Influence of arbuscular mycorrhizae and phosphorus fertilization on growth, nodulation and N2 fixation (15N) inMedicago sativa at four salinity levels

The rose of an isolate of the arbuscular mycorrhizal (AM) fungusGlomus mosseae in the protection ofMedicago sativa (+Rhizobium meliloti) against salt stress induced by the addition of increasing levels of soluble salts was studied. The interactions between soluble P in soil (four levels), mycorrhizal inoculum and degree of salinity in relation to plant growth, nutrition and infective parameters were evaluated. Salt stress was induced by sequential irrigation with saline water having four concentrations of three salts (NaCl, CaCl₂, and MgCl₂).¹⁵N-labelled ammonium sulphate was added to provide a quantitative estimate of N₂ fixation under moderate to high salinity levels. N and P concentration and nodule formation increased with the amount of plant-available P or mycorrhizal inoculum in the soil and generally declined as the salinity in the solution culture increased from a moderate to a high level. The mycorrhizal inoculation protected the plants from salt stress more efficiently than any amount of plant-available P in soil, particularly at the highest salinity level applied (43.5 dS m^–1). Mycorrhizal inoculation matched the effect on dry matter and nutrition of the addition in the soil of 150 mg P kg^–1. Nevertheless the highest saline solution assayed (43.5 dS m^–1) affected more severely plants supplemented with phosphorus than those with the addition of mycorrhizal inoculum. Such a saline-depressing effect was 1.5 (biomass), 1.4 (N) and 1.5 (P) times higher in plants supplied with soluble phosphate than with AM inoculum. Mechanisms beyond those mediated by P must be involved in the AM-protectioe effect against salinity. The¹⁵N methodology used allowed the determination of N₂ fixation as influenced by different P applications compared to mycorrhizal inoculation. A lack of correlation between nodule formation and function (N₂ fixation) was evidenced in mycorrhizal-inoculated plants. In spite of the reduced activity per nodule in mycorrhizal-inoculated In spite of the reduced activity per nodule in mycorrhizal-inoculated plants, the N contents determined indicated the highest acquisition of N occurred in plants with the symbiotic status. Moreover, N and P uptake increased while Ca and Mg decreased in AM-inoculated plants. Thus P/Ca ratios and cation/anion balance in general were altered in mycorrhizal treatments. This study therefore confirms previous findings that AM-colonized plants have optional and alternative mechanisms available to satisfy their nutritive requirements and to maintain their physiological status in stress situations and in disturbed ecosystems.     

Contribution of denitrification and autotrophic and heterotrophic nitrification to nitrous oxide production in andosols

To quantify the contribution of denitrification and autotrophic and heterotrophic nitrification to N₂O production in Andosols with a relatively high organic matter content, we first examined the effect of C₂H₂ concentrations on N₂O production and on changes in mineral N contents. The optimum C₂H₂ concentration for inhibiting autotrophic nitrification was 10 Pa. Secondly, and Andosol taken from an arable field was incubated for 32 days at 30°C at 60, 80, and 100% water-holding capacity with or without the addition of NH ₄ ⁺ or NO _inf3 ^sup- (200 mg N kg^-1), and subsamples collected every 4–8 days were further incubated for 24 h with or without C₂H₂ (10 Pa). At 60 and 80% water-holding capacity with NH ₄ ⁺ added, 87–92% of N₂O produced (200–250 g N₂O–N kg^-1) was derived from autotrophic nitrification. In contrast, at 100% water-holding capacity with or without added NO _inf3 ^sup- , enormous amounts of N₂O (29–90 mg N₂O–N kg^-1) were produced rapidly, mostly by denitrification (96–98% of total production). Thirdly, to examine N₂O production by heterotrophic nitrification, the Andosol was amended with peptone or NH ₄ ⁺ (both 1000 mg N kg^-1)+citric acid (20 g C kg^-1) and with or without dicyandiamide (200 mg N kg^-1). Treatment with citric acid alone or with citric acid+dicyandiamide suppressed N₂O production. In contrast, peptone increased N₂O production (5.66 mg N₂O–N kg^-1) mainly by denitrification (80% of total production). However, dicyandiamide reduced N₂O production to 1.1 mg N₂O–N kg^-1. These results indicate that autotrophic nitrification was the main process for N₂O production except at 100% water-holding capacity where denitrification became dominant and that heterotrophic nitrification had a lesser importance in the soils examine.Dedicated to Professor J. C. G. Ottow on the occasion of his 60th birthday     

