Estimating N2 fixation by Sesbania rostrata and S. cannabina (syn. S. aculeata) in lowland rice soil by the 15N dilution method

Summary A field experiment in concrete-based plots was conducted to estimate the contribution of N derived from air (Ndfa) or biological N₂ fixation in Sesbania rostrata and S. cannabina (syn. S. aculeata), using various references, by the ¹⁵N dilution method. The two Sesbania species as N₂-fixing reference plants and four aquatic weed species as non-N₂-fixing references were grown for 65 days after sowing in two consecutive crops, in the dry and the wet seasons, under flooded conditions. Soil previously labeled with ¹⁵N at 0.26 atom % ¹⁵N excess in mineralizable N was further labeled by ammonium sulfate with 3 and 6 atom % ¹⁵N excess. The results showed that ¹⁵N enrichment of soil NH ₄ ⁺ -N dropped exponentially in the first crop to half the original level in 50 days while in the second crop, it declined gradually to half the level in 130 days. The decline in ¹⁵N enrichment, in both N₂-fixing and non-fixing species, was also steeper in the first crop than in the second crop. Variations in ¹⁵N enrichment among non-fixing species were smaller in the second crop. The ratio of the uptake of soil N to that of fertilizer N in N₂-fixing and non-fixing species was estimated by the technique of varying the ¹⁵N level. In the second crop, this ratio in non-fixing species was higher than that in N₂-fixing species. Comparable estimates of % Ndfa were obtained by using ¹⁵N enrichment of various non-fixing species. There was also good agreement between the estimates obtained by using ¹⁵N enrichment of non-fixing species and those by using soil NH ₄ ⁺ -N, particularly in the second crop. By 25 days after sowing, the first crop of both Sesbania spp. had obtained 50% of total N from the atmosphere and the second crop had obtained 75%. The contribution from air increased with the age of the plant and ranged from 70% to 95% in 45–55 days. S. rostrata fixed substantially higher amounts of N₂ due to its higher biomass production compared with S. cannabina. Mathematical considerations in applying the ¹⁵N dilution method are discussed with reference to these results.     

Influence of mycorrhiza vs. soluble phosphate on growth,nodulation, and N2 fixation (15N) in alfalfa under different levels of water potential

Summary The legume Medicago sativa (+Rhizobium melilott) was grown under controlled conditions to study the interactions between soluble P in soil (four levels), or a mycorrhizal inoculum, and the degree of water potential (four levels) in relation to plant development and N₂ fixation. ¹⁵N-labelled ammonium sulphate was added to each pot for a qualitative estimate of N₂ fixation, in order to rank the effects of the different treatments.Dry-matter yield, nutrient content and nodulation increased with the amount of plant-available P in the soil, and decreased as the water stress increased, for each P-level. The mycorrhizal effect on dry matter, N yield, and on nodulation was little affected by the water potential. Since P uptake was affected by the water content in mycorrhizal plants, additional mechanisms, other than those mediated by P, must be involved in the mycorrhizal activity.There was a positive correlation between N yield and nodulation for the different P levels and the mycorrhizal treatment at all water levels. A high correlation between plant unlabelled N content and atom% ¹⁵N excess was also found for all levels of P. In mycorrhizal plants, however, the correlation between unlabelled N yield and ¹⁵N was lower. This suggests that mycorrhiza supply plants with other N sources in addition to those derived from the improvement on N₂ fixation.     

Nitrogen fixation in paddy soils

The 4 long-term experimental plots (Umbric haplaquept) with different fertilizer treatment at Cent. Agric. Exp. Sta. in Konosu City, Saitama Prefecture, were used for the sites of investigation. The 4 plots were NF (applied with no fertilizer), IF (applied with inorganic fertilizers), GM (applied with green manure and CaCO₂), and OM (applied with manure and inorganic fertilizers). Flooded water, floating weed, upper (0-2cm) and lower (2-10cm) parts of Apg horizon and rhizosphere were collected from each plot before flooding, during flooding, and after drainage. These samples were analyzed for N₂-fixing activity by acetylene reduction method, pH, Eh, and contents of Fe²⁺, NH₄ ⁺, chlorophyll-type compounds, and water-soluble carbohydrates.

