Field evaluation of N2 fixation and N partitioning in climbing bean (Phaseolus vulgaris L.) using 15N

Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N₂ fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha^-1 of ¹⁵N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha^-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the ¹⁵N fertilizer. N₂ fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N₂ fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N₂ fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N₂-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.     

Ontogenic variation in nitrogen fixation and accumulation of nitrogen in mungbean,blackgram, cowpea,and groundnut

Ontogenic variations in N₂ fixation and accumulation of N by the mungbean (Vigna radiata L. Wilczek), blackgram (Vigna mungo L. Hepper), cowpea (Vigna unguiculata L. Walp.), and groundnut (Arachis hypogaea L.) were studied by a ¹⁵N-dilution technique. Pots filled with 7 kg of red yellow podzolic soil were used. Samples were taken 20, 40, 60, and 80 days after emergence which approximately corresponded to preflowering, flowering, early/mid-pod filling and late pod filling stages, respectively. During early growth (up to 40 days after emergence), the carryover of seed N accounted for a considerable fraction of the total plant N in the legumes, the highest being in the groundnut. With a correction for carryover, the groundnut derived over 45% of its N content from the atmosphere 20 days after emergence whereas the corresponding figures were 33% for the blackgram and about 28% for the cowpea and mungbean. Between flowering and early pod fill, there was a rapid increase in N₂ fixation in all legumes except in groundnut which showed highest fixation from 60 to 80 days after emergence. In the mungbean, N₂ fixation and uptake of soil N were insignificant 60 days after emergence while in other legumes these processes continued beyond this time. All legumes derived about 90% of their N from atmosphere by 80 days after emergence. However, due to considerable interspecific differences in total N yield the final amount of N₂ fixed showed an appreciable variation among legumes. It was highest in the groundnut (443 mg N plant^-1) followed by the cowpea (385), blackgram (273), and mungbean (145), respectively. The groundnut maintained nodules until the late pod filling stage while in other legumes, nodules senesced progressively following the mid-pod filling stage. During pod filling there was a net mobilization of N from vegetative tissues to developing pods in the mungbean, which amounted to about 20% of N in seeds. This mobilization was not evident in other legumes.     

Quantitative analysis of the initial transport of fixed nitrogen in nodulated soybean plants using 15N as a tracer

The quantitative analysis of the initial transport of fixed isotope 15-nitrogen (¹⁵N) in intact nodulated soybean plants (Glycine max [L.] Merr. cv. Williams) was investigated at the vegetative stage (36 days after planting, DAP) and pod-filling stage (91 DAP) by the ¹⁵N pulse-chase experiment. The nodulated roots were exposed to N₂ gas labeled with a stable isotope ¹⁵N for 1 h, followed by 0, 1, 3 and 7 h of exposure with normal air. Plant roots and shoots were separated into three sections (basal, middle and distal parts) with the same length of the main stem or primary root. Approximately 80 and 92% of fixed N was distributed in the basal part of the nodulated roots at the vegetative and pod-filling stages by the end of 1 h of ¹⁵N₂ exposure, respectively. In addition, about 90% of fixed ¹⁵N was retained in the nodules and 10% was exported to root and shoot after 1 h of ¹⁵N₂ exposure at 91 DAP. The percentage distribution of ¹⁵N in the nodules at the pod-filling stage decreased from 90% to 7% during the 7 h of the chase period, and increased in the roots (14%), stems (54%), leaves (12%), pods (10%) and seeds (4%). The ¹⁵N distribution was negligible in the distal root segment, suggesting that N fixation activity was negligible and recycling fixed N from the shoot to the roots was very low in the initially short time of the experiment.     

Sulphur fertilizer effect on cotton: I. Nitrogen and sulphur status and petiole nitrate‐nitrogen

Abstract

The effect of S fertilization on S and N status and petiole NO₃ ^?‐N in cotton was observed during the growing seasons of 1980 and 1981. Four sites representing 2 soil subgroups were studied using a randomized complete block design with 4 replications. Leaf and petiole sampling began one week prior to bloom initiation and continued weekly for eight weeks. Leaf samples were analyzed for S and N and the petioles for NO₃ ^?‐N. Levels of leaf‐S varied directly with amounts of applied S. Leaf‐N and petiole NO₃ ^?‐N varied directly with amounts of applied N. Though not always significant, petiole NO₃ ^?‐N and leaf‐N showed negative correlations with leaf‐S. These results suggest that knowledge of the cotton plant S status may be necessary to interpret petiole NO₃ ^?‐N for N fertilization of cotton.     

