The uptake and distribution of 15N2 into the various organs of Typha latifolia L.

Direct evidence of heterotrophic dinitrogen fixation associated with the emergent aquatic angiosperm, Typha latifolia L., was obtained through the exposure of actively growing plants to ¹⁵N₂ gas for 7 days in a gas-tight exposure vessel. Highest enrichments of ¹⁵N were found in roots/rhizomes and leaf bases. Slight enrichments were also found in the leaves due to translocation from the roots, rhizomes and leaf bases. Total fixed ¹⁵N values were 71.8 μg for the plant and 49.1 μg for the soil.Plants growing in silica sand, which received a nutrient solution containing combined nitrogen, exhibited higher enrichments and fixed 86% more ¹⁵N after exposure to ¹⁵N₂ gas than plants which received a nutrient solution lacking combined nitrogen. It is hypothesized that the concentration of combined nitrogen added was insufficient to repress nitrogen fixation and resulted in an increase in nitrogen fixation by associated microorganisms.Propane was used to trace the loss and movement of gases from the ¹⁵N₂ vessel and between the upper leaf chamber and the lower root chamber. Gas was rapidly exchanged between the upper and lower chambers through the leaves and roots of T. latifolia. Further investigations showed that propane moved at a rate of 1223 μmol day⁻¹ from the leaves to the roots and 2652 μmol day⁻¹ from the roots to the leaves. These data demonstrated that gases diffuse rapidly through the plant body of T. latifolia.     

Evidence against associative N2 fixation as a significant N source in long-term wheat plots

In monocropped cereal systems, annual N inputs from non-fertilizer sources may be more than 30 kg ha^-1. We examined the possibility that these inputs are due to biological N₂ fixation (BNF) associated with roots or decomposing residues. Wheat was grown under greenhouse conditions in pots (34 cm long by 10 cm diameter) containing soil from a plot cropped to spring wheat since 1911 without fertilization. The roots and soil were sealed from the atmosphere and exposed to a¹⁵N₂-enriched atmosphere for three to four weeks during vegetative, reproductive or post-reproductive stages. This technique permitted detection of as little as 1 μg fixed N plant^-1 in plant material and 40 μg fixed N plant^-1 in soil. No fixation of¹⁵N₂ occurred during either of the first two labelling periods. In the final labelling period, straw returned to the soil was significantly enriched in¹⁵N, especially in a pot with a higher soil moisture content. Total BNF in this pot was 13 μg N plant^-1, or about 30 g N ha^-1. In a separate experiment with soil from the same plot, we detected BNF only when soil was amended with glucose at a high soil moisture content. Measured associative BNF was insufficient to account for observed N gains under field conditions. Lethbridge Research Centre contribution no. 3879488. Lethbridge Research Centre contribution no. 3879488.     

Carbon and nitrogen fluxes between beech and their ectomycorrhizal assemblage

To determine the exchange of nitrogen and carbon between ectomycorrhiza and host plant, young beech (Fagus sylvatica) trees from natural regeneration in intact soil cores were labelled for one growing season in a greenhouse with ¹³CO₂ and ¹⁵NO₃ ¹⁵NH₄. The specific enrichments of ¹⁵N and ¹³C were higher in ectomycorrhizas (EMs) than in any other tissue. The enrichments of ¹³C and ¹⁵N were also higher in the fine-root segments directly connected with the EM (mainly second-order roots) than that in bulk fine or coarse roots. A strict, positive correlation was found between the specific ¹⁵N enrichment in EM and the attached second-order roots. This finding indicates that strong N accumulators provide more N to their host than low N accumulators. A significant correlation was also found for the specific ¹³C enrichment in EM and the attached second-order roots. However, the specific enrichments for ¹⁵N and ¹³C in EM were unrelated showing that under long-term conditions, C and N exchange between host and EMs are uncoupled. These findings suggest that EM-mediated N flux to the plant is not the main control on carbon flux to the fungus, probably because EMs provide many different services to their hosts in addition to N provision in their natural assemblages.     