The N₂-fixing activity of all samples showed almost the same pattern of change with time: very low before flooding, rapidly increased after flooding, the maximum value at the maximum tillering stage of rice plant, declined afterwards and reached a very low value after drainage.

Rough estimation of the “N₂-fixing capacity” of each part of the paddy field revealed that the most important site of the N₂ fixation was the reduced Apg horizon, that the importance of flooded water and/or the oxidized layer in the N₂ fixation was rather low except in infertile soil, and that the role of rhizosphere in the N₂ fixation could not be neglected also in Japan.

Reduced condition and content of easily decomposable organic substances were judged to be main factors which control the N₂-fixing activity in the flooded soil on the basis of correlations between the Nt-fixing activity and several analytical data of the paddy soils.     

Use of 15N isotope dilution for quantification of N2 fixation associated with roots of kallar grass (Leptochloa fusca (L.))

Summary Leptochloa fusca (L.) Kunth (kallar grass) has previously been found to exhibit high rates of nitrogen fixation. A series of experiments to determine the level of biological nitrogen fixation using ¹⁵N isotopic dilution were carried out in nutrient solution and saline soil. In the nutrient solution, E. coli inoculated plants were taken as non-nitrogen-fixing control. It was observed that nearly 60%–80% of the plant N was derived from atmospheric fixation. Estimations based on the N difference method gave much lower values (18%–35%). In experiments with saline soil which was initially sterilized with chloroform fumigation, a mixed culture of N₂-fixing rhizospheric isolates from kallar grass roots was inoculated and planted to kallar grass. Uninoculated treatments were regarded as controls. The soil was previously labelled with ¹⁵N by adding cellulose and (¹⁵NH₄)₂SO₄. The results of these studies showed fixation values of 6%–32% when estimated by ¹⁵N dilution, whereas by the N difference method 54% of the plant N was estimated to be derived from fixation. This discrepancy is due to the increase in root proliferation due to inoculation, which results in greater uptake of soil N. The distribution of ¹⁵N in different fractions of the soil-N indicted isotopic dilution due to bacterial fixation of atmospheric N₂.     

Evaluation of N2 fixation by applying 15N labeled plant material and ammonium sulfate

Effect of different ¹⁵N labeled sources on the estimation of N₂ fixation was investigated. The combination of ¹⁵N labeled ammonium sulfate, ¹⁵N labeled plant material, and ¹⁵N labeled ammonium sulfate with unlabeled plant material, was examined in pot experiments. Two cultivars of soybean (Glycine max) and one of mungbean (Vigna radiata) were used. No significant difference was observed among the treatments for the estimation of N₂ fixation. This was due to the homogeneity and stability of the ¹⁵N abundance in soil which resulted in a similar N uptake from the soil by the N² fixing and reference crops. The plant yield, total N uptake and amount of N₂ fixed were higher in the Yellow Soil than in the Andosol. The amount of N₂ fixed was strongly influenced by the plant growth and consequently it affected the plant yield. The slow decomposition of plant material in the Andosol resulted in a low yield in both the N₂ fixing and reference crops. Thus, the artificial decrease of the available N content in soil, by application of plant material, did not stimulate N, fixation but suppressed plant growth and N₂ fixation.     

Free-living nitrogen-fixing bacteria in temperate cropping systems: Influence of nitrogen source