Trace gas emissions from soil of the central highlands of Mexico as affected by natural vegetation: a laboratory study

In the central highlands of Mexico, mesquite (Prosopis laevigata) and huisache (Acacia schaffneri), N₂-fixing trees or shrubs, dominate the vegetation and are currently used in a reforestation program to prevent erosion. We investigated how natural vegetation or cultivation of soil affected oxidation of CH₄, and production of N₂O. Soil was sampled under the canopy of mesquite (MES treatment) and huisache trees (HUI treatment), outside their canopy (OUT treatment) and from fields cultivated with maize (ARA treatment) at three different sites while production of CO₂, and dynamics of CH₄, N₂O and inorganic N (NH₄⁺, and NO₃^–) were monitored in an aerobic incubation. The production of CO₂ was 2.3 times higher and significantly greater in the OUT treatment, 3.0 times higher in the MES treatment and 4.0 times higher in the HUI treatment compared to the ARA treatment. There was no significant difference in oxidation of CH₄ between the treatments, which ranged from 0.019 g CH₄–C kg^–1 day^–1 for the HUI treatment to 0.033 CH₄–C kg^–1 day^–1 for the MES treatment. The production of N₂O was 30 g N₂O–N kg^–1 day^–1 in the MES treatment and >8 times higher compared to the other treatments. The average concentration of NO₃^– was 2 times higher and significantly greater in the MES treatment than in the HUI treatment, 3 times greater than in the OUT treatment and 10 times greater than in the ARA treatment. It was found that cultivation of soil decreased soil organic matter content, C and N mineralization, but not oxidation of CH₄ or production of N₂O.     

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.     

Role of Azospirillum brasilense nitrate reductase in nitrate assimilation by wheat plants

Summary The nitrogen metabolism of wheat plants inoculated with various Azospirillum brasilense strains and nitrate reductase negative (NR^–) mutants was studied in two monoxenic test tube experiments. The spontaneous mutants selected with chlorate under anaerobic conditions with nitrite as terminal electron acceptor fixed N₂ in the presence of 10 mM NO₃ ^– and were stable after the plant passage. One strain (Sp 245) isolated from surface-sterilized wheat roots produced significant increases in plant weight at both NO₃ levels (1 and 10 mM) which were not observed with the NR^– mutants or with the two other strains. Similar effects were observed in a pot experiment with soil on dry weight and total N incorporation but only at the higher N fertilizer level. In the monoxenic test tube experiments plants inoculated with the mutants showed lower nitrogenase activities than NR⁺ strains at the low NO₃ ^– level (1 = mM) but maintained the same level of activity with 10 mM NO₃ ^– where the activity of all NR⁺ strains was completely repressed. The nitrate reductase activity of roots increased with the inoculation of the homologous strains and with the mutants at both NO₃ ^– levels. At the low NO₃ ^– level this also resulted in increased activity in the shoots, but at the high NO₃ ^– level the two homologous strains produced significantly lower nitrate reductase activity in shoots while the mutants more than doubled it. The possible role of the bacterial nitrate reductase in NO₃ ^– assimilation by the wheat plant is discussed.     

