N<Subscript>2</Subscript> fixation in three perennial <Emphasis Type="Italic">Trifolium</Emphasis> species in experimental grasslands of varied plant species richness and composition

This study is the first to investigate quantitative effects of plant community composition and diversity on N₂ fixation in legumes. N₂ fixation in three perennial Trifolium species grown in field plots with varied number of neighbouring species was evaluated with the ¹⁵N natural abundance method (two field sites, several growing seasons, no N addition) and the isotope dilution method (one site, one growing season, 5 g N m⁻²). The proportion of plant N derived from N₂ fixation, pNdfa, was generally high, but the N addition decreased pNdfa, especially in species-poor communities. Also following N addition, the presence of grasses in species-rich communities increased pNdfa in T. hybridum and T. repens L., while legume abundance had the opposite effect. In T. repens, competition for light from grasses appeared to limit growth and thereby the amount of N₂ fixed at the plant level, expressed as mg N₂ fixed per sown seed. We conclude that the occurrence of diversity effects seems to be largely context dependent, with soil N availability being a major determinant, and that species composition and functional traits are more important than species richness regarding how neighbouring plant species influence N₂ fixation in legumes.     

N<Subscript>2</Subscript> fixation estimates in real-time by cavity ring-down laser absorption spectroscopy

The most common currency for estimating N₂ fixation is acetylene reduction to ethylene. Real-time estimates of nitrogen fixation are needed to close the global nitrogen budget and these remain a critical gap in both laboratory and field experiments. We present a new method for continuous real-time measurements of ethylene production: Acetylene Reduction Assays by Cavity ring-down laser Absorption Spectroscopy (ARACAS). In ARACAS, air in the headspace of an incubation chamber is circulated with a diaphragm pump through a cavity ring-down ethylene spectrometer and back to the incubation chamber. This paper describes the new approach and its benefits compared to the conventional detection of ethylene by flame ionization detector gas chromatography. First, the detection of acetylene reduction to ethylene is non-intrusive and chemically non-destructive, allowing for real-time measurements of nitrogenase activity. Second, the measurements are made instantaneously and continuously at ppb levels, allowing for observation of real-time kinetics on time intervals as short as a few seconds. Third, the instrument can be automated for long time periods of measurement. Finally, the technique will be widely accessible by the research community as it can be readily adapted to most existing acetylene reduction protocols and is based on a modestly priced, commercially available instrument. We illustrate its use for measuring N₂ fixation using two species, the diazotrophic bacterium Azotobacter vinelandii and the lichen Peltigera praetextata. We also discuss potential limitations of the approach, primarily the implications of leaks in the analyzer, as well as future improvements.     

Hydrothermal Stability of Adenine Under Controlled Fugacities of N<Subscript>2</Subscript>, CO<Subscript>2</Subscript> and H<Subscript>2</Subscript>

An experimental study has been carried out on the stability of adenine (one of the five nucleic acid bases) under hydrothermal conditions. The experiments were performed in sealed autoclaves at 300 degrees C under fugacities of CO(2), N(2) and H(2) supposedly representative of those in marine hydrothermal systems on the early Earth. The composition of the gas phase was obtained from the degradation of oxalic acid, sodium nitrite and ammonium chloride, and the oxidation of metallic iron. The results of the experiments indicate that after 200 h, adenine is still present in detectable concentration in the aqueous phase. In fact, the concentration of adenine does not seem to be decreasing after approximately 24 h, which suggests that an equilibrium state may have been established with the inorganic constituents of the hydrothermal fluid. Such a conclusion is corroborated by independent thermodynamic calculations.     

Trade-off between C and N recycling and N<Subscript>2</Subscript>O emissions of soils with summer cover crops in subtropical agrosystems

Aims

Biological soil crusts (biocrusts) are soil-surface communities in drylands, dominated by cyanobacteria, mosses, and lichens. They provide key ecosystem functions by increasing soil stability and influencing soil hydrologic, nutrient, and carbon cycles. Because of this, methods to reestablish biocrusts in damaged drylands are needed. Here we test the reintroduction of field-collected vs. greenhouse-cultured biocrusts for rehabilitation.

Methods

We collected biocrusts for 1) direct reapplication, and 2) artificial cultivation under varying hydration regimes. We added field-collected and cultivated biocrusts (with and without hardening treatments) to bare field plots and monitored establishment.