Sustainable cropping systems rely on a minimum of external inputs. In these systems N is largely acquired in animal manures and leguminous green manures. Little is known of how these organic forms of N fertilizer influence the presence and activity of free-living N₂-fixing bacteria. High concentrations of inorganic N in soil inhibit N₂-fixation in cyanobacteria and Azotobacter spp. It is likely that manure and fertilizer applications would result in concentrations of inorganic N capable of inhibiting N₂ fixation and, ultimately, the presence of these organisms. We investigated the effect of synthetic and organic N fertilizer sources on the populations and N₂-fixation potential of free-living N₂-fixing bacteria in the Farming Systems Trial at the Rodale Research Institute. Field plots received the following N treatments prior to corn (Zea mays L.) production: (1) Legume rotations and green manures supplying about 165 kg N ha^-1; (2) beef cattle manure applied at a rate of 220 kg N ha^-1 (plus 60 kg N ha^-1 from 1994 hay plow-down); or (3) fertilizer N (urea and NH₄NO₃) applied at a rate of 145 kg N ha^-1. Soil samples were collected at two depths from corn plots four times during the growing season, and analyzed for soil moisture, soil pH, numbers of N₂-fixing cyanobacteria and Azotobacter spp., extractable NH _inf4 ^sup+ and NO _inf3 ^sup- , and potentially mineralizable N. Soil samples collected in mid-July were analyzed for nitrogenase activity (by C₂H₂ reduction) and total C and N. Populations of Azotobacter spp. and cyanobacteria were influenced only slightly by treatment; however, cyanobacteria species composition was notably influenced by treatment. Nitrogenase activity in surface soils was greatest in legume-N plots and in subsurface plots levels were greatest in fertilizer-N plots. Populations and activity of free-living N-fixing bacteria appeared to be somewhat reduced in all plots as a result of low soil pH levels and high concentrations of inorganic N across all treatments. Annual applications of N to all plots resulted in high levels of potentially mineralizable N that in turn may have reduced non-symbiotic N₂-fixation in all plots.     

Sources of CO2 efflux from soil and review of partitioning methods

Five main biogenic sources of CO₂ efflux from soils have been distinguished and described according to their turnover rates and the mean residence time of carbon. They are root respiration, rhizomicrobial respiration, decomposition of plant residues, the priming effect induced by root exudation or by addition of plant residues, and basal respiration by microbial decomposition of soil organic matter (SOM). These sources can be grouped in several combinations to summarize CO₂ efflux from the soil including: root-derived CO₂, plant-derived CO₂, SOM-derived CO₂, rhizosphere respiration, heterotrophic microbial respiration (respiration by heterotrophs), and respiration by autotrophs. These distinctions are important because without separation of SOM-derived CO₂ from plant-derived CO₂, measurements of total soil respiration have very limited value for evaluation of the soil as a source or sink of atmospheric CO₂ and for interpreting the sources of CO₂ and the fate of carbon within soils and ecosystems. Additionally, the processes linked to the five sources of CO₂ efflux from soil have various responses to environmental variables and consequently to global warming. This review describes the basic principles and assumptions of the following methods which allow SOM-derived and root-derived CO₂ efflux to be separated under laboratory and field conditions: root exclusion techniques, shading and clipping, tree girdling, regression, component integration, excised roots and insitu root respiration; continuous and pulse labeling, ¹³C natural abundance and FACE, and radiocarbon dating and bomb-¹⁴C. A short sections cover the separation of the respiration of autotrophs and that of heterotrophs, i.e. the separation of actual root respiration from microbial respiration, as well as methods allowing the amount of CO₂ evolved by decomposition of plant residues and by priming effects to be estimated. All these methods have been evaluated according to their inherent disturbance of the ecosystem and C fluxes, and their versatility under various conditions. The shortfalls of existing approaches and the need for further development and standardization of methods are highlighted.     

Isotope 15N enrichment of soil-ammonium N as a reference to estimate N2 fixation by stem and root-inoculated S. rostrata

