Ammonium fixation in selected California decomposed granites

Revegetation on disturbed, low organic matter content, decomposed granite (DG) substrates are limited by low plant-available moisture and nitrogen. Data from a single DG site in northern California, USA, showed that a significant fraction of the ammonium from fertilizers or organic matter mineralization was fixed into silicate interlayer positions. To evaluate the broader relevance of NH₄⁺ fixation, the NH₄⁺ fixation capacities of 11 other drastically disturbed DG substrates throughout California were evaluated. The fixation capacities of the substrates were quite varied and increased as added NH₄⁺ application levels increased (124–1,670 kg NH₄⁺ ha⁻¹). When amended with 124 kg NH₄⁺ ha⁻¹, 7 of the 11 substrates fixed between 14 and 78% of the added NH₄⁺. Analysis of particle size fractions of a typical material indicated that the very fine sand fraction had the highest fixation capacity and the clays and very coarse sands had the lowest, on a gravimetric basis. The overall fixation capacities showed no significant relation to potential predictive characteristics, including extractable K⁺, NH₄⁺, or total N levels. Three methods of cation exchange capacity (CEC) measurement were tested for their ability to predict NH₄⁺ fixation. The Ba method which utilizes an indicator cation that is not subject to interlayer fixation was not a reliable indicator of NH₄⁺ fixation. The NH₄⁺ method had the strongest relation to NH₄⁺ fixation in the DG materials. The difference between the measured CEC of the NH₄⁺ method and the Ba method was found to be most predictive of NH₄⁺ fixation.     

Gaseous nitrogen losses and ammonia volatilization measurement following land application of cattle slurry in the mid-Atlantic region of the USA

To provide locally-determined field data for extension and environmental management purposes, gaseous N losses were measured following cattle slurry application to an arable silty-loam soil in the mid-Atlantic region of the USA. The field had been cropped to no-till maize. NH₃ volatilization was measured with the micro-meteorological, integrated horizontal flux (IHF) method, and denitrification with a core incubation method using acetylene inhibition. An early-winter surface application (5 December 1996; 88 m³ ha⁻¹ supplying 91 kg NH ₄ ⁺ -N ha⁻¹) was either unincorporated or immediately incorporated. NH₃ volatilization was measured from the unincorporated application, and denitrification from both slurry treatments and appropriate control soils. Total NH₃ loss from the unincorporated slurry application was 19% of applied NH ₄ ⁺ -N; temperatures were cool (4–6 °C), and 25 mm of rain fell within 24 h of application. For 3 months, enhanced denitrification occurred from the two slurry treatments, with generally higher rates from the incorporated slurry. Total net denitrification loss from the surface-applied and incorporated slurry treatments was, respectively, 11 and 17% of applied NH ₄ ⁺ -N. Denitrification loss over the winter/early-spring period was appreciable but not substantial, even where NH₃ volatilization was restricted by immediate incorporation. From the spring application (30 April 1997, 39 m³ ha⁻¹ supplying 51 kg NH ₄ ⁺ -N ha⁻¹), total NH₃ loss was 71% of applied NH ₄ ⁺ -N. These NH₃ volatilization loss data and the similarity of climate suggest that NH₃ loss factors from recent NW European work are likely to be generally applicable in the mid-Atlantic region. NH₃ volatilization from the spring application was also measured using the Z-instrument (Z_INST) approach, and with a system of small wind tunnels. A comparative assessment of the three methods is reported.     

Effects of the glutamine synthetase inhibitor methionine sulfoximine on CO2 fixation inLemna gibba

Summary Net CO₂ fixation inLemna gibba L. was inhibited by 0.5 mM L-methionine D,L-sulfoximine (MSX) both under photorespiratory conditions (21% O₂) and in 2% O₂. The inhibition was noticeably delayed by addition of 5 mM glutamine. Glutamine also delayed MSX-induced inactivation of glutamine synthetase. An increase in intracellular NH ₄ ⁺ concentration was noted in the presence of MSX only, and in the presence of 10 mM NH ₄ ⁺ only. However, presence of 10 mM NH ₄ ⁺ did not cause any inhibition of CO₂ fixation.     

