Growth patterns of common bean cultivars affect the ‘B’ value required to quantify biological N2 fixation using the 15N natural abundance technique

Plant and Soil - While soil organic matter 13C isotope helped evaluate vegetation-related change in soil organic carbon (SOC), less is understood about δ13CSOC and SOC in reforested...     

Evidence from field nitrogen balance and 15N natural abundance data for the contribution of biological N2 fixation to Brazilian sugarcane varieties

Aims

In Brazil N fertilization of sugarcane (Saccharum spp.) is low compared to most other countries. ¹⁵N-aided studies and the occurrence of many N₂-fixing bacteria associated with cane plants suggest significant contributions from biological N₂ fixation (BNF). The objective of this study was to evaluate BNF contributions to nine cane varieties under field conditions using N balance and ¹⁵N natural abundance techniques.

Methods

The field experiment was planted near Rio de Janeiro in 1989, replanted in 1999 and harvested 13 times until 2004. Soil total N was evaluated at planting and again in 2004. Samples of cane leaves and weeds for the evaluation of ¹⁵N natural abundance were taken in 2000, 2003 and 2004.

Results

N accumulation of the commercial cane varieties and a variety of Saccharum spontaneum were persistently high and N balances (60 to 107?kg?N ha^?1?yr^?1) significantly (p?<?0.05) positive. The δ¹⁵N of leaf samples were lower than any of the weed reference plants and data obtained from a greenhouse study indicated that this was not due to the cane plants tapping into soil of lower ¹⁵N abundance at greater depth.

Conclusion

The results indicate that the Brazilian varieties of sugarcane were able to obtain at least 40?kg?N ha^?1?yr^?1 from BNF.     

Genotypic variation in biological nitrogen fixation by common bean

Field experiments were performed in Austria, Brazil, Chile, Colombia, Guatemala, Mexico and Peru as part of an FAO/IAEA Co-ordinated Research Programme to investigate the nitrogen fixing potential of cultivars and breeding lines of common bean (Phaseolus vulgaris L.). Each experiment included approximately 20 bean genotypes which were compared using the ¹⁵N isotope dilution method. Great differences in nitrogen fixation were observed between and within experiments, with average values of 35% N derived from atmosphere (% Ndfa) and highest values of 70% Ndfa being observed. These values which were larger than had been reported previously for common bean, were observed only when environmental factors were favorable. Therefore, common bean lines are available, which can support high biological nitrogen fixation. These can be used either directly as cultivars for production or in breeding programmes to enhance nitrogen fixation in other cultivars.     

Estimates of seasonal nitrogen fixation of annual subterranean clover-based pastures using the15N natural abundance technique

Annual pasture legumes play a key role in ley farming systems of southern Australia, providing biologically fixed nitrogen (N) to drive the production of the pastures as well as subsequent crops grown in rotation. Seasonal inputs of biologically fixed N in shoot biomass of the subterranean clover (Trifolium subterraneum) component of grazed annual pastures were assessed using the¹⁵N natural abundance technique and appropriately timed sampling of herbage dry matter (DM) for N accumulation. At three study sites spanning a gradient across the Western Australian wheatbelt from 300 to 600 mm annual rainfall the performance of the clover and non-legume herbs and grasses was examined as paired comparisons involving two management treatments expected to give contrasting effects on pasture productivity, botanical composition and N₂ fixation. The proportion of clover N derived from atmospheric N₂ fixation (%Ndfa) ranged from 65 to 95% across sites, treatments and sampling times. Amounts of fixed N accumulated in clover shoot biomass ranged from 50 to 125 kg ha⁻¹, and paralleled trends in clover production. Substantial increases in pasture production in high yielding treatments generally occurred without decrease in %Ndfa, suggesting that N₂ fixation was essentially non-limiting to performance of the clover component. Seasonal profiles for accumulation of fixed N were skewed towards the late winter and spring period, particularly in low plant density pastures following a cereal crop. There were seasonal, site and treatment-specific effects on the proportion of clover and non-legume pasture components and consequently clover yield and N₂ fixation were variably affected by competition from non-legume species.     