Summary A pot experiment in the greenhouse was conducted to compare the contribution of N derived from the atmosphere or from biological N₂ fixation by Sesbania rostrata inoculated with Azorhizobium caulinodans, applied either to roots or to roots and stems (single or multiple stem inoculation). Two subsequent crops were grown for 50 days under flooded conditions. N derived from air was estimated by ¹⁵N dilution using ¹⁵N enrichment of soil NH _inf4 ^sup+ -N and of Echinochloa crusgalli as the non-N₂-fixing reference datum and compared with estimates obtained by the N-difference method. The first crop was grown to stabilize the ¹⁵N into the soil organic N fraction. The ¹⁵N enrichment of soil NH _inf4 ^sup+ -N in the second crop declined slowly. The extractability ratio (¹⁵N enrichment of extractable soil N to ¹⁵N enrichment of total soil N) decreased from 4.8 to 4.1 50 days after planting. The enrichment of soil NH _inf4 ^sup+ -N was comparable to that of E. crus-galli, resulting in similar estimates of N derived from air when either soil NH _inf4 ^sup+ -N or enrichment of E. crus-galli was used as a non-fixing reference. The N-difference method did not always provide reliable estimates of N derived from air; percentages ranged from 75 to more than 80 by 50 days after planting in both crops and did not differ among treatments. The study demonstrates the potential of using ¹⁵N enrichment of soil NH _inf4 ^sup+ -N as a non-N₂-fixing reference for reliable BNF estimates of crops in lowland puddled soil.     

Nitrogen fixation in paddy soils

Several important features of the N. fixation in paddy fields which were reported previously were confirmed and some new additional results regarding the evaluation of the N₂ fixation in the rhizosphere were obtained by reinvestigation in the fields. In addition, rice plants were cultivated in the submerged soil in pots and various parts of the soil were analyzed for the N₂-fixing activity as well as several other properties. The results of the pot experiments were found to be fairly similar to those observed in the field investigations, indicating the validity of the submerged soil in a pot as a rather simulated model for the actual paddy field. By using this model system, the following facts were ascertained: (1) Water-percolation had almost no effect on the N₂-fixing activities of both the rhizosphere and the non-rhizosphere soils. (2) Suppressing effect of washing the root of rice plant on the N₂-fixing activity was slight in the seedling stage and marked in the tillering and flowering stages. (3) The N₂-fixing activity of a single rice root varied from tip to base.     

Fate of nitrogen and carbon in the vadose zone: in situ and laboratory measurements of seasonal variations in aerobic respiratory and denitrifying activities

In situ and laboratory measurements of aerobic respiratory and denitrifying activities were studied in the vadose zone (almost 2.5 m thick) of a fluvic hypercalcaric cambisol characterized by transitory anaerobic conditions. A field experiment was conducted in a bare soil, over a 7-month period starting just after maize harvest and incorporation of maize crop residues. Weather variables (air and soil temperature, rainfall), soil water content, soil solutes (NO₃⁻ and dissolved organic carbon) and soil gases (CO₂ and N₂O), were recorded throughout the experiment. Four soil layers were defined. Bacterial counts were performed in each layer using the most probable number (MPN) method. Aerobic respiratory and denitrifying activities were estimated from laboratory measurements. In situ microbial activity, as revealed by CO₂ and N₂O measurements in the soil atmosphere, was strongly influenced by weather. Laboratory measurements showed that potential aerobic respiratory activity (ARA) occurred throughout the soil profile, whereas semi-potential denitrifying activities SPDA (i.e. measured under organic-C limiting condition) occurred mainly in the top 30 cm soil layer. In the soil profile, the CO₂ concentration gradient was stronger than the N₂O concentration gradient. Seasonal variations in microbial activities increased with depth, whereas DOC concentrations, and variations in those concentrations, decreased with depth, suggesting that DOC quality investigations are necessary in the deep vadose zone to understand microbial activities seasonal variations. Laboratory measurements of potential activities agreed well with in situ microbial activity in natural environmental conditions. NO₃⁻ was a stronger limiting factor for SPDA than was denitrifier density in the soil profile.     