Dinitrogen fixation and soil n uptake by soybean as affected by phosphorus availability

The effects of phosphorus supply (0, 30, and 90 mg P kg^‐1) on growth, N₂ fixation, and soil N uptake by soybean (Glycine max (L.) Merr.) were studied in a pot experiment using the ¹⁵N isotope technique. Phosphorus supply increased the top dry matter production at flowering and the dry matter production of seeds, straw, pod shells, and roots at late pod filling of inoculated soybeans. Phosphorus supply reduced the N concentration of plant tops at flowering, but increased the amount of N accumulated at both flowering and late pod filling. In inoculated soybeans total N accumulation paralleled the dry matter production. The P concentration in above‐ground plant parts of nodulated soybeans was not affected by P application. At flowering only 18 to 34% of total N was derived from N₂ fixation, whereas as much as 74% was derived from N₂ fixation at late pod filling. Only the addition of 90 mg P kg^‐1 soil significantly increased the amount of N₂ fixed at the late pod filling stage. Phosphorus supply did not influence the uptake of fertilizer or soil N in soybeans, even if the root mass was increased up to 60% by the P supply.     

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.     

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.     

Modelling nitrogen fixation of pea ( Pisum sativum L.)

Abstract

Nitrogen fixation was simulated for a leafless variety (Delta) of pea (Pisum sativum L.) in central Sweden. It is assumed that N₂ fixation is basically proportional to root biomass, but limited by high root N or low substrate carbon concentrations. Input data on root carbon and nitrogen were estimated from observations of above-ground biomass and nitrogen. The simulated N₂ fixation was compared with estimated values from observations using the ¹⁵N labelling technique. Test data were taken from pea monocultures and pea-oat mixtures with varying pea biomass levels during 1999. Simulated within-season accumulated N₂ fixation correlated to the estimated N₂ fixation with a correlation coefficient (R ²) of 0.74. For seasonal simulations, the predictability was higher (R ²=0.93). Two alternative non-dynamic models, estimating seasonal N₂ fixation as proportional to above-ground biomass and above-ground N, respectively, gave lower predictability (R ²=0.83 and 0.80, respectively). The models were also applied to a second year (1998) and two other sites by comparison with accumulated N₂ fixation estimated by the Difference method. A halved specific N₂ fixation rate (expressed per unit of root biomass) in 1999, compared with 1998, corresponded to essentially dryer and warmer soil conditions during 1999. It was indicated that the variations in soil moisture were more important than soil temperature. It was concluded that the abiotic responses might be of great importance for modelling N₂ fixation rate under different soil conditions.     

Quantification of N2 fixation and annual N benefit from N2 fixation in soybean accrued to the soil under soybean‐wheat continuous rotation

A computational exercise was undertaken to quantify the percent N derived from atmosphere %Ndfa) in soybean and consequent N benefit from biological N₂‐fixation process annually accrued to the soil by the soybean crop using average annual N‐input/‐output balance sheet from a 7 yr old soybean‐wheat continuous rotational experiment on a Typic Haplustert. The experiment was conducted with 16 treatments comprised of combinations of four annual rates of farmyard manure (FYM ? 0, 4, 8, and 16 t ha^–1) and four annual rates of fertilizer N (? 0, 72.5, 145, and 230 kg N ha^–1) applications. The estimated N contributed through residual biomass of soybean (RBN_S) consisting of leaf fall, root, nodules, and rhizodeposition varied in the ranges of 7.02–16.94, 11.65–28.83, 3.31–8.91, and 11.3–23.8 kg N ha^–1 yr^–1, respectively. A linear relationship was observed between RBN_S and harvested biomass N (HBN_S) of soybean in the form of RBN_S = 0.461 × HBN_S – 20.67 (r = 0.989, P < 0.01), indicating that for each 100 kg N assimilated by the harvested biomass of soybean, 25.4 kg N was added to the soil through residual biomass. The Ndfa values ranged between 13% and 81% depending upon the annual rates of application of fertilizer N and FYM. As per the main effects, the %Ndfa declined from 76.4 to 26.0 with the increase in annual fertilizer‐N application from 0 to 230 kg N ha^–1, whereas %Ndfa increased from 40.8 to 65.8 with the increase in FYM rates from 0 to 16 t ha^–1, respectively. The N benefit from biological N₂ fixation accrued to the soil through residual biomass of soybean ranged from 7.6 to 53.7 kg N ha^–1 yr^–1. The treatments having %Ndfa values higher than 78 showed considerable annual contribution of N from N₂ fixation to the soil which were sufficient enough to offset the quantity of N removed from the soil (i.e., native soil N / FYM‐N / fertilizer‐N) with harvested biomass of soybean.     