Effects of Rhizobium and vesicular-arbuscular mycorrhiza inoculations on nodulation,symbiotic nitrogen fixation and soybean yield in subtropical-tropical fields

Summary Field experiments were carried out to determine the effects of single and mixed inoculations with Rhizobium and vesicular-arbuscular mycorrhiza (VAM) on nodulation, symbiotic N₂ fixation and yield of soybeans in six Taiwan subtropical-tropical sites. Inoculation with Rhizobium alone significantly increased nodulation, nodule weight and nitrogenase activity of nodules in three out of six experimental fields, and affected soybean yields in the range –13% to + 134%. Inoculation with VAM fungi alone did not have a significant effect on nodulation and nitrogenase activity. Mycorrhiza inoculation affected soybean yields in the range –13% to + 65%, but only the yield increases at one out of six sites with N application were statistically significant. Mixed inoculation with Rhizobium and mycorrhiza affected yields in the range –8% to + 145% A synergistic effect from mixed inoculation of Rhizobium-mycorrhiza on soybean yields was found in one out of six experimental fields. The yield response to N application (40 kg N ha^–1) in these six paddy-field trials was not significant. These results suggest that single or mixed inoculation of rhizobia can greatly assist soybean grain yields and can replace N fertilizers.     

Nitrogen-fixing potential of Acacia mangium and Acacia auriculiformis seedlings inoculated with Bradyrhizobium and Rhizobium spp.

Summary Two Australian Acacia species, A. mangium and A. auriculiformis were inoculated in vitro with eight strains of Bradyrhizobium spp. and two strains of Rhizobium spp. On the two plant species, only Bradyrhizobium spp. strains formed effective N₂-fixing nodules. A. mangium, which nodulates effectively with a restricted range of Bradyrhizobium spp. strains, is a specific host compared to A. auriculiformis. A. auriculiformis is assumed to be a promiscuous host because it nodulates effectively with a wide range of Bradyrhizobium spp. strains. Nodule efficiency as expressed by the ratio of N₂ fixed to nodule dry weight appeared to be higher in A. auriculiformis (0.44–0.81) than in A. mangium (0.23–0.55).     

A comparison of nitrogen fixation in genotypes of groundnut (Arachis hypogaea L.) using 15N-isotope dilution

Summary Nitrogen fixation in seven groundnut genotypes was measured by ¹⁵N-isotope dilution using a non-nodulating cultivar of groundnut as the nonfixing reference plant. Nitrogen fixation varied between 100 kg N ha^–1 in genotype J-11 and 153 kg N ha^–1 in Robut 33-1. The amount of plant-available soil N was small, so that 86%–92% of plant nitrogen was derived from N₂-fixation. Thus differences in N₂-fixation between genotypes closely reflected differences in their total N accumulation.ICRISAT Journal Article no. 600     

Evaluation of 15N-isotope dilution for measurement of nitrogen fixation in chickpea (Cicer arietinum L.)

Summary N accumulation, nodulation, and acetylene reduction activity were measured at frequent intervals during the growth of two chickpea genotypes, and N₂ fixation was estimated by an isotope-dilution method, using safflower as a non-N₂-fixing reference. Safflower was more efficient at N uptake than both the chickpea genotypes for at least the first 50 days and thus could not be used as an accurate reference control. We recommend that further work should employ non-nodulatiog genotypes of chickpea as reference plants and use slow-release forms of 15N fertilizer. Direct genotype comparison by isotope dilution estimated that genotype K 850 fixed 16–18 kg ha^–1 more N than G 130, and this difference was supported by the greater nodule mass and acetylene reduction activity in the K 850 cultivar. Inoculation with an ineffective chickpea Rhizobium sp. led to 69% nodulation on cultivar K 850 but only 33% on G 130. While nodule weight, N uptake, and acetylene reduction activity decreased with inoculation in K 850, the isotope dilutions were similar for both inoculation treatments. The lack of a significant effect on N₂ fixation was ascribed to the partial success of inoculant establishment.Published as Journal Article No. JA 692 of the International Crops Research Institute for the Semi Arid Tropics, Patancheru, A.P. 502324, India     