Photoproduction of ammonium ion from N2 inRhodospirillum rubrum

NH ₄ ⁺ excretion was undetectable in N₂-fixing cultures ofRhodospirillum rubrum (S-1) and nitrogenase activity in these cultures was repressed by the addition of 10 mM NH ₄ ⁺ to the medium. The glutamate analog,l-methionine-dl-sulfoximine (MSX), derepressed N₂ fixation even in the presence of 10 mM extracellular NH ₄ ⁺ . When 10 mg MSX/ml was added to cultures just prior to nitrogenase induction they developed nitrogenase activity (20% of the control activities) and excreted most of their fixed N₂ as NH ₄ ⁺ . Nitrogenase activities and NH ₄ ⁺ production from fixed N₂ were increased considerably when a combined nitrogen source, NH ₄ ⁺ (>40 moles NH ₄ ⁺ /mg cell protein in 6 days) orl-glutamate (>60 moles NH ₄ ⁺ /mg cell protein in 6 days) was added to the cultures together with MSX.Biochemical analysis revealed thatR. rubrum produced glutamine synthetase and glutamate synthase (NADP-dependent) but no detectable NADP-dependent glutamate dehydrogenase. The specific activity of glutamine synthetase was observed to be maximal when nitrogenase activity was also maximal. Nitrogenase and glutamine synthetase activities were repressed by NH ₄ ⁺ as well as by glutamate.The results demonstrate that utilization of solar energy to photoproduce large quantities of NH ₄ ⁺ from N₂ is possible with photosynthetic bacteria by interfering with their regulatory control of N₂ fixation.     

CO2 fixation in halobacteria

Seven strains of extremely halophilic bacteria (Halobacterium spp., Halococcus spp., and Haloarcula sp.) fixed CO₂ under light and dark conditions. Light enhanced CO₂ fixation in Halobacterium halobium but inhibited it in Halobacterium volcanii and Haloarcula strain GN-1. Propionate stimulated ¹⁴CO₂ incorporation in some strains, but inhibited it in others. Semi-starvation in basal salts plus glycerol induced enhanced CO₂ fixation rates. ¹⁴CO₂ fixation in semi-starved cells was stimulated by NH ₄ ⁺ or pyruvate and inhibited by succinate and acetate in most strains. No possible reductant was found. In cell-free extracts of H. halobium, NH ₄ ⁺ but not propionate stimulated ¹⁴CO₂ fixation. No RuBP carboxylase activity was detected. The main ¹⁴C-labeled -keto acid detected after a 2-min incubation with ¹⁴CO₂ and pyruvate was pyruvate. Little or no -ketobutyrate was detected among the early products of propionate-stimulated CO₂ fixation. Glycine was the major amino acid synthesized during a 2-min incubation with NH ₄ ⁺ , propionate, and ¹⁴CO₂. Propionate-stimulated CO₂ fixation was sensitive to trimethoprim and insensitive to avidin. A novel pathway for non-reductive CO₂ fixation involving a glycine synthase reaction with CO₂, NH ₄ ⁺ , and a methyl carbon derived from the -carbon cleavage of propionate is tentatively proposed.Abbreviations used BBS buffered basal salts - HEPES N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid - MOPS 3-(N-morpholino)propanesulfonic acid - DNPH 2,4-dinitrophenylhydrazine - DNP dinitrophenyl - TLC thin-layer chromatography - FH₄ tetrahydrofolate This work was supported by National Science Foundation grant PCM-8116330 and Petroleum Research Fund grant PRF 13704-AC2     

Methylamine,an inhibitor of ammonium oxidation and chemoautotrophic growth in the marine nitrifying bacteriumNitrosococcus oceanus

Methylamine (CH₃NH ₃ ⁺ ) appeared to utilize the same transport mechanism as ammonium (NH ₄ ⁺ ) to enter cells ofNitrosococcus oceanus. Methylamine uptake did not show clear evidence of saturable kinetics and was not fully saturated at 20 mM. Assimilated CH₃NH ₃ ⁺ was not incorporated into macromolecular constituents, but inhibited rates of nitrification, chemoautotrophic CO₂ fixation and growth. The degree of inhibition was dependent on the relative concentrations of NH ₄ ⁺ and CH₃NH ₃ ⁺ . Rates of CO₂ fixation and growth were inhibited four times more than the rate of nitrification.     

