Nitrogen fixation by Nodularia spumigena Mertens (Cyanobacteriaceae). 2: Laboratory studies

The effects of changes in diurnal light patterns, salinity, and phosphorus on nitrogen fixation (as measured by acetylene reduction) by Nodularia spumigena Mertens were examined. As well, the effects of added inorganic nitrogen on growth, nitrogen fixation and heterocyt frequencies, and changes in nitrogen fixation and heterocyst frequencies during the growth cycle of Nodularia in cultures were determined.The diurnal pattern of nitrogenase activity in Nodularia was primarily light-induced, though dark activity did occur. Nitrogenase activity following a period of darkness exceeded the normal light rate (> 90 compared to 50 nmol · C₂H₂ reduced · ml^–1 · h^–1). Nitrogen fixation was reduced by high and very low salinities (5 to 10 was the optimum range), and added phosphorus stimulated nitrogenase in P-starved cells. Added nitrogen (ammonium or nitrate) had no effect on the growth of Nodularia, but in short term studies, ammonium completely inhibited nitrogenase activity. Heterocyst frequencies were greatest in the log phase of growth (to 40 per mm). During stationary phase, nitrogenase activity was negligable.     

Nitrogenase activity of two genotypes of Acacia mangium as affected by phosphorus nutrition

The specific nodulation, nitrogenase activity (acetylene reduction) and budgets of carbon allocation to respiration by nodulated roots were examined in two provenances of Acacia mangium Willd. grown in a glasshouse for 17 weeks to investigate the effects of soil phosphorus and genotypes of the host plant on symbiotic nitrogen fixation. Application of phosphorus (0–80 mg P kg^-1 soil) increased specific nodulation (g nodule dry weight g^-1 plant dry weight) of provenance Ma11 by two-fold and the percentage of nodulated root respiration allocated to nitrogenase by 50%, but had no effect on specific activity of nitrogenase or specific respiration coupled with nitrogenase activity. Improved phosphorus nutrition increased the specific nitrogenase activity of provenance Ma9 by 2-fold, the percentage of nodulated root respiration allocated to nitrogenase, and specific nitrogenase-linked respiration by 50%, respectively, but had no effect on the specific nodulation. The percentage of respiration coupled with nitrogenase activity in nodulated root respiration by provenance Ma9 was 60–70% higher than that in provenance Ma11, regardless of phosphorus levels applied. At the optimal level of phosphorus addition (10 mg P kg^-1 soil), provenance Ma9 had a lower dry mass than provenance Ma11. This was accompanied by a lower nodulated root respiration and a higher percentage of nodulated root respiration allocated to nitrogenase activity in provenance Ma9.     

Effects of fertilization rate and crop rotation on diazotrophic cyanobacteria in paddy field

Application of chemical fertilizers at the recommended level (medium fertility) or lower stimulated growth of the diazotrophic cyanobacterial population and nitrogenase activity in a paddy field. High fertilizer levels proved to be inhibitory to nitrogen-fixing cyanobacteria indicating that indiscriminate use of chemical fertilizers for a longer period drastically disturbed the natural ecological balance. The rice–mustard–moong (RMM) crop rotation was observed to be more suitable for cyanobacterial nitrogen fixation than rice–wheat–maize rotation. The cropped plots had higher nitrogenase activity than fallow plots. The low fertility coupled with RMM rotation were found to be best suited for promoting nitrogen fixation by cyanobacteria to supply the rice plants. A top dressing of chemical nitrogenous fertilizer drastically suppressed the cyanobacterial nitrogenase activity (ARA) within 12 h; the magnitude of inhibition varied with respect to the cropping system. The inhibition was overcome by the 10th day and the ARA value reached the preapplication value or even higher in the case of low fertility and medium fertility level plots. A regression equation was established to predict nitrogen fixation in a given soil ecosystem.     