Results

Both field-collected and cultivated cyanobacteria increased cover dramatically during the experimental period. Cultivated biocrusts established more rapidly than field-collected biocrusts, attaining ~82% cover in only one year, but addition of field-collected biocrusts led to higher species richness, biomass (as assessed by chlorophyll a) and level of development. Mosses and lichens did not establish well in either case, but late successional cover was affected by hardening and culture conditions.

Conclusions

This study provides further evidence that it is possible to culture biocrust components from later successional materials and reestablish cultured organisms in the field. However, more research is needed into effective reclamation techniques.

     

Study of Plasma Induced Chemistry by DC Discharges in CO<Subscript>2</Subscript>/N<Subscript>2</Subscript>/H<Subscript>2</Subscript>O Mixtures Above a Water Surface

The chemistry induced by atmospheric pressure DC discharges above a water surface in CO(2)/N(2)/H(2)O mixtures was investigated. The gaseous mixtures studied represent a model prebiotic atmosphere of the Earth. The most remarkable changes in the chemical composition of the treated gas were the decomposition of CO(2) and the production of CO. The concentration of CO increased logarithmically with the increasing input energy density and an increasing initial concentration of CO(2) in the gas. The highest achieved concentration of CO was 4.0 +/- 0.6 vol. %. The production of CO was crucial for the synthesis of organic species, since reactions of CO with some reactive species generated in the plasma, e. g. H* or N* radicals, were probably the starting point in this synthesis. The presence of organic species (including the tentative identification of some amino acids) was demonstrated by the analysis of solid and liquid samples by high-performance liquid chromatography, infrared absorption spectroscopy and proton-transfer-reaction mass spectrometry. Formation of organic species in a completely inorganic CO(2)/N(2)/H(2)O atmosphere is a significant finding for the theory of the origins of life.     

Simulation and optical spectroscopy of a DC discharge in a CH<Subscript>4</Subscript>/H<Subscript>2</Subscript>/N<Subscript>2</Subscript> mixture during deposition of nanostructured carbon films

Two-dimensional numerical simulations of a dc discharge in a CH₄/H₂/N₂ mixture in the regime of deposition of nanostructured carbon films are carried out with account of the cathode electron beam effects. The distributions of the gas temperature and species number densities are calculated, and the main plasmachemical kinetic processes governing the distribution of methyl radicals above the substrate are analyzed. It is shown that the number density of methyl radicals above the substrate is several orders of magnitude higher than the number densities of other hydrocarbon radicals, which indicates that the former play a dominant role in the growth of nanostructured carbon films. The model is verified by comparing the measured optical emission profiles of the H(n ≡ 3), C ₂ ^* , CH*, and CN* species and the calculated number densities of excited species, as well as the measured and calculated values of the discharge voltage and heat fluxes onto the electrodes and reactor walls. The key role of ion–electron recombination and dissociative excitation of H₂, C₂H₂, CH₄, and HCN molecules in the generation of emitting species (first of all, in the cold regions adjacent to the electrodes) is revealed.     

Temporal Variability of CO<Subscript>2</Subscript> and N<Subscript>2</Subscript>O Flux Spatial Patterns at a Mowed and a Grazed Grassland

Spatial patterns of ecosystem processes constitute significant sources of uncertainty in greenhouse gas flux estimations partly because the patterns are temporally dynamic. The aim of this study was to describe temporal variability in the spatial patterns of grassland CO₂ and N₂O flux under varying environmental conditions and to assess effects of the grassland management (grazing and mowing) on flux patterns. We made spatially explicit measurements of variables including soil respiration, aboveground biomass, N₂O flux, soil water content, and soil temperature during a 4-year study in the vegetation periods at grazed and mowed grasslands. Sampling was conducted in 80 × 60 m grids of 10 m resolution with 78 sampling points in both study plots. Soil respiration was monitored nine times, and N₂O flux was monitored twice during the study period. Altitude, soil organic carbon, and total soil nitrogen were used as background factors at each sampling position, while aboveground biomass, soil water content, and soil temperature were considered as covariates in the spatial analysis. Data were analyzed using variography and kriging. Altitude was autocorrelated over distances of 40–50 m in both plots and influenced spatial patterns of soil organic carbon, total soil nitrogen, and the covariates. Altitude was inversely related to soil water content and aboveground biomass and positively related to soil temperature. Autocorrelation lengths for soil respiration were similar on both plots (about 30 m), whereas autocorrelation lengths of N₂O flux differed between plots (39 m in the grazed plot vs. 18 m in the mowed plot). Grazing appeared to increase heterogeneity and linkage of the spatial patterns, whereas mowing had a homogenizing effect. Spatial patterns of soil water content, soil respiration, and aboveground biomass were temporally variable especially in the first 2 years of the experiment, whereas spatial patterns were more persistent (mostly significant correlation at p < 0.05 between location ranks) in the second 2 years, following a wet year. Increased persistence of spatial patterns after a wet year indicated the recovery potential of grasslands following drought and suggested that adequate water supply could have a homogenizing effect on CO₂ and N₂O fluxes.     

