Electron transport-linked nitrous oxide synthesis and reduction by Paracoccusdenitrificans monitored with an electrode

The nitrous oxide reductase activity of $\text{Paracoccus}\text{denitrificans}$ can be conveniently measured using an electrochemical method for determining N₂O. Introduction of this procedure has shown that (i) N₂O reductase activity is reversibly inhibited by oxygen; (ii) antimycin strongly inhibits electron flow to N₂O and that the inhibition is bypassed by tetramethyl-p-phenylenediamine; (iii) ascorbate plus tetramethyl-p-phenylenediamine, presumably by donating electrons to cytochrome c, is an effective reductant for nitrous oxide reductase; (iv) in the presence of the nitrous oxide reductase inhibitor, acetylene, N₂O is promptly produced from nitrite, consistent with the product of nitrite reductase being N₂O.     

The DCCD-binding polypeptide alone is insufficient for proton translocation through F0 in membranes of Escherichia, coli

In contrast to membrane vesicles of wild-type strains which become leaky to protons on removal of the F₁ ATPase, those of the mutant $\text{Escherichia}\text{,}\text{coli}\text{,}\text{N}{}_{\mathrm{I44}}$, which lacks the F₁ ATPase, can maintain a proton gradient. A normal N,N′-dicyclohexylcarbodiimide (DCCD)-binding polypeptide is present in the F₀ portion of the ATPase complex of the mutant. However, the 19000 molecular weight component of F₀ is absent. We conclude that the latter polypeptide, in addition to the DCCD-binding polypeptide, is required for a functional proton channel in F₀.     

Effect of H2 evolution on 15N2 fixation, C2H2 reduction and relative efficiency of leguminous symbionts

The (C₂H₄+ H_2(C2H₂))/¹⁵N₂ ratios of 15 clover- Rhizobium symbionts. soybean, and black medick symbionts were measured. Relative efficiency based on the C2H4 production and on ¹⁵N₂ incorporation were compared, and in most symbionts there was little difference between the two measures of relative efficiency. Total measurable electron flux through nitrogenase during acetylene reduction and ¹⁵N₂ incorporation were nearly equal for most symbionts studied. The relative efficiency and the (C₂H₄+ H_2(C2H₂))/¹⁵N₂ ratio showed an inverse correlation. Use of this ratio appears preferable to use of the ratio of C2H2 reduction/N₂ reduction. Some evolution of H₂ was observed in the presence of C₂H₂.     

Biohydroxylation of N,N-dialkylarylamines by the isolated topsoil bacterium Bacillus megaterium

Topsoil microorganisms were screened for their acceptability of the standard substrate N,N-dimethylaniline in bacterial ‘whole-cell’ incubations. One bacterium converted N,N-dimethylaniline and was identified as Bacillus megaterium by 16S rDNA analysis and DNA/DNA-hybridization. In contrast to the well-known C-hydroxylation by liver microsomes, leading to p-hydroxylation, B. megaterium formed o- and p-monohydroxylated products, i.e. N,N-dimethyl-2-aminophenol and N,N-dimethyl-4-aminophenol, both identified by gas chromatography–mass spectrometry (GC–MS) using synthesized reference compounds. The observed hydroxylation showed slight regioselectivity in favour of the o-hydroxylated product. Two further substrates, N,N-diethylaniline and N-ethyl-N-methylaniline, were also successfully biohydroxylated by B. megaterium with corresponding regioselectivity. Interestingly, aniline, known to be transformed easily by cytochrome P-450meg into p-aminophenol, was not accepted as substrate.     

Glucosides of ionone-related compounds in several nicotiana species

The β-glucosides of 3-oxo-α-ionol and 5,6-epoxy-5,6-dihydro-3-hydroxy-β-ionol were isolated from fresh leaves of Nicotiana rustica. Two or more of the glucosides of 3-oxo-α-ionol, 5,6-epoxy-5,6-dihydro-3-hydroxy-β-ionol, 3-hydroxy-β-damascone, blumenol A, 4-(3-hydroxybutylidene)-3,5,5-trimethyl-2-cyclohexenl-one and blumenol C were shown to be present and the amounts measured in N. alata, N. repanda, N. rustica, N. undulata, N. accuminata, N. sylvestris and N. tabacum. No glucosides were detected in N. paniculata.     