Field measurement of symbiotic nitrogen fixation in an established lucerne ley using15N and an acetylene reduction method

Summary Lucerne is an important forage legume in the south and south-east of Sweden on well-drained soils. However, data is lacking on the apparent amount of nitrogen derived through N₂ fixation by field-grown lucerne. This report provides basic information on the subject. The experiment was performed in a lucerne ley grown 40 km north of Uppsala. The input of nitrogen through fixation to the above-ground plant material of an established lucerne (Medicago sativa L.) ley was estimate by¹⁵N methodology during two successive years. The amount of fixed N was 242 kg N ha^–1 in 1982 and 319 kg N ha^–1 in 1983. The proportion of N derived from the atmosphere (%Ndfa) was 70% and 80% for the two years respectively. The first harvest in both years contained a lower proportion fixed N. Both N₂ fixation and dry matter production were enhanced during the second year, particularly in the first harvest. The Ndfa was 61% in the first harvest in 1982, compared to 72% Ndfa during the same period in 1983. This demonstrates the strong influence of environment on both dry matter production and N₂ fixation capacity of the lucerne.In addition anin situ acetylene reduction assay was used in 1982 to measure the seasonal distribution of the N₂ fixation and in 1983 to study the effect of soil moisture on the N₂ fixation process. The seasonal pattern showed great dependence on physiological development and harvest pattern of the lucerne ley. The maximum rate of N₂ fixation occurred at the bud or early flower stage of growth and was followed by a rapid decline as flowering proceeded. After harvest the nitrogenase activity markedly decreased and remained low during at least two weeks until regrowth of new shoots began. Irrigation doubled the nitrogenase activity of the lucerne in late summer 1983, when soil moisture content in the top soil was near wilting point. No changes in nitrogenase activity did occur in response to watering earlier during the summer, when the soil matric potential was around –0.30 MPa.     

Variation among common-bean accessions (Phaseolus vulgaris L.) from the Iberian Peninsula for N2-dependent growth and phosphorus requirement

The common-bean often faces phosphorus deficiency in soils where it is grown. Such a deficiency is a major limitation to yield improvement, especially as the common-bean depends upon nitrogen fixation. Screening for symbiotic nitrogen fixation under phosphorus deficiency was performed with 33 common-bean accessions representing the diversity of 15 European market classes from the Iberian Peninsula. These accessions were inoculated withRhizobium tropici CIAT899 and grown in an aerated nitrogen-free nutrient solution at deficientversus sufficient phosphorous supplies (75 vs. 250 μmol plan⁻¹ week⁻¹) in a glasshouse. A large variability in N₂-dependent growth under P deficiency was found with most tolerance to P deficiency among late type IV Andean landraces, with the exception of 3 early type I Andean landraces. From this screening four contrasting landraces were selected for their high efficiency in the use of P for their symbiotic N nutrition, and compared with the cultivar Linex in fields of a reference production area over 3 years. The landraces from the Iberian Peninsula expressed a higher growth than the cultivar Linex, although they showed a lower nodulation. We hypothesize that the identified P tolerance among Iberian Peninsula accessions may be useful for improving symbiotic nitrogen fixation in the common-bean when growth is limited by available soil-P and could contribute to sustainable farming systems by reducing farmers’ dependence on fertilizers.     

Uptake of phosphate from soils and fertilizers as affected by soil P availability and solubility of phosphorus fertilizers

Fertilizers labelled with ³²P were used to measure amounts of phosphorus, P_s and P_F, taken up by Lolium perenne from available soil P and from P fertilizer respectively, when applied at a rate of 66 mg P·(kg soil^–1) in greenhouse experiments. The quantity P_s of phosphorus taken up from soil in the presence of P fertilizer was compared to the quantity P_o taken up from soil without P fertilizer. The quantity (P_s–P_o) is positive for low P_o values, i.e. in soils poor in available phosphorus, but is negative for high P_o values indicating that an input of P fertilizer can induce a decrease in the utilization of available soil phosphorus. Moreover, for a given soil, the quantity (P_s–P_o) depends on the chemical form of the fertilizer. The standard method of evaluation of P fertilizer efficiency is based on the assumption that P_s=P_o, but P_s can differ from P_o. This result can explain the contradictory data published from field experiments about the efficiency of the various P fertilizers.     

Changes in Asymbiotic, Heterotrophic Nitrogen Fixation on Leaf Litter of Metrosideros polymorpha with Long-Term Ecosystem Development in Hawaii

We measured nitrogenase activity (acetylene reduction) of asymbiotic, heterotrophic, nitrogen-fixing bacteria on leaf litter from the tree Metrosideros polymorpha collected from six sites on the Hawaiian archipelago. At all sites M. polymorpha was the dominant tree, and its litter was the most abundant on the forest floor. The sites spanned a soil chronosequence of 300 to 4.1 million y. We estimated potential nitrogen fixation associated with this leaf litter to be highest at the youngest site (1.25 kg ha^-1 y^-1), declining to between 0.05 and 0.22 kg ha^-1 y^-1 at the oldest four sites on the chronosequence. To investigate how the availability of weathered elements influences N fixation rates at different stages of soil development, we sampled M. polymorpha leaf litter from complete, factorial fertilization experiments located at the 300-y, 20,000-y and 4.1 million–y sites. At the youngest and oldest sites, nitrogenase activity on leaf litter increased significantly in the plots fertilized with phosphorus and “total” (all nutrients except N and P); no significant increases in nitrogenase activity were measured in leaf litter from treatments at the middle-aged site. The results suggest that the highest rates of N fixation are sustained during the “building” or early phase of ecosystem development when N is accumulating and inputs of geologically cycled (lithophilic) nutrients from weathering are substantial. Received 4 February 1999; accepted 29 March 2000.     