Ontogenetic variations on absorption and utilization of phosphorus in common bean cultivars under biological nitrogen fixation

Salinity is a major yield-reducing factor in coastal and arid, irrigated rice production systems. Salt tolerance is a major breeding objective. Three rice cultivars with different levels of salt tolerance were studied in the field for growth, sodium uptake, leaf chlorophyll content, specific leaf area (SLA), sodium concentration and leaf CO₂ exchange rates (CER) at photosynthetic active radiation (PAR)-saturation. Plants were grown in Ndiaye, Senegal, at a research station of the West Africa Rice Development Association (WARDA), during the hot dry season (HDS) and the wet season (WS) 1994 under irrigation with fresh or saline water (flood water electrical conductivity = 3.5 mS cm^-1). Relative leaf chlorophyll content (SPAD method) and root, stem, leaf blade and panicle dry weight were measured at weekly intervals throughout both seasons. Specific leaf area was measured on eight dates, and CER and leaf sodium content were measured at mid-season on the first (topmost) and second leaf. Salinity reduced yields to nearly zero and dry-matter accumulation by 90% for the susceptible cultivar in the HDS, but increased leaf chlorophyll content and CER at PAR- saturation. The increase in CER, which was also observed in the other cultivars and seasons, was explained by a combination of two hypotheses: leaf chlorophyll content was limited by the available N resources in controls, but not in salt-stressed plants; and the sodium concentrations were not high enough to cause early leaf senescence and chlorophyll degradation. The growth reductions were attributed to loss of assimilates (mechanisms unknown) that must have occurred after export from the sites of assimilation. The apparent, recurrent losses of assimilates, which were between 8% and 49% according to simulation with the crop model for potential yields in irrigated rice, ORYZA S, might be partly due to root decomposition and exudation. Possibly more importantly, energy-consuming processes, such as osmoregulation, interception of sodium and potassium from the transpiration stream in leaf sheaths and their subsequent storage, drained the assimilate supply. This revised version was published online in August 2006 with corrections to the Cover Date.     

Response to inoculation and N fertilization for increased yield and biological nitrogen fixation of common bean (Phaseolus vulgaris L.)

Common bean (Phaseolus vulgaris L.) is able to fix 20–60 kg N ha^–1 under tropical environments in Brazil, but these amounts are inadequate to meet the N requirement for economically attractive seed yields. When the plant is supplemented with N fertilizer, N₂ fixation by Rhizobium can be suppressed even at low rates of N. Using the ¹⁵N enriched method, two field experiments were conducted to compare the effect of foliar and soil applications of N-urea on N₂ fixation traits and seed yield. All treatments received a similar fertilization including 10 kg N ha^–1 at sowing. Increasing rates of N (10, 30 and 50 kg N ha^–1) were applied for both methods. Foliar application significantly enhanced nodulation, N₂ fixation (acetylene reduction activity) and yield at low N level (10 kg N ha^–1). Foliar nitrogen was less suppressive to nodulation, even at higher N levels, than soil N treatments. In the site where established Rhizobium was in low numbers, inoculation contributed substantially to increased N₂ fixation traits and yield. Both foliar and soil methods inhibited nodulation at high N rates and did not significantly increase bean yield, when comparing low (10 kg N ha^–1) and high (50 kg N ha^–1) rates applied after emergence. In both experiments, up to 30 kg N ha^–1 of biologically fixed N₂ were obtained when low rates of N were applied onto the leaves.     

Limitations of the N natural abundance method for estimating biological nitrogen fixation in Amazonian forest legumes