Effect of nitrogen levels and Rhizobium strains on symbiotic N2 fixation and grain yield of Phaseolus vulgaris L. genotypes in normal and saline-sodic soils

Summary Following screening, selection, characterization, and symbiotic N₂ fixation with 12,5, 25.0, and 40.0 mg N kg^–1 in normal and saline-sodic soils, only two Phaseolus vulgaris genotypes (HUR 137 and VL 63) and two Rhizobium spp. strains (ND 1 and ND 2) produced maximum nodulation, nitrogenase activity, plant N contents, and grain yields in saline-sodic soil, with 12.5 mg N kg^–1, compared with the other strains. However, interactions between strains (USDA 2689, USDA 2674, and ND 5) and genotypes (PDR 14, HUR 15, and HUR 138) were significant and resulted in more nodulation, and greater plant N contents, nitrogenase activity, and grain yields in normal soils with 12.5 mg N kg^–1 compared with salt-tolerant strains. Higher levels of N inhibited nodulation and nitrogenase activity without affecting grain yields. To achieve high crop yields from saline-sodic and normal soils in the plains area, simultaneous selection of favourably interacting symbionts is necessary for N economy, so that bean yields can be increased by the application of an active symbiotic system.     

Transitioning from standard to minimum tillage: Trade-offs between soil organic matter stabilization, nitrous oxide emissions, and N availability in irrigated cropping systems

Few studies address nutrient cycling during the transition period (e.g., 1–4 years following conversion) from standard to some form of conservation tillage. This study compares the influence of minimum versus standard tillage on changes in soil nitrogen (N) stabilization, nitrous oxide (N₂O) emissions, short-term N cycling, and crop N use efficiency 1 year after tillage conversion in conventional (i.e., synthetic fertilizer-N only), low-input (i.e., alternating annual synthetic fertilizer- and cover crop-N), and organic (i.e., manure- and cover crop-N) irrigated, maize–tomato systems in California. To understand the mechanisms governing N cycling in these systems, we traced ¹⁵N-labeled fertilizer/cover crop into the maize grain, whole soil, and three soil fractions: macroaggregates (>250 μm), microaggregates (53–250 μm) and silt-and-clay (<53 μm). We found a cropping system effect on soil N_new (i.e., N derived from ¹⁵N-fertilizer or -¹⁵N-cover crop), with 173 kg N_new ha⁻¹ in the conventional system compared to 71.6 and 69.2 kg N_new ha⁻¹ in the low-input and organic systems, respectively. In the conventional system, more N_new was found in the microaggregate and silt-and-clay fractions, whereas, the N_new of the organic and low-input systems resided mainly in the macroaggregates. Even though no effect of tillage was found on soil aggregation, the minimum tillage systems showed greater soil fraction-N_new than the standard tillage systems, suggesting greater potential for N stabilization under minimum tillage. Grain-N_new was also higher in the minimum versus standard tillage systems. Nevertheless, minimum tillage led to the greatest N₂O emissions (39.5 g N₂O–N ha⁻¹ day⁻¹) from the conventional cropping system, where N turnover was already the fastest among the cropping systems. In contrast, minimum tillage combined with the low-input system (which received the least N ha⁻¹) produced intermediate N₂O emissions, soil N stabilization, and crop N use efficiency. Although total soil N did not change after 1 year of conversion from standard to minimum tillage, our use of stable isotopes permitted the early detection of interactive effects between tillage regimes and cropping systems that determine the trade-offs among N stabilization, N₂O emissions, and N availability.     