Nitrogen rhizodeposition assessed by a NH3 shoot pulse-labelling of Lolium perenne L. grown on soil exposed to 9 years of CO2 enrichment

The effects of elevated CO₂ concentration upon rhizodeposition of nitrogen were investigated on field-grown Lolium perenne planted in soil cores set into the resident soil of a intensively managed ryegrass sward treated with elevated CO₂ for nine consecutive years, under two contrasted N fertilisation regimes (Swiss FACE Experiment). The planted cores were excavated from the ambiant (35 Pa pCO₂) and enriched (60 Pa pCO₂) rings at two dates during the growing season (spring and early autumn). The cores were brought back to the laboratory for a pulse-labelling of ryegrass shoots with ¹⁵NH₃, in order to quantify ¹⁵N-rhizodeposition.A recovery of 10–16% of the total ¹⁵N administred to the plant was recovered in the plant–soil system 48 h after the pulse-labelling; significant amounts of ¹⁵N were released into the soil adhering (adhering soil: AS) to the roots (0.44 μg ¹⁵N g AS⁻¹ and 0.60 μg g AS⁻¹ in the spring and the autumn samplings, respectively).In the spring sampling, there was no effect of atmospheric CO₂ concentration on N rhizodeposition. In the autumn sampling, elevated CO₂ stimulated N rhizodeposition that amounted to 7.2 and 5.2 mg ¹⁵N m⁻², under elevated and ambient CO₂, respectively. Nitrogen rhizodeposition was higher at high N (56 gN m⁻²) than at low N fertilisation (14 gN m⁻²), whatever the sampling date investigated.The mechanisms by which elevated atmospheric CO₂ leads to a stimulation of the net root-released N flux remains to be investigated: was it caused by a higher nitrogen immobilisation by the microbial biomass and a reduced re-assimilation of mineralized N and/or by a stimulation of N efflux from roots? Concomitant to the observed reduction of C rhizodeposition, the stimulation of net N efflux suggests that the quality of root released compounds was modified under elevated CO₂ concentration.     

Effects of elevated CO2, temperature and N fertilization on nitrogen fluxes in a temperate grassland ecosystem

The soil nitrogen cycle was investigated in a pre‐established Lolium perenne sward on a loamy soil and exposed to ambient and elevated atmospheric CO₂ concentrations (350 and 700 μL L^?1) and, at elevated [CO₂], to a 3 °C temperature increase. At two levels of mineral nitrogen supply, N– (150 kgN ha^?1 y^?1) and N+ (533 kgN ha^?1 y^?1), ¹⁵N‐labelled ammonium nitrate was supplied in split applications over a 2.5‐y period. The recovery of the labelled fertilizer N was measured in the harvests, in the stubble and roots, in the macro‐organic matter fractions above 200 μm in size (MOM) and in the aggregated organic matter below 200 μM (AOM). Elevated [CO₂] reduced the total amount of N harvested in the clipped parts of the sward. The harvested N derived from soil was reduced to a greater extent than that derived from fertilizer. At both N supplies, elevated [CO₂] modified the allocation of the fertilizer N in the sward, in favour of the stubble and roots and significantly increased the recovery of fertilizer N in the soil macro‐organic matter fractions. The increase of fertilizer N immobilization in the MOM was associated with a decline of fertilizer N uptake by the grass sward, which supported the hypothesis of a negative feedback of elevated [CO₂] on the sward N yield and uptake. Similar and even more pronounced effects were observed for the native N mineralized in the soil. At N–, a greater part of the fertilizer N organized in the root phytomass resulted in an underestimation of N immobilized in dead roots and, in turn, an underestimation of N immobilization in the MOM. The 3 °C temperature increase alleviated the [CO₂] effect throughout much of the N cycle, increasing soil N mineralization, N derived from soil in the harvests, and the partitioning of the assimilated fertilizer N to shoots. In conclusion, at ambient temperature, the N cycle was slowed down under elevated [CO₂], which restricted the increase in the aboveground production of the grass sward, and apparently contributed to the sequestration of carbon belowground. In contrast, a temperature increase under elevated [CO₂] stimulated the soil nitrogen cycle, improved the N nutrition of the sward and restricted the magnitude of the soil C sequestration.     