Composition of <Emphasis Type="Italic">nifH</Emphasis> in a wastewater treatment system reliant on N<Subscript>2</Subscript> fixation

High levels of nitrogen fixation have been observed in the wastewaters of pulp and paper mills. In this study, we show that nitrogen fixation in a model pulp and paper wastewater treatment system is supported by a high density of nifH sequences that are of low diversity. Quantitative PCR revealed a ratio of nifH to 16S rDNA of 1.14 +/- 0.76 which shows that very high levels of the nifH gene were enriched to support the high rates of nitrogen fixation that occur in this wastewater. Changes in wastewater composition and dissolved oxygen levels did not affect the nifH levels and allowed stable wastewater treatment. The nifH sequences identified display a similar profile to those seen in forest soil environments where nifH sequences derived from alpha-proteobacteria and beta-proteobacteria are also prevalent.     

Microbial N Turnover and N-Oxide (N<Subscript>2</Subscript>O/NO/NO<Subscript>2</Subscript>) Fluxes in Semi-arid Grassland of Inner Mongolia

Gross rates of N mineralization and nitrification, and soil–atmosphere fluxes of N₂O, NO and NO₂ were measured at differently grazed and ungrazed steppe grassland sites in the Xilin river catchment, Inner Mongolia, P. R. China, during the 2004 and 2005 growing season. The experimental sites were a plot ungrazed since 1979 (UG79), a plot ungrazed since 1999 (UG99), a plot moderately grazed in winter (WG), and an overgrazed plot (OG), all in close vicinity to each other. Gross rates of N mineralization and nitrification determined at in situ soil moisture and soil temperature conditions were in a range of 0.5–4.1 mg N kg⁻¹ soil dry weight day⁻¹. In 2005, gross N turnover rates were significantly higher at the UG79 plot than at the UG99 plot, which in turn had significantly higher gross N turnover rates than the WG and OG plots. The WG and the OG plot were not significantly different in gross ammonification and in gross nitrification rates. Site differences in SOC content, bulk density and texture could explain only less than 15% of the observed site differences in gross N turnover rates. N₂O and NO _x flux rates were very low during both growing seasons. No significant differences in N trace gas fluxes were found between plots. Mean values of N₂O fluxes varied between 0.39 and 1.60 μg N₂O-N m⁻² h⁻¹, equivalent to 0.03–0.14 kg N₂O-N ha⁻¹ y⁻¹, and were considerably lower than previously reported for the same region. NO _x flux rates ranged between 0.16 and 0.48 μg NO _x -N m⁻² h⁻¹, equivalent to 0.01–0.04 kg NO _x -N ha⁻¹ y⁻¹, respectively. N₂O fluxes were significantly correlated with soil temperature and soil moisture. The correlations, however, explained only less than 20% of the flux variance.     

