Nitrogen fixation by nodulated soybean under tropical field conditions estimated by the 15N isotope dilution technique

The contribution of biological N₂ fixation to the N nutrition of nodulated soybean was estimated using the ¹⁵N isotope dilution technique and a non-nodulating soybean isoline as a non-fixing control plant. The plants were grown in the field in concrete cylinders (60 cm dia) and harvested at seven stages of plant growth. Labelled N was added to the soil either as labelled organic matter before planting or in seven small additions (2kg N ha^?1) of (NH₄)₂SO₄ during the growing period.There was good agreement between isotope dilution estimates of nitrogen fixation for the two labelling methods. Acetylene reduction assays on intact root systems greatly underestimated N₂ fixing activity. The difference in total N between nodulated and non-nodulated plants generally gave higher estimates compared with the isotope technique. The data indicate that this was because nodulated plants recovered more N from the soil than the non-nodulated plants. After 92 days of growth, the soybean derived approximately 250kg N ha^?1 from biological N₂ fixation.     

Assessment of the relative uptake of added and indigenous soil nitrogen by nodulated legumes and reference plants in the 15N dilution measurement of N2 fixation: Glasshouse application of method

A method for calculating the relative uptake (R) of added N and indigenous soil N by a legume (Trifolium subterraneum) and non-legume (Lolium rigidum), growing together, was investigated in two pot experiments. In the first experiment, ¹⁵N-labelled sodium nitrate was applied to the soil surface at rates equivalent to 0.3 or 1.0kg N ha^?1. Twenty one days later, the legume had fixed about 95% of its total N and this was unaffected by N addition. There was no difference in R values between legume and non-legume at both N rates.In the second experiment using a soil of higher total N, sodium nitrate or ammonium sulphate were surface-applied at a rate equivalent to 1 kg N ha^?1 and harvests were made at 3, 6, 12 and 27 days after N addition. Fixation of atmospheric N₂ by the legume did not begin until day 12 but accounted for about 40% of the total N assimilated by the legume by day 27. There was no difference in R values between legume and non-legume throughout the growth period when sodium nitrate was applied. However, when ammonium sulphate was added to label to soil N, the uptake of added N relative to indigenous soil N was greater for the non-legume than the legume. This caused an overestimation (51 vs 43%) of the proportion on N fixed by the legume when compared with that for the control or sodium nitrate treatments.     

Growth and nitrogen fixation by Medicago littoralis in pot experiments: Effect of plant density and competition from Lolium multiflorum

The legume Medicago littoralis cv. Harbinger, was grown either alone (1–4 plants per pot) or with Lolium multiflorum (ryegrass) at a total of 4 plants per pot, using two soils of contrasting N status. An ¹⁵N dilution technique was used to distinguish the amounts of plant N due to N₂ fixation and to N uptake from soil. Medic outyielded (dry weight and total plant N) ryegrass in a soil which released low amounts of inorganic N (Roseworthy) but ryegrass outyielded medic in a soil of higher N availability (Avon).For both soils, all combinations of medic and ryegrass plants utilized 70–73% of the inorganic N released on incubation. Competition from ryegrass invariably reduced yields of dry matter, total N, and fixed N of the medic plants, especially in the Avon soil. For both soils, the percentage reduction in the amounts of fixed N resulting from competition from ryegrass was directly proportional to the percentage increase of plant dry matter due to ryegrass. Medic plants grown in Roseworthy soil contained much higher proportions of N due to N₂-fixation than did medic plants grown in Avon soil. The amounts of plant N, fixed N and plant dry weight increased with increasing numbers of medic plants, when grown alone in Roseworthy soil, but not in the Avon soil containing more than two plants per pot. Nevertheless, irrespective of the soil used, medic numbers per pot, or competition from ryegrass, the amounts of fixed N correlated well with total N and with dry matter yields of medic plants. The proportions of fixed N to total N varied consistently in each of the medic plant parts (roots < = leaves < stems < pods).     