Soil 13C–15N dynamics in an N2-fixing clover system under long-term exposure to elevated atmospheric CO2

Reduced soil N availability under elevated CO₂ may limit the plant's capacity to increase photosynthesis and thus the potential for increased soil C input. Plant productivity and soil C input should be less constrained by available soil N in an N₂‐fixing system. We studied the effects of Trifolium repens (an N₂‐fixing legume) and Lolium perenne on soil N and C sequestration in response to 9 years of elevated CO₂ under FACE conditions. ¹⁵N‐labeled fertilizer was applied at a rate of 140 and 560 kg N ha^?1 yr^?1 and the CO₂ concentration was increased to 60 Pa pCO₂ using ¹³C‐depleted CO₂. The total soil C content was unaffected by elevated CO₂, species and rate of ¹⁵N fertilization. However, under elevated CO₂, the total amount of newly sequestered soil C was significantly higher under T. repens than under L. perenne. The fraction of fertilizer‐N (f_N) of the total soil N pool was significantly lower under T. repens than under L. perenne. The rate of N fertilization, but not elevated CO₂, had a significant effect on f_N values of the total soil N pool. The fractions of newly sequestered C (f_C) differed strongly among intra‐aggregate soil organic matter fractions, but were unaffected by plant species and the rate of N fertilization. Under elevated CO₂, the ratio of fertilizer‐N per unit of new C decreased under T. repens compared with L. perenne. The L. perenne system sequestered more ¹⁵N fertilizer than T. repens: 179 vs. 101 kg N ha^?1 for the low rate of N fertilization and 393 vs. 319 kg N ha^?1 for the high N‐fertilization rate. As the loss of fertilizer‐¹⁵N contributed to the ¹⁵N‐isotope dilution under T. repens, the input of fixed N into the soil could not be estimated. Although N₂ fixation was an important source of N in the T. repens system, there was no significant increase in total soil C compared with a non‐N₂‐fixing L. perenne system. This suggests that N₂ fixation and the availability of N are not the main factors controlling soil C sequestration in a T. repens system.     

P effects on N uptake and remobilization during regrowth of Italian ryegrass (Lolium multiflorum)

The influence of P deficiency on the utilization of two sources of N, mineral N (exogenous N) and reserved N (endogenous N), for regrowth of Italian ryegrass (Lolium multiflorum) was studied. P-sufficient (+P) or P-free (−P) nutrition solution was applied from 7 days before defoliation to 24 days of regrowth and the N flows derived from two different N sources within the plant were quantified by ¹⁵N pulse-chase labeling. Shoot regrowth significantly reduced by 12 days of regrowth, while root growth was more in −P plants. Inorganic P (P_i) concentration was highly reduced by P deprivation more in the stubble and regrowing shoots and less in the roots. At defoliation, P deprivation had induced a higher accumulation for all N compounds in the stubble and for amino acids in the roots. The previously incorporated ¹⁵N in stubble and roots as nitrate and amino acids was much decreased in −P plants especially for the first 6 days of regrowth. Total N content in the regrowing leaves was not significantly different between +P and −P plants, but percentage contribution of remobilized N for total leaf N formation was significantly higher in −P plants (78%) than in +P plants (69%) at 6 days of regrowth. From day 12, the utilization of both endogenous and exogenous N was apparently inhibited in −P plants.     