This study evaluates the ¹⁵N natural abundance method for estimating biological nitrogen fixation (BNF) in 12 secondary and five primary forest sites in central Amazonia. Investigations were based on spatially systematic sampling of 20 potentially N₂-fixing legume tree and liana species, and of a range of differing references representing the ‘soil-derived nitrogen’.The absence of systematic differences in the δ¹⁵N-signals of potential N₂-fixers and reference plants resulted in many invalid % Ndfa-estimates (i.e., the proportion of foliar N derived from BNF).Whereas the % Ndfa-estimates based on non-legumes were randomly distributed around zero, the use of non-N₂-fixing legumes resulted in largely positive % Ndfa-estimates. Grouping of reference species into species types was a valid strategy to reduce the risk of atypical values. Leaf litter was an inadequate measure of soil-derived nitrogen. The impact of the B-value was low or moderate at best, the percentage of negative % Ndfa-estimates remained unaltered. The manner of individually pairing putative N₂-fixers with reference plants only marginally affected the % Ndfa-estimates, the assumption of an overall site mean reference δ¹⁵N-signal was sufficient.Since legumes were aggregated in clusters, BNF locally affected the surrounding soil nitrogen in secondary regrowth. Negative correlations between the vegetation shares of potential N₂-fixers and leaf litter δ¹⁵N-signals indicate ¹⁵N-dilution from the input side. Interpolated reference plant δ¹⁵N-signals appear low in the surroundings of legume clusters in some of the older regrowth sites, confirming such ¹⁵N-dilution.We conclude that, with our current state of knowledge, the ¹⁵N natural abundance method fails to quantitatively estimate BNF in heterogeneous forests. Future research should focus on the search for a more direct measure of the plant-available δ¹⁵N-signal of the soil, as well as on plant–soil interactions and the resulting small-scale isotopic heterogeneity.

Zusammenfassung

Diese Arbeit untersucht die Eignung der ¹⁵N natürlichen Abundanz Methode zur Schätzung der Biologischen Stickstoffixierung (BSF) in 12 Sekundär- und 5 Primärwaldflächen Zentralamazoniens. Die Studie ist räumlich systematisch und umfasst 20 potentiell N₂-fixierende Leguminosen Baum- und Lianenarten, sowie verschiedene Referenzpflanzen die den ,,bodenbürtigen Stickstoff” repräsentieren.Da die δ¹⁵N-Signale der potentiellen N₂-Fixierer sich nicht systematisch von denen der Referenzpflanzen unterschieden, waren viele % Ndfa-Schätzwerte (Anteil des luftbürtigen Stickstoffs im Blatt) ungültig.Während die auf nicht-Leguminosen basierenden % Ndfa-Schätzwerte zufällig um Null verteilt waren, ergaben die nicht N₂-fixierenden Leguminosen überwiegend positive % Ndfa. Unsere Referenzpflanzengruppierung in Art-Typen war erfolgreich. Der Einfluss des B-Wertes war nur gering, der Anteil der ungültigen Schätzwerte blieb unbeeinflusst. Die Art in der potentielle N₂-Fixierer und Referenzpflanzen gepaart wurden, beeinflusste nur unwesentlich die % Ndfa-Schätzwerte; die Annahme eines gemittelten Referenz δ¹⁵N-Signals für jede Fläche war ausreichend.Wegen ihrer geklumpten Verteilung können N₂-fixierende Leguminosen den Bodenstickstoff in ihrer Umgebung beeinflussen. Die negative Korrelation zwischen dem Biomassenanteil potentieller N₂-Fixierer und den δ¹⁵N-Signalen der Blattstreu deutet auf eine ¹⁵N-Verdünnung des N-Eintrages. Visuelle Karteninterpretation bestätigt eine solche ¹⁵N-Verdünnung auf einigen älteren Sekundärwaldflächen.Zusammenfassend folgern wir, dass die ¹⁵N natürliche Abundanz Methode bei gegenwärtigem Wissensstand zur quantitativen BSF-Schätzung in heterogenen Wäldern ungeeignet ist. Zukünftige Forschung sollte sich auf eine schlüssige Methode zur Bestimmung des bodenbürtigen δ¹⁵N-Signals, sowie auf die Interaktionen zwischen Vegetation und Boden und der daraus resultierenden kleinräumigen δ¹⁵N-Variabilität konzentrieren.     