Effects of various nitrogen fertilizers on emission of nitrous oxide from soils

Summary Field studies of the effects of different N fertilizers on emission of nitrous oxide (N20) from three Iowa soils showed that the N₂O emissions induced by application of 180 kg ha^–1 fertilizer N as anhydrous ammonia greatly exceeded those induced by application of the same amount of fertilizer N as aqueous ammonia or urea. On average, the emission of N₂O-N induced by anhydrous ammonia was more than 13 times that induced by aqueous ammonia or urea and represented 1.2% of the anhydrous ammonia N applied. Experiments with one soil showed that the N₂O emission induced by anhydrous ammonia was more than 17 times that induced by the same amount of N as calcium nitrate. These findings confirm indications from previous work that anhydrous ammonia has a much greater effect on emission of N₂O from soils than do other commonly used N fertilizers and merits special attention in research relating to the potential adverse climatic effect of N fertilization of soils.Laboratory studies of the effect of different amounts of NH₄OH on emission of N₂O from Webster soil showed that the emission of N₂O-N induced by addition of 100 g NH₄OH-N g^–1 soil represented only 0.18% of the N applied, whereas the emissions induced by additions of 500 and 1 000 g NH4OH-N g^–1 soil represented 1.15% and 1.19%, respectively, of the N applied. This suggests that the exceptionally large emissions of N₂O induced by anhydrous ammonia fertilization are due, at least in part, to the fact that the customary method of applying this fertilizer by injection into soil produces highly alkaline soil zones of high ammonium-N concentration that do not occur when urea or aqueous ammonia fertilizers are broadcast and incorporated into 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.     

Time-course of nitrogen fixation in two bambara groundnut (Vigna subterranea L. Verdc.) cultivars

TheA-value method, involving the application of a higher¹⁵N rate to a reference non-N₂-fixing plant, was used to assess the magnitude of N₂ fixation in two bambara groundnut cultivars at four growth stages [vegetative, 0–47 days after planting (DAP); early pod-filling, 47–99 DAP; mid-pod-filling, 99–120 DAP; physiological maturity, 120–148 DAP). The cultivars were Ex-Ada, a bunchy type, and CS-88-11, a slightly spreading type. They were grown on a loamy sand. Uninoculated Ex-Ada and CS-88-11 were used as reference plants to measure the N₂ fixed in the inoculated bambara groundnuts. In this greenhouse study, soil was the major source of N in bambara groundnuts during vegetative growth, and during this period it accounted for over 80% of the N accumulaed in the plants. However, N₂ fixation became the major source of plant N during reproductive growth. There were significant differences between the two cultivars in the ability to fix N₂, and at physiological maturity, almost 75% of the N in CS-88-11 was derived from the atmosphere compared to 55% in Ex-Ada. Also, the total N fixed in CS-88-11 at physiological maturity was almost double that in Ex-Ada. Our data indicate that the higher N₂ fixation in CS-88-11 was due to two factors, a higher intensity of N₂ fixation and a longer active period of N₂ fixation. The results also suggest that bambara groundnut genotypes could be selected for higher N₂ fixation in farining systems.     

Acetylene reduction,hydrogen evolution and nodule respiration in Phaseolus vulgaris

Summary In an experiment performed under greenhouse conditions, four cultivars of Phaseolus vulgaris L. (Venezuela-350; Aroana; Moruna; Carioca) were inoculated with three Rhizobium leguminosarum biovar phaseoli strains (C-05; C-40 = CIAT 255; C-89 = CIAT 55) and were fertilized with an N-free mineral nutrient solution. The plants were harvested 25, 40, and 55 days after emergence and the following paramenters were evaluated: Nitrogenase activity of nodulated roots, H₂ evolution by the nodules; relative efficiency of nitrogenase; respiration rates of nodulated roots and detached nodules; dry weight and total N of stems, leaves, pods, roots, and nodules. Generally the bean cultivar, Rhizobium strain, had an effect and there was an interaction effect with both symbiotic partners, on all parameters. On average, nodules represented 23% of total root respiration but the best symbiotic combinations showed lower ratios of C respired to N fixed. The maximum N-assimilation rate (between 40 and 55 days after emergence) of 11.93 mg N plant^–1 day^–1 occurred with the symbiotic combination of Carioca × C-05, while the poorest rate of 0.55 mg N plant^–1 day^–1 was recorded with Venezuela-350 × C-89. The best symbiotic combinations always showed the highest relative nitrogenase efficiency, but the differences in N₂-fixation rates cannot be explained solely in terms of conservation of energy by recycling of H₂. This requires further investigation.     

