Improvement of protoplast regeneration by growing yeast cells hypertonically

N2O was produced during the reduction of NO2- by resting cells of Lactobacillus lactis TS4. At an initial NO2- concentration of 69 micrograms/ml, the rate of N2O production was 1.97 nmol/min per mg of protein, and the recovery of reduced NO2- -N as N2O-N after 24 h was 77%. Higher initial NO2- concentrations decreased both the rate of production of N2O and the recovery of reduced NO2- -N. CO2 production increased during NO2- reduction.     

减量施氮对冬小麦-夏玉米种植体系中氮利用与平衡的影响

研究了冬小麦-夏玉米种植体系中减量施N对作物N利用与平衡的影响,结果表明,与原有高量施N处理(N240和N360)相比,在冬小麦季减半施N未引起产量和吸N量的变化。但在原有低量施N处理(N120)下减半施N显著降低了小麦产量和吸N量;在夏玉米季,在上季减半施N的基础上停止施N后作物产量和吸N量均比原固定施N处理显著下降,N平衡计算结果表明,减量施N条件下0～1m土壤N残留和表观损失的数量均显著低于原有施N量处理,作物N利用率显著提高,但在1～2m层次中累积的硝态氮却不因减量施N而下降,说明这一土层的硝态氮可能难以被作物吸收利用,由此可见,在前茬高施N量下减少氮肥用量有利于提高作物的氮肥利用率、减少N残留与表观损失。     

Effect of sulfur compounds on biological reduction of nitric oxide in aqueous Fe(II)EDTA2- solutions.

Biological reduction of nitric oxide (NO) in aqueous solutions of EDTA chelated Fe(II) is one of the main steps in the BioDeNOx process, a novel bioprocess for the removal of nitrogen oxides (NOx) from polluted gas streams. Since NOx contaminated gases usually also contain sulfurous pollutants, the possible interferences of these sulfur compounds with the BioDeNOx process need to be identified. Therefore, the effect of the sulfur compounds Na2SO4, Na2SO3, and H2S on the biological NO reduction in aqueous solutions of Fe(II)EDTA2- (25 mM, pH 7.2, 55 degrees C) was studied in batch experiments. Sulfate and sulfite were found to not affect the reduction rate of Fe(II)EDTA2- complexed NO under the conditions tested. Sulfide, either dosed externally or formed during the batch incubation out of endogenous sulfur sources or the supplied sulfate or sulfite, influences the production and consumption of the intermediate nitrous oxide (N2O) during Fe(II)EDTA2- bound NO reduction. At low concentrations (0.2 g VSS/l) of denitrifying sludge, 0.2 mM free sulfide completely inhibited the nitrosyl-complex reduction. At higher biomass concentrations (1.3-2.3 g VSS/l), sulfide (from 15 microM to 0.8 mM) induced an incomplete NO denitrification with N2O accumulation. The reduction rates of NO to N2O were enhanced by anaerobic sludge, presumably because it kept FeEDTA in the reduced state.     

Production of nitrous oxide from nitrite in Klebsiella pneumoniae: mutants altered in nitrogen metabolism.

Under anaerobic conditions, Klebsiella pneumoniae reduced nitrite (NO2-), yielding nitrous oxide (N2O) and ammonium ions (NH4+) as products. Nitrous oxide formation accounted for about 5% of the total NO2- reduced, and NH4+ production accounted for the remainder. Glucose and pyruvate were the electron donors for NO2- reduction to N2O by whole cells, whereas glucose, NADH, and NADPH were found to be the electron donors when cell extracts were used. On the one hand, formate failed to serve as an electron donor for NO2- reduction to N2O and NH4+, whereas on the other hand, formate was the best electron donor for nitrate reduction in either whole cells or cell extracts. Mutants that are defective in the reduction of NO2- to NH4+ were isolated, and these strains were found to produce N2O at rates comparable to that of the parent strain. These results suggest that the nitrite reductase producing N2O is distinct from that producing NH4+. Nitrous oxide production from nitric oxide (NO) occurred in all mutants tested, at rates comparable to that of the parent strain. This result suggests that NO reduction to N2O, which also uses NADH as the electron donor, is independent of the protein(s) catalyzing the reduction of NO2- to N2O.     

