Fungal control of nitrous oxide production in semiarid grassland

Fungi are capable of both nitrification and denitrification and dominate the microbial biomass in many soils. Recent work suggests that fungal rather than bacterial pathways dominate N transformation in desert soils. We evaluated this hypothesis by comparing the contributions of bacteria and fungi to N₂O production at control and N fertilized sites within a semiarid grassland in central New Mexico (USA). Soil samples were taken from the rhizosphere of blue grama (B. gracilus) and the microbiotic crusts that grow in open areas between the bunch grasses. Soils incubated at 30% or 70% water holding capacity, were exposed to one of three biocide treatments (control, cycloheximide or streptomycin). After 48 h, N₂O and CO₂ production were quantified along with the activities of several extracellular enzymes. N₂O production from N fertilized soils was higher than that of control soils (165 vs. 41 pmol h⁻¹ g⁻¹), was higher for crust soil than for rhizosphere soil (108 vs. 97 pmol h⁻¹ g⁻¹), and increased with soil water content (146 vs. 60 pmol h⁻¹ g⁻¹). On average, fungicide (cycloheximide) addition reduced N₂O production by 85% while increasing CO₂ production by 69%; bactericide (streptomycin) reduced N₂O by 53% with mixed effects on CO₂ production. N₂O production was significantly correlated with C and N mineralization potential as measured by assays for glycosidic and proteolytic enzymes, and with extractable nitrate and ammonium. Our data indicate that fungal nitrifier denitrification and bacterial autotrophic nitrification dominate N transformation in this ecosystem and that N₂O production is highly sensitive to soil cover, N deposition and moisture.     

Identifying sources of nitrous oxide emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) acclimated in different aeration rates

Both long term and batch experiments were carried out to identify the sources of the N₂O emission in anoxic/aerobic sequencing batch reactors (A/O SBRs) under different aeration rates. The obtained results showed that aeration rate has an important effect on the N₂O emission of A/O SBR and most of the N₂O was emitted during the aerobic phase. During the anoxic phase, nitrate ammonification was the major source of N₂O emission while denitrification performed as a sink of N₂O, in all three bioreactors. The N₂O emission mechanisms during the aerobic phase differed with the aeration rate. At low and high aeration rates (Run 1 and Run 3), both coupled-denitrification and nitrifier denitrification were ascribed to be the source of N₂O emission. At mild aeration rate (Run 2), nitrifier denitrification by Nitrosomonas-like ammonia oxidizing-bacterial (AOB) was responsible for N₂O emission while coupled-denitrification turned out to be a sink of N₂O because of the presence of inner anaerobic region in sludge flocs.     

寒温带兴安落叶松林土壤温室气体通量的时间变异

采用静态箱/气相色谱(GC)法,对寒温带兴安落叶松林区6-9月生长季土壤CO2、CH4和N2O通量进行原位测定,研究了土壤温室气体通量的季节和昼夜变化及其与环境因子的关系.结果表明:在生长季,兴安落叶松林土壤为大气CH4的汇,吸收通量为22.3～107.8 μg CH4-C·m-2·h-1,6-9月月均甲烷吸收通量为(34.0±7.1)、(71.4±9.4)、(86.3±7.9)和(40.7-±6.2) μg·m-2·h-1;不同季节土壤CH4昼夜通量的变化规律相同,一天中均在10:00达到最大吸收高峰.土壤CO2日通量呈明显的双峰曲线,月均CO2通量大小顺序为7月＞8月＞6月＞9月.土壤N2O通量变异较大,在-9.1 ～31.7μg·m-2·h-1之间.土壤温度和湿度是影响CO2和CH4通量的重要因子,N2O通量主要受温度的影响.在兴安落叶松林区,10:00左右观测获得的温室气体地-气交换通量,经矫正后可以代表当日气体通量.     

Challenges for simultaneous nitrification, denitrification, and phosphorus removal in microbial aggregates: mass transfer limitation and nitrous oxide production

The microbial community composition and activity was investigated in aggregates from a lab-scale bioreactor, in which nitrification, denitrification and phosphorus removal occurred simultaneously. The biomass was highly enriched for polyphosphate accumulating organisms facilitating complete removal of phosphorus from the bulk liquid; however, some inorganic nitrogen still remained at the end of the reactor cycle. This was ascribed to incomplete coupling of nitrification and denitrification causing NO(3)(-) accumulation. After 2 h of aeration, denitrification was dependent on the activity of nitrifying bacteria facilitating the formation of anoxic zones in the aggregates; hence, denitrification could not occur without simultaneous nitrification towards the end of the reactor cycle. Nitrous oxide was identified as a product of denitrification, when based on stored PHA as carbon source. This observation is of critical importance to the outlook of applying PHA-driven denitrification in activated sludge processes.     