Nitrogen gas (N2+N2O) flux from urea applied to lowland rice as affected by green manure

A field study was conducted on a clay soil (Andaqueptic Haplaquoll) in the Philippines to directly measure the evolution of (N₂+N₂O)−¹⁵N from 98 atom %¹⁵N-labeled urea broadcast at 29 kg N ha⁻¹ into 0.05-m-deep floodwater at 15 days after transplanting (DT) rice. The flux of (N₂+N₂O)−¹⁵N during the 19 days following urea application never exceeded 28 g N ha⁻¹ day⁻¹. The total recovery of (N₂+N₂O)−¹⁵N evolved from the field was only 0.51% of the applied N, whereas total gaseous¹⁵N loss estimated from unrecovered¹⁵N in the¹⁵N balance was 41% of the applied N. Floodwater (nitrate+nitrite)−N in the 5 days following urea application never exceeded 0.14 g N m⁻³ or 0.3% of the applied N. Prior cropping of cowpea [Vigna unguiculata (L.) Walp.] to flowering with subsequent incorporation of the green manure (dry matter=2.5 Mg ha⁻¹, C/N=15) at 15 days before rice transplanting had no effect on fate of urea applied to rice at 15 DT. The recovery of (N₂+N₂O)−¹⁵N and total¹⁵N loss during the 19 days following urea application were 0.46 and 40%, respectively. Direct recovery of evolved (N₂+N₂O)−¹⁵N and total¹⁵N loss from 27 kg applied nitrate-N ha⁻¹ were 20% and 53% during the same 19-day period. The failure of directly-recovered (N₂+N₂O)−¹⁵N to match total¹⁵N loss from added nitrate-¹⁵N might be due to entrapment of denitrification end products in soil or transport of gaseous end products to the atmosphere through rice plants. The rapid conversion of added nitrate-N to (N₂+N₂O)−N, the apparently sufficient water soluble soil organic C for denitrification (101 μg C g⁻¹ in the top 0.15-m soil layer), and the low floodwater nitrate following urea application suggested that denitrification loss from urea was controlled by supply of nitrate rather than by availability of organic C.     

Measurement of N2-fixation in field-grown pigeonpea [Cajanus cajan (L.) Millsp.] using15N-labelled fertilizer

Two experiments were carried out from 1981 to 1983 in Vertisol field at ICRISAT Center, Patancheru, India to measure N₂-fixation of pigeonpea [Cajanus cajan (L.) Millsp.] using the¹⁵N isotope dilution technique. One experiment examined the effect of control of a nodule-eating insect on fixation while another in vestigated the effect of intercroping with cereals on fixation and the residual effect of pigeonpea on a succeeding cereal crop. Although both experiments indicated that at least 88% of the N in pigeonpea was fixed from the atmosphere, one result is considered fortuitous in view of the differential rates of growth of the legume and the control, sorghum [Sorghum bicolor (L.) Moench]. The difference method of calculation in dieated negative fixation and the results emphasized the problem of finding a suitable nonfixing control. In a second experiment, when all plants were confined to a known volume of soil to which¹⁵N fertilizer was added in the field, these problems were overcome, and isotope dilution and difference methods gave similar results of N₂-fixation of about 90%. In intercropped pigeonpea 96% of the total N was derived from the atmosphere. This estimate might be an artifact. There was no evidence of benefit from N fixed by pigeonpea to intercropped sorghum plants. Plant tissue¹⁵N enrichments of cereal crops grown after pigeonpea indicated that the cereal derived some N fixed by the previous pigeonpea. Thus residual benefits to cereals are not only an effect of ‘sparing’ of soil N.     