施氮对水稻土N2O释放及反硝化功能基因(narG/nosZ)丰度的影响

以紫潮泥和红黄泥两种不同质地的水稻土壤作为研究对象,通过室内培养试验,分析施用硝态氮肥对N2O释放和反硝化基因(narG/nosZ)丰度的影响,并探讨反硝化基因丰度与N2O释放之间的关系。结果表明,施用硝态氮显著增加两种水稻土的N2O释放量。在72h培养过程中,施氮改变了紫潮泥反硝化基因(narG/nosZ)的丰度,但并未明显影响红黄泥反硝化基因(narG/nosZ)丰度。通过双变量相关分析发现,除了紫潮泥narG基因外,其它的反硝化基因丰度和N2O释放之间并没有显著相关性。     

Australian mulga ecosystems –13C and 15N natural abundances of biota components and their ecophysiological significance

Samples of recently produced shoot material collected in winter/spring from common plant species of mulga vegetation in eastern and Western Australia were assayed for ¹³C and ¹⁵N natural abundance. ¹³C analyses showed only three of the 88 test species to exhibit C₄ metabolism and only one of seven succulent species to be in CAM mode. Non-succulent winter ephemeral C₃ species showed significantly lower mean δ¹³C values (– 28·0‰) than corresponding C₃-type herbaceous perennials, woody shrubs or trees (– 26·9, – 25·7 and – 26·2‰, respectively), suggesting lower water stress and poorer water use efficiency in carbon acquisition by the former than latter groups of taxa. Corresponding values for δ¹⁵N of the above growth and life forms lay within the range 7·5–15·5‰. δ¹⁵N of soil NH₄⁺ (mean 19·6‰) at a soft mulga site in Western Australia was considerably higher than that of NO₃^– (4·3‰). Shoot dry matter of Acacia spp. exhibited mean δ¹⁵N values (9·10 ± 0·6‰) identical to those of 37 companion non-N₂-fixing woody shrubs and trees (9·06 ± 0·5‰). These data, with no evidence of nodulation, suggested little or no input of fixed N₂ by the legumes in question. However, two acacias and two papilionoid legumes from a dune of wind-blown, heavily leached sand bordering a lake in mulga in Western Australia recorded δ¹⁵N values in the range 2·0–3·0‰ versus 6·4–10·7‰ for associated non-N₂-fixing taxa. These differences in δ¹⁵N, and prolific nodulation of the legumes, indicated symbiotic inputs of fixed N in this unusual situation. δ¹⁵N signals of lichens, termites, ants and grasshoppers from mulga of Western Australia provided evidence of N₂ fixation in certain termite colonies and by a cyanobacteria-containing species of lichen. Data are discussed in relation to earlier evidence of nitrophily and water availability constraints on nitrate utilization by mulga vegetation.     

土壤硝态氮时空变异与土壤氮素表观盈亏研究Ⅰ.冬小麦

不同氮肥用量下对冬小麦生育期间土壤硝态氮时空变化特征及土壤氮素表观盈亏量的研究结果表明,氮肥用量不同,硝态氮分布特征有差异,并且随着冬小麦的生长,其变化也不同。在冬小麦快速生长阶段,作物吸收可在一定深度的土层出现硝态氮亏缺区。由于灌溉的影响,土壤表层硝态氮向深层淋洗严重,即使在低氮肥水平,土壤深层仍可观察到硝态氮含量升高现象,存在淋出2m土体的可能性。并且氮肥用量越高,土壤硝态氮含量越高,硝酸盐向深层淋洗也越严重,淋出2m土体的可能性和也相应增大;在冬小麦生长前期（播种－拔节）,即使在不施氮肥处理也有土壤氮素的表观盈余,随着施肥量的增加,在拔节－扬花也出现了土壤氮素表观盈余,而扬花后各个氮肥处理均出现土壤氮素的表观亏缺,氮肥用量越高,小麦一生中土壤表观氮盈余量越大,1m土体内平均最大盈余量达199.8kgN/hm^2。研究表明,土壤氮损失是盈余氮素的一个主要去向,而硝态氮淋洗是冬小麦生育期间土壤氮素损失的一个重要的途径。     