Minimizing the effect of mineral nitrogen on biological nitrogen fixation in common bean by increasing nutrient levels

Although common bean (Phaseolus vulgaris L.) has good potential for N₂ fixation, some additional N provided through fertilizer usually is required for a maximum yield. In this study the suppressive effect of N on nodulation and N₂ fixation was evaluated in an unfertile soil under greenhouse conditions with different levels of soil fertility (low=no P, K and S additions; medium = 50, 63 and 10 mg kg^–1 soil and high = 200, 256 and 40 mg kg^–1 soil, respectively) and combined with 5, 15, 60 and 120 mg N kg^–1 soil of ¹⁵N-labelled urea. The overall average nodule number and weight increased under high fertility levels. At low N applications, nitrogen had a synergistic effect on N₂ fixation, by stimulating nodule formation, nitrogenase activity and plant growth. At high fertility and at the highest N rate (120 mg kg^–1 soil), the stimulatory effect of N fertilizer on N₂ fixation was still observed, increasing the amounts of N₂ fixed from 88 up to 375 mg N plant^–1. These results indicate that a suitable balance of soil nutrients is essential to obtain high N₂ fixation rates and yield in common beans.     

Symbiotic nitrogen fixation in a tropical rainforest: 15N natural abundance measurements supported by experimental isotopic enrichment

* Leguminous trees are very common in the tropical rainforests of Guyana. Here, species-specific differences in N(2) fixation capability among nodulating legumes growing on different soils and a possible limitation of N(2) fixation by a relatively high nitrogen (N) and low phosphorus (P) availability in the forest were investigated. * Leaves of 17 nodulating species and 17 non-nodulating reference trees were sampled and their delta(15)N values measured. Estimates of N(2) fixation rates were calculated using the (15)N natural abundance method. Pot experiments were conducted on the effect of N and P availability on N(2) fixation using the (15)N-enriched isotope dilution method. * Nine species showed estimates of > 33% leaf N derived from N(2) fixation, while the others had low or undetectable N(2) fixation rates. High N and low P availability reduced N(2) fixation substantially. * The results suggest that a high N and low P availability in the forest limit N(2) fixation. At the forest ecosystem level, N(2) fixation was estimated at c. 6% of total N uptake by the tree community. We conclude that symbiotic N(2) fixation plays an important role in maintaining high amounts of soil available N in undisturbed forest.     

Application of 15N natural abundance technique for evaluating biological nitrogen fixation in oil palm ecotypes at nursery stage in pot experiments and at mature plantation sites

A range of different species of diazotrophic bacteria has been found in tissues and the rhizosphere of oil palm plants, suggesting a potential to benefit from biological nitrogen fixation (BNF). A few studies have confirmed that plantlets at nursery stage can benefit significantly from BNF after inoculation with Azospirillum spp. but no data are available regarding the benefit from naturally-occurring diazotrophic bacteria in oil palm. The results described here were derived from two pot trials laid out under controlled conditions with plantlets from two important regions for palm oil production in Brazil, as well as from different field sites of mature oil palm plantations. The ¹⁵N natural abundance technique was employed to estimate plant dependence on BNF (%Ndfa) by the different ecotypes grown in soil and previously characterized as hosting diazotrophic bacteria. From both pot trials it was possible to identify some ecotypes of high potential for N₂-fixation that reached in some cases approximately 50%Ndfa. However, the accuracy of measurement still needs to be improved using more suitable reference plants for pot experiments. Values of δ ¹⁵N signals from oil palm and reference plants in the field were inconclusive concerning any benefit from BNF to oil palm, owing to apparently high temporal and spatial variability of δ ¹⁵N of the plant-available N in the heterogeneous soil matrix for the different palm and reference plant tested.     

Measurement of biological dinitrogen fixation in grassland: Comparison of the enriched 15N dilution and the natural 15N abundance methods at different nitrogen application rates and defoliation frequencies