Soil potassium mobility and uptake by corn under differential soil moisture regimes

This study examined the effects of soil moisture on soil K mobility, dynamics of soil K, soil K fixation, plant growth and K uptake. A pot experiment, with and without corn (Zea maysL.), was conducted over a 16-d duration using a Yolo silt loam treated with two soil moisture regimes, i.e. constant moisture vs. wetting–drying (W–D) cycles. Soil K dynamics were determined using both ion exchange resin and direct extraction of soil solution. Soil K mobility increased significantly with soil moisture content (θ_v) and there was a positive curvilinear relationship between θ_v and effective diffusion coefficient (D_e), suggesting that more K⁺ can diffuse to the plant roots at sufficient soil moistures. Increase in D_e could be attributed to the decrease of impedance factor. During W–D cycles, soil solution K concentration increased as soil solution volume decreased, but soil solution K and NH₄ ⁺-extractable K pools decreased. In the constant moisture regime, available K pools decreased over the 16-d duration, but to a lesser extent than in W–D regime. The W–D cycles significantly enhanced K fixation and reduced available K pools in the soil in contrast to the constant moisture regime. Potassium fixation by the soil showed a biphasic pattern under the W–D regime, with a rapid fixation within the first 2 d after re-wetting, followed by a slower fixation. In the soil with constant moisture, K fixation was rapid during the first 8 h after wetting the soil, and then proceeded so slowly that no significant K fixation was observed after 4 d. The W–D cycles decreased root and shoot growth and K uptake by corn compared to constant moisture condition. Our results support the hypothesis that W–D cycles enhance soil K fixation, reduce soil K mobility and plant growth, and therefore reduce plant K⁺ uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

15N-Determined effect of inoculation with N2 fixing bacteria on nitrogen assimilation in Western Canadian wheats

Summary A two-year field study was undertaken using¹⁵N isotope techniques to differentiate between stimulation of N uptake and N₂ fixation in Western Canadian cultivars of spring wheat (Triticum aestivum L. emend Thell) and durum (T. turgidum L. emend Bowden) in response to inoculation with N₂-fixing bacteria. Bacterial inoculation either had no effect or lowered the % N derived from the fertilizer and the fertilizer use efficiency. Despite the depression of fertilizer uptake, inoculants did not alter the relative uptake from soil and fertilizer-N pools indicating that bacterial inoculation did not alter rooting patterns. Nitrogen-15 isotope dilution indicated that N₂ fixation did occur. In 1984, % plant N derived from the atmosphere (% Ndfa) due to inoculation with Bacillus C-11-25 averaged 23.9% while that withAzospirillum brasilense ATCC 29729 (Cd) averaged 15.5%. In 1985, higher soil N levels reduced these values by approximately one-half. Cultivar x inoculant interactions, while significant, were not consistent across years. However, these interactions did not affect cultivars ‘Cadet’ and ‘Rescue’. In agreement with previous results, ‘Cadet’ performed well with all inoculants in both years while ‘Rescue’ performed poorly. Among 1984 treatments, the N increament in inoculated plants was positively correlated with % Ndfa but no such correlation existed in 1985. N₂ fixation averaged over all cultivars and strains was 17.9 and 6.7 kg N fixed ha⁻¹ in 1984 and 1985, respectively. Highest rates of N₂ fixation were estimated at 52.4 kg N ha⁻¹ for ‘Cadet’ in 1984 and 31.3 kg N ha⁻¹ for ‘Owens’ in 1985, both inoculated with Bacillus C-11-25, an isolate from southern Alberta soils. Inoculation with either ofAzospirillum brasilense strain Cd (ATCC29729) or 245 did not result in as consistent or as high N₂ fixation, suggesting that these wheats had not evolved genetic compatability with this exogenous microorganism. These agronomically significant amounts of N₂ fixation occurred under optimally controlled experimental conditions in the field. It is yet to be determined if N₂ fixation would occur in response to bacterial inoculation under dryland conditions commonly occurring in Western Canada. Contribution from Agriculture Canada Research Station, Lethbridge, Alberta, Canada.     