Isotopic fractionation during N2 fixation by four tropical legumes

To quantify the contribution of biological nitrogen fixation (BNF) to legume crops using the ¹⁵N natural abundance technique, it is necessary to determine the ¹⁵N abundance of the N derived from BNF—the B value. In this study, we used a technique to determine B whereby both legume and non-N₂-fixing reference plants were grown under the same conditions in two similar soils, one artificially labelled with ¹⁵N, and the other not. The proportion of N derived from BNF (%Ndfa) was determined from the plants grown in the ¹⁵N-labelled soil and it was assumed that the %Ndfa values of the legumes grown in the two soils were the same, hence the B value of the legumes could be calculated. The legumes used were velvet bean (Mucuna pruriens), sunnhemp (Crotalaria juncea), groundnut (Arachis hypogaea) and soybean (Glycine max) inoculated, or not, with different strains of rhizobium. The values of %Ndfa were all over 89%, and all the legumes grown in unlabelled soil showed negative δ¹⁵N values even though the plant-available N in this soil was found to be approximately +6.0‰. The B values for the shoot tissue (B_s) were calculated and ranged from approximately −1.4‰ for inoculated sunnhemp and groundnut to −2.4 and −4.5‰ for soybean inoculated with Bradyrhizobium japonicum strain CPAC 7 and Bradyrhizobium elkanii strain 29W, respectively. The B (B_wp) values for the whole plants including roots, nodules and the original seed N were still significantly different between the soybean plants inoculated with CPAC 7 (−1.33‰) and 29W (−2.25‰). In a parallel experiment conducted in monoxenic culture using the same soybean variety and Bradyrhizobium strains, the plants accumulated less N from BNF and the values were less negative, but still significantly different for soybean inoculated with the two different Bradyrhizobium strains. The results suggest that the technique utilized in this study to determine B with legume plants grown in soil in the open air, yields B values that are more appropriate for use under field conditions.     

Slow-release 15N fertilizer formulations to measure N2-fixation by isotope dilution

Pot experiments were carried out to examine the effects of slow-release fertilizer formulations on estimates of N₂-fixation determined by the isotope dilution method. Soybeans were used as the N₂-fixing plants, with non-nodulated soybeans and maize as the non-fixing controls. The ¹⁵N-fertilizer formulations used were (¹⁵NH₄)SO₄, K¹⁵NO₃, gypsum-pelleted K¹⁵NO₃, (¹⁵NH₄)₂SO₄ + glucose, ground plant material enriched with ¹⁵N or ¹⁵N-oxamide. The estimate of the amount of N₂ fixed by the nodulated soybean plants depended upon both the control plant and the fertilizer formulation used. Maize took up N later than non-nodulated soybean and estimates of soil N-pool (soil “A” value + fertilizer N added) calculated from the enrichment of this control were about twice as large as those calculated from the enrichment of non-nodulated soybean receiving the same fertilizer treatment. As a consequence, estimates of N₂-fixation relative to this control were lower than those relative to non-nodulating soybean (mean 140 mg N per pot compared with 292 mg N per pot). With unstablilized ¹⁵N salts errors were sufficient to produce negative estimates of fixation relative to maize. Even with a “well-matched” control (non-nodulated soybean) estimates of fixation varied with fertilizer formulation.     

Nitrogen rhizodeposition in agricultural crops: Methods,estimates and future prospects