A plant mixture of white clover (Trifolium repens L.), red clover (Trifolium pratense L.), and ryegrass (Lolium perenne L.) was established in the spring of 1991 under a cover-crop of barley. Treatments were two levels of nitrogen (400 and 20 kg N ha^-1) and two cutting intensities (3 and 6 cuts per season). Fixation of atmospheric derived nitrogen was estimated by two ¹⁵N dilution methods, one based on application of ¹⁵N to the soil, the other utilising small differences in natural abundance of ¹⁵N.Both methods showed that application of 400 kg N ha^-1 significantly reduced dinitrogen fixation, while cutting frequency had no effect. Atmospheric derived nitrogen constituted between 50 and 64% of harvested clover nitrogen in the high-N treatment, while between 73% and 96% of the harvested clover nitrogen was derived from the atmosphere in the low-N treatment. The amounts of fixed dinitrogen varied between 31–72 kg N ha^-1 and 118–161 kg N ha^-1 in the high-N and low-N treatment, respectively. The highest values for biological dinitrogen fixation were estimated by the enriched ¹⁵N dilution method.Estimates of transfer of atmospheric derived nitrogen from clover to grass obtained by the natural ¹⁵N abundance method were consistently higher than those obtained by the enriched ¹⁵N dilution method. Neither mineral nitrogen application nor defoliation frequency affected transfer of atmospheric derived nitrogen from clover to grass.Isotopic fractionation of ¹⁴N and ¹⁵N (B value) was estimated by comparing results for nitrogen fixation obtained by the enriched ¹⁵N dilution and the natural ¹⁵N abundance method, respectively. B was on average +1.20, which was in agreement with a B value determined by growing white clover in a nitrogen free media.     

Pattern of natural 15N abundance in lakeside forest ecosystem affected by cormorant-derived nitrogen

Waterbirds are one of the most important groups of organisms inhabiting the land–water interface, especially with regard to mediating the transport of materials from the aquatic to the terrestrial environment. The great cormorant (Phalacrocorax carbo) is a colonial piscivorous bird that transports nutrients from fresh water to forest. We measured cormorant-derived nitrogen at two nesting colonies on the Isaki Peninsula and Chikubu Island at Lake Biwa, Japan, and analyzed the long-term effects of cormorant colonization on the forest nitrogen cycle, and the mechanisms of nitrogen retention. Three sites were examined in each colony: a currently occupied area, a previously occupied but now abandoned area, and a control area never colonized by cormorants. High nitrogen stable isotope ratios of cormorant excreta, the forest floor, mineral soil, and living plants showed cormorant-derived nitrogen in both occupied and abandoned areas. The relationship between δ¹⁵N and N content showed that the high δ¹⁵N of the excreta and N turnover in the soil were important at the occupied sites, whereas high δ¹⁵N of litter was important at the abandoned sites. Physiological changes of various organisms are also important for the N decomposition process. In conclusion, cormorant-derived nitrogen remains in the forest ecosystem as a result of two cormorant activities: heavy deposition of excreta and collection of nitrogen-rich nest material. Colony stage (occupied, abandoned, or never inhabited) and historical change of N decomposition process of an area can be identified from the relationship between δ¹⁵N and N content.     

15N自然丰度法在陆地生态系统氮循环研究中的应用

随着氮沉降的不断增加以及人们对全球变化问题的日益关注, 稳定同位素技术在全球变化研究中得到广泛的应用。因为植物和土壤的氮同位素组成记录了氮循环影响因子的综合作用, 并且具有测量简单以及不受取样时间和空间限制的优点, 所以氮同位素自然丰度法被用于氮循环的研究中。该文从氮循环过程中植物和土壤的氮分馏入手, 总结国内外相关文献, 阐述了植物和土壤氮自然丰度在预测生态系统氮饱和和氮循环长期变化趋势中的应用; 总结了利用树轮δ ¹⁵N法研究氮循环过程中应该注意的事项以及目前尚未解决的问题。     

Symbiotic N2 fixation by soybean in organic and conventional cropping systems estimated by 15N dilution and 15N natural abundance