AMMONIUM STIMULATION OF DARK CARBON FIXATION AS AN INDICATOR OF NITROGEN DEFICIENCY IN PHYTOPLANKTON: POTENTIAL ERRORS CAUSED BY AMMONIUM-OXIDIZING BACTERIA1

Stimulation of dark fixation of carbon by NH₄⁺ is often used as an indicator of phytoplankton N deficiency. This assay is based on the influence of available NH₄⁺ on anaplerotic CO₂ fixation by algae. However, carbon fixation by chemoautotrophic NH₄⁺-oxidizing bacteria may also be stimulated by NH₄⁺ enrichment, a process that can mask the algal response in natural communities. NH₄⁺ addition enhanced dark carbon fixation up to 300%, relative to unamended controls, in organisms collected on a 0.7-μm retention filter in oligotrophic Flathead Lake, Montana, but the effect was not detectable in the presence of nitrapyrin, an inhibitor of NH₄⁺-oxidizing bacteria. Dark carbon fixation was enhanced with addition of NH₄⁺ in organisms retained on 2-μm filters (which should allow passage of most bacteria). NH₄⁺ stimulated dark carbon fixation in N-deficient axenic cultures of Chlamydomonas reinhardtii Dang but not in N-replete cultures in both the presence and absence of nitrapyrin. Application of nitrapyrin or size fractionation treatments, to separate the processes of dark carbon fixation by nitrifiers and phytoplankton, may improve the efficacy of assays using NH₄⁺ stimulation of dark carbon fixation to specifically indicate N deficiency in natural algal communities.     

Comparative study of N uptake and distribution in three lines of common bean (Phaseolus vulgaris L.) at early pod filling stage

Summary The uptake and distribution of¹⁵NH ₄ ⁺ ,¹⁵NO ₃ ⁻ and¹⁵N₂ was studied in greenhouse-grown beans (Phaseolus vulgaris L.) with a commercial cultivar and 2 recombinant inbred backcross lines;¹⁵N was supplied in the nutrient solution at the R3 (50% bloom) stage. Plants were harvested 1, 5 and 10 days after treatment, and were separated into nodules, roots, stems, mature leaflets, immature leaflets, and flowers/fruits. All 3 lines showed rapid increases in the N content of flowers/fruits after the R3 stage. However, the percentage N in these tissues decreased after the R3 stage. One of the recombinant lines showed a greater uptake of NH ₄ ⁺ than the other 2 lines. Rates of¹⁵N₂ fixation and NO ₃ ⁻ uptake were similar for all 3 lines, N₂ fixation estimated from total N content showed the 2 recombinant lines with 24 and 34 percent greater activity than the commercial cultivar. Distribution of¹⁵N at the whole plant level was similar for all 3 lines for a similar N source.¹⁵NO ₃ ⁻ was transported first to leaflets and the label then moved into flowers/fruits. Transport of fixed N₂ was from the nodules to roots, stems and into flowers/fruits; usually less than 10 percent entered the leaflets. This indicates that N₂ fixation furnishes N directly to flowers/fruits with over 50 percent of the fixed N being deposited into flowers/fruits within 5 days after treatment.     

Ammonium versus Nitrate Nutrition of Plants Stimulates Microbial Activity in the Rhizosphere