The role of N<Subscript>2</Subscript> fixation in alleviating N limitation in wetland metaphyton: enzymatic,isotopic, and elemental evidence

The role of nitrogen (N₂) fixation in balancing N supply to wetland metaphyton was assessed by comparing primary production with enzymatic, isotopic, and elemental correlates. Primary production, N₂ fixation (acetylene reduction, AR), phosphatase activity, C:N:P ratio, and N isotopic composition of metaphyton were measured along a nutrient gradient in a freshwater marsh during May through September 2004. N₂ fixation and phosphatase activity in metaphyton were negatively correlated with inorganic N and P concentrations, respectively. Although metaphyton N₂ fixation demonstrated a clear spatial pattern along the nutrient gradient, N₂ fixation rates varied monthly and decreased sharply in September. However, the percent contribution of N₂ fixation to N uptake by metaphyton consistently decreased throughout the summer. Furthermore, the decreased contribution of N₂ fixation to N uptake corresponded with an increase in metaphyton N content during the growing season. Nitrogen isotopic data suggested the sustained importance of an atmospheric N₂ source through September at the most downstream (nutrient poor) site even though the percent contribution of N₂ fixition to N uptake was lowest in that month. This suggests that metaphyton were efficiently accumulating and recycling fixed N₂ in support of primary production. Over the course of the summer, metaphyton primary production showed a weak inverse correlation with metaphyton phosphatase activity (r ² = 0.58). The largest residuals in this regression corresponded to the largest vartiation in metaphyton N content. When metaphyton primary production was normalized to metaphyton N content, production rates for the entire growing season were more strongly inversely correlated with metaphyton phosphatase activity (r ² = 0.78). Results of the study suggest that N₂ fixation in N poor areas may adequately supplement community N requirements in metaphyton, thereby causing limitation by other elemental resources such as P.     

Emissions of CO<Subscript>2</Subscript>, CH<Subscript>4</Subscript> and N<Subscript>2</Subscript>O from undisturbed,drained and mined peatlands in Estonia

The aim of this study is to estimate emissions of greenhouse gases CO₂, CH₄ and N₂O, and the effects of drainage and peat extraction on these processes, in Estonian transitional fens and ombrotrophic bogs. Closed-chamber-based sampling lasted from January to December 2009 in nine peatlands in Estonia, covering areas with different land-use practices: natural (four study sites), drained (six sites), abandoned peat mining (five sites) and active peat mining areas (five sites). Median values of soil CO₂ efflux were 1,509, 1,921, 2,845 and 1,741 kg CO₂-C ha^?1 year^?1 from natural, drained, abandoned and active mining areas, respectively. Emission of CH₄-C (median values) was 85.2, 23.7, 0.07 and 0.12 kg ha^?1 year^?1, and N₂O-N ?0.05, ?0.01, 0.18 and 0.19 kg ha^?1 year^?1, respectively. There were significantly higher emissions of CO₂ and N₂O from abandoned and active peat mining areas, whereas CH₄ emissions were significantly higher in natural and drained areas. Significant Spearman rank correlation was found between soil temperature and CO₂ flux at all sites, and CH₄ flux with high water level at natural and drained areas. Significant increase in CH₄ flux was detected for groundwater levels above 30 cm.     

Theoretical investigation of reactivities of amines in the <Emphasis Type="Italic">N</Emphasis>-nitrosation reactions by N<Subscript>2</Subscript>O<Subscript>3</Subscript>

N-nitrosamine is a class of carcinogenic, mutagenic, and teratogenic compounds, which can be produced from N-nitrosation of amine by nitrosating agents. N-nitrosation of 19 amines (eight acyclic amines, five heterocyclic amines, and six amines with unsaturated groups) by N₂O₃ was investigated at the CBS-QB3 level of theory. The results indicate that generally the heterocyclic amines have the highest reactivities among the three kinds of amines, whereas the reactivities of the amines with unsaturated and electron-withdrawing groups are relatively low. Frontier molecular orbital analysis indicates that the energy gap between the HOMO of an amine and the LUMO of N₂O₃ has a close connection with the reactivity of an amine. A structure-reactivity relationship of amines in the N-nitrosation reactions by N₂O₃ was established using the stepwise multivariate linear regression. The results indicate that the reactivity of an amine has a definite relationship (R_adj² = 0.947) with the heterolytic bond dissociation energy of R₁R₂N-H bond, energy of HOMO, NBO occupancy of the natural lone pair orbital of N atom, the NBO charge of the N atom, and the pyramidalization angle of an amine. This work will be helpful to gain more insight into the N-nitrosation reactions.     