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.     

Benefits of inoculation of the common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis>) crop with efficient and competitive<Emphasis Type="Italic"> Rhizobium tropici</Emphasis> strains

Cropping in low fertility soils, especially those poor in N, contributes greatly to the low common bean (Phaseolus vulgaris L.) yield, and therefore the benefits of biological nitrogen fixation must be intensively explored to increase yields at a low cost. Six field experiments were performed in oxisols of Paraná State, southern Brazil, with a high population of indigenous common bean rhizobia, estimated at a minimum of 10³ cells g^–1 soil. Despite the high population, inoculation allowed an increase in rhizobial population and in nodule occupancy, and further increases were obtained with reinoculation in the following seasons. Thus, considering the treatments inoculated with the most effective strains (H 12, H 20, PRF 81 and CIAT 899), nodule occupancy increased from an average of 28% in the first experiment to 56% after four inoculation procedures. The establishment of the selected strains increased nodulation, N₂ fixation rates (evaluated by total N and N-ureide) and on average for the six experiments the strains H 12 and H 20 showed increases of 437 and 465 kg ha^–1, respectively,in relation to the indigenous rhizobial population. A synergistic effect between low levels of N fertilizer and inoculation with superior strains was also observed, resulting in yield increases in two other experiments. The soil rhizobial population decreased 1 year after the last cropping, but remained high in the plots that had been inoculated. DGGE analysis of soil extracts showed that the massive inoculation apparently did not affect the composition of the bacterial community.     

Gaseous and leaching nitrogen losses from no-tillage and conventional tillage systems following surface application of cattle manure

Previous studies have demonstrated inconsistent results on the impact of tillage systems on nitrogen (N) losses from field-applied manure. This study assessed the impact of no-tillage (NT) and conventional tillage (CT) systems on gaseous N losses, N₂O:N₂O + N₂ ratios and NO₃⁻-N leaching following surface application of cattle manure. The study was undertaken during the 2003/2004 and 2004/2005 seasons at two field sites in Nova Scotia namely, Streets Ridge (SR) in Cumberland County and the Bio-environmental Engineering Centre (BEEC) in Truro. Results showed that the NT system had higher (p < 0.05) NH₃ losses than CT. Over the two seasons, manure incorporation in CT reduced NH₃ losses on average by 86% at SR and 78% at BEEC relative to NT. At both sites and during both seasons, denitrification rates and N₂O fluxes in NT were generally higher than in CT plots, presumably due to higher soil water and organic matter content in NT. Over the two seasons, mean denitrification rates at SR were 239 and 119 g N ha⁻¹ d⁻¹, while N₂O fluxes were 120 and 64 g N ha⁻¹ d⁻¹ under NT and CT, respectively. At BEEC mean denitrification rates were 114 and 71 g N ha⁻¹ d⁻¹, while N₂O fluxes were 52 and 27 g N ha⁻¹ d⁻¹ under NT and CT, respectively. Conversely, N₂O:N₂O + N₂ ratios were lower in NT than CT suggesting more complete reduction of N₂O to N₂ under NT. When averaged across all soil depths, NO₃⁻-N was higher (p < 0.05) in CT than NT. Nitrate-N decreased with depth at both sites regardless of tillage. In most cases, NO₃⁻-N was higher under CT than NT at all soil depths. Similarly, flow-weighted average NO₃⁻-N concentrations in drainage water were generally higher under CT. This may be partly attributed to higher denitrification rates under NT. Therefore, NT may be a viable strategy to remove NO₃⁻-N from the soil, and thus, reduce NO₃⁻-N contamination of groundwater. However, it should be noted that while the use of NT reduces NO₃⁻-N leaching it may come with unintended environmental tradeoffs, including increased NH₃ and N₂O emissions.     