Effect of nitrogen application on the A N value of soil

Pot experiments were conducted with two soils, from Rottenhaus and Seibersdorf in Austria, to ascertain whether the rate of fertilizer N application and the test crop would influence the amount of N available in the soil as assessed by the A-value method. ¹⁵N-labelled fertilizer was applied at rates of 10, 25, 40, 60, and 100 mg N kg^-1 soil, corresponding approximately to 20, 50, 80, 120 and 200 kg N ha^-1 respectively, and two crop species, barley (Hordeum vulgareL.) and non-nodulating soybean (Glycine max L.) were used to determine the soil A _N value under the various fertilizer regimes. The results showed that the Rottenhaus soil had a higher A _N value than the Seibersdorf soil, suggesting that the former was more fertile than the latter. The A _N values of both soils were significantly affected by the level of N application. When grown in the same soil, the two test crops showed significantly different fertilizer use efficiency and per cent N derived from fertilizer when the rate of N application exceeded 20 kg ha^-1. Thus, the A _N value as determined by the two test crops differed significantly for the same soil when the rate of N application was greater than 20 kg/ha. The difference was greater when the soil fertility level was high. The dependence of the A _N value on the level of N application and the species of crop seriously compromises the suitability of this method for determining plant-associated N₂ fixation. Hence, considerable caution is required when using this method to estimate plant-associated N₂ fixation.     

Landscape-scale estimates of dinitrogen fixation by Pisum sativum by nitrogen-15 natural abundance and enriched isotope dilution

Topography and slope position influence the soil and environmental factors that affect N₂ fixation by legumes. The present study was conducted to (1) estimate N₂ fixation by field peas in a gently rolling farm field using the natural ¹⁵N abundance and the ¹⁵N-enriched isotope dilution techniques and (2) identify soil and environmental factors that influence N₂ fixation at the landscape scale. Whereas soil available water capacity, available NH _inf4 ^sup+ , total crop yield, and percent N derived from N₂ fixation (% Ndfa) estimated using enriched N were significantly affected by landform patterns, soil NO _inf3 ^sup- levels, seed yield, and the % Ndfa estimated using natural abundance did not follow landform patterns. The % Ndfa using natural abundance was correlated with NH _inf4 ^sup+ but not with available soil water, pH, electrical conductivity, NO _inf3 ^sup- , or particle size. Estimates of the % Ndfa using enriched ¹⁵N ranged from 0 to 92.8%. The highest median value (68.6%) for % Ndfa using enriched N occurred on the divergent footslopes, with the lowest value (28.1%) on the convergent shoulders. Estimates of % Ndfa using natural abundance ranged from 13.2% to 96.9%. Smaller fluctuations during the growing season in the ¹⁵N of the available N pool may have resulted in less variability for % Ndfa using natural abundance compared to enriched ¹⁵N. Despite similar mean values for % Ndfa using natural abundance (44.5) and enriched ¹⁵N (49.6), no significant correlation between the two estimates was found. These results suggest that although topography may exert gross controls on N₂ fixation, large variations in N₂ fixation at the microsite level may preclude correlations between individual estimates and limit detection of landscape scale patterns of N₂ fixation.Contribution No. R754 of the Saskatchewan Center of Soil Research     

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.     