Comparison of two 15N labelling methods for assessing nitrogen rhizodeposition of pea

Two ¹⁵N labelling methods for assessing net rhizodeposition of nitrogen (N) in pea crop (Pisum sativum L.) were compared in the greenhouse and in the field: the cotton-wick (CW) and the split-root (SR) methods. Rhizodeposition is defined as the organic material lost from roots during their growth through the soil. CW is a method in which ¹⁵N urea was supplied to the plant in pulses via a wick threaded through the stem. In SR, the root system was divided between a hydroponic labelling compartment (LC) containing the labelling nutrient solution (1 or 5 mM ¹⁵NO₃–¹⁵NH₄) and a compartment filled with soil in which the amount of ¹⁵N rhizodeposition was assessed. The percentage of N derived from rhizodeposition (%Ndfr), was used to calculate the amount of N rhizodeposition which was obtained from the ratio of atom % ¹⁵N excess of the soil : atom % ¹⁵N excess of the roots. Above ground parts in the field accumulated markedly more dry matter and N than in the greenhouse, regardless of the labelling method. ¹⁵N enrichments of above ground parts were higher than those of roots recovered from the soil. Results indicated that amount of ¹⁵N applied to plants were lower in SR than in CW. Additionally, LC roots of SR tended to retain large amounts of ¹⁵N. As a consequence, atom % ¹⁵N excess of roots was less than 1% in SR, whereas most values varied from 1% to 4% in CW. However, relationships between enrichments of the soil and of the roots were different in SR and CW. It was not possible to compare the Ndfr:root-N ratio between the two methods, but the ratio of Ndfr:plant-N was found to be 10% higher in SR than in CW. Finally, relative to total plant-N, the total contribution of below ground parts to the N pool of the soil reached 22–25% at maturity for the two methods. From our experiments, we could not conclude that one method is better than the other for estimating either net rhizodeposition of N or the contribution of a pea plant to the soil N pool. However, CW is easier to adapt and monitor under field conditions than SR.     

Nitrogen fixation (C2H2 reduction) in soil samples from rhizosphere of rice grown under alternate flooded and nonflooded conditions

Summary In a greenhouse study the influence of alternate flooded and nonflooded conditions on the N₂-ase activity of rice rhizosphere soil was investigated by C₂H₂ reduction assay. The soil fraction attached to roots represent the rhizosphere soil. Soil submergence always accelerated N₂-ase and this effect was more pronounced in planted system. Moreover, rice plant exhibited phase-dependent N₂-ase with a maximum activity at 60 days after transplanting. The alternate flooded and nonflooded regimes resulted in alterations of the N₂-ase activity. Thus, the N₂-ase activity increased following a shift from nonflooded to flooded conditions, but the activity decreased when the flooded soil was returned to nonflooded condition by draining. However, the differential influence of the water regime on N₂-ase was not marked in prolonged flooded-nonflooded cycles. Microbial analysis indicated the stimulation of different groups of free-living and associative N₂-fixing microorganisms depending on the water regime.     

Influence of light intensity on the distribution of carbon and consequent effects on mineralization of soil nitrogen in a barley (Hordeum vulgare L.)-soil system

A pot experiment was conducted in a ¹⁴C-labelled atmosphere to study the influence of living plants on organic-N mineralization. The soil organic matter had been labelled, by means of a 200-days incubation, with ¹⁵N. The influence of the carbon input from the roots on the formation of microbial biomass was evaluated by using two different light intensities (I). Mineralization of ¹⁵N-labelled soil N was examined by following its fate in both the soil biomass and the plants. Less dry matter accumulated in shoots and roots at the lower light intensity. Furthermore, in all the plant-soil compartments examined, with the exception of rhizosphere respiration, the proportion of net assimilated ¹⁴C was lower in the low-I treatment than in the high-I treatment. The lower rates of ¹⁴C and ¹⁵N incorporation into the soil biomass were associated with less root-derived ¹⁴C. During the chamber period (¹⁴CO₂-atmosphere), mineralized amounts of ¹⁵N (measured as plant uptake of ¹⁵N) were small and represented about 6.8 to 7.8% of the initial amount of organic ¹⁵N in the soil. Amounts of unlabelled N found in the plants, as a percentage of total soil N, were 2.5 to 3.3%. The low availability of labelled N to microorganisms was the result of its stabilization during the 210 days of soil incubation. Differences in carbon supply resulted in different rates of N mineralization which is consistent with the hypothesis that roots induce N mineralization. N mineralization was higher in the high-I treatment. On the other hand, the rate of mineralization of unlabelled stable soil N was lower than labelled soil ¹⁵N which was stabilized. The amounts of ¹⁵N mineralized in planted soil during the chamber period (43 days) which were comparable with those mineralized in unplanted soil incubated for 210 days, also suggested that living plants increased the turnover rate of soil organic matter.     