淹水条件下添加有机物料对蔬菜地土壤硝态氮及氮素气体排放的影响

蔬菜地大量施用氮肥可以引起土壤硝态氮积累,导致土壤退化,快速消除土壤积累的硝态氮,可以提高蔬菜地土壤质量,延长其使用时间.在硝态氮(360 mg N·kg^-1)积累的蔬菜地土壤中,分别加入0、2500、5000和7500 kg C·hm^-2黑麦草(记为CK、C₂₅₀₀、C₅₀₀₀和C₇₅₀₀),淹水条件下,30 ℃恒温室内培养240 h,研究土壤硝态氮含量及氮素气体排放量变化.结果表明：培养结束时,CK处理中土壤硝态氮含量高达310 mg N·kg^-1,添加黑麦草能有效地消除土壤中积累的硝态氮,C₂₅₀₀、C₅₀₀₀和C₇₅₀₀处理中土壤硝态氮含量降低至10 mg N·kg^-1以下需要的时间分别为240、48和24 h.添加黑麦草显著提高了土壤pH,降低了土壤电导率,其变化幅度随黑麦草添加量的增加而增大.添加黑麦草处理的土壤N₂O和N₂累积排放量为270～378 mg N·kg^-1,N₂O/N₂为0.6～1.5.淹水条件下添加黑麦草可快速消除蔬菜地土壤积累的硝态氮,但应充分重视N₂O在这一过程中的大量排放.     

Dissimilatory Reduction of NO(2) to NH(4) and N(2)O by a Soil Citrobacter sp

Dissimilatory reduction of NO(2) to N(2)O and NH(4) by a soil Citrobacter sp. was studied in an attempt to elucidate the physiological and ecological significance of N(2)O production by this mechanism. In batch cultures with defined media, NO(2) reduction to NH(4) was favored by high glucose and low NO(3) concentrations. Nitrous oxide production was greatest at high glucose and intermediate NO(3) concentrations. With succinate as the energy source, little or no NO(2) was reduced to NH(4) but N(2)O was produced. Resting cell suspensions reduced NO(2) simultaneously to N(2)O and free extracellular NH(4). Chloramphenicol prevented the induction of N(2)O-producing activity. The K(m) for NO(2) reduction to N(2)O was estimated to be 0.9 mM NO(2), yet the apparent K(m) for overall NO(2) reduction was considerably lower, no greater than 0.04 mM NO(2). Activities for N(2)O and NH(4) production increased markedly after depletion of NO(3) from the media. Amendment with NO(3) inhibited N(2)O and NH(4) production by molybdate-grown cells but not by tungstate-grown cells. Sulfite inhibited production of NH(4) but not of N(2)O. In a related experiment, three Escherichia coli mutants lacking NADH-dependent nitrite reductase produced N(2)O at rates equal to the wild type. These observations suggest that N(2)O is produced enzymatically but not by the same enzyme system responsible for dissimilatory reduction of NO(2) to NH(4).     

含硝化抑制剂DMPP复合肥对日光温室芹菜生长和营养品质的影响

通过田间试验研究了含硝化抑制剂DMPP复合肥对日光温室芹菜生长和品质的影响. 结果表明,与普通复合肥相比,一次基施DMPP复合肥67.5和54.0 kg·hm^-2 氮处理分别使芹菜增产5.78%和10.14%; DMPP复合肥可降低芹菜可食部分硝酸盐含量,提高Vc、游离氨基酸、可溶性糖及氮、磷含量. 与分次施用相比,适当减少DMPP复合肥施用次数和用量可提高芹菜产量并改善其品质,降低生产成本.DMPP复合肥在施入土壤中后具有显著的硝化抑制作用,延缓了菜地土壤铵态氮向硝态氮的转化,降低了氮素向水体迁移的风险.芹菜收获后土壤中全氮、铵态氮、硝态氮残留较多,有利于保持地力.     