Using an alkaline calcareous soil, pot experiments were conducted to elucidate the effects of NH ₄ ⁺ vs. NO ₃ ⁻ nutrition (50 or 100 mg kg⁻¹ soil) of wheat and maize on microbial activity in the rhizosphere and bulk soils. Dicyandiamide was used as nitrification inhibitor to maintain NH ₄ ⁺ as the predominant N source for plants grown in NH ₄ ⁺ -treated soil. While maize grew equally well on both N sources, root and shoot growth of wheat was higher under NH ₄ ⁺ than under NO ₃ ⁻ nutrition. Bacterial population density on roots, but not in the rhizosphere soil, was higher under NH ₄ ⁺ than under NO ₃ ^– supplied at 150 mg N kg⁻¹ soil; whereas at both N levels applied, NH ₄ ⁺ compared to NO ₃ ⁻ nutrition of wheat and maize significantly increased microbial biomass in the rhizosphere soil. Under both plant species, NH ₄ ⁺ vs. NO ₃ ⁻ nutrition also increased aerobic and anaerobic respiration, and dehydrogenase activity in the rhizosphere. As microbial activity in the planted bulk and unplanted soils was hardly affected by the N-source, we hypothesize that the stimulation by NH ₄ ⁺ of the rhizosphere microbial activity was probably due to higher availability of root exudates under NH ₄ ⁺ than under NO ₃ ⁻ nutrition.     

Seasonal accumulation and partitioning of nitrogen by lentil

Although wheat (Triticum aestivum L.) is the dominant crop of the semi-arid plains of Canada and the western United States, lentil (Lens culinaris Medik.) has become an important alternative crop. Sources and seasonal accumulation of N must be understood in order to identify parameters that can lead to increased N₂-fixing activity and yield. Inoculated lentil was grown in a sandy-loam soil at an irrigated site in Saskatchewan, Canada. Wheat was used as the reference crop to estimate N₂ fixation by the A-value approach. Lentil and wheat received 10 and 100 kg N ha⁻¹ of ammonium nitrate, respectively. Crops were harvested six times during the growing season and plant components analyzed. During the first 71 days after planting the wheat had a higher daily dry matter and N accumulation compared to lentil. However, during the latter part of the growing season, daily dry matter and N accumulation were greater for lentil. The maximum total N accumulation for lentil at maturity was 149 kg ha⁻¹. In contrast, wheat had a maximum N accumulation of 98 kg ha⁻¹ in the Feekes 11.1 stage, or 86 days after planting. The maximum daily rates of N accumulation were 3.82 kg N ha⁻¹ day⁻¹ for lentil and 2.21 kg N ha⁻¹ day⁻¹ for wheat. The percentage of N derived from N₂ fixation (% Ndfa) ranged from 0 at the first harvest to 92 % at final harvest. Generative plant components had higher values for % Ndfa than the vegetative components which indicates that N in the reproductive plant parts was derived largely from current N₂ fixation and lentil continued to fix N until the end of the pod fill stage. At final harvest, lentil had derived 129 kg N ha⁻¹ from N₂ fixation with maximum N₂-fixing activity (4.4 kg N ha⁻¹ day⁻¹) occurring during the early stages of pod fill. Higher maximum rates of N₂-fixing activity than net N accumulation (3.82 kg N ha⁻¹ day⁻¹) may have been caused by N losses like volatilization. In addition, lentil provided a net N contribution to the soil of 59 kg ha⁻¹ following the removal of the grain.     

Delayed inoculation and starter nitrogen for enhancing early growth and nitrogen status ofLespedeza cuneata

Summary Two growth chamber experiments were conducted to determine the response ofLespedeza cuneata (Dumont) G. Don. (sericea lespedeza) to delayed inoculation and low levels of nitrogen fertilization. Nitrogen was supplied either as NH ₄ ⁺ or as NO ₃ ^– in solution. At 0.5 and 5.0 ppm nitrogen early growth and N₂(C₂H₂) fixation was inhibited by NH ₄ ⁺ and promoted by NO ₃ ^– . Inoculation at seeding did not negatively affect growth prior to the onset of N₂(C₂H₂) fixation. Delayed inoculation until the trifoliate stage thus did not increase growth or N₂ fixation during the first 40 days of growth. After 40 days, specific nitrogenase activity was highest for plants inoculated at the first trifoliate stage of growth. In contrast, growth and total shoot nitrogen accumulation were higher in plants inoculated at planting. The experimental results suggest that delaying inoculation is not a useful technique for improving early growth ofL. cuneata for surface mine reclamation.     