A function of cyanobacterial mats in phosphorus-limited tropical wetlands

Cyanobacterial mats are important components of oligotrophic wetland ecosystems in the limestone-based regions of the Caribbean. Our goals were to: (1) Estimate the biomass and primary production of cyanobacterial mats, quantify the extent of nitrogen fixation and measure the activity of alkaline phosphatase (APA) in representative marshes of northern Belize; (2) Record changes in these variables following nutrient additions. The mat biomass ranged from 200 to 700 g m^–2 AFDM, with the epipelon contributing up to 87% of the total. Tissue nitrogen was similar in all marshes (1.1–1.5%), while tissue phosphorus was extremely low (0.0055–0.0129%) and well correlated with the N:P ratio in water. Nitrogen fixation expressed as nitrogenase activity was high in some marshes (17.5 nmol C₂H₄ cm^–2 h^–1) and low (< 5 nmol C₂H₄ cm^–2 h^–1) in others depending mainly on the proportion of heterocyst-forming cyanobacteria (Nostocales, Stigonematales) in the mat. Alkaline phosphatase activity was positively correlated with the N:P ratio of the mat. Experimental addition of phosphorus resulted in significant increase in primary production and nitrogen fixation while it suppressed the APA activity. The presented data clearly showed that oligotrophic marshes of northern Belize are strongly P limited. Increased input of phosphorus would profoundly change their structure and functions.     

Effects of plant species on phosphorus availability in a range of grassland soils

Vegetative conversion from grass to forest may influence soil nutrient dynamics and availability. A short-term (40 weeks) glasshouse experiment was carried out to investigate the impacts of ryegrass (Lolium perenne) and radiata pine (Pinus radiata) on soil phosphorus (P) availability in 15 grassland soils collected across New Zealand using ³³P isotopic exchange kinetics (IEK) and chemical extraction methods. Results from this study showed that radiata pine took up more P (4.5–33.5 mg P pot^–1) than ryegrass (1.1–15.6 mg pot^–1) from the soil except in the Temuka soil in which the level of available P (e.g., E _1min P_i, bicarbonate extractable P_i) was very high. Radiata pine tended to be better able to access different forms of soil P, compared with ryegrass. There were no significant differences in the level of water soluble P (Cp, intensity factor) between soils under ryegrass and radiata pine, but the levels of Cp were generally lower compared with original soils due to plant uptake. The growth of both ryegrass and radiata pine resulted in the redistribution of soil P from the slowly exchangeable P_i pool (E _{> 10m} P_i, reduced by 31.8% on the average) to the rapidly exchangeable P_i (E _1min-1d P_i, E _1d-10m P_i) pools in most soils. The values of R/r ₁ (the capacity factor) were also generally greater in most soils under radiata pine compared with ryegrass. Specific P mineralisation rates were significantly greater for soils under radiata pine (8.4–21.9%) compared with ryegrass (0.5–10.8%), indicating that the growth of radiata pine enhanced mineralisation of soil organic P. This may partly be ascribed to greater root phosphatase activity for radiata pine than for ryegrass. Plant species × soil type interactions for most soil variables measured indicate that the impacts of plant species on soil P dynamics was strongly influenced by soil properties.     