The effects on N2 fixation (C2H2 reduction), bacterial population and rice plant growth of two modes of straw application to a wetland rice field

Summary The effects of incorporation and surface application of straw to a wetland rice field on nitrogen fixation (C₂H₂ reduction), bacterial population and rice plant growth were studied. Rice straw (5 t ha^–1) was chopped (10- to 15-cm pieces) and applied to the field 2 weeks before transplanting IR42, a long-duration variety, and IR50, a short-duration variety. The acetylene-reducing activity (ARA) of IR42 and IR50 measured at heading stage for 3 consecutive days showed significantly higher ARA in IR42 as a result of the 2 straw application methods. Mostly up to 20 days after straw surface application and incorporation, the dark ARA in the soil, total and N₂-fixing heterotrophs, and photoorganotrophic purple nonsulphur bacteria (POPNS) in the soil and in association with degrading straw were stimulated. Higher bacterial populations were associated with straw on the surface than with straw incorporated. The POPNS counts, in particular, were increased hundreds fold in the surface-applied straw treatment. Straw applications also increased the root, shoot and total plant biomass at heading stage and the total dry matter yield at harvest in both varieties. The data show the potentials of straw as a source of substrate for the production of microbial biomass and for the non-symbiotic N₂ fixation to improve soil fertility and plant nutrition.     

Denitrification potential in a grassland subsoil: effect of carbon substrates

We examined the potential of a subsoil to denitrify nitrate under optimal anaerobic conditions in a laboratory-based incubation when supplied with a range of C substrates of increasing recalcitrance. Both topsoil and its associated subsoil were supplied with nitrate and either glucose, starch or cellulose. Microbial respiration and the evolution of N₂O and N₂ were measured. The subsoil supported low amounts of microbial activity and responded only to the glucose treatment; with less than one-fifth of the N₂O production measured in the top soil. Overall, our findings demonstrated that the denitrification potential of this particular subsoil is relatively low and that only simple carbohydrates could be utilised readily by the resident microorganisms.     

Effects of land-use type and nitrogen addition on nitrous oxide and carbon dioxide production potentials in Japanese Andosols

Land-use type and nitrogen (N) addition strongly affect nitrous oxide (N₂O) and carbon dioxide (CO₂) production, but the impacts of their interaction and the controlling factors remain unclear. The aim of this study was to evaluate the effect of both factors simultaneously on N₂O and CO₂ production and associated soil chemical and biological properties. Surface soils (0–10 cm) from three adjacent lands (apple orchard, grassland and deciduous forest) in central Japan were selected and incubated aerobically for 12 weeks with addition of 0, 30 or 150 kg N ha^–1 yr^–1. Land-use type had a significant (p < 0.001) impact on the cumulative N₂O and CO₂ production. Soils from the apple orchard had higher N₂O and CO₂ production potentials than those from the grassland and forest soils. Soil net N mineralization rate had a positive correlation with both soil N₂O and CO₂ production rates. Furthermore, the N₂O production rate was positively correlated with the CO₂ production rate. In the soils with no N addition, the dominant soil properties influencing N₂O production were found to be the ammonium-N content and the ratio of soil microbial biomass carbon to nitrogen (MBC/MBN), while those for CO₂ production were the content of nitrate-N and soluble organic carbon. N₂O production increased with the increase in added N doses for the three land-use types and depended on the status of the initial soil available N. The effect of N addition on CO₂ production varied with land use type; with the increase of N addition doses, it decreased for the apple orchard and forest soils but increased for the grassland soils. This difference might be due to the differences in microbial flora as indicated by the MBC/MBN ratio. Soil N mineralization was the major process controlling N₂O and CO₂ production in the examined soils under aerobic incubation conditions.     

Estimating N2 fixation by field-grown lupins (Lupinus angustifolius L.) using soil and plant 15N enrichment