The objective of the present review was to present the current knowledge on nitrogen (N) rhizodeposition, including techniques for ¹⁵N labelling of agricultural plants, amounts of N rhizodeposition and its fate in soil. Rhizodeposition is the process of release of organic and inorganic compounds from living plant roots. It is often quantified in terms of carbon (C) and less often as N derived from rhizodeposition (NdfR). Rhizodeposition of N can be estimated by labelling plants with ¹⁵N and following its fate in soil. Most methods used for labelling plants with ¹⁵N can only be applied after appearance of the first leaf and only allow pulse or multiple pulse labelling. Only the split-root technique and the application of gaseous ¹⁵N allow continuous labelling. All methods available at present have their flaccidities mostly due to the fact that the application of N is not following its physiological pathway of assimilation or by using artificial conditions. In the studies reviewed, amounts of N rhizodeposits ranged from 4% to 71% of total assimilated plant N. In legumes the median was 16% and in cereals it was 14%. Rhizodeposits were 15–96% of the below-ground plant biomass (BGP). In legumes the median was 73% and in cereal it was 57%. The high variability of these results shows the need for more investigations on N rhizodeposition looking especially on the factors influencing the amounts released in different plant species under field conditions.     

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

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

The 15N-isotope dilution technique applied to the estimation of biological nitrogen fixation associated with Paspalum notatum cv. batatais in the field

The contribution of associated biological nitrogen fixation to the nitrogen nutrition of Paspaulum notatum cv. batatais was estimated using the ¹⁵N-isotope dilution technique. The plants were grown in the field in concrete cylinders (60 cm dia) filled with soil, to which were added small quantities of ¹⁵N-labelled fertilizer at frequent intervals over 12 months. The pensacola cultivar of P. notatum was used as a non-N₂-fixing control plant. This was justified by the observation that nitrogenase (intact core C₂H₂ reduction) activity associated with the pensacola cultivar was consistently much lower than that of the batatais cultivar.At the first harvest, no evidence for N₂ fixation associated with the batatais cultivar was obtained, probably because of slow establishment of the N₂-fixing association. However, at the subsequent three harvests the batatais cultivar exhibited a lower ¹⁵N-enrichment and yielded more N than the pensacola cultivar. These data together suggested that 8–25% of the N in the batatais cultivar originated from N₂ fixation.The grass Paspalum maritimum was also included in the experiment and exhibited low nitrogenase activity similar to that of the pensacola cultivar of P. notatum. However, the total N and ¹⁵N data of these two grasses were not in good agreement indicating that it is important for the use of the isotope dilution technique that control plants are of very similar physiology and growth habit.     

Effects of Multiple Reference Plants,Season, and Irrigation on Biological Nitrogen Fixation by Pasture Legumes using the Isotope Dilution Method

Abstract

The popular and widely used ¹⁵nitrogen (N)–isotope dilution method for estimating biological N fixation (BNF) of pasture and tree legumes relies largely on the ability to overcome the principal source of error due to the problem of selecting appropriate reference plants. A field experiment was conducted to evaluate the suitability of 12 non‐N₂‐fixing plants (i.e., nonlegumes) as reference plants for estimating the BNF of three pasture legumes (white clover, Trifolium repens L.; lucerne, Medicago sativa; and red clover, Trifolium pratense L.) in standard ryegrass–white clover (RWC) and multispecies pastures (MSP) under dry‐land and irrigation systems, over four seasons in Canterbury, New Zealand. The ¹⁵N‐isotope dilution method involving field ¹⁵N‐microplots was used to estimate BNF. Non‐N₂‐fixing plants were used either singly or in combination as reference plants to estimate the BNF of the three legumes. Results obtained showed that, on the whole, ¹⁵N‐enrichment values of legumes and nonlegumes varied significantly according to plant species, season, and irrigation. Grasses and herb species showed higher ¹⁵N‐enrichment than those of legumes. Highest ¹⁵N‐enrichment values of all plants occurred during late summer under dry‐land and irrigation conditions. Based on single or combined non‐N₂‐fixing plants as reference plants, the proportion of N derived from the atmosphere (% Ndfa) values were high (50 to 90%) and differed between most reference plants in the MSP pastures, especially chicory (Cichorium intybus), probably because it is different in phenology, rooting depth, and N‐uptake patterns compared to those of legumes. The percent Ndfa values of all plants studied also varied according to plant species, season, and irrigation in the MSP pastures. Estimated daily amounts of BNF varied according to pasture type, time of plant harvest, and irrigation, similar to those shown by percent Ndfa results as expected. Irrigation increased daily BNF more than 10‐fold, probably due to increased dry‐matter yield of pasture under irrigation compared to dry‐land conditions. Seasonal and irrigation effects were more important in affecting estimates of legume BNF than those due to the appropriate matching of N₂‐fixing and non‐N₂‐fixing reference plants.     