Nitrous oxide and dinitrogen emissions from urine-affected soil under controlled conditions

A ¹⁵N labelling technique was used to measure N₂O and N₂ emissions from an undisturbed grassland soil treated with cow urine and held at 30 cm water tension and 20°C in a laboratory. Large emissions of dinitrogen were detected immediately following urine application to pasture. These coincided with a rapid and large increase in soil water-soluble carbon levels, some of this increase being attributed to solubilization of soil organic matter by high pH and ammonia concentrations. Emissions of nitrous oxide generally increased with time in contrast to dinitrogen fluxes which decreased as time progressed. Estimated losses of N₂O and N₂ over a 30 day period were between 1 to 5% and 30 to 65% of the urine N applied plus N mineralized from soil organic matter, respectively. Most of the N₂ and N₂O originated from denitrification with nitrification-denitrification being of minor significance as a source of N₂O. Comparisons of the ¹⁵N enrichments in the soil mineral N pools and the evolved N₂O suggested that much of the N₂O was produced in the 5–8 cm zone of the soil. It is concluded that established grassland soils contain large amounts of readily-oxidizable organic carbon which may be used by soil denitrifying organisms when nitrate is non-limiting and soil redox potential is lowered due to high rates of biological activity and high soil moisture contents. ei]{gnR}{fnMerckx}     

Distribution of fixed-N and nitrate-N in cowpea during pod development

If the quality and quantity of yields from cowpea (Vigna unguiculata [L.] Walp.) are to be maximised, a complete understanding of the N nutrition of the plant must be achieved. The N requirement for developing pods of this species may come from mobilization of N in vegetative tissue, biological N fixation and uptake of N from soil. In this study, the fate of a pulse of fixed ¹⁵N₂ or of ¹⁵NO₃-given to different cowpea plants during pod development was determined. The plants were grown in vermiculite in plastic pots that were able to be sealed with silicone adhesive and equipped with a rubber septum so that ¹⁵N₂ gas could be injected into the air space above the vermiculite, and gas losses would be eliminated. Nineteen days after injection of ¹⁵N₂ the pods, leaves, nodules and roots contained 65%, 15%, 9%, and 4%, respectively of the quantity of ¹⁵N₂ fixed. When ¹⁵NO₃-¹⁵N was taken up by other plants during this period, these plant parts contained 40%, 26%, 3% and 19%, respectively, of the total plant ¹⁵N. The percentage ¹⁵N in roots was greater, and that of ¹⁵N in nodules was lower, when ¹⁵NO₃-¹⁵N was applied than when ¹⁵N₂ was utilised by plants. These results indicate that, while a high percentage of fixed-N or NO₃-N given to cowpea plants moved to the developing pods, other sinks were competing for this newly-aquired N.     

Sealed lysimeters for the direct estimation of dinitrogen fixation by grain legume crops

Abstract A new technique has been devised for the direct estimation of the contribution of N₂-fixation to the total nitrogen of a legume crop. Sealed lysimeters and ancillary equipment are described by which it is possible to enclose in a gas-tight system the roots of some of the plants within the crop, together with their associated core of soil. The normal soil atmosphere can then be replaced by one containing ¹⁵N₂, thus allowing, from the ¹⁵N content of the resulting plants direct calculation of the N₂-fixation. Regular monitoring is necessary to ensure that soil O₂, CO₂ and moisture contents are maintained at normal field levels. The results indicate that the technique is capable of achieving its objectives and, provided the seedlings establish well initially, the resultant plants fully match the field average at final harvest. It has been possible to maintain the labelling of the soil atmosphere sufficiently constant to ensure that reliable and highly reproducible estimates of N₂-fixation are obtained. Using Pisum sativum cv. Meteor at densities of 160 plants m^?2, fixation accounted for about 90% of the total nitrogen uptake. The limitations and merits of the method are compared with those of the ¹⁵N-fertilizer dilution method.     