Summary Biological N₂ fixation was estimated in a field experiment following the addition of NH₄Cl or KNO₃ to unconfined microplots (1.5 m²) at 2.5 g N m^-2 (10 atom% ¹⁵N). A model of total N and ¹⁵N accumulation in lupins and decreasing ¹⁵N enrichment in the KCl-extractable soil-N pool (0–0.15 m depth) was used to estimate the proportion of N in lupins derived from biological N₂ fixation. Estimates of N₂ fixation derived from the model were compared with ¹⁵N isotope-dilution estimates obtained using canola, annual ryegrass, and wheat as nonfixing reference plants. Biomass, total N accumulation, or ¹⁵N enrichment in the lupin and reference crops did not differ whether NH _inf4 ^sup+ or NO _inf3 ^sup- was added as the labelled inorganic-N source. The decrease in soil ¹⁵N enrichment was described by first-order kinetics, whereas total N and ¹⁵N accumulation in the lupins were described by logistical equations. Using these equations, the uptake of soil N by lupins was estimated and was then used to calculate fixed N₂. Estimates of N₂ fixation derived from the model increased from 0 at 50 days after sowing to a maximum of 0.79 at 190 days after sowing. Those based on the ¹⁵N enrichment of the NO _inf3 ^sup- pool were 10% higher than those based on the mineral-N pool. ¹⁵N isotope-dilution estimates of N₂ fixation ranged from 0.37 to 0.55 at 68 days after sowing and from 0.71 to 0.77 at 190 days after sowing. Reference plant-derived values of N₂ fixation were all higher than modelled estimates during the early states of growth, but were similar to modelled estimates at physiological maturity. The use of the model to estimate N₂ derived from the atmosphere has the intrinsic advantage that the need for a non-fixing reference plant is avoided.     

Soil N Losses by Denitrification Evaluated Using the 15N Tracer Method

Molecular nitrogen (N₂) and nitrous oxide (N₂O) generated by denitrification increase N losses in the soil–plant system. This study aimed to quantify N₂ and N₂O from potassium nitrate (K¹⁵NO₃) applied to soils with different textures and moisture contents in the absence and presence of a source of carbon (C) using the ¹⁵N tracer method. In the three soils used (sandy texture (ST), sandy clay loam texture (SCLT), and clayey texture (CT)), three moisture contents were evaluated (40%, 60%, and 80% of the water holding capacity (WHC)) with (D⁺) and without (D^?) dextrose added. The treatments received 100 mg N kg^?1 (KNO₃ with 23.24 atom% ¹⁵N). N₂ emissions occurred in all of the treatments, but N₂O emissions only occurred in the D⁺ treatment, showing increases with increasing moisture content. SCLT with 80% WHC in the D⁺ treatment exhibited the highest accumulated N emission (48.26 mg kg^?1). The ¹⁵N balance suggested trapping of the gases in the soil.     

Quantification and potential availability of non-symbiotically fixed 15N in soil

Summary Non-symbiotic N₂ fixation was studied under laboratory conditions in two soils from Pakistan (Hafizabad silt loam and Khurrarianwala silt loam) and one from Illinois, USA (Drummer silty clay loam) incubated in a ¹⁵N-enriched atmosphere. N₂ fixation was greatest with the Drummer soil (18–122 g g^–1 soil, depending upon the soil treatment) and lowest with the Khurrarianwala soil (4–81 g g^–1 soil). Fixation was increased by the addition of glucose, a close correlation being observed between the amount of glucose added and the amount of N₂ fixed in the three soils (r = 0.96). Efficiency of N₂ fixation varied with soil type and treatment and was greatest in the presence of added inorganic P. Application of Mo apparently had a negative effect on the amount and efficiency of N₂ fixation in all the soils. The percentage of non-symbiotically fixed ¹⁵N in potentially mineralizable form (NH ₄ ⁺ -N released in soil after a 15-day incubation period under anaerobic conditions) was low (2%–18%, depending upon the soil treatment), although most of the fixed N (up to 90%) was recovered as forms hydrolysable with 6N HCl. Recovery in hydrolysable forms was much greater for the fixed N than for the native soil N, indicating that the former was more available for uptake by plants.     

Phytotron studies to compare nitrogen losses from corn-planted soil by the 15-N balance or direct dinitrogen and nitrous oxide measurements