Environmental controls on nitric oxide emission from northern Chihuahuan desert soils

A survey of nitric oxide (NO) emission from Chihuahuan desert soils found mean NO fluxes <0.1 ng NO-N cm^–2h^–1 during the dry season. These fluxes were at thelower end of the range reported for temperate grassland and woodlandecosystems. NO fluxes from wet or watered soils were higher(0.1–35 ng NO-N cm^–2 h^–1).Watering of black grama grassland soils produced an initial pulse of 12ng cm^–2 h^–1 (12-h after 1-cm watering)with high fluxes sustained over 4 days with repeated watering. Initialpulses from shrubland soils were lower (maximum 5 ngcm^–2 h^–1), and fluxes declined withrepeated watering. Repeated watering of creosotebush soils depleted thesoil NH ₄ ⁺ pool, and NO emissions weredirectly related to soil NH ₄ ⁺ concentrationsat the end of the experiment. In watered andNH ₄ ⁺ -fertilized creosotebush soils, NO fluxeswere positively related to potential net nitrification rates.NH ₄ ⁺ -fertilization boosted the initial NOpulse 15 times in the shrubland and 5 times in black grama grasslandrelative to watered controls. These experimental results point towardgreater substrate limitation in shrublands. In this desert basin, NOemission averaged 0.12 kg N ha^–1 y^–1in untreated soil and 0.76 kg N ha^–1y^–1 in watered soil. We multiplied these averages bythe distribution of grassland and shrubland vegetation within a58,600-ha area of the Jornada del Muerto basin to estimate regionallosses of 0.15–0.38 kg NO-N ha^–1y^–1 for this area of the Chihuahuan desert.     

Release of non-exchangeable {}^{{{\text{15}}}}{\text{NH}}^{{\text{ + }}}_{{\text{4}}} from subgrade, decomposed granite substrates and uptake by non-mycorrhizal and mycorrhizal California native annual grass, Vulpia microstachys

Release rates of recently fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ from non-exchangeable interlayer sites in 2:1 silicate minerals were determined for decomposed granite (DG) saprolites from three locations in California, USA. Recently-fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release from the DG substrate was quantified by extracting diffused $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ with H-resin, as well as a native, annual grass Vulpia microstachys. The $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release data varied with via the method of extraction, which included H-resin pre-treatments (Na⁺ or H⁺) and V. microstachys uptake (mycorrhizal inoculated or uninoculated). After 6 weeks (1008 h), more $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ was recovered from fixed interlayer positions by the H-resins as compared to uptake by V. microstachys. The H⁺ treated H-resins recovered more released $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ (≈94 mg ${\text{NH}}^{{\text{ + }}}_{{\text{4}}} - {\text{N}}\;{\text{kg}}^{1} $ or (12%) of total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ ) in two of the three DG samples as compared to the Na⁺ treated resins, (which recovered ≈70–78 mg ${\text{NH}}^{{\text{ + }}}_{{\text{4}}} - {\text{N}}\;{\text{kg}}^{{{\text{ - 1}}}} $ (or 9–10%) of the total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ ). The V. microstachys assimilated 8–9% of the total fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ with mycorrhizal inoculum as compared to only 2% without a mycorrhizal inoculum, over the same time period. The fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release kinetics from the H-resin experiments were most accurately described by first order and power function models, and can be characterized as biphasic using a heterogeneous diffusion model. Uptake of both the ¹⁵N and ambient, unlabelled N from the soils was closely related to plant biomass. There was no significant difference in percent of N per unit of biomass between the control and mycorrhizal treatments. The findings presented here indicate that observed, long-term $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ release rates from DG in studies utilizing resins, may overestimate the levels of fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ made available to plants and microorganisms. Additionally, the study suggested that mycorrhizae facilitate the acquisition and plant uptake of fixed $ {\text{NH}}^{{\text{ + }}}_{{\text{4}}} $ , resulting in markedly increased plant biomass production.     