Natural 15N abundance of presumed N2-fixing and non-N2-fixing plants from selected ecosystems

Summary The ¹⁵N/¹⁴N ratios of plant and soil samples from Northern California ecosystems were determined by mass spectrometry. The ¹⁵N abundance of 176 plant foliar samples averaged 0.0008 atom % ¹⁵N excess relative to atmospheric N₂ and ranged from-0.0028 to 0.0064 atom % ¹⁵N excess relative to atmospheric N₂. Foliage from reported N₂-fixing species had significantly lower mean ¹⁵N abundance (relative to atmospheric N₂ and total soil N) and significantly higher N concentration (% N dry wt.) than did presumed non-N₂-fixing plants growing on the same sites. The mean difference between N₂-fixing species and other plants was 0.0007 atom % ¹⁵N. N₂-fixing species had lower ¹⁵N abundance than the other plants on most sites examined despite large differences between sites in vegetation, soil, and climate. The mean ¹⁵N abundance of N₂-fixing plants varied little between sites and was close to that of atmospheric N₂. The ¹⁵N abundance of presumed non-N₂-fixing species was highest at coastal sites and may reflect an input of marine spray N having relatively high ¹⁵N abundance. The ¹⁵N abundance of N₂-fixing species was not related to growth form but was for other plants. Annual herbaceous plants had highest ¹⁵N abundance followed in decreasing order by perennial herbs, shrubs, and trees. Several terrestrial ferns (Pteridaceae) had ¹⁵N abundances comparable to N₂-fixing legumes suggesting N₂-fixation by these ferns. On sites where the ¹⁵N abundance of soil N differs from that of the atmosphere, N₂-fixing plants can be identified by the natural ¹⁵N abundance of their foliage. This approach can be useful in detecting and perhaps measuring N₂-fixation on sites where direct recovery of nodules is not possible.     

Soil carbon and nitrogen across a chronosequence of woody plant expansion in North Dakota

In annual crops, the partitioning of photosynthates to support root growth, respiration and rhizodeposition should be greater during early development than in later reproductive stages due to source/sink relationships in the plant. Therefore, seasonal fluctuations in carbon dioxide (CO₂) and nitrous oxide (N₂O) production from roots and root-associated soil may be related to resource partitioning by the crop. Greenhouse studies used ¹³C and ¹⁵N stable isotopes to evaluate the carbon (C) partitioning and nitrogen (N) uptake by corn and soybean. We also measured the CO₂ and N₂O production from planted pots as affected by crop phenology and N fertilization. Specific root-derived respiration was related to the ¹³C allocated to roots and was greatest during early vegetative growth. Root-derived respiration and rhizodeposition were greater for corn than soybean. The ¹⁵N uptake by corn increased between vegetative growth, tasseling and milk stages, but the ¹⁵N content in soybean was not affected by phenology. A peak in N₂O production was observed with corn at the milk stage, suggesting that the corn rhizosphere supported microbial communities that produced N₂O. Most of the ¹⁵N-NO₃ applied to soybean was not taken up by the plant and negative N₂O production during vegetative growth and floral initiation stages suggests that soybean roots supported the reduction of N₂O to dinitrogen (N₂). We conclude that crop phenology and soil N availability exert important controls on rhizosphere processes, leading to temporal variation in CO₂ and N₂O production.     

Transformations of fertilizer nitrogen in soil

Summary Five crops of oats were grown over a 14-month period on a Chester silt loam soil fertilized with N¹⁵-labelled (NH₄)₂SO₄. All plant material from the first four crops was returned to the soil. Following a fifth crop, oat tops and roots were harvested, and the soil was subjected to repeated extractions by autoclaving in 0.01M CaCl₂. The distribution of N¹⁵ and of indigenous soil N among chemical fractions of the extracts, and in the acid-soluble and acid-soluble and acid-insoluble portions of the soil residues following 0.01M CaCl₂ extraction, was remarkably similar. Since appreciable equilibrations between added N¹⁵ and the more resistant forms of soil organic N is unlikely, it is postulated that fertilizer N became incorporated in newly-formed complexes, similar to those already present in the soil. This view is in harmony with the finding that percentage removals of total and N¹⁵-labelled N remained almost the same, even with recovery of approximately 55 per cent of the amounts originally present. N mineralization capacity of the soil was reduced appreciably as a result of extraction.     