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

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

Nitrate removal from drained and reflooded fen soils affected by soil N transformation processes and plant uptake

Knowledge about nitrate transformation processes and how they are affected by different plants is essential in order to reduce the loss of valuable N fertiliser as well as to prevent environmental pollution due to nitrate leaching or N₂O emission after fertilisation or the reflooding of degraded fens with nitrate-containing municipal sewage. Therefore four microcosm ¹⁵N tracer experiments were performed to evaluate the effect of common wetland plants (Phalaris arundinacea, Phragmites australis) combined with different soil moisture conditions (from dry to reflooded) on nitrate turnover processes. At the end of experiment, the total formation of gaseous N compounds was calculated using the ¹⁵N balance method. In two experiments (wet and reflooded soil conditions) the N₂O and N₂ emissions were also directly determined.Our results show that in degraded fen soils, which process mainly takes place—denitrification or transformation into organic N compounds—is determined by the soil moisture conditions. Under dry soil moisture conditions (water filled pore space: 31%) up to 80% of the ¹⁵N nitrate added was transformed into organic N compounds. This transformation process is not affected by plant growth. Under reflooded conditions (water filled pore space: 100%), the total gaseous N losses were highest (77-95% of the ¹⁵N-nitrate added) and the transformation into organic N compounds was very low (1.8% of ¹⁵N nitrate added). Under almost all soil conditions plant growth reduced the N losses by 20-25% of the ¹⁵N nitrate added due to plant uptake. The N₂ emissions exceeded the N₂O emissions by a factor of 10-20 in planted soil, and as much as 30 in unplanted soil. In the treatments planted with Phragmites australis, N₂O emission was about two times higher than in the corresponding unplanted treatment. 15% of the N₂O and N₂ formed was transported via the Phragmites shoots from the soil into the atmosphere. By contrast, Phalaris arundinacea did not affect N₂O emissions and no emission via the shoots was observed.     

Nitrous oxide emissions from grain legumes as affected by wetting/drying cycles and crop residues

Grain legume production with rhizobial inoculation has drawn attention not only because of the economic value of nitrogen (N) fixation by grain legumes, but also because of the concern that N₂ fixation by grain legumes may enhance emissions of nitrous oxide (N₂O), a powerful greenhouse gas. However, the relationship between N₂O emissions and biological N₂ fixation by grain legumes is not well understood. The objective of this study was to quantify N₂O emissions associated with N₂ fixation by grain legumes as affected by wetting/drying cycles and crop residues. Two grain legumes, lentil (Lens esculenta Moench) and pea (Pisum sativum L.), either inoculated with two Rhizobium leguminosarum biovar viciae strains, 99A1 and RGP2, respectively, or fertilized with ¹⁵N-labeled fertilizer were grown in a controlled environment under three wetting/drying cycles. Profile N₂O concentrations and surface N₂O emissions were measured from the soil–plant systems, which were compared with those from a cereal, spring wheat (Triticum aestivum L. ac. Barrie). After harvest, crop residues were incorporated into soils that were seeded to spring wheat to evaluate the effect of crop residues on N₂O emissions. Results indicated that: (1) inoculating grain legumes with non-denitrifying rhizobia did not enhance N₂O emissions and the presence of grain legumes did not increase N₂O emissions compared with the cereal crop, and (2) profile N₂O accumulation and surface emissions were not related to the type of crop residues added to the soil, but related to the residual N applied previously as N fertilizer. This suggests that N₂O emissions are not directly related to biological N₂ fixation by grain legumes, and on a short time scale, N rich residues of N₂-fixing crops have a limited impact on N₂O emissions compared with N fertilization.     