Antagonism of nitrous oxide antinociception in mice by intrathecally administered antisera to endogenous opioid peptides

Previously it was demonstrated that nitrous oxide antinociception in the mouse abdominal constriction test is mediated by kappa-opioid receptors. Since nitrous oxide is thought to cause the neuronal release of endogenous opioid peptide to stimulate opioid receptors, this study was designed to identify the opioid peptides involved, especially in the spinal cord, by determining whether nitrous oxide antinociception can be differentially inhibited by intrathecally (i. t.) administered antisera to different opioid peptides. Male NIH Swiss mice were pretreated i.t. with rabbit antisera to opioid peptides then exposed 24 h later to one of three different concentrations of nitrous oxide in oxygen. Dose-response curves constructed from the data indicated that the antinociceptive effect of nitrous oxide was significantly antagonized by antisera to various dynorphins (DYNs) and methionine-enkephalin (ME), but not by antiserum to beta-endorphin (beta-EP). The AD(50) values for nitrous oxide antinociception were significantly elevated by antisera to DYNs and ME but not beta-EP. These findings of this study support the hypothesis that nitrous oxide antinociception in the mouse abdominal constriction test involves the neuronal release of DYN and ME in the spinal cord.     

Mechanism of nitrite reduction to nitrous oxide in a photodenitrifier,Rhodopseudomonas sphaeroide forma sp. denitrificans

The mechanism of anaerobic reduction of NO₂^? to N₂O in a photodenitrifier, Rhodopseudomonas sphaeroides forma sp. denitrificans, was investigated. With ascorbate-reduced phenazine methosulfate (PMS) as the electron donor, the nitrite reductase of this photodenitrifier reduced NO₂^? to NO and a trace amount of N₂O. With dithionite-reduced benzyl viologen as the electron donor, the major product of NO₂^? reduction was NH₂OH, and a trace amount of N₂O was also produced. The nitrate reductase itself had no NO reductase activity with ascorbate-reduced PMS. It was concluded that the essential product of NO₂^? reduction by the purified nitrite reductase is NO. Chromatophore membranes stoichiometrically produced N₂O from NO₂^? with any electron donor, such as dithionite-redduced benzyl viologen, ascorbate-reduced PMS or NADH/FMN. The membranes also contrained activity of NO reduction of N₂O with either ascorbate-reduced PMS or duroquinol. The NO reductase activity with duroquinol was inhibited by antimycin A. Stoichiometric production of N₂O from N₂^? also was observed in the reconstituted NO₂^? reduction system which contained the cytochrome bc₁ complex, cytochrome c₂, the nitrite reductase and duroquinol as the electron donor. The preparation of the cytochrome bc₁ complex itself contianed NO reductase activity. From these results the mechanism of NO₂^? reduction to N₂O in this photodenitrifier was determined as the nitrite reductase reducing NO₂^? to NO with electrons from the cytochrome bc₁ complex, and NO subsequently being reduced, without release, to N₂O with electrons from the cytochrome bc₁ complex by the NO reductase, which is closely associated with the complex.     

Copper-sulfur complexes supported by N-donor ligands: Towards models of the CuZ site in nitrous oxide reductase

The distinctive structure of the [(his)₇Cu₄(μ-S)]ⁿ⁺ cluster in the “Cu_Z” active site of nitrous oxide reductase and the intriguing mechanistic hypotheses for its catalytic reactivity provide inspiration for synthetic model studies aimed at characterizing relevant copper-sulfur compounds and obtaining fundamental insights into structure and bonding. In this brief review, we summarize such studies that have focused on the synthesis and characterization of a range of copper-sulfur complexes supported by N-donor ligands. Compounds with variable nuclearities and sulfur redox levels have been isolated, with the nature of the species obtained being dependent on the supporting ligand, sulfur source, and the reaction conditions. Spectroscopic data and theoretical calculations, often performed with a view toward drawing comparisons to oxygen analogs, have provided insight into the nature of the copper-sulfur bonding interactions in the complexes.     