Decreased N2O reduction by low soil pH causes high N2O emissions in a riparian ecosystem

Quantification of harmful nitrous oxide (N(2)O) emissions from soils is essential for mitigation measures. An important N(2)O producing and reducing process in soils is denitrification, which shows deceased rates at low pH. No clear relationship between N(2)O emissions and soil pH has yet been established because also the relative contribution of N(2)O as the denitrification end product decreases with pH. Our aim was to show the net effect of soil pH on N(2)O production and emission. Therefore, experiments were designed to investigate the effects of pH on NO(3)(-) reduction, N(2)O production and reduction and N(2) production in incubations with pH values set between 4 and 7. Furthermore, field measurements of soil pH and N(2)O emissions were carried out. In incubations, NO(3)(-) reduction and N(2) production rates increased with pH and net N(2)O production rate was highest at pH 5. N(2)O reduction to N(2) was halted until NO(3)(-) was depleted at low pH values, resulting in a built up of N(2)O. As a consequence, N(2)O:N(2) production ratio decreased exponentially with pH. N(2)O reduction appeared therefore more important than N(2)O production in explaining net N(2)O production rates. In the field, a negative exponential relationship for soil pH against N(2)O emissions was observed. Soil pH could therefore be used as a predictive tool for average N(2)O emissions in the studied ecosystem. The occurrence of low pH spots may explain N(2)O emission hotspot occurrence. Future studies should focus on the mechanism behind small scale soil pH variability and the effect of manipulating the pH of soils.     

硝态氮和铵态氮对墨兰生长发育的影响

以无土栽培的方法研究不同氮水平的硝态氮和铵态氮对墨兰［Ｃｙｍｂｉｄｉｕｍｓｉｎｅｎｓｅ（Ａｎｄｒ．）Ｗｉｌｌｄ．］生长发育和某些生理特性的影响。１ｍｍｏｌ／Ｌ和１０ｍｍｏｌ／Ｌ的硝态氮和铵态氮处理都促进叶芽和叶片正常生长,前者似乎比后者好一些。５０ｍｍｏｌ／Ｌ的两种形态的氮均使叶片生长缓慢,尤其是铵态氮。１和１０ｍｍｏｌ／Ｌ硝态氮处理则正常开花,而５０ｍｍｏｌ／Ｌ则例外。铵态氮处理各种浓度均不形成花芽。建议墨兰营养生长期施用１ｍｍｏｌ／Ｌ的硝态氮或铵态氮,但生长后期则应施１—１０ｍｍｏｌ／Ｌ硝态氮,以利花芽分化。作者认为,墨兰生长缓慢与光合速率极慢和硝酸还原酶活性极低有关,墨兰开花与否和Ｅ／Ｎ比大小有关。     

Nitrous oxide production by Escherichia coli is correlated with nitrate reductase activity

Nitrous oxide production by Escherichia coli seems to result from the reduction of NO2- by NO3- reductase. This hypothesis is consistent with previous observations and with the observation that molybdenum was required for both NO3- reduction and N2O production. Several E. coli NO3- reductase mutants were assayed for both N2O-producing and NO3--reducing activity. The hypothesized role of NO3- reductase is supported by the correlation of these two activities. Nitrate reduction to NH4+ enhanced growth, but NO2- reduction to N2O apparently did not. Therefore, this process differs significantly from respiratory denitrification.     

Inhibition by sulfide of nitric and nitrous oxide reduction by denitrifying Pseudomonas fluorescens.

The influence of low redox potentials and H2S on NO and N2O reduction by resting cells of denitrifying Pseudomonas fluorescens was studied. Hydrogen sulfide and Ti(III) were added to achieve redox potentials near -200 mV. The control without reductant had a redox potential near +200 mV. Production of 13NO, [13N]N2O, and [13N]N2 from 13NO3- and 13NO2- was followed. Total gas production was similar for all three treatments. The accumulation of 13NO was most significant in the presence of sulfide. A parallel control with autoclaved cells indicated that the 13NO production was largely biological. The sulfide inhibition was more dramatic at the level of N2O reduction; [13N]N2O became the major product instead of [13N]N2, the dominant product when either no reductant or Ti(III) was present. The results indicate that the specific action of sulfide rather than the low redox potential caused a partial inhibition of NO reduction and a strong inhibition of N2O reduction in denitrifying cells.     

Nitrous oxide production by Escherichia coli is correlated with nitrate reductase activity.