Short-term measurements of uptake of nitrogen fixed in the rhizospheres of sorghum (Sorghum bicolor) and millet (Pennisetum americanum)

Summary In a series of short-term experiments root systems of young sorghum and millet plants inoculated with N₂-fixing bacteria were exposed to ¹⁵N₂-enriched atmospheres for 72 h. The plants were grown in a normal atmosphere for up to 22 days after the end of the exposure to allow them to take up the fixed N₂. Environmental conditions and genotypes of sorghum and millet were selected to maximise N₂-fixation in the rhizosphere. Detectable amounts of fixed N (> 16 g/plant) were rapidly incorporated into sorghum plants grown in a sand/farmyard manure medium, but measurable fixation was found on only one occasion in plants grown in soil. N₂ fixation was detectable in some experiments with soil-grown millet plants but the amounts were small (2–4 g/plant) and represented less than 1 % of plant N accumulated over the same period. In many cases there was no detectable ¹⁵N₂ incorporation despite measurable increases in ethylene concentration found during an acetylene reduction assay.Published as ICRISAT Journal Article No. JA 740     

Competition between inoculated and indigenous Rhizobium/Bradyrhizobium spp. strains for nodulation of grain and fodder legumes in Pakistan

Summary The competitive ability of inoculated and indigenous Rhizobium/Bradyrhizobium spp. to nodulate and fix N₂ in grain legumes (Glycine max, Vigna unguiculata, Phaseolus vulgaris) and fodder legumes (Vicia sativa, Medicago sativa, and Trifolium subterraneum) was studied in pots with two local soils collected from two different fields on the basis of cropping history. The native population was estimated by a most-probable-number plant infectivity test in growth pouches and culture tubes. The indigenous rhizobial/bradyrhizobial population ranged from 3 to 2×10⁴ and 0 to 4.4×10³ cells g^-1 in the two soils (the first with, the second without a history of legume cropping). Inoculated G. max, P. vulgaris, and T. subterraneum plants had significantly more nodules with a greater nodule mass than uninoculated plants, but N₂ fixation was increased only in G. max and P. vulgaris. A significant response to inoculation was observed in the grain legume P. vulgaris in the soil not previously used to grow legumes, even in the presence of higher indigenous population (>10³ cells g^-1 soil of Rhizobium leguminosarum bv phaseoli). No difference in yield was observed with the fodder legumes in response to inoculation, even with the indigenous Rhizobium sp. as low as <14 cells g^-1 soil and although the number and weight of nodules were significantly increased by the inoculation in T. subterraneum. Overall recovery of the inoculated strains was 38–100%, as determined by a fluorescent antibody technique. In general, the inoculation increased N₂ fixation only in 3 out of 12 legume species-soil combinations in the presence of an indigenous population of rhizobial/bradyrhizobial strains.     

The use of nitrogen-15 natural abundance and nitrogen yield of non-nodulating isolines to estimate nitrogen fixation by soybeans (Glycine max L.) across three elevations

Summary Dissimilarities in soil N uptake between N₂-fixing and reference non-N₂-fixing plants can lead to inaccurate N₂ fixation estimates by N difference and ¹⁵N enrichment methods. The natural ¹⁵N abundance ( ¹⁵N) method relies on a stabilized soil ¹⁵N pool and may provide reliable estimates of N₂ fixation. Estimates based on the ¹⁵N and differences in N yield of nodulating and non-nodulating isolines of soybean were compared in this study. Five soybeans from maturity groups 00, IV, VI, and VIII and their respective non-nodulating isolines were grown at three elevations differing in ambient temperature and soil N availability. Despite large differences in phenological development and N yield between the non-nodulating isolines, the ¹⁵N values measured on seeds were relatively constant within a site. The ¹⁵N method consistently produced lower N₂ fixation estimates than the N difference method, but only in three of the 15 observations did they differ significantly. The average crop N derived from N₂ fixation across sites and maturity groups was 81% by N difference compared to 71% by ¹⁵N. The magnitude of difference between the two methods increased with increasing proportions of N derived from N₂ fixation. These differences between the two methods were not related to differences in total N across sites or genotypes. The low N₂ fixation estimates based on ¹⁵N might indicate that the nodulating isolines had assimilated more soil N than the non-nodulating ones. A lower variance indicated that the estimates by N difference using non-nodulating isolines were more precise than those by ¹⁵N. Since the differences between the estimates were large only at high N₂ fixation levels (low soil N availability), either method may be used in most situations when a non-nodulating isoline is used as the reference plant. The ¹⁵N method may have a comparative advantage over N difference and ¹⁵N enrichment methods in the absence of a suitable non-N₂-fixing reference plant such as a non-nodulating isoline.     