Short-term ammonium inhibition of nitrate uptake by Azotobacter chroococcum

Addition of NH₄Cl at low concentrations to Azotobacter chroococcum cells caused an immediate cessation of nitrate uptake activity, which was restored when the added NH ₄ ⁺ was exhausted from the medium or by adding an NH ₄ ⁺ assimilation inhibitor, l-methionine-dl-sulfoximine (MSX) or l-methionine sulfone (MSF). In the presence of such inhibitors the newly-reduced nitrate was released into the medium as NH ₄ ⁺ . When the artificial electron donor system ascorbate/N-methylphenazinium methylsulfate (PMS), which is a respiratory substrate that was known to support nitrate uptake by A. chroococcum while inhibiting glutamine synthetase activity, was the energy source, externally added NH ₄ ⁺ had no effect on nitrate uptake. It is concluded that, in A. chroococcum cells, NH ₄ ⁺ must be assimilated to exert its short-term inhibitory effect on nitrate uptake. A similar proposal was previously made to explain the short-term ammonium inhibition of N₂ fixation in this bacterium.Abbreviations MOPS morpholinopropanesulfonic acid - MSX l-methionine-dl-sulfoximine - PMS N-methylphenazinium methylsulfate - MSF l-methionine sulfone     

Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method

Dinitrogen fixation in white clover (Trifolium repens L.) grown in pure stand and mixture with perennial ryegrass (Lolium perenne L.) was determined in the field using ¹⁵N isotope dilution and harvest of the shoots. The apparent transfer of clover N to perennial ryegrass was simultaneously assessed. The soil was labelled either by immobilizing ¹⁵N in organic matter prior to establishment of the sward or by using the conventional labelling procedure in which ¹⁵N fertilizer is added after sward establishment. Immobilization of ¹⁵N in the soil organic matter has not previously been used in studies of N₂ fixation in grass/clover pastures. However, this approach was a successful means of labelling, since the ¹⁵N enrichment only declined at a very slow rate during the experiment. After the second production year only 10–16% of the applied ¹⁵N was recovered in the harvested herbage. The two labelling methods gave, nonetheless, a similar estimate of the percentage of clover N derived from N₂ fixation. In pure stand clover, 75–94% of the N was derived from N₂ fixation and in the mixture 85–97%. The dry matter yield of the clover in mixture as percentage of total dry matter yield was relatively high and increased from 59% in the first to 65% in the second production year. The average daily N₂ fixation rate in the mixture-grown clover varied from less than 0.5 kg N ha⁻¹ day⁻¹ in autumn to more than 2.6 kg N ha⁻¹ day⁻¹ in June. For clover in pure stand the average N₂ fixation rate was greater and varied between 0.5 and 3.3 kg N ha⁻¹ day⁻¹, but with the same seasonal pattern as for clover in mixture. The amount of N fixed in the mixture was 23, 187 and 177 kg N ha⁻¹ in the seeding, first and second production year, respectively, whereas pure stand clover fixed 28, 262 and 211 kg N ha⁻¹ in the three years. The apparent transfer of clover N to grass was negligible in the seeding year, but clover N deposited in the rhizosphere or released by turnover of stolons, roots and nodules, contributed 19 and 28 kg N ha⁻¹ to the grass in the first and second production year, respectively. This revised version was published online in June 2006 with corrections to the Cover Date.     

Nodulation,nitrogen fixation and nitrogen uptake in pigeonpea (Cajanus cajan (L.) Millsp) of different maturity groups

Summary The seasonal patterns of nodulation, acetylene reduction, nitrogen uptake and nitrogen fixation were studies for 11 pigeonpea cultivars belonging to different maturity groups grown on an Alfisol at ICRISAT Center, Patancheru, India. In all cultivars the nodule number and mass increased to a maximum around 60–80 days after sowing and then declined. The nodule number and mass of medium- and late-maturing cultivars was greater than that of early-maturing cultivars. The nitrogenase activity per plant increased to 60 days after sowing and declined thereafter, with little activity at 100 days when the crop was flowering. At later stages of plant growth nodules formed down to 90 cm below the soil surface but those at greater depth appeared less active than those near the surface. All the 11 cultivars continued to accumulate dry matter until 140 days, with most biomass production by the late-maturing cultivars (up to 11 t ha⁻¹) and least by the early-maturing determinate cultivars (4 t ha⁻¹). Total nitrogen uptake ranged from 69 to 134 kg ha⁻¹. Nitrogen fixation by pigeonpea was estimated as the difference in total nitrogen uptake between pigeonpea and sorghum and could amount to 69 kg N ha⁻¹ per season, or half the total nitrogen uptake. Fixation by pigeonpea increased with crop duration, but there were differences within each maturity group. The limitations of the methods used for estimating N₂ fixation by pigeonpea are discussed. Submitted as J.A. No. 552 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Is KCl a reliable extractant of15NH 4 + added to coastal marine sediments?