华北平原冬小麦/夏玉米轮作体系土壤硝态氮的适宜含量

采用冬小麦季不同施氮处理(夏玉米季不施氮)研究了华北平原冬小麦/夏玉米轮作体系夏玉米季土壤硝态氮的适宜含量.结果表明:在播前土壤无机氮含量较高的条件下,冬小麦季施用150kgN.hm-2即可满足冬小麦/夏玉米两季作物的氮素需求;各氮肥处理在冬小麦季的氮肥施用当季的利用率仅为11%～23%,在夏玉米季氮肥残效利用率则高达30%～52%.当夏玉米播前0～90cm土层硝态氮含量达到82kg.hm-2时,无需施氮即可保证夏玉米十叶期的生长,达到151kg.hm-2时,无需施氮即可保证整个生育期的生长.夏玉米十叶期和收获后0～90cm土层硝态氮含量低于46和65kg.hm-2时,则影响作物正常生长.综合考虑产量和环境效应,冬小麦/夏玉米轮作体系中0～90cm土层硝态氮含量应控制在65～151kg.hm-2之间.     

Solid state conformational study of complexes containing the (o-P-C6H4-CN-κN,P) palladium six-membered chelated ring

The Cambridge Structural Database (CSD) V. 5.23 has been searched for all the structures containing the fragment Pd{o-P-C₆H₄-CN-κ²N,P}. Bond lengths, bond angles and the conformations adopted by the PdPCCCN chelated ring have been studied statistically. It has been found Pd-P and Pd-N distances in the mentioned compounds are, respectively, shorter and longer than the mean value found for these parameters when the CSD was searched irrespective of the ligands. The conformation of the chelated ring has been characterized by means of two torsion angles, and can be described in most cases as a plane containing the P and carbon atoms with Pd and N atoms situated at the same side out of this plane. Molecular mechanics calculations have been employed to justify the conformational preferences found. The calculated strain energy suggests a path for the movement of Pd and N atoms from one side of the PCCC plane to the other through a planar conformation. MM calculations in complex (η³-allyl)-(N-(2-(diphenylphosphino)benzylidene)-4-methoxyphenylamine)-palladium(II) indicate that planar conformation is energetically accessible, and the extent of preorganization for the free ligand in this compound has been evaluated of 74%.     

Nitrate and ammonium influxes in soybean (Glycine max) roots: direct comparison of 13N and 15N tracing

We compared influxes and internal transport in soybean plants (Glycine max cv. Kingsoy) of labelled N from external solutions where either ammonium or nitrate was labelled with the stable isotope¹⁵N and the radioactive isotope¹³N. The objective was to see whether mass spectrometric determinations of tissue ¹⁵N content were sufficiently sensitive to measure influxes accurately over short time periods. Our findings were as follows. (1) There was a close quantitative correspondence between estimates of N influx of individual plants using ¹⁵N or ¹³N measurements with either NO_3/^? or NH₄⁺ at 4 or 2 mol^?3, respectively in the external solution. (2) Transport to the shoot of N from NO₃ absorbed over a 5–15 min period could be monitored when the external NO₃^? concentration ranged from 0–05 to 4 mol m^?3. NH₄⁺ as the N source labelled shoot tissue more slowly, and estimates of the transport between root and shoot could be made only with ¹³N. (3) Influx of NO₃^? into root tissue could be measured by ¹⁵N enrichment after 5–10 min at concentrations approaching the probable K_M of the high-affinity transport system. (4) There was some indication of isotope discrimination, especially with respect to the movement of labelled N to the shoot, when NO₃^? is the N source. For many purposes, ¹⁵N tracing can be used satisfactorily to estimate influxes of both NO₃^? and NH₄⁺ in soybean roots. Use of the short-lived radio nuclide ¹³N remains the method of choice for more refined measurements of internal distribution and assimilation.     

Plant and microbial N acquisition under elevated atmospheric CO2 in two mesocosm experiments with annual grasses