N form and concentration: Effects on reproductive development,growth and N content of southern peas 1

Southern peas [Vigna unguiculata, (L.) Walp.] cultured with 100% NH⁺ ₄ produced no viable flowers, while treatments in which NO^‐ ₃ composed 50% or more of the N form were not significantly different in the number of flowers formed. Flower abortion was least with 100% NO^‐ ₃ at the lower N concentration and with 75% and 100% NO^‐ ₃ at the higher N concentration. Further increments of NH⁺ ₄ resulted in greater flower abortion. The trends in flower survival were reflected in the number of pods and number of seed/plant. At the lower N concentration, the addition of NH⁺ ₄ slowed pod maturity, while at the higher N concentration pod maturity was hastened with the addition of up to 50% NH⁺ ₄. The dry weight and N content of tissues were generally greater with the higher N concentration and with N combinations containing predominantly NO^‐ ₃, but trends varied with the plant part being analyzed. Ammonium appears to adversely influence reproductive development and/or NO^‐ ₃ is essential to complete the reproductive development of southern peas. The observed differences in the response of southern peas to N form may account for previously reported discrepancies concerning the effectiveness of N fertilization on growth and yield parameters. Also, vegetative growth and vegetative N content appear to be poor indicators of final seed yields of southern peas if NH⁺ ₄ supplies a significant portion of the N form utilized by the plant.     

Glyphosate- and imazethapyr-induced effects on yield, nodule mass and biological nitrogen fixation in field-grown glyphosate-resistant soybean

Over half of the 21 Mha of soybean planted in Brazil is now transgenic glyphosate-resistant (GM_RR). A field experiment was carried out to investigate whether the application of glyphosate or imazethapyr to the GM_RR variety reduced the input of N₂ fixation (BNF). No effects on yield, total N accumulation, nodulation and BNF (δ¹⁵N) could be assigned to the genetic modification of the plant. Imazethapyr reduced soybean yield but had no significant effect on BNF. Even though yields were not affected by glyphosate, the significant reduction of nodule mass and BNF to the GM_RR suggests that the use of this herbicide could lead to an increased dependence on soil N and consequently an eventual decrease of SOM reserves.     

Effect of nitrogen fertilization on yield component distribution and assimilate translocation of determinate and indeterminate soybean lines

Studies were conducted to compare the main‐stem and branch yield component distribution of two soybean [Glycine max (L.) Merr.] lines differing in architecture and growth habit, and to relate the partitioning of carbon assimilates during pod filling and seed filling stages. An indeterminate line (MI) and a determinate line (MD) were planted in early May on a fine, mixed mesic Udic Haplustalf soil in southwestern France. For the N treatment, N fertigation was applied at the end of the vegetative phase and the early stages of reproductive growth. Pod production on branches was stimulated in response to N application for the determinate line, and N also enhanced the weight of seeds located on branches for both lines. At different stages during pod setting and seed filling, mid‐canopy‐level leaves were allowed to assimilate ¹⁴CO₂ and 24 h after exposure, radioactivity was measured in different organs and levels of the whole plant. ¹⁴C‐assimilates appeared to be preferentially transferred to the lower level of the main‐stem in MI and to the lower level and branches in the MD line. For N‐treated plants, radiocarbon accumulation was particularly marked in pods located on the lower level and on branches; this was consistent with the yield component distribution patterns seen at harvest.     

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Abstract

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean (Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97?min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10?min with protons at an energy of 18.3?MeV and an electric current of 5?μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35?mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10?mL of O₂ and 5?mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17?±?0.10?μmol N₂?h^?1 from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012?±?0.011?μmol?N₂?h^?1. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

NATURAL ABUNDANCES OF 15NITROGEN AND 13CARBON INDICATIVE OF GROWTH AND N2 FIXATION IN POTASSIUM FED LENTIL GROWN UNDER WATER STRESS

Dual natural abundance analysis of ¹⁵Nitrogen (N) and ¹³Carbon (C) isotopes in lentil plants subjected to different soil moisture levels and rates of potassium (K) fertilizer were determined to assess crop performance variability in terms of growth and N₂-fixation (Ndfa). The δ¹⁵N values in lentils ranged from +0.67 to +1.36‰; whereas, those of the N₂-fixed and reference plant were ?0.45 and +2.94‰, respectively. Consequently, the Ndfa% ranged from 45 and 65% of total plant N uptake. Water stress reduced Δ¹³C values. However, K fertilization enhanced whole plant Δ¹³C along with dry matter yield and N₂-fixation. The water stressed plants amended with K fertilizer seemed to be the best treatment because of its highest pod yield, high N balance, and N₂-fixation with low consumption of irrigation water. This illustrates the ecological and economical importance of K fertilizer in alleviating water stress occurring during the post-flowering period of lentil.     