Nitrous oxide production by Escherichia coli seems to result from the reduction of NO2- by NO3- reductase. This hypothesis is consistent with previous observations and with the observation that molybdenum was required for both NO3- reduction and N2O production. Several E. coli NO3- reductase mutants were assayed for both N2O-producing and NO3--reducing activity. The hypothesized role of NO3- reductase is supported by the correlation of these two activities. Nitrate reduction to NH4+ enhanced growth, but NO2- reduction to N2O apparently did not. Therefore, this process differs significantly from respiratory denitrification.     

Perivascular nitric oxide and superoxide in neonatal cerebral hypoxia-ischemia

Decreased cerebral blood flow (CBF) has been observed following the resuscitation from neonatal hypoxic-ischemic injury, but its mechanism is not known. We address the hypothesis that reduced CBF is due to a change in nitric oxide (NO) and superoxide anion O(2)(-) balance secondary to endothelial NO synthase (eNOS) uncoupling with vascular injury. Wistar rats (7 day old) were subjected to cerebral hypoxia-ischemia by unilateral carotid occlusion under isoflurane anesthesia followed by hypoxia with hyperoxic or normoxic resuscitation. Expired CO(2) was determined during the period of hyperoxic or normoxic resuscitation. Laser-Doppler flowmetry was used with isoflurane anesthesia to monitor CBF, and cerebral perivascular NO and O(2)(-) were determined using fluorescent dyes with fluorescence microscopy. The effect of tetrahydrobiopterin supplementation on each of these measurements and the effect of apocynin and N(omega)-nitro-L-arginine methyl ester (L-NAME) administration on NO and O(2)(-) were determined. As a result, CBF in the ischemic cortex declined following the onset of resuscitation with 100% O(2) (hyperoxic resuscitation) but not room air (normoxic resuscitation). Expired CO(2) was decreased at the onset of resuscitation, but recovery was the same in normoxic and hyperoxic resuscitated groups. Perivascular NO-induced fluorescence intensity declined, and O(2)(-)-induced fluorescence increased in the ischemic cortex after hyperoxic resuscitation up to 24 h postischemia. L-NAME treatment reduced O(2)(-) relative to the nonischemic cortex. Apocynin treatment increased NO and reduced O(2)(-) relative to the nonischemic cortex. The administration of tetrahydrobiopterin following the injury increased perivascular NO, reduced perivascular O(2)(-), and increased CBF during hyperoxic resuscitation. These results demonstrate that reduced CBF follows hyperoxic resuscitation but not normoxic resuscitation after neonatal hypoxic-ischemic injury, accompanied by a reduction in perivascular production of NO and an increase in O(2)(-). The finding that tetrahydrobiopterin, apocynin, and L-NAME normalized radical production suggests that the uncoupling of perivascular NOS, probably eNOS, due to acquired relative tetrahydrobiopterin deficiency occurs after neonatal hypoxic-ischemic brain injury. It appears that both NOS uncoupling and the activation of NADPH oxidase participate in the changes of reactive oxygen concentrations seen in cerebral hypoxic-ischemic injury.     

Proliferation of maize (Zea mays L.) roots in response to localized supply of nitrate

Maize (Zea mays L.) plants with two primary nodal root axes were grown for 8 d in flowing nutrient culture with each axis independently supplied with NO3-. Dry matter accumulation by roots was similar whether 1.0 mol m-3 NO3- was supplied to one or both axes. When NO3- was supplied to only one axis, however, accumulation of dry matter within the root system was significantly greater in the axis supplied with NO3-. The increased dry matter accumulation by the +N-treated axis was attributable entirely to increased density and growth of lateral branches and not to a difference in growth of the primary axis. Proliferation of lateral branches for the +N axis was associated with the capacity for in situ reduction and utilization of a portion of the absorbed NO3-, especially in the apical region where lateral primordia are initiated. Although reduced nitrogen was translocated to the -N axis, concentrations in the -N axis remained significantly lower than in the +N axis. The concentration of reduced nitrogen, as well as in vitro NO3- reductase activity, was greater in apical than in more basal regions of the +N axis. The enhanced proliferation of lateral branches in the +N axis was accompanied by an increase in total respiration rate of the axis. Part of the increased respiration was attributable to increased mass of roots. The specific respiration rate (micromoles CO2 evolved per hour per gram root dry weight) was also greater for the +N than for the -N axis. If respiration rate is taken as representative of sink demand, stimulation of initiation and growth of laterals by in situ utilization of a localized exogenous supply of NO3- establishes an increased sink demand through enhanced metabolic activity and the increased partitioning of assimilates to the +N axis responds to the difference in sink demand between +N and -N axes.     