Nutrient Limitation to Nitrogen Fixation in Young Volcanic Sites

I used measures of ¹⁵N natural abundance and of nitrogenase activity (acetylene reduction) to examine whether the supply of non-N nutrients limits rates of N₂ fixation on young volcanic substrates in Hawaii. Leaves of the dominant tree (Metrosideros polymorpha, a nonfixer) were strongly depleted in ¹⁵N in control plots (–10.8 to –11.1⁰/₀₀). More than 5 y of repeated fertilization with P increased δ¹⁵N to –8.9 to –9.9⁰/₀₀, and the addition of all other essential plant nutrients (except N) together with P further increased ¹⁵N to –8.1 to –9.3⁰/₀₀. This pattern is consistent with enhanced N₂ fixation, because newly fixed N would have a δ¹⁵N near 0⁰/₀₀. Assays of nitrogenase activity in the experimental plots demonstrated that potential N fixation associated with nonvascular plants and with tree and fern litter were increased significantly by additions of P and by the combined nutrient treatment; when these were added together, the increase in nitrogenase activity was 6- to 11-fold over control plots. The supply of P and other weathering-derived nutrients constrains rates of N₂ fixation in these young volcanic sites and thereby contributes to the maintenance of N limitation to primary production and other ecosystem processes. Received 7 January 1999; accepted 3 May 1999.     

A multiple-trait breeding program for improving the symbiosis for N2 fixation betweenMedicago sativa L. andRhizobium meliloti

Summary The goal of breeding alfalfa for increased N₂ fixation potential is addressed. A chronological progression of breeding, physiological, microbiological, and plant pathological research is described. Studies describing the interrelationships among plant morphological, plant physiological, andRhizobium effectiveness traits are summarized. It was concluded that N₂ fixation in alfalfa is affected by coordinated responses among many physiological and biochemical traits. The simultaneous improvement of many factors in the symbiosis requires a comprehensive multiple-step breeding program. The current program includes selection in the glasshouse for seedling vigor,Rhizobium preference, shoot growth, nodule mass, root growth, nitrogenase (as measured by acetylene reduction), and nodule enzyme activity. The inclusion of additional selection traits is anticipated. Field evaluations of N₂ fixation potential of alfalfa populations are made with¹⁵N isotope dilution techniques. Plant germplasm sources used in the breeding program include several heterogeneous populations which have good combining ability and pest resistance when they are intercrossed. Significant progress has been made in achieving the goal of breeding alfalfa for improved N₂ fixation.     

Long-term changes in extractable soil phosphorus (P) in organic dairy farming systems

On five farms that have been managed organically for several years, all cultivated soils were sampled on two occasions. The time span between the first and second soil sampling varied from 6 to 12 years. At the first sampling the farms had been managed organically for 3, 4, 6, 11 or 53 years. The average phosphorus (P) concentrations in topsoil (0-20 cm) extracted by ammonium-acetate lactate solution (P-AL) decreased from the first to the second sampling on all farms. At the second soil sampling, the average topsoil P-AL concentrations on the five farms were 50, 64, 65, 75 and 119 mg P kg^–1, which is characterised as medium (26–65 mg P kg^–1) or high (66–150 mg P kg^–1). The decrease occurred mostly in soils with high and very high (>150 mg P kg^–1) P-AL concentrations at the first sampling. In these samples, the average value decreased from 100 to 87 and from 188 to 151 mg P kg^–1, respectively. In subsoil (20–40 cm), an increase from 15 to 27 mg P kg^–1 (P<0.01) in P-AL concentration was found in subsoil samples with low P-AL concentrations (0–25 mg P kg^–1) at the first sampling. This indicates P transfer from topsoil to subsoil. The pattern of decrease in topsoil was fairly well explained by farm level P balances. The average topsoil concentrations of P-AL were well below values for comparable conventional farms, but still at a level acceptable for crop production. Crop yields were acceptable, but the general pattern of decrease shows that in the future, some P should be supplied from external sources to avoid a further decrease, especially on the fields with lowest P-AL concentrations.     

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.     

Effect of NPK on growth and nitrogen fixation of Sesbania rostrata as a green manure for lowland rice (Oryza sativa L.)

The stem-nodulating tropical legume Sesbania rostrata is a promising green manure species for low input rice-farming systems in lowland areas. However, its success as biofertilizer depends on its biomass production and N₂ fixation. Nutrient imbalances and soils low in available nutrients can considerably affect biofertilizer production. Use of mineral N, P, and K fertilizers in growing S. rostrata as biofertilizer for lowland rice was therefore evaluated in pot experiments, and in the fields in Central Luzon, Philippines. Two soils low in Olsen P (3–7.3 mg kg^–1) and exchangeable K (0.05–0.08 meq 100g^-1) were used. Increasing amounts of N (0, 10, 20, 30, and 40 mg kg^-1), P (0, 50, and 100 mg kg^-1), and K (0, 100, 200, and 300 mg kg^-1) were applied to S. rostrata grown in the greenhouse, whereas small amounts of N, P, and K fertilizers (30, 15, and 33 kg ha^-1, respectively) were applied in the field.Mineral N application depressed nodulation and N₂ fixation in roots. It however, stimulated nodulation and N₂ fixation in stems. Applying 30 kg N ha^-1 as urea increased total N accumulation by S. rostrata and yield of the subsequent rice crop (IR64). Applied P and K both stimulated growth, nodulation, and N₂ fixation of S. rostrata. Nitrogen accumulation in P- and K-fertilized S. rostrata was about 40% higher than that in nonfertilized green manure. Thus integration of mineral N, P, and K fertilizers in a green manure-based rice-farming system can considerably improve biofertilizer production and increase rice grain yield.     