Evolution of dinitrogen and nitrous oxide from the soil to the atmosphere through rice plants

Summary It is commonly assumed that a large fraction of fertilizer N applied to a rice (Oryza sativa L.) field is lost from the soil-water-plant system as a result of denitrification. Direct evidence to support this view, however, is limited. The few direct field, denitrification gas measurements that have been made indicate less N loss than that determined by ¹⁵N balance after the growing season. One explanation for this discrepancy is that the N₂ produced during denitrification in a flooded soil remains trapped in the soil system and does not evolve to the atmosphere until the soil dries or is otherwise disturbed. It seems likely, however, that N₂ produced in the soil uses the rice plants as a conduit to the atmosphere, as does methane. Methane evolution from a rice field has been demonstrated to occur almost exclusively through the rice plants themselves. A field study in Cuttack, India, and a greenhouse study in Fort Collins, Colorado, were conducted to determine the influence of rice plants on the transport of N₂ and N₂O from the soil to the atmosphere. In these studies, plots were fertilized with 75 or 99 atom % ¹⁵N-urea and ¹⁵N techniques were used to monitor the daily evolution of N₂ and N₂O. At weekly intervals the amount of N₂+N₂O trapped in the flooded soil and the total-N and fertilized-N content of the soil and plants were measured in the greenhouse plots. Direct measurement of N₂+N₂O emission from field and greenhouse plots indicated that the young rice plant facilitates the efflux of N₂ and N₂O from the soil to the atmosphere. Little N gas was trapped in the rice-planted soils while large quantities were trapped in the unplanted soils. N losses due to denitrification accounted for only up to 10% of the loss of added N in planted soils in the field or greenhouse. The major losses of fertilizer N from both the field and greenhouse soils appear to have been the result of NH₃ volatilization.     

Copper,nitrogen, and Rhizobium japonicum relationships in determinate soybean

A relationship among Cu, N, and Rhizobium japonicum was hypothesized because previous research had shown that (a) 35% or more legumes in the Atlantic Coastal Plain have Cu concentrations of 6 mg kg^‐1 or less, (b) Cu influences N fixation in some legumes, and (c) irrigated soybean (Glycine max L. Merr.) can accumulate most of its N through fixation. Soybean were grown on a Cu‐deficient Norfolk (fine‐loamy, siliceous, thermic Typic Paleudult) loamy sand with 3 fertilizer sources of Cu, 2 strains of R. japonicum, and with or without 336 kg ha^‐1 of N fertilizer. Application of Cu significantly increased the number of pods plant^‐1 suggesting pod abortion in determinate soybean may be caused by low Cu, but seed yield was not increased. Fertilization with N increased vegetative growth, but not total biomass or seed yield. Inoculation with R. japonicum strain 110 significantly increased seed yield by 0.3 Mg ha^‐1 compared to strain 587. The yield increase was similar with or without fertilizer N application indicating strain response was not totally caused by improved N efficiency. There was no relationship between seed yield and nodule occupancy as measured by the ELISA technique.     

Incorporation of 15N into various nitrogenous compounds in intact soybean nodules after exposure to 15N2 gas

The intact nodules attached to the upper part of soybean roots were exposed to ¹⁵N₂ and the incorporation of ¹⁵N into various soluble nitrogen constituents was investigated. Results indicated that ammonia, a primary product of N₂ fixation, was located in more than two compartments. Ammonia reduced from N₂ gas seemed to be incorporated firstly into glutamine especially amido-group nitrogen. Newly fixed nitrogen was secondly incorporated into glutamic acid and alanine in this sequence. These results suggested that fixed ammonia was assimilated by glutamine synthetase/glutamate synthase pathway. Turn-over rate of allantoin plus allantoic acid and serine was relatively high, although apparently these compounds were not primary products of newly fixed ammonia. ¹⁵N content of allantoin was always higher than that of allantoic acid. ¹⁵N incorporation to aspartic acid and asparagine was relatively slow, especially in early period. In bacteroid fraction there is much amount of ammonia comparing with other compounds, while allantoin and asparagine were presented exclusively in cytosol. ¹⁵N was incorporated into nitrate within a few minutes especially in bacteroids.