锡林郭勒生物圈保护区草原生态系统服务间接价值动态评估

草地是我国面积最大的陆地生态系统类型,对于维持我国北方的生态安全具有重要价值。北方草原的大面积退化已引起人们的极大关注。以内蒙古锡林郭勒生物圈保护区为研究区域．利用2个年份的遥感数据,结合地面样方调查,在GIS的支持下,绘制出草原退化空间分布图。选取草原生态系统在控制土壤侵蚀、减少土壤肥力流失、营养物质循环、CO2储存和O2释放方面的功能指标,对草原生态系统服务间接价值进行了动态评估。1985年和1999年草原退化面积分别为7190．9和7689．3km^2,占研究区总面积的67．19％和71．86％。2个年份草原生态系统间接服务总价值分别为28．24和26．75亿元．下降了5．28％,单位面积间接服务价值分别为2990和2830元。下降了5．35％。人类的过度放牧是导致草原大面积退化,生态系统功能降低和服务价值下降的主要原因。     

Prevalence of nitrous oxide reducing capacity in denitrifiers from a variety of habitats

Summary A total of 81 strains isolated by T. N. Gamble from soils from eight countries, fresh water lake sediments and nitrified poultry manure were examined for their ability to grow on N₂O as their electron acceptor, as well as for their tendency to produce N₂O from NO ₃ ⁻ in the absence and presence of acetylene. Seventy-seven of the 81 strains were confirmed as denitrifiers. Fifty-nine of the 77 strains grew on N₂O, while 12 strains produced N₂O but could not utilize it. Six strains reduced NO ₃ ⁻ to N₂ but could not grow on N₂O, suggesting that even if N₂O is always an intermediate product of denitrification, it is not always a freely diffusible intermediate. The organisms, however, would consume N₂O that accumulated early in growth and accumulated N₂O in the presence of acetylene. Thus the total number of N₂O users was 65 strains or 83% of the total tested. This implies that the N₂O reducing capacity of denitrifiers occur widely in nature. A high proportion ofPseudomonas fluorescens biotype II reduced N₂O. The accumulation of N₂O from NO ₃ ⁻ in the presence of acetylene provides strong evidence that N₂O is generally an intermediate in denitrification as well as provides additional support for the usefulness of this chemical as a general inhibitor of N₂O reduction.     

Photosynthetic and morphological functional types from different steppe communities in Inner Mongolia, North China

Morphological functional types and photosynthetic pathway types were identified for the forage species from steppe communities in Inner Mongolia, China, using the data of both field survey and published papers. Seven typical steppe communities were selected to investigate the morphological functional type and photosynthetic pathway type compositions and plant functional type (PFT) diversity in steppe communities at regional scale. Morphological functional types, based on plant height and leaf type combined with life span, were optimal for comparing the community differences in the region, while photosynthetic pathway types were fairly coarse for such studies. Of the seven morphological functional types in the steppe communities, perennial forbs (PEF) were the dominant type, and 60 % of species belonged to this type. Each of the high perennial grass (HPG), short perennial grass (SPG), and annual grass (ANG) types represented less than 10 % of the total, even though the grass species were dominant in the seven steppe communities. The differences of PFTs between the steppe communities were remarkable, and the PFT richness and diversity increased from the communities with moist conditions to the ones with dry environments.This revised version was published online in March 2005 with corrections to the page numbers.     

Effects of sulfide and low redox potential on the inhibition of nitrous oxide reduction by acetylene in Pseudomonas nautica

Membrane introduction mass spectrometry was used to investigate the inhibitory effect of acetylene on the nitrous oxide reductase activity of intact cells of Pseudomonas nautica. We studied the effects of the concentrations of nitrate and sulfide, and the redox potential, which have all been implicated in causing a decrease in the inhibitory effects of acetylene during measurements of denitrification in natural environments. There was no evidence that the concentration of nitrate influenced the effect of acetylene. Lowering the redox potential with the reductant Ti(III)-nitrilotriacetate caused a slight alleviation of acetylene inhibition. Much greater effects at the same redox potential were obtained with concentrations of sulfide in the range 1-10 microM.     

Nutrient resorption responses to water and nitrogen amendment in semi-arid grassland of Inner Mongolia, China