Nitrogen (N) is often the most limiting nutrient in organic cropping systems. N₂ fixing crops present an important option to improve N supply and to maintain soil fertility. In a field experiment, we investigated whether the lower N fertilization level and higher soil microbial activity in organic than conventional systems affected symbiotic N₂ fixation by soybean (Glycine max, var. Maple Arrow) growing in 2004 in plots that were since 1978 under the following systems: bio-dynamic (DYN); bio-organic (ORG); conventional with organic and mineral fertilizers (CON); CON with exclusively mineral fertilizers (MIN); non-fertilized control (NON). We estimated the percentage of legume N derived from the atmosphere (%Ndfa) by the natural abundance (NA) method. For ORG and MIN we additionally applied the enriched ¹⁵N isotope dilution method (ID) based on residual mineral and organic ¹⁵N labeled fertilizers that were applied in 2003 in microplots installed in ORG and MIN plots. These different enrichment treatments resulted in equal %Ndfa values. The %Ndfa obtained by NA for ORG and MIN was confirmed by the ID method, with similar variation. However, as plant growth was restricted by the microplot frames the NA technique provided more accurate estimates of the quantities of symbiotically fixed N₂ (Nfix). At maturity of soybean the %Ndfa ranged from 24 to 54%. It decreased in the order ORG > CON > DYN > NON > MIN, with significantly lowest value for MIN. Corresponding Nfix in above ground plant material ranged from 15 to 26 g N m^-2, with a decreasing trend in the order DYN = ORG > CON > MIN > NON. For all treatments, the N withdrawal by harvested grains was greater than Nfix. This shows that at the low to medium %Ndfa, soybeans did not improve the N supply to any system but removed significant amounts of soil N. High-soil N mineralization and/or low-soil P availability may have limited symbiotic N₂ fixation.     

Soil moisture and potassium affect the performance of symbiotic nitrogen fixation in faba bean and common bean

Potassium (K) is reported to improve plant's resistance against environmental stress. A frequently experienced stress for plants in the tropics is water shortage. It is not known if sufficient K supply would help plants to partially overcome the effects of water stress, especially that of symbiotic nitrogen fixation which is often rather low in the tropics when compared to that of temperate regions. Thus, the impact of three levels of fertilizer potassium (0.1, 0.8 and 3.0 mM K) on symbiotic nitrogen fixation was evaluated with two legumes under high (field capacity to 25% depletion) and low (less than 50% of field capacity) water regimes. Plants were grown in single pots in silica sand under controlled conditions with 1.5 mM N (¹⁵N enriched NH₄NO₃). The species were faba bean (Vicia faba L.), a temperate, amide producing legume and common bean (Phaseolus vulgaris L.), a tropical, ureide producing species. In both species, 0.1 mM K was insufficient for nodulation at both moisture regimes, although plant growth was observed. The supply of 0.8 or 3.0 mM K allowed nodulation and subsequent nitrogen fixation which appeared to be adequate for respective plant growth. High potassium supply had a positive effect on nitrogen fixation, on shoot and root growth and on water potential in both water regimes. Where nodulation occurred, variations caused by either K or water supply had no consequences on the percentage of nitrogen derived from the symbiosis. The present data indicate that K can apparently alleviate water shortage to a certain extent. Moreover it is shown that the symbiotic system in both faba bean and common bean is less tolerant to limiting K supply than plants themselves. However, as long as nodulation occurs, N assimilation from the symbiotic source is not selectively affected by K as opposed to N assimilation from fertilizer.     

Measurement of nitrogen fixation by soybean in the field using the ureide and natural N abundance methods

Nitrogen fixation by field-grown soybean (Glycine max [L.] Merrill) was assessed by the natural ¹⁵N abundance and ureide methods. The field sites (five) and genotypes (six, plus two levels of inoculation on Bragg) were chosen to provide a range of proportions of plant N derived from nitrogen fixation (P). Genotypes K466, K468, nts1007, and nts1116 and Davis were included on the basis of their reported tolerance of the suppressive effects of nitrate on nodulation and nitrogen fixation. Bragg was included as a `nitrate-sensitive' genotype. Seeds of all genotypes were inoculated at sowing with Bradyrhizobium japonicum CB1809 (USDA136). Amounts of nitrate in the soil profile (0-1.2 meter depth) at sowing ranged from 70 (site 3) to 278 kilograms per hectare (site 5), resulting in large effects on plant nodulation, on the δ¹⁵N values of nodulated plants, on the relative abundance of ureide-N in vacuum-extracted sap (VES) and stem extracts, and finally on the estimates of P. There was no relationship between amount of soil nitrate at sowing and the δ¹⁵N of the plant-available soil N. Correlation matrices of the measured and calculated parameters indicated generally weak correlations between crop growth (dry matter and N) and the parameters of symbiotic activity (nodule weight, δ¹⁵N, relative ureide-N); correlations were strong and highly significant between nodulation and the measures of nitrogen fixation (δ¹⁵N, relative ureide-N; r = 0.79-0.92). Estimates of P ranged between 0 and 68% (δ¹⁵N) and between 6 and 56% (ureide) and were highly correlated (r = 0.97). Results indicated that the ureide method can be used with confidence to assess P by field-grown crops of soybean.     