The impact of elevated CO₂ on terrestrial ecosystem C balance, both in sign or magnitude, is not clear because the resulting alterations in C input, plant nutrient demand and water use efficiency often have contrasting impacts on microbial decomposition processes. One major source of uncertainty stems from the impact of elevated CO₂ on N availability to plants and microbes. We examined the effects of atmospheric CO₂ enrichment (ambient+370 μmol mol^?1) on plant and microbial N acquisition in two different mesocosm experiments, using model plant species of annual grasses of Avena barbata and A. fatua, respectively. The A. barbata experiment was conducted in a N‐poor sandy loam and the A. fatua experiment was on a N‐rich clayey loam. Plant–microbial N partitioning was examined through determining the distribution of a ¹⁵N tracer. In the A. barbata experiment, ¹⁵N tracer was introduced to a field labeling experiment in the previous year so that ¹⁵N predominantly existed in nonextractable soil pools. In the A. fatua experiment, ¹⁵N was introduced in a mineral solution [(¹⁵NH₄)₂SO₄ solution] during the growing season of A. fatua. Results of both N budget and ¹⁵N tracer analyses indicated that elevated CO₂ increased plant N acquisition from the soil. In the A. barbata experiment, elevated CO₂ increased plant biomass N by ca. 10% but there was no corresponding decrease in soil extractable N, suggesting that plants might have obtained N from the nonextractable organic N pool because of enhanced microbial activity. In the A. fatua experiment, however, the CO₂‐led increase in plant biomass N was statistically equal to the reduction in soil extractable N. Although atmospheric CO₂ enrichment enhanced microbial biomass C under A. barbata or microbial activity (respiration) under A. fatua, it had no significant effect on microbial biomass N in either experiment. Elevated CO₂ increased the colonization of A. fatua roots by arbuscular mycorrhizal fungi, which coincided with the enhancement of plant competitiveness for soluble soil N. Together, these results suggest that elevated CO₂ may tighten N cycling through facilitating plant N acquisition. However, it is unknown to what degree results from these short‐term microcosm experiments can be extrapolated to field conditions. Long‐term studies in less‐disturbed soils are needed to determine whether CO₂‐enhancement of plant N acquisition can significantly relieve N limitation over plant growth in an elevated CO₂ environment.     

Mixed S/N and S/P/N ligands from carbohydrates: Synthesis and application in palladium-catalyzed allylic alkylation

A modular synthetic approach to bidentate and tridentate imino-thioglycoside ligands is reported. In only 5 steps from the known glucosamine derivative 1, a conveniently functionalized thioglycoside 5 is obtained, which after imination afforded the desired ligands in excellent yields. The tridentate phosphine imine thioglycoside ligand 10 was found to be a highly efficient catalyst precursor for palladium-(0)-catalysed asymmetric alkylation of 1,3-diphenylpropenylacetate (8) with dimethyl malonate. The study of a Pd(II) complex shows that there is an efficient stereochemical control of the sulfur configuration upon coordination to the palladium.     

Drought turns a Central European Norway spruce forest soil from an N2O source to a transient N2O sink

Based on current climate scenarios, a higher frequency of summer drought periods followed by heavy rainfall events is predicted for Central Europe. It is expected that drying/rewetting events induce an increased matter cycling in soils and may contribute considerably to increased emissions of the greenhouse gas N₂O on annual scales. To investigate the influence of drying/rewetting events on N₂O emissions in a mature Norway spruce forest in the Fichtelgebirge area (NE Bavaria, Germany), a summer drought period of 46 days was induced by roof installations on triplicate plots, followed by a rewetting event of 66 mm experimental rainfall in 2 days. Three nonmanipulated plots served as controls. The experimentally induced soil drought was accompanied by a natural drought. During the drought period, the soil of both the throughfall exclusion and control plots served as an N₂O sink. This was accompanied by subambient N₂O concentrations in upper soil horizons. The sink strength of the throughfall exclusion plots was doubled compared with the control plots. We conclude that the soil water status together with the soil nitrate availability was an important driving factor for the N₂O sink strength. Rewetting quickly turned the soil into a source for atmospheric N₂O again, but it took almost 4 months to turn the cumulative soil N₂O fluxes from negative (sink) to positive (source) values. N₂O concentration and isotope analyses along soil profiles revealed that N₂O produced in the subsoil was subsequently consumed during upward diffusion along the soil profile throughout the entire experiment. Our results show that long drought periods can lead to drastic decreases of N₂O fluxes from soils to the atmosphere or may even turn forest soils temporarily to N₂O sinks. Accumulation of more field‐scale data on soil N₂O uptake as well as a better understanding of underlying mechanisms would essentially advance our knowledge of the global N₂O budget.