia, preventing nodule formation by inoculated strains, and N fertilizer may be necessary for maximum yields. A significant interaction effect between N fertilizer and inoculation was detected for serogroup distribution only at site 2, with N fertilizer decreasing nodule occupancy by the inoculated strain and increasing the occurrence of indigenous strains. Consequently, although no benefits were obtained by the combination of inoculation and N fertilizer, this practice may be feasible with the selection of appropriate N-tolerant strains from the indigenous rhizobial population. Received: 26 May 1999     

土壤氮气排放研究进展

自20世纪初人类发明并掌握工业合成氨的技术以来,氮肥施用量迅速增长。在一部分国家或地区,氮肥的施入量已经超过作物对氮素的需求,导致大量氮素损失到环境中,造成氨挥发、氧化亚氮排放、地下水硝酸盐污染等环境问题。土壤在微生物的作用下可以通过反硝化、厌氧氨氧化等过程将活性氮素转化为惰性氮气,达到清除过多活性氮的目的。由于大气中氮气背景浓度太高,因此很难直接准确测定土壤的氮气排放速率,导致土壤氮气排放通量、过程与调控机制研究远远落后于土壤氮循环的其他方面。本文综述了土壤氮气排放主要途径(反硝化、厌氧氨氧化与共反硝化)及其对土壤氮气排放的贡献;测定土壤氮气排放速率的方法(乙炔抑制法、氮同位素示踪法、N2/Ar比率-膜进样质谱法、氦环境法与N2O同位素自然丰度法)及其优缺点;调控土壤氮气排放通量的主要因素(氧气、可溶性有机碳、硝酸盐、微生物群落结构与功能基因表达等)及其相关作用机制。最后指出研发新的测定原位无扰动土壤氮气通量的方法是推进本领域相关研究的关键;定量典型生态系统(如旱地农田、稻田、森林、草地与湿地)土壤氮气排放通量,阐明其中的微生物学机制,模拟并预测土壤氮气排放对全球变化的响应规律是本领域的研究热点与发展方向。     

Dinitrogen emissions and the N2:N2O emission ratio of a Rendzic Leptosol as influenced by pH and forest thinning

Reduction of nitrous oxide (N₂O) to dinitrogen (N₂) by denitrification in soils is of outstanding ecological significance since it is the prevailing natural process converting reactive nitrogen back into inert molecular dinitrogen. Furthermore, the extent to which N₂O is reduced to N₂ via denitrification is a major regulating factor affecting the magnitude of N₂O emission from soils. However, due to methodological problems in the past, extremely little information is available on N₂ emission and the N₂:N₂O emission ratio for soils of terrestrial ecosystems. In this study, we simultaneously determined N₂ and N₂O emissions from intact soil cores taken from a mountainous beech forest ecosystem. The soil cores were taken from plots with distinct differences in microclimate (warm-dry versus cool-moist) and silvicultural treatment (untreated control versus heavy thinning). Due to different microclimates, the plots showed pronounced differences in pH values (range: 6.3–7.3). N₂O emission from the soil cores was generally very low (2.0 ± 0.5–6.3 ± 3.8 μg N m⁻² h⁻¹ at the warm-dry site and 7.1 ± 3.1–57.4 ± 28.5 μg N m⁻² h⁻¹ at the cool-moist site), thus confirming results from field measurements. However, N₂ emission exceeded N₂O emission by a factor of 21 ± 6–220 ± 122 at the investigated plots. This illustrates that the dominant end product of denitrification at our plots and under the given environmental conditions is N₂ rather than N₂O. N₂ emission showed a huge variability (range: 161 ± 64–1070 ± 499 μg N m⁻² h⁻¹), so that potential effects of microclimate or silvicultural treatment on N₂ emission could not be identified with certainty. However, there was a significant effect of microclimate on the magnitude of N₂O emission as well as on the mean N₂:N₂O emission ratio. N₂:N₂O emission ratios were higher and N₂O emissions were lower for soil cores taken from the plots with warm-dry microclimate as compared to soil cores taken from the cool-moist microclimate plots. We hypothesize that the increase in the N₂:N₂O emission ratio at the warm-dry site was due to higher N₂O reductase activity provoked by the higher soil pH value of this site. Overall, the results of this study show that the N₂:N₂O emission ratio is crucial for understanding the regulation of N₂O fluxes of the investigated soil and that reliable estimates of N₂ emissions are an indispensable prerequisite for accurately calculating total N gas budgets for the investigated ecosystem and very likely for many other terrestrial upland ecosystems as well.     

Denitrification in grass swards is increased under elevated atmospheric CO2

Emissions of N₂O and N₂ were measured from Lolium perenne L. swards under ambient (36 Pa) and elevated (60 Pa) atmospheric CO₂ at the Swiss free air carbon dioxide enrichment experiment following application of 11.2 g N m⁻² as ¹⁵NH₄¹⁵NO₃ or ¹⁴NH₄¹⁵NO₃ (1 at.% excess ¹⁵N). Total denitrification (N₂O+N₂) was increased under elevated pCO₂ with emissions of 6.2 and 19.5 mg ¹⁵N m⁻² measured over 22 d from ambient and elevated pCO₂ swards, respectively, supporting the hypothesis that increased belowground C allocation under elevated pCO₂ provides the energy for denitrification. Nitrification was the predominant N₂O producing process under ambient pCO₂ whereas denitrification was predominant under elevated pCO₂. The N₂-to-N₂O ratio was often higher under elevated pCO₂ suggesting that previous estimates of gaseous N losses based only on N₂O emissions have greatly underestimated the loss of N by denitrification.