Litterfall and fine root production is a major pathway for carbon and nutrient cycling in forest ecosystems. We investigated leaf litterfall, fine-root mass, production and turnover rate in the upper soil (0–30 cm) under four major tree species (Leucaena leucocephala, Acacia nilotica, Azadirachta indica, Prosopis juliflora) of the semi-arid region of India. All the four tree species showed an unimodal peak of leaf litterfall with distinct seasonality. Leucaena leucocephala and Acacia nilotica had maximum leaf litterfall between September and December while Azadirachta indica and Prosopis juliflora shed most of their leaves between February and May. Annual leaf litterfall of the four species ranged from 3.3 Mg ha^?1 (Leucaena leucocephala) to 8.1 Mg ha^?1 (Prosopis juliflora). Marked seasonal variations in amount of fine root biomass were observed in all the four tree species. Fine root production was maximum in Prosopis juliflora (171 g m^?2 y^?1) followed by Azadirachta indica (169 g m^?2 y^?1), Acacia nilotica (106 g m^?2 y^?1) and Leucaena leucocephala (79 g m^?2 y^?1). Fine root biomass showed a seasonal peak after the rainy season but fell to its lowest value during the winter and dry summer season. Fine root turnover rate ranged from 0.56 to 0.97 y^?1 and followed the order Azadirachta indica > Leucaena leucocephala > Prosopis juliflora > Acacia nilotica. The results of this study demonstrated that Prosopis juliflora and Azadirachta indica had greater capability for maintaining site productivity as evidenced from greater leaf litterfall and fine root production.     

The expression of redox proteins of denitrification inThiosphaera pantotropha grown with oxygen,nitrate, and nitrous oxide as electron acceptors

The redox proteins and enzymes involved in denitrification inThiosphaera pantotropha exhibited a differential expression in response to oxygen. Pseudoazurin was completely repressed during batch or continuous culture under oxic conditions. Cytochromecd ₁ nitrite reductase was also heavily repressed after aerobic growth. Nitrite, nitric oxide, and nitrous oxide reductase activities were detected in intact cells under some conditions of aerobic growth, indicating that aerobic denitrification might occur in some circumstances. However, the rates of denitrification were much lower after aerobic growth than after anaerobic growth. Growth with nitrous oxide as sole electron acceptor mimicked aerobic growth in some respects, implying that expression of parts of the denitrification apparatus might be controlled by the redox state of a component of the electron transport chain rather than by oxygen itself. Nevertheless, the regulation of expression of nitrous oxide reductase was linked to the oxygen concentration.     

Effect of experimental nitrogen load on methane and nitrous oxide fluxes on ombrotrophic boreal peatland

Methane (CH₄) and nitrous oxide (N₂O) dynamics were studied in a boreal Sphagnum fuscum pine bog receiving annually (from 1991 to 1996) 30 or 100 kg NH₄NO₃-N ha^–1. The gas emissions were measured during the last three growing seasons of the experiment. Nitrogen treatment did not affect the CH₄ fluxes in the microsites where S. fuscum and S. angustifolium dominated. However, addition of 100 kg NH₄NO₃-N ha^–1 yr^–1 increased the CH₄ emission from those microsites dominated by S. fuscum. This increase was associated with the increase in coverage of cotton grass (Eriophorum vaginatum) induced by the nitrogen treatment. The differences in the CH₄ emissions were not related to the CH₄ oxidation and production potentials in the peat profiles. The N₂O fluxes were negligible from all microsites. Only minor short-term increases occurred after the nitrogen addition.     

Nitrous oxide and methane fluxes of a pristine slope mire in the German National Park Harz Mountains

Pristine peatlands covered by Histosols (bogs and fens) with high water table and a restricted oxygen (O₂) availability are known to have low emissions of nitrous oxide (N₂O) but may be a significant source for atmospheric methane (CH₄) which are both important greenhouse gases. For the first time N₂O and CH₄ fluxes of a pristine slope mire in the German Harz Mountains have been monitored. Previously reported peatlands are characterised by anaerobic conditions due to high water table levels. Slope mires monitored here receive O₂ through slope water inflow. Gas fluxes have been monitored deploying closed chamber method on a central non-forested area and a forested area at the periphery of the slope mire. By means of groundwater piezometers water table levels, ammonium and nitrate contents as well as hydro-chemical variables like oxygen content and redox potential of the mire pore water have been concurrently measured with trace gas fluxes at both monitoring sites of the slope mire. The slope mire took up small amounts of atmospheric methane at a rate of −0.02 ± 0.01 kg C ha⁻¹ year⁻¹ revealing no significant difference between the forested and non-forested site. Higher uptake rates were observed during low water table level. In contrast to pristine peatlands influx of oxygen containing pore water into slope mire does limit reduction processes and resultant CH₄ emission. N₂O fluxes of the forested and non-forested sites of the slope mire did not differ and amounted to 0.25 ± 0.44 kg N ha⁻¹ year⁻¹. Higher emissions were observed at low water table levels and during thawing periods. In spite of favourable conditions N₂O fluxes of the slope mire have been comparable to those of pristine peatlands.     