Isolation and characterization of a <Emphasis Type="Italic">Mastigocladus</Emphasis> species capable of growth,N<Subscript>2</Subscript>-fixation and N-assimilation at elevated temperature

A Mastigocladus species was isolated from the hot spring of Jakrem (Meghalaya) India. Uptake and utilization of nitrate, nitrite, ammonium and amino acids (glutamine, asparagine, arginine, alanine) were studied in this cyanobacterium grown at different temperatures (25°C, 45°C). There was 2–3 fold increase in the heterocyst formation and nitrogenase activity in N-free medium at higher temperature (45°C). Growth and uptake and assimilation of various nitrogen sources were also 2–3 fold higher at 45°C indicating that it is a thermophile. The extent of induction and repression of nitrate uptake by NO₃ ⁻ and NH₄ ⁺, respectively, differed from that of nitrite. It appeared that Mastigocladus had two independent nitrate/nitrite transport systems. Nitrate reductase and nitrite reductase activitiy was not NO₃ ⁻-inducible and ammonium or amino acids caused only partial repression. Presence of various amino acids in the media partially repressed glutamine synthetase activity. Ammonium (methylammonium) and amino acid uptake showed a biphasic pattern, was energy-dependent and the induction of uptake required de novo protein synthesis. Ammonium transport was substrate (NH₄ ⁺)-repressible, while the amino acid uptake was substrate inducible. When grown at 25°C, the cyanobacterium formed maximum akinetes that remained viable upto 5 years under dry conditions.     

The adsorption of NO,NH<Subscript>3</Subscript>, N<Subscript>2</Subscript> on carbon surface: a density functional theory study

To explore the adsorption mechanism of NO, NH₃, N₂ on a carbon surface, and the effect of basic and acidic functional groups, density functional theory was employed to investigate the interactions between these molecules and carbon surfaces. Molecular electrostatic potential, Mulliken population analyses, reduced density gradient, and Mayer bond order analyses were used to clarify the adsorption mechanism. The results indicate that van der Waals interactions are responsible for N₂ physisorption, and N₂ is the least likely to adsorb on a carbon surface. Modification of carbon materials to decorate basic or acidic functional groups could enhance the NH₃ physisorption because of hydrogen bonding or electrostatic interactions, however, NO physisorption on a carbon surface is poor. Zig-zag sites are more reactive than armchair sites when these gas molecules absorb on the edge sites of carbon surface.

Open image in new window

Graphical abstract NH₃, N₂, NO adsortion on carbon surface

     

The encapsulated lithium effect on the first hyperpolarizability of C<Subscript>60</Subscript>Cl<Subscript>2</Subscript> and C<Subscript>60</Subscript>F<Subscript>2</Subscript>

In this paper, we report a study on the structure and first hyperpolarizability of C₆₀Cl₂ and C₆₀F₂. The calculation results show that the first hyperpolarizabilities of C₆₀Cl₂ and C₆₀F₂ were 172 au and 249 au, respectively. Compared with the fullerenes, the first hyperpolarizability of C₆₀Cl₂ increased from 0 au to 172 au, while the first hyperpolarizability of C₆₀F₂ increased from 0 au to 249 au. In order to further increase the first hyperpolarizability of C₆₀Cl₂ and C₆₀F₂, Li@C₆₀Cl₂ and Li@C₆₀F₂ were obtained by introducing a lithium atom to C₆₀Cl₂ and C₆₀F₂. The first hyperpolarizabilities of Li@C₆₀Cl₂ and Li@C₆₀F₂ were 2589 au and 985 au, representing a 15-fold and 3.9-fold increase, respectively, over those of C₆₀Cl₂ and C₆₀F₂. The transition energies of four molecules (C₆₀Cl₂, Li@C₆₀Cl₂, C₆₀F₂, Li@C₆₀F₂) were calculated, and were found to be 0.17866 au, 0.05229 au, 0.18385 au, and 0.05212 au, respectively. A two-level model explains why the first hyperpolarizability increases for Li@C₆₀Cl₂ and Li@C₆₀F₂.     