Dinitrogen emissions and the N2:N2O emission ratio of a Rendzic Leptosol as influenced by pH and forest thinning

Reduction of nitrous oxide (N₂O) to dinitrogen (N₂) by denitrification in soils is of outstanding ecological significance since it is the prevailing natural process converting reactive nitrogen back into inert molecular dinitrogen. Furthermore, the extent to which N₂O is reduced to N₂ via denitrification is a major regulating factor affecting the magnitude of N₂O emission from soils. However, due to methodological problems in the past, extremely little information is available on N₂ emission and the N₂:N₂O emission ratio for soils of terrestrial ecosystems. In this study, we simultaneously determined N₂ and N₂O emissions from intact soil cores taken from a mountainous beech forest ecosystem. The soil cores were taken from plots with distinct differences in microclimate (warm-dry versus cool-moist) and silvicultural treatment (untreated control versus heavy thinning). Due to different microclimates, the plots showed pronounced differences in pH values (range: 6.3–7.3). N₂O emission from the soil cores was generally very low (2.0 ± 0.5–6.3 ± 3.8 μg N m⁻² h⁻¹ at the warm-dry site and 7.1 ± 3.1–57.4 ± 28.5 μg N m⁻² h⁻¹ at the cool-moist site), thus confirming results from field measurements. However, N₂ emission exceeded N₂O emission by a factor of 21 ± 6–220 ± 122 at the investigated plots. This illustrates that the dominant end product of denitrification at our plots and under the given environmental conditions is N₂ rather than N₂O. N₂ emission showed a huge variability (range: 161 ± 64–1070 ± 499 μg N m⁻² h⁻¹), so that potential effects of microclimate or silvicultural treatment on N₂ emission could not be identified with certainty. However, there was a significant effect of microclimate on the magnitude of N₂O emission as well as on the mean N₂:N₂O emission ratio. N₂:N₂O emission ratios were higher and N₂O emissions were lower for soil cores taken from the plots with warm-dry microclimate as compared to soil cores taken from the cool-moist microclimate plots. We hypothesize that the increase in the N₂:N₂O emission ratio at the warm-dry site was due to higher N₂O reductase activity provoked by the higher soil pH value of this site. Overall, the results of this study show that the N₂:N₂O emission ratio is crucial for understanding the regulation of N₂O fluxes of the investigated soil and that reliable estimates of N₂ emissions are an indispensable prerequisite for accurately calculating total N gas budgets for the investigated ecosystem and very likely for many other terrestrial upland ecosystems as well.     

Effects of rate and depth of fertilizer application on emission of nitrous oxide from soil fertilized with anhydrous ammonia

Summary Field studies to determine the effect of different rates of fertilization on emission of nitrous oxide (N₂O) from soil fertilized with anhydrous ammonia showed that the fertilizer-induced emission of N₂O-N in 116 days increased from 1.22 to 4.09 kg ha^–1 as the rate of anhydrous ammonia N application was increased from 75 to 450 kg ha^–1. When expressed as a percentage of the N applied, the fertilizer-induced emission of N₂O-N in 116 days decreased from 1.6% to 0.9% as the rate of fertilizer N application was increased from 75 to 450 kg N ha^–1. The data obtained showed that a 100% increase in the rate of application of anhydrous ammonia led to about a 60% increase in the fertilizer-induced emission of N₂O.Field studies to determine the effect of depth of fertilizer injection on emission of N₂O from soil fertilized with anhydrous ammonia showed that the emission of N₂O-N in 156 days induced by injection of 112 kg anhydrous ammonia N ha^–1 at a depth of 30 cm was 107% and 21 % greater than those induced by injection of the same amount of N at depths of 10 cm and 20 cm, respectively. The effect of depth of application of anhydrous ammonia on emission of N₂O was less when this fertilizer was applied at a rate of 225 kg N ha^–1.