Symbiotically-fixed and soil-derived nitrogen in legumes grown in pots in soils with different amounts of available nitrate

Symbiotically-fixed and soil-derived nitrogen have been measured in pot experiments for Medicago littoralis (medic), grown alone or with Lolium multiflorum (ryegrass) and for Pisum sativum (field pea). The four soils used contained organic matter labelled with ¹⁵N, and differed in their capacities to release available N. During a 4–12-week incubation each released inorganic N (NO^?₃) of approximately constant ¹⁵N atom% enrichment. In one soil, the mineralized N was supplemented by ¹⁵NO^?₃ of similar ¹⁵N atom% enrichment. Incubation of soils under intermittently moist and dry conditions increased N mineralization rates, but did not affect the ¹⁵N atom% enrichments of the released N.For all soils and treatments the amounts of soil-derived N taken up by plants equalled the amounts of available N in moist incubated, unplanted soils. The enrichment of ryegrass root N grown alone or with medic was slightly but consistently less than that of top N. Nitrogen of the legume nodules and pods (peas) was least enriched, followed by N of legume stems, leaves and roots; the ¹⁵N atom% enrichments of root N were 4–5 times those of nodule N.Peas generally outyielded and fixed more N than medic grown alone. Medic grown with ryegrass yielded least and fixed least N.For unamended soils, yields of legume dry matter and amounts of N fixed were greatest in Roseworthy or Avon sandy loam soils and least in Northfield clay loam. Addition of ¹⁵NO^?₃ to Avon soil decreased N fixed by peas and by medic grown alone or with ryegrass. For this soil, soil-derived N of plant tops exceeded fixed N of roots, even for unamended soil where fixation by legumes was relatively high. Thus, complete removal of plant tops would have produced a net loss of N from the soil, the net loss increasing with increasing amounts of ¹⁵NO^?₃ added.     

Nitrogen accumulation in three legumes and two cereals with emphasis on estimation of N2 fixation in the legumes by the natural 15N-abundance technique

Summary N accumulation and natural ¹⁵N abundance in three legumes (groundnuts, cowpeas, and soybeans) and in two cereals (sorghum and maize) were investigated over two seasons in Alfisols with and without N fertilization. Using the N uptake and natural ¹⁵N abundance of non-nodulating plants as the indication of N derived from soil and fertilizer, the per cent N derived from atmospheric N₂ was calculated for nodulated plants. In the first experiment, the groundnut genotype contained 85% atmosphere-derived N, but the percentage decreased with N application. Estimates of atmosphere-derived N by the N-difference and ¹⁵N-abundance techniques gave identical results. The percentages of atmosphere-derived N estimated by the two methods at different stages of groundnut growth were also similar. In the second experiment, atmosphere-derived N was estimated in plants grown with 0–200 kg ha^-1 applied N. The estimated atmosphere-derived N ranged from 42% to 61% for groundnuts from 33% to 77% for cowpeas, and from 24% to 48% for soybeans, depending on the amount of N applied. Inoculation with a Bradyrhizobium strain increased the percentage of atmospherederived N in soybean plants grown without any fertilizer N. The natural ¹⁵N abundance of sorghum and maize was very close to that of the non-nodulating groundnut, suggesting that these cereals can be used as reference plants in the estimation of atmosphere-derived N by the natural ¹⁵N-abundance method.ICRISAT Journal Article No. 876     