Biological N2 Fixation in wetland rice fields: Estimation and contribution to nitrogen balance

This paper 1) reviews improvements and new approaches in methodologies for estimating biological N₂ fixation (BNF) in wetland soils, 2) summarizes earlier quantitative estimates and recent data, and 3) discusses the contribution of BNF to N balance in wetland-rice culture.Measuring acetylene reducing activity (ARA) is still the most popular method for assessing BNF in rice fields. Recent studies confirm that ARA measurements present a number of problems that may render quantitative extrapolations questionable. On the other hand, few comparative measures show ARA's potential as a quantitative estimate. Methods for measuring photodependent and associative ARA in field studies have been standardized, and major progress has been made in sampling procedures. Standardized ARA measurements have shown significant differences in associative N₂ fixation among rice varieties.The ¹⁵N dilution method is suitable for measuring the percentage of N derived from the atmosphere (% Ndfa) in legumes and rice. In particular, the ¹⁵N dilution technique, using available soil N as control, appears to be a promising method for screening rice varieties for ability to utilize biologically fixed N. Attempts to adapt the ¹⁵N dilution method to aquatic N₂ fixers (Azolla and blue-green algae [BGA]) encountered difficulties due to the rapid change in ¹⁵N enrichment of the water.Differences in natural ¹⁵N abundance have been used to show differences among plant organs and species or varieties in rice and Azolla, and to estimate Ndfa by Azolla, but the method appears to be semi-quantitative.Recent pot experiments using stabilized ¹⁵N-labelled soil or balances in pots covered with black cloth indicate a contribution of 10–30 kg N ha^-1 crop^-1 by heterotrophic BNF in flooded planted soil with no or little N fertilizer used.Associative BNF extrapolated from ARA and ¹⁵N incorporation range from 1 to 7 kg N ha^-1 crop^-1. Straw application increases heterotrophic and photodependent BNF. Pot experiments show N gains of 2–4 mg N g^-1 straw added at 10 tons ha^-1.N₂ fixation by BGA has been almost exclusively estimated by ARA and biomass measurements. Estimates by ARA range from a few to 80 kg N ha^-1 crop^-1 (average 27 kg). Recent extensive measurements show extrapolated values of about 20 kg N ha^-1 crop^-1 in no-N plots, 8 kg in plots with broadcast urea, and 12 kg in plots with deep-placed urea.Most information on N₂ fixed by Azolla and legume green manure comes from N accumulation measurements and determination of % Ndfa. Recent trials in an international network show standing crops of Azolla averaging 30–40 kg N ha^-1 and the accumulation of 50–90 kg N ha^-1 for two crops of Azolla grown before and after transplanting rice. Estimates of % Ndfa in Azolla by ¹⁵N dilution and delta ¹⁵N methods range from 51 to 99%. Assuming 50–80% Ndfa in legume green manures, one crop can provide 50–100 kg N ha^-1 in 50 days. Few balance studies in microplots or pots report extrapolated N gains of 150–250 kg N ha^-1 crop^-1.N balances in long-term fertility experiments range from 19 to 98 kg N ha^-1 crop^-1 (average 50 kg N) in fields with no N fertilizer applied. The problems encountered with ARA and ¹⁵N methods have revived interest in N balance studies in pots. Balances are usually highest in flooded planted pots exposed to light and receiving no N fertilizer; extrapolated values range from 16 to 70 kg N ha^-1 crop^-1 (average 38 kg N). A compilation of balance experiments with rice soil shows an average balance of about 30 kg N ha^-1 crop^-1 in soils where no inorganic fertilizer N was applied.Biological N₂ fixation by individual systems can be estimated more or less accurately, but total BNF in a rice field has not yet been estimated by measuring simultaneously the activities of the various components in situ. As a result, it is not clear if the activities of the different N₂-fixing systems are independent or related. A method to estimate in situ the contribution of N₂ fixed to rice nutrition is still not available. Dynamics of BNF during the crop cycle is known for indigenous agents but the pattern of fixed N availability to rice is known only for a few green manure crops.     