Nitrogen fixation,transfer and turnover in upland and lowland grass-clover swards,using15N isotope dilution

The stable isotope¹⁵N is particularly valuable in the field for measuring N fixation by isotope dilution. At the same time other soil-plant processes can be studied, including¹⁵N recovery, and nitrogen transfer between clover and grass. Three contrasting sites and soils were used in the present work: a lowland soil, an upland soil, and an upland peat. Nitrogen fixation varied from 12 gm^–2 on lowland soil to 2.7 gm^–2 on upland peat. Most N transfer occurred on upland soil (4.2 gm^–2) which, added to nitrogen fixed, made a total of 8.7 gm² input during summer 1985.¹⁵N recovery for the whole experiment was small, around 25%.Measurement of dead and dying leaves, stubble and roots, suggests that plant organ death is the first stage in N transfer from white clover to ryegrass, through the decomposer cycle. Decomposition was fastest on lowland soils, slowest on peat. On lowland soil this decomposer nitrogen is apparently subverted before transfer, probably by soil microbes.Variations in natural abundance of¹⁵N in plants were found in the two species on the different soils. These might be used to measure nitrogen fixation without adding isotope, but the need for many replicates and repeat samples would limit throughput.     

The influence of drainage and soil phosphorus on the vegetation of Douala-Edea Forest Reserve,Cameroun

All living trees (30 cm gbh) were enumerated in 104 80×80 m plots arranged along four transects in the Douala-Edea Forest Reserve Cameroun, a system of low-lying ancient coastal sand dunes interspersed by numerous streams and swamps. The extent of permanent and seasonal swamps was recorded for each plot. Two hundred thirty taxa were recognized of which 63% were identified to species. Mean tree density was 376 ha^–1, basal area 31.0 m² ha^–1 and number of species per plot 39. The Olacaceae were the most abundant family in terms of basal area, but the Euphorbiaceae the most frequently represented. The most abundant species wasCoula edulis (Olacaceae). Twenty-two plots had most of their area permanently or seasonally swamped. Percentage sand, silt and clay ranged between 32–100, 0–64, 0–21% respectively. The ranges for other variables recorded were: pH (2.7–5.4), organic carbon (1.5–12.4%), available phosphorus (7–90 ppm) and potassium (28–188 ppm), and nitrogen (ammonium 4–40 ppm, nitrate 1–12 ppm).Classification of the plots on the basis of six soil variables provided three large distinct groups: swamp plots and non-swamp plots, the latter divided into plots of low and high available soil phosphorus. Swamp plots were distinguished by high abundances ofProtomegabaria stapfiana andLibrevillea klainei, though correspondence ordination of plots in these groups showedP. stapfiana associated with more clayey soils andLibrevillea klainei (andGluema ivorensis) on the very sandy soils. Direct gradient analysis highlighted several species associated with these lower phosphorus soils. Available soil phosphorus is not as low at Douala-Edea as in parts of Korup, and the association of these Douala-Edea soils with the Caesalpinioideae is correspondingly weaker.Nomenclature follows Aubréville (1963–1983).The field work was supported by grant numbers RR00167-14, RR00167-15 and RR0167-16 from the National Institutes of Health for the operation of the Wisconsin Regional Primate Research Center, and N.A.T.O. Scientific Affairs grant number 1748 (to PGW and JSG). It was greatly facilitated by the skill and dedication of Ferdinand Namata. R. M. Polhill and D. W. Thomas assisted considerably in the identification of plant species. Sue Gartlan collected and collated the meteorological data, besides other field support. In the field phase J.S.G., P.G.W. and D.B.McK. were researches attached to the National Office of Scientific and Technical Research (ONAREST), Yaoundé. We thank M. D. Swaine for comments on earlier drafts, R. Letouzey for checking species nomenclature, the Computer Unit at Stirling University for facilities, M. Burnett for the typing at Stirling, and the Department of Soil Science, University of Wisconsin, for undertaking the soil chemical analyses.Reprint requests to D.McC.N. at Stirling.Publication No. 23-025 of the Wisconsin Regional Primate Center.     