An estimation of CO<Subscript>2</Subscript> fixation capacity in mangrove forest using two methods of CO<Subscript>2</Subscript> gas exchange and growth curve analysis

In many coastal areas of South-East Asia, attempts have been made to revive coastal ecosystem by initiating projects that encourage planting of mangrove trees. Compared to the terrestrial trees, mangrove trees possess a higher carbon fixation capacity. It becomes a very significant option for clean development mechanism (CDM) program. However, a reliable method to estimate CO₂ fixation capacity of mangrove trees has not been established. Acknowledging the above fact, we decided to set up an estimation method for the CDM program, using gas exchange analysis to estimate mangrove productivity, we put into consideration the net CO₂ fixation of reforested Kandelia candel (5-, 10-, and 15-year-old stand). This was estimated by gas exchange analysis and growth curve analysis. In growth curve analysis, we drew a growth curve of a single stand using data of above- and below-ground biomass. In the gas exchange analysis, we calculated CO₂ fixation capacity by (1) measuring respiration rate of each organ of stand and calculating respiratory CO₂ emission from above- to below-ground biomass. (2) Measuring the single-leaf photosynthetic rate in response to light intensity and calculating the photosynthetic CO₂ absorption. (3) We also developed a model for the diurnal changes in temperature, and monthly averages based on one-day estimation of CO₂ absorption and emission, which we corrected by this model in order to estimate the net CO₂ fixation capacity in response to temperature. Comparing the biomass accumulation of the two methods constructed for the same forest, the above-ground biomass accumulation of 10-year-old forest (34.3 ton ha⁻¹ yr⁻¹) estimated by gas exchange analysis was closely compared to those of growth curve analysis (26.6 ton ha⁻¹ yr⁻¹), suggesting that the gas exchange analysis was capable of estimating mangrove productivity. The validity of the estimated CO₂ fixation capacity by the gas exchange analysis and the growth curve analysis was also discussed.     

The interactions of plant growth regulators and H<Subscript>2</Subscript>O<Subscript>2</Subscript> during germination improvement of sweet corn seed through spermidine application

Low seed vigor was the main constraint on the production of sweet corn in China. Spermidine (Spd) was proved to enhance sweet corn seed germination. However, little was known about the metabolisms and interactions of plant growth regulators (PGRs) and H₂O₂ in the enhancement of Spd upon sweet corn seed germination. Spd, GA, C₂H₄ and H₂O₂ soaking treatments significantly enhanced seed vigor; while their respective biosynthesis inhibitors and ABA significantly declined seed vigor. Besides, as compared with control, seed vigor showed no significant difference in Spd+ProG (prohexadione-calcium, the inhibitor of GA), however it decreased significantly in Spd+ABA. The seed vigor treated by Spd+AVG (aviglycine hydrochloride, the inhibitor of C₂H₄) and Spd+NAC (n-acetyl-l-cysteine, a scavenger of H₂O₂) were significantly lower than those soaked in Spd solution, but still significantly higher than the control. Spd+NAC with significantly lower H₂O₂ content still up-regulated GA and C₂H₄ contents and down-regulated ABA content during seed germination. The results suggested that it was Spd rather than H₂O₂ (produced through Spd) made a direct effect on PGRs metabolism regulation in seed germination enhancement by Spd. The metabolism of GA and ABA played crucial rolesas compared with C₂H₄ and H₂O₂. Besides, complicated PGRs interactions and crosstalk between H₂O₂ and PGRs existed during sweet corn seed germination after Spd soaking, and ABA might be a key hormone in this process.     

Nitrosation of malononitrile by HONO,ClNO and N<Subscript>2</Subscript>O<Subscript>3</Subscript>: A theoretical study

Nitrosation reactions of malononitrile by three nitrosating agents, HONO, ClNO, and N₂O₃, have been theoretically investigated at the B3LYP/cc-pVTZ and MP2/cc-pVDZ levels. Two possible competitive paths for nitrosation of malononitrile to give 2-nitroso-malononitrile were proposed: (a) direct C-nitrosation and (b) N-nitrosation and subsequent nitroso transfer from N to C atom. The calculations show that at both B3LYP and MP2 levels, path b is kinetically favored over path a for nitrosations by HONO and N₂O₃. In the case of ClNO, the B3LYP predicts preference of path b, while the MP2 calculations suggest that both paths have similar rate-determining barriers. The data suggest that N₂O₃ is the preferred nitrosating agent for the nitrosation of malononitrile in aqueous solution. Transformation of 2-nitroso-malononitrile to form malononitrileoxime via intramolecular proton transfer has also been explored, and it is found that inclusion of an assistant water molecule can drastically accelerate the tautomerization.