Detergent inhibition of nitric-oxide reductase activity

Gas chromatography revealed that exposure of extracts of the denitrifiers 'Achromobacter cycloclastes', Paracoccus denitrificans, Pseudomonas aeruginosa and Pseudomonas perfectomarina to Triton X-100 inhibited reduction of NO to N2O, and thus concomitantly inhibited reduction of NO2- to N2O. After exposure of extracts to Triton X-100, the ratio of H+ consumed to NO2- added decreased from approx. 2.0 (for untreated extracts) to approx. 1.5, which indicated that NO2- was reduced to NO by the treated extracts. Addition of a CHAPS-soluble extract (devoid of nitrite reductase activity but rich in nitric-oxide reductase activity) to the Triton X-100-treated extract of P. denitrificans restored capacity for reduction of NO2- on to N2O. Exposure to either the NO that accumulated from reduction of NO2- or to enthetic NO transiently inhibited rates of NO2- reduction in Triton X-100-treated extracts. Use of an Oxides of Nitrogen analyzer indicated that only 5-33% of NO2- reduced by untreated extracts appeared in the stripping gas as NO, whereas 80-95% of NO2- reduced by Triton X-100-treated extracts was recovered as NO.     

Chemical and biological interactions during nitrate and goethite reduction by Shewanella putrefaciens 200

Although previous research has demonstrated that NO(3)(-) inhibits microbial Fe(III) reduction in laboratory cultures and natural sediments, the mechanisms of this inhibition have not been fully studied in an environmentally relevant medium that utilizes solid-phase, iron oxide minerals as a Fe(III) source. To study the dynamics of Fe and NO(3)(-) biogeochemistry when ferric (hydr)oxides are used as the Fe(III) source, Shewanella putrefaciens 200 was incubated under anoxic conditions in a low-ionic-strength, artificial groundwater medium with various amounts of NO(3)(-) and synthetic, high-surface-area goethite. Results showed that the presence of NO(3)(-) inhibited microbial goethite reduction more severely than it inhibited microbial reduction of the aqueous or microcrystalline sources of Fe(III) used in other studies. More interestingly, the presence of goethite also resulted in a twofold decrease in the rate of NO(3)(-) reduction, a 10-fold decrease in the rate of NO(2)(-) reduction, and a 20-fold increase in the amounts of N(2)O produced. Nitrogen stable isotope experiments that utilized delta(15)N values of N(2)O to distinguish between chemical and biological reduction of NO(2)(-) revealed that the N(2)O produced during NO(2)(-) or NO(3)(-) reduction in the presence of goethite was primarily of abiotic origin. These results indicate that concomitant microbial Fe(III) and NO(3)(-) reduction produces NO(2)(-) and Fe(II), which then abiotically react to reduce NO(2)(-) to N(2)O with the subsequent oxidation of Fe(II) to Fe(III).     

Nitrate reduction in a groundwater microcosm determined by N gas chromatography-mass spectrometry

Aerobic and anaerobic groundwater continuous-flow microcosms were designed to study nitrate reduction by the indigenous bacteria in intact saturated soil cores from a sandy aquifer with a concentration of 3.8 mg of NO(3)-N liter. Traces of NO(3) were added to filter-sterilized groundwater by using a Darcy flux of 4 cm day. Both assimilatory and dissimilatory reduction rates were estimated from analyses of N(2), N(2)O, NH(4), and N-labeled protein amino acids by capillary gas chromatography-mass spectrometry. N(2) and N(2)O were separated on a megabore fused-silica column and quantified by electron impact-selected ion monitoring. NO(3) and NH(4) were analyzed as pentafluorobenzoyl amides by multiple-ion monitoring and protein amino acids as their N-heptafluorobutyryl isobutyl ester derivatives by negative ion-chemical ionization. The numbers of bacteria and their [methyl-H]thymidine incorporation rates were simultaneously measured. Nitrate was completely reduced in the microcosms at a rate of about 250 ng g day. Of this nitrate, 80 to 90% was converted by aerobic denitrification to N(2), whereas only 35% was denitrified in the anaerobic microcosm, where more than 50% of NO(3) was reduced to NH(4). Assimilatory reduction was recorded only in the aerobic microcosm, where N appeared in alanine in the cells. The nitrate reduction rates estimated for the aquifer material were low in comparison with rates in eutrophic lakes and coastal sediments but sufficiently high to remove nitrate from an uncontaminated aquifer of the kind examined in less than 1 month.     