Production of nitrous oxide in artificially flooded and drained soils

Production of nitrous oxide (N₂O) was studied in one peaty and one sandy soil undergoing wetting and drying cycles. The background concentration of N₂O in the soil was compared with the N₂O produced during 4 hours of incubation with and without addition of acetylene. The concentration of N₂O in the soil under flooded conditions was relatively stable, and net consumption of N₂O was observed as often as net production. The reference area and drained soils showed somewhat different patterns compared to the flooded soils, which was probably an effect of intermediate soil water conditions. During flooding, the nitrous oxide made up less than 1% of total denitrification on 50% and 54% of the sampling occasions for the peaty and the sandy soil, respectively, and N₂O/(N₂O+N₂)-ratios exceeded 0.2 on only 6% and 3% of the sampling occasions. Under drained conditions and in the reference areas, the ratios showed a more even frequency distribution. Grouping the nitrous oxide production data for different seasons and field conditions, we found few seasonal trends. At the sandy site, mean production of N₂O was larger during the winter months. There were weak correlations between N₂O production and floodwater nitrate concentration, and between N₂O production and soil temperature. N₂O production in the reference area varied between consumption and 4.6 kg N ha^–1 month^–1 and in flooded and drained soil between consumption and 2.6 kg N ha^–1 month^–1.     

Estimation of biological nitrogen fixation associated withBrachiaria andPaspalum grasses using15N labelled organic matter and fertilizer

Summary Six pasture grasses,Paspalum notatum cv batatais,P. notatum cv pensacola,Brachiaria radicans, B. ruziziensis, B. decumbens andB. humidicola, were grown in concrete cylinders (60 cm diameter) in the field for 31 months. The soil was amended with either a single addition of¹⁵N labelled organic matter or frequent small (2 kg N. ha^–1) additions of¹⁵N enriched (NH₄)₂SO₄. In the labelled fertilizer treatment soil analysis revealed that there was a very drastic change in¹⁵N enrichment in plant-available nitrogen (NO ₃ ^– +NH ₄ ⁺ ) with depth. The different grass cultivars recovered different quantities of applied labelled N, and evidence was obtained to suggest that the roots exploited the soil to different depths thus obtaining different¹⁵N enrichments in soil derived N. This invalidated the application of the isotope dilution technique to estimate the contribution of nitrogen fixation to the grass cultivars in this treatment. In the labelled organic matter treatment the¹⁵N label in the plant-available N declined at a decreasing rate during the experiment until in the last 12 months the decrease was only from 0.274 to 0.222 atom % excess. There was little change in¹⁵N enrichment of available N with depth, hence it was concluded that although the grasses recovered different quantities of labelled N, they all obtained virtually the same¹⁵N enrichment in soil derived N. Data from the final harvests of this treatment indicated thatB. humidicola andB. decumbens obtained 30 and 40% respectively of their nitrogen from N₂ fixation amounting to an input of 30 and 45 kg N.ha^–1 year^–1 respectively.     

Waterlogging and fire impacts on nitrogen availability and utilization in a subtropical wet heathland (wallum)

Protein, amino acids and ammonium were the main forms of soluble soil nitrogen in the soil solution of a subtropical heathland (wallum). After fire, soil ammonium and nitrate increased 90- and 60-fold, respectively. Despite this increase in nitrate availability after fire, wallum species exhibited uniformly low nitrate reductase activities and low leaf and xylem nitrate. During waterlogging soil amino acids increased, particularly γ-aminobutyric acid (GABA) which accounted for over 50% of amino nitrogen. Non-mycorrhizal wallum species were significantly (P < 0.05) ¹⁵N-enriched (0.3–4.3‰) compared to species with mycorrhizal associations (ericoid-type, ecto-, va-mycorrhizal) which were strongly depleted in ¹⁵N (-6.3 to -1.8‰). Lignotubers and roots had δ¹⁵N signatures similar to that of the leaves of respective species. The exceptions were fine roots of ecto-, ecto/va-, and ericoid type mycorrhizal species which were enriched in ¹⁵N (0.1–2.4‰). The 5¹⁵N signatures of δ¹⁵N_{total soil N} and δ¹⁵N_{soil NH4+} were in the range 3.7–4.5‰, whereas δ¹⁵N_{soil NO3?} was significantly (P < 0.05) more enriched in ¹⁵N (9.2–9.8‰). It is proposed that there is discrimination against ¹⁵N during transfer of nitrogen from fungal to plant partner. Roots of selected species incorporated nitrogen sources in the order of preference: ammonium > glycine > nitrate. The exception were proteoid roots of Hakea (Proteaceae) which incorporated equal amounts of glycine and ammonium.     