The release of NH ₄ ⁺ and¹⁵N-labelled NH ₄ ⁺ by one-step KCl extraction was assessed in different types of coastal marine sediments. KCl was efficient to extract NH ₄ ⁺ from sandy sediments and less efficient in silt sediments, where an extended extraction period was required for obtaining a maximum NH ₄ ⁺ yield. Extraction at 0 or 20 °C had only a little effect on the efficiency of KCl. KCl gave always non complete recovery of¹⁵NH ₄ ⁺ in silt sediments. However, the added label could be fully recovered by addition of 80 mol·cm^–3 exogenous NH ₄ ⁺ prior to KCl, or when NaCl or ASW replaced KCl.¹⁵NH ₄ ⁺ was added to non-biological silt sediment, which was incubated at 0 °C up to 16 hours, to see the effect of physical processes on the partition of¹⁵NH₄ among porewater (29–49%) exchangeable (9–30%) and non-extractable, organic bound pools (24–42%). Total¹⁵N recovery was approximately 100%. KCl failed to remove¹⁵NH₄ which entered to unknown, bound pools in sediment. Only shortly after addition of¹⁵N (0.1 h), the extraction period resulted in significantly different¹⁵N recoveries (P < 0.05) in KCl extractable NH ₄ ⁺ , 17% versus 9% of label was recovered after 1 min or 60 min extraction of sediment, respectively. Two hours of incubation time were required for complete equilibrium of¹⁵NH ₄ ⁺ among porewater, exchangeable and organic bound pools. Sediments (silt) to which¹⁵NH ₄ ⁺ has been added in order to measure NH ₄ ⁺ turn-over and KCl is used as extractant, should be incubated for at least 2 hours, before taking a zero-time sample.     

Plant and microbial nitrogen use and turnover: Rapid conversion of nitrate to ammonium in soil with roots

Immobilization of ammonium (NH ₄ ⁺ ) by plants and microbes, a controlling factor of ecosystem nitrogen (N) retention, has usually been measured based on uptake of¹⁵NH ₄ ⁺ solutions injected into soil. To study the influence of roots on N dynamics without stimulating consumption of NH ₄ ⁺ , we estimated gross nitrification in the presence or absence of live roots in an agricultural soil. Tomato (Lycopersicon esculentum var. Peto76) plants were grown in microcosms containing root exclosures. When the plants were 7 weeks old,¹⁵N enriched nitrate (NO ₃ ⁻ ) was applied in the 0–150 mm soil layer. After 24 h, > 30 times more¹⁵NH ₄ ⁺ was found in the soil with roots than in the soil of the root exclosures. At least 18% of the NH ₄ ⁺ -N present at this time in the soil with roots had been converted from NO ₃ ⁻ . We estimated rates of conversion of NO ₃ ⁻ to NH ₄ ⁺ , and rates ofNH ₄ ⁺ immobilization by plants and microbes, by simulating N-flow of¹⁴⁺¹⁵N and¹⁵N in three models representing mechanisms that may be underlying the experimental data: Dissimilatory NO ₃ ⁻ reduction to NH ₄ ⁺ (DNRA), plant N efflux, and microbial biomass nitrogen (MBN) turnover. Compared to NO ₃ ⁻ uptake, plant NH ₄ ⁺ uptake was modest. Ammonium immobilization by plants and microbes was equal to at least 35% of nitrification rates. The rapid recycling of NO ₃ ⁻ to NH ₄ ⁺ via plants and/or microbes contributes to ecosystem N retention and may enable plants growing in agricultural soils to capture more NH ₄ ⁺ than generally assumed.