浮床植物系统对富营养化水体中氮、磷净化特征的初步研究

以浮床空心菜(Ipomoea aquatica)、水芹(Oenanthe javanica)和无植物系统为对象,研究了其在富营养化水体中对N、P的去除及其N₂O的排放情况.结果表明,浮床植物系统对水体中N、P具有良好的净化效果,植物组织所累积的N、P量分别占各自系统去除量的40.32%～63.87%,说明植物的同化吸收作用是N、P去除的主要途径.换水周期内浮床植物系统中硝化反应进行充分,而反硝化反应相对缓慢,导致系统具有较高的NH₄⁺-N去除率,而产生NO₃--N累积.植物的存在降低了系统中N₂O的排放通量.生长较好的空心菜系统在换水前后平均N₂O排放量最低,为17.14μgN·m^-2h^-1,空白高达8.08μgN·m^-2h^-1,水芹为37.38μg N·m^-2·h^-1.     

Identification of the sources of nitrous oxide produced by oxidative and reductive processes in Nitrosomonas europaea

1. Cells of Nitrosomonas europaea produced N(2)O during the oxidation of ammonia and hydroxylamine. 2. The end-product of ammonia oxidation, nitrite, was the predominant source of N(2)O in cells. 3. Cells also produced N(2)O, but not N(2) gas, by the reduction of nitrite under anaerobic conditions. 4. Hydroxylamine was oxidized by cell-free extracts to yield nitrite and N(2)O aerobically, but to yield N(2)O and NO anaerobically. 5. Cell extracts reduced nitrite both aerobically and anaerobically to NO and N(2)O with hydroxylamine as an electron donor. 6. The relative amounts of NO and N(2)O produced during hydroxylamine oxidation and/or nitrite reduction are dependent on the type of artificial electron acceptor utilized. 7. Partially purified hydroxylamine oxidase retained nitrite reductase activity but cytochrome oxidase was absent. 8. There is a close association of hydroxylamine oxidase and nitrite reductase activities in purified preparations.     

Early determinants of H2O2-induced endothelial dysfunction

Reactive oxygen species (ROS) can stimulate nitric oxide (NO(*)) production from the endothelium by transient activation of endothelial nitric oxide synthase (eNOS). With continued or repeated exposure, NO(*) production is reduced, however. We investigated the early determinants of this decrease in NO(*) production. Following an initial H(2)O(2) exposure, endothelial cells responded by increasing NO(*) production measured electrochemically. NO(*) concentrations peaked by 10 min with a slow reduction over 30 min. The decrease in NO(*) at 30 min was associated with a 2.7-fold increase in O(2)(*-) production (p < 0.05) and a 14-fold reduction of the eNOS cofactor, tetrahydrobiopterin (BH(4), p < 0.05). Used as a probe for endothelial dysfunction, the integrated NO(*) production over 30 min upon repeated H(2)O(2) exposure was attenuated by 2.1-fold (p = 0.03). Endothelial dysfunction could be prevented by BH(4) cofactor supplementation, by scavenging O(2)(*-) or peroxynitrite (ONOO(-)), or by inhibiting the NADPH oxidase. Hydroxyl radical (()OH) scavenging did not have an effect. In summary, early H(2)O(2)-induced endothelial dysfunction was associated with a decreased BH(4) level and increased O(2)(*-) production. Dysfunction required O(2)(*-), ONOO(-), or a functional NADPH oxidase. Repeated activation of the NADPH oxidase by ROS may act as a feed forward system to promote endothelial dysfunction.