Direct assessment of symbiotically fixed nitrogen in the rhizosphere of alfalfa

Rhizodeposition has been proposed as one mechanism for the accumulation of significant amounts of N in soil during legume growth. The objective of this experiment was to directly quantify losses of symbiotically fixed N from living alfalfa (Medicago sativa L.) roots to the rhizosphere. We used ¹⁵N-labeled N₂ gas to tag recently fixed N in three alfalfa lines [cv. Saranac, Ineffective Saranac (an ineffectively nodulated line), and an unnamed line in early stages of selection for apparent N excretion] growing in 1-m long polyvinylchloride drainage lysimeters in loamy sand soil in a greenhouse. Plants were in the late vegetative to flowering growth stage during the 2-day labelling period. We determined the fate of this fixed N in various plant organs and soil after a short equilibration period (2 to 4 days) and after one regrowth period (35 to 37 days). Extrapolated N₂ fixation rates (46 to 77g plant^–1 h^–1) were similar to rates others have measured in the field. Although there was significant accretion of total N in rhizosphere compared to bulk soil, less than 1% was derived from newly fixed N and there were no differences between the excreting line and Saranac. Loss of N in percolate water was small. These results provide the first direct evidence that little net loss of symbiotically-fixed N occurs from living alfalfa roots into surrounding soil. In addition, these results confirm our earlier findings, which depended on indirect ¹⁵N labelling techniques. Net N accumulation in soil during alfalfa growth is likely due to other processes, such as decomposition of roots, nodules, and above ground litter, rather than to N excretion from living roots and nodules.     

Short-term Nitrogen Fixation by Legume Seedlings and Resprouts After Fire in Mediterranean Old-fields

Fires may greatly alter the N budget of a plant community. During fire nitrogen is lost to the atmosphere. Although high light availability after fire promotes N₂-fixation, the presumably high soil N availability could limit N₂-fixation activity. The latter limitation might be particularly acute in legume seedlings compared with resprouts, which have immediate access to belowground stored carbon. We wished to learn whether early post-fire conditions were conducive to N₂-fixation in leguminous seedlings and resprouts in two types of grassland and in a shrubland and whether seedlings and resprouts incurred different amounts of N₂-fixation after fire. We set 18 experimental fires in early autumn on 6 plots, subsequently labelling 6 subplots (2 × 2 m²) in each community with ¹⁵NH₄⁺-N (99 atom % excess). For 9 post-fire months we measured net N mineralisation in the top 5 cm of soil and we calculated the fraction of legume N derived from the atmosphere (%Ndfa) in seedlings and resprouts. We used two independent estimates of the amounts of N derived from non-atmospheric sources in potentially N₂-fixing plants: mean soil pool abundance and the ¹⁵N-enrichment of non-legumes. Despite substantial soil net N mineralisation in all burned community types (about 2.6 g Nm⁻² during the first nine months after fire), the %Ndfa of various legume species was 52–99%. Legumes from both grasslands showed slightly higher N₂-fixation values than shrubland legumes. As grassland legumes grew in more belowground dense communities than shrubland legumes, we suggest that higher competition for soil resources in well established grass-resprouting communities may enhance the rate of N₂-fixation after fire. In contrast to our hypothesis, legume seedlings and resprouts from the three plant communities studied, had similar %Ndfa and apparently acquired most of their N from the atmosphere rather than from the soil.