Estimation of symbiotic dinitrogen fixation in alder forest by the method based on natural15N abundance

Annual N₂-fixation in virgin forest ecosystems has been measured using a¹⁵N natural abundance (¹⁵N) procedure. This method was compared to a¹⁵N labelled fertilizer isotopic dilution method. For young alders (5–6 years old), ¹⁵N of leaves gave results in good agreement with the isotopic dilution of fertilizer method. Since ¹⁵N variability was expected according to plant physiology, for alder trees, leaves were collected at various heights after the end of the growing season, and, to take account of isotopic variations coming from derived inputs, ¹⁵N of leaves of a large number of other plants in the same are were measured to give control values. Following this procedure, the ¹⁵N method gave reliable evaluation of the nitrogen supply, by through N₂-fixation, to alders, which were found to maintain high nitrogen fixing capacity in a sequence ranging from first stage of establishment of climactic formation. Moreover, the same method is reported to discriminate various origins ofAlnus glutinosa grown in natural conditions, possibly in relation to the genetic diversity of this species.     

Symbiotic nitrogen fixation in eight Acacia senegal provenances in dryland clays of the Blue Nile Sudan estimated by the 15N natural abundance method

The symbiotic biological N₂fixation by Acacia senegal was estimated using the ¹⁵N natural abundance (δ ¹⁵N) procedure on eight provenances collected from different environments and soil types grown in a clay soil in the Blue Nile region, Sudan. Balanites aegyptiaca (a non-legume) was used as a non-N₂-fixing reference plant to allow ¹⁵N-based estimates of the proportion of the Acacia N derived from atmospheric N₂ (N_dfa) to be calculated. Results show variation in leaf δ ¹⁵N between A. senegal and the reference plant and among years. The relative δ ¹⁵N values (‰) were higher in B. aegyptiaca than in the N₂-fixing acacia provenances. Provenances originally collected from clay soils fixed little N in the first year, but the amount fixed increased as the trees aged. All provenances showed a decrease in δ ¹⁵N with age. The N_dfa varied between 24% (Mazmoom provenance) and 61% (Rahad provenance) 4 years after planting. There was no significant difference in δ ¹⁵N between provenance groups based on soil type or rainfall at original growing site. The amount of N_dfa increased significantly with age in all provenances. The above-ground contribution of fixed N to foliage growth in a 4-year-old A. senegal was highest in the Rahad sand–soil provenance (46.7 kg N ha⁻¹) and lowest in the Mazmoom clay-soil provenance (28.7 kg N ha⁻¹). Our study represents the first use of the δ ¹⁵N method for estimating the N input by A. senegal to the clay plain soils of the gum belt in the Sudan.     

15N natural abundance in plants of the Amazon River floodplain and potential atmospheric N2 fixation

Summary The¹⁵N natural abundance values of various Amazon floodplain (várzea) plants was investigated. Samples of young leaf tissues were collected during three different periods of the river hydrography (low water, mid rising water and high water) and during one period in the Madeira River (high water). A large variation of¹⁵N abundance was observed, both among the different plant types and between the different flood stages. This variation probably, reflected, in part, the highly variable nature of the floodplain, sometimes dry and oxygenated and at other times inundated and anaerobic and, in part, changes in plant nitrogen metabolism. Comparison of the nitrogen isotopic composition of leguminous plants with that of non-leguminous plants showed that, on average, the¹⁵N abundance was lower in the legumes than non-legumes, suggesting active N-fixation. Also, the¹⁵N natural abundance in aquatic grasses of the generaPaspalum, was in general, lower than the¹⁵N abundance of aquatic grasses of the generaEchinochloa. As both of these grasses grow in the same general habitat, it appears thatPaspalum grasses may also be nitrogen fixers.