Summary Containers filled with soil mixed with potassium nitrate highly enriched in ¹⁵N were planted with corn (Zea mays L.) and kept in a phytotron under controlled conditions for 79 days. Soil water content was normally maintained at exactly 60% water-holding capacity (–33 kPa), but it was increased several times to 85% (–5 kPa) for short periods to favour denitrification. The soil headspace was sealed from the phytotron atmosphere and aerated by a continuous stream of air. Nitrous oxide emission was measured by estimating the N₂O concentration differences in the air entering and leaving the containers. Emission of N₂ was estimated by mass spectroscopy from changes in the N₂ composition in the temporarily enclosed soil headspace. Both methods were carefully checked for accuracy by different tests. At specific times during the experiment the distribution of ¹⁵N between plants and soil was determined and a ¹⁵N balance established. Emission of N gases peaked at times of increased water content and reached maxima of 149 and 142 g N pot^–1 day^–1 for N₂O and N₂, respectively. While N losses of 5% ± 2% were indicated by the ¹⁵N balance, only 1.1% ± 0.3% loss from 2.7 g applied N was estimated from the N₂O and N₂ measurements after 79 days. Possible reasons for these differences are discussed.     

Processes responsible for the nitrous oxide emission from a Costa Rican Andosol under a coffee agroforestry plantation

We used the inhibitor acetylene (C₂H₂) at partial pressures of 10 Pa and 10 kPa to inhibit autotrophic nitrification and the reduction of nitrous oxide (N₂O) to N₂, respectively. Soils (Andosol) from a Coffea arabica plantation shaded by Inga densiflora in Costa Rica were adjusted to 39, 58, 76 and 87% water-filled pore space (WFPS) and incubated for 6 days in the absence or presence of C₂H₂. Soil respiration, nitrification rates and N₂O emissions by both processes were measured in relation to soil moisture conditions. At all WFPS studied, rates of N₂O and N₂ productions were small (4.8; 14.7; 23 and 239.6 ng N–N₂O g⁻¹ d.w. d⁻¹ at 39, 58, 76 and 87% WFPS, respectively), and despite a low soil pH (4.7), N₂O was mainly produced by nitrification, which was responsible for 85, 91, 84 and 87% of the total N₂O emissions at 39, 58, 76 and 87% WFPS, respectively. At the three smaller values of WFPS, a linear relationship was established between WFPS, soil respiration, nitrification and N₂O released by nitrification; no N₂ was produced by denitrification. At more anaerobic conditions achieved by a WFPS of 87%, a large rate of N₂O production was measured during nitrification, and N₂ production accounted for 84% of the gaseous N fluxes caused by denitrification.     

Background level of nitrous oxide in the atmosphere in relation to flux from soil

According to Broadbent and Clark (3), there are numerous data indicating that denitrification leads to the emission of N₂O together with N₂, whereby loss of N is developed from soils. Nitrous oxide is also released from soils to the atmosphere during the nitrification of ammonium and ammonium-producing fertilizers under aerobic conditions (1). Relatively few attempts have been made to directly measure N₂O evolution under field conditions (6, 7, 10–12), although a number of laboratory studies have been reported. These studies are essential for determining the N balance between additions and losses of soil N.     

Effects of greenhouse intensive cultivation and organic amendments on greenhouse gas emission according to a soil incubation study

Soil cultivation changes and usage of agricultural wastes can have profound impacts on greenhouse gas (GHG) emission from soil. In this study, the effects of soil cultivation and organic amendment on GHG emission were investigated using aerobic incubation. Surface soil (0–20 cm) from (1) rice–legume consecutive rotation (Rice) and (2) recently (<3 years) converted from rice field to plastic-covered intensive vegetable and flower production (VegC) were collected in Kunming, P.R. China. Rose (Rosa rugosa Thunb.) residues and cattle manure were applied at 5% by weight. Results indicated that N₂O and CO₂ fluxes were significantly influenced by soil cultivation, organic amendment, incubation time and their interaction (p <0.05). Applying cattle manure increased, while rose residue decreased, cumulative N₂O emissions from soil (84 days). Rose residue application significantly increased cumulative CO₂ emissions with peak values of 6371 (Rice) and 7481 mg kg^?1 (VegC), followed by cattle manure addition figure of 2265 (VegC) and 3581 mg kg^?1 (Rice). Both were significantly higher (p <0.05) than the un-amended Control at 709 (VegC) and 904 mg kg^?1 (Rice). Our study demonstrates that a low C/N ratio in cattle manure is better than a high C/N ratio in rose residue in regard to reducing the global warming potential of agricultural soil.