Impact of COD/N ratio on nitrous oxide emission from microcosm wetlands and their performance in removing nitrogen from wastewater

Constructed wetlands (CWs) are considered to be important sources of nitrous oxide (N₂O). In order to investigate the effect of influent COD/N ratio on N₂O emission and control excess emission from nitrogen removal, free water surface microcosm wetlands were used and fed with different influent. In addition, the transformation of nitrogen was examined for better understanding of the mechanism of N₂O production under different operating COD/N ratios. It was found that N₂O emission and the performance of microcosm wetlands were significantly affected by COD/N ratio of wastewater influent. Strong relationships exist between N₂O production rate and nitrite (r = 0.421, p < 0.01). During denitrification process, DO concentration crucially influences N₂O production rate. An optimal influent COD/N ratio was obtained by adjusting external carbon sources for most effective N₂O emission control and best performance of the CWs in nitrogen removal from wastewater. It is concluded that under the operating condition of COD/N ratio = 5, total N₂O emission is minimum and the microcosm wetland is most effective in wastewater nitrogen removal.     

内蒙古典型草原马粪分解过程中氮素组分的变化

2008年6月至2009年9月,在野外条件下,采用堆置于地表和埋入地下2种处理方式,研究了内蒙古典型草原马粪分解过程中氮素组分的变化特征.结果表明:2种处理残留马粪中,氨态氮、氨基酸态氮和氨基糖态氮在分解前期(0～90 d)维持较高浓度,后期(330～ 450d)浓度显著降低;酸解未知氮和非酸解未知氮浓度随分解呈升高趋势,分解后期升高幅度更为明显.鲜马粪中,铵态氮是无机氮的主要存在形态,随分解呈逐渐降低趋势;鲜马粪中的硝态氮浓度较低,其在残留马粪中的淋溶损失较低,随分解逐渐累积.马粪埋入地下,对铵态氮以气态氨的挥发过程有显著影响,对其他氮素组分的影响不明显.马粪分解前期,氮素矿化的主要有机氮源为氨态氮、氨基酸态氮和氨基糖态氮,后期主要为酸解未知氮和非酸解未知氮.铵态氮的生物有效性主要体现在马粪分解前期,硝态氮则体现在分解后期.     

Mechanism of action of nitrous oxide gas applied as a polyploidizing agent during meiosis in lilies

Nitrous oxide gas (N₂O) can be used to produce polyploid plants, but the mechanism of action is unknown. The actin and microtubule cytoskeleton was observed in N₂O-treated microsporocytes of Lilium spp ‘Asiatic hybrid lilies’ using fluorescence microscopy after staining with DAPI, FITC-conjugated tubulin antibody, and phalloidin-conjugated Alexa Fluor 546. Additionally, microsporocytes of L. longiflorum were observed with acetocarmine staining following N₂O treatment. A typical metaphase I microtubule distribution was observed in control microsporocytes. After treatment with N₂O for 24 h, microtubules were effectively depolymerized; this prevented chromosomes from moving to the poles, resulting in chromosome retention in the center of N₂O-treated cells. Cell plate formation took place without delay, however, yielding one daughter cell with a diploid genome and another daughter without chromosomes. In addition, N₂O treatment often induced micronuclei due to aberrant chromosome separation during cytokinesis. Actin filaments in microsporocytes are insensitive to N₂O. These findings indicate that N₂O mediates polyploidization by inhibiting microtubule polymerization, but not actin filament formation, during microsporocyte meiosis.     

Change in fluxes of carbon dioxide,methane and nitrous oxide due to forest drainage of mire sites of different trophy

Northern peatlands accumulate atmospheric CO₂ thus counteracting climate warming. However, CH₄ which is more efficient as a greenhouse gas than CO₂, is produced in the anaerobic decomposition processes in peat. When peatlands are taken for forestry their water table is lowered by ditching. We studied long-term effects of lowered water table on the development of vegetation and the annual emissions of CO₂, CH₄ and N₂O in an ombrotrophic bog and in a minerotrophic fen in Finland. Reclamation of the peat sites for forestry had changed the composition and coverage of the field and ground layer species, and increased highly the growth of tree stand at the drained fen. In general, drainage increased the annual CO₂ emissions but the emissions were also affected by the natural fluctuations of water table. In contrast to CO₂, drainage had decreased the emissions of CH₄, the drained fen even consumed atmospheric CH₄. CO₂ and CH₄ emissions were higher in the virgin fen than in the virgin bog. There were no N₂O emissions from neither type of virgin sites. Drainage had, however, highly increased the N₂O emissions from the fen. The results suggest that post-drainage changes in gas fluxes depend on the trophy of the original mires.