利用15NO3-标记法研究土壤微生物量氮的化学及生物有效性

采用加入含15N的硝态氮培养方法标记了土壤微生物量氮 ,然后利用碱解扩散法测定了标记土壤有效氮含量 ,温室培养法评价了小麦对标记的土壤微生物量氮的吸收情况。结果表明 ,碱解扩散法对土壤微生物量固持的15N的提取比率 (即提取液中15N原子百分超 /土壤15N原子百分超 )在 1 47～ 2 83之间(平均 2 0 1 ) ,碱解氮中约有 3 0 1 %～ 61 6% (平均 42 9% )来自土壤微生物固持氮。植物体15N丰度在0 749%～ 1 1 62 %之间 ,明显高于15N的自然丰度 ,表明土壤微生物固持的15N在小麦生长期间发生释放 ,为植物利用。土壤微生物固持氮对植物的有效性比率 (植物地上部分15N原子百分超 /土壤15N原子百分超 )在 3 7～ 7 1之间。可见 ,土壤微生物量固持氮有较高的化学及生物有效性     

Growth and Nitrogen Fixation in Dhaincha/Sorghum and Dhaincha/Sunflower Intercropping Systems Using 15Nitrogen and 13Carbon Natural Abundance Techniques

A field experiment on dhaincha, sunflower, and sorghum plants grown in monocropping and intercropping systems was conducted to evaluate growth and nitrogen (N₂) fixation using ¹³carbon (C) and ¹⁵N natural abundance techniques. Intercropping of sesbania/sorghum showed a greater efficiency than monocropping in producing dry matter during the entire growth period, whereas the efficiency of producing dry matter in the sesbania/sunflower intercropping was similar to that in the monocropping system. Moreover, sorghum plants (C4) were more competitive than sesbania (C3) for soil N uptake, whereas sesbania seemed to be more competitive than its associated sunflower (C3). Nitrogen uptake in the mixed stand of sesbania/sorghum was improved as a result of the increase in soil N uptake by the component sorghum and the greater root nodule activity of component sesbania without affecting the amount of N₂ fixed. The Δ ¹³C in plant materials was affected by plant species and the cropping system.     

高氮和NO2-对中亚热带森林土壤N2O和NO产生的影响

利用15N同位素标记方法,研究在两种水分条件即60％和90％ WHC下,添加硝酸盐(NH4NO3,N 300 mg kg-1)和亚硝酸盐(NaNO2,N 1 mg kg-1)对中亚热带天然森林土壤N2O和NO产生过程及途径的影响.结果表明,在含水量为60％ WHC的情况下,高氮输入显著抑制了N2O和NO的产生(p＜0.01);但当含水量增为90％ WHC后,实验9h内抑制N2O产生,之后转为促进.所有未灭菌处理在添加NO2-后高氮抑制均立即解除并大量产生N2O和NO,与对照成显著差异(p＜0.01),在60％ WHC条件下,这种情况维持时间较短(21 h),但如果含水量高(90％ WHC)这种情况会持续很长时间(2周以上),说明水分有效性的提高和外源NO2-在高氮抑制解除中起到重要作用.本实验中N2O主要来源于土壤反硝化过程,而且加入未标记NO2-后导致杂合的N2O(14N15NO)分子在实验21 h内迅速增加,表明这种森林土壤的反硝化过程可能主要是通过真菌的“共脱氮”来实现,其贡献率可多达80％以上.Spearman秩相关分析表明未灭菌土壤NO的产生速率与N2O产生速率成显著正相关性(p＜0.05),土壤含水量越低二者相关性越高.灭菌土壤添加NO2-能较未灭菌土壤产生更多的NO,但却几乎不产生N2O,表明酸性土壤的化学反硝化对NO的贡献要大于N2O.