Phosphorus fertilizer recovery from calcareous soils amended with humic and fulvic acids

Precipitation of Ca phosphates negatively affects recovery by plants of P fertilizer applied to calcareous soils, but organic matter slows the precipitation of poorly soluble Ca phosphates. To study the effect of high molecular weight organic compounds on the recovery of applied P, a mixture of humic and fulvic acids was applied to calcareous soils with different levels of salinity and Na saturation which were fertilized with 200 and 2000 mg P kg^–1 as NH₄H₂PO₄. Recovery was measured as the ratio of increment in Olsen P-to-applied P after 30, 60 and 150 days, and associated P forms were studied using sequential chemical fractionation and ³¹P NMR spectroscopy. Application of the humic-fulvic acid mixture (HFA) increased the amount of applied P recovered as Olsen P in all the soils except in one soil with the highest Na saturation. In soils with high Ca saturation and high Olsen P, recovery increased from < 15% in the absence of amendment to > 40% at a 5 g HFA kg^–1 amendment rate (30 days incubation and 200 mg P kg^–1 fertilizer rate). This is ascribed to inhibition of the precipitation of poorly soluble Ca phosphates, consistent with the sequential chemical extraction (reduction of the HCl extractable P) and P concentration in 0.01 M CaCl₂ (1:10 soil:solution ratio) extracts. ³¹P NMR spectra revealed that in non-amended samples, most spectral shifts were due to poorly soluble P compounds (carbonate apatite); on the other hand, at the 5 g HFA kg^–1 rate, significant amounts of amorphous Ca phosphate and dicalcium phosphate dihydrate (DCDP) were identified. The increase in the recovery of applied P due to HFA reveals a positive effect of the application of organic matter as soil amendments on the efficiency of P fertilizers and also explains that manures and other organic sources of P were more efficient increasing available P than inorganic P fertilizers in calcareous soils.     

Symbiotic effectiveness of Hup+ Rhizobium,VAM fungi and phosphorus levels in relation to nitrogen fixation and plant growth ofCajanus cajan

Effect of hydrogen uptake positive (Hup⁺) strain ofRhizobium sp. (pigeon pea) and VAM fungus (Glomus fasciculatum) was studied on the symbiotic parameters of pigeon pea (Cajanus cajan) cv. AL-15 at various levels of phosphorus. The Hup⁺ Rhizobium strain showed more nodulation, plant biomass and plant nitrogen content than its Hup⁻ counterpart. VAM infection in pigeon pea roots helped in translocating phosphorus from the soil and improved nitrogen fixation. Similarly, addition of phosphorus was found to play a positive role in enhancing all these parameters. Dual inoculation of Hup⁺ Rhizobium strain and VAM significantly increased nodulation, nitrogenase activity, plant nitrogen and phosphorus content and plant biomass compared to single inoculation of either organism and dual inoculation with Hup⁻ and VAM fungus.     

Response of Leucaena/Rhizobium symbiosis to mineral nutrients in Southwestern Nigeria

Pot and field experiments carried out at the International Institute of Tropical Agriculture (IITA) and at Fashola, Southwestern Nigeria, examined the effect of inoculation and N, P and micronutrients on nodulation and growth ofLeucaena leucocephala (Lam.) de Wit. In pot studies all parameters measured, except the percentage of nitrogen in shoots, were improved by inoculation, nitrogen and phosphorus. Micronutrients increased only nitrogen and allantoin contents. Interactions between inoculation and P, N and micronutrients on nodulation and growth of leucaena were observed. The effect of inoculation and fertilization with phosphorus or micronutrients was further investigated in field experiments. Establishment of uninoculated and unfertilized leucaena was poor at both locations due to low soil fertility and the presence of only a few native leucaena rhizobia. At one site, only inoculated plants were nodulated, while at the other, all plants produced nodules. Shoot dry weight, total nitrogen and phosphorus of inoculated plants were statistically equal to nitrogen-fertilized plants. Uninoculated plants were stunted. Generally, micronutrients did not influence nodulation, total nitrogen or growth of leucaena. They had only a positive effect on nitrogenase activity. Phosphorus increased total nitrogen and phosphorus uptake and plant growth. A 75% increase in shoot dry weight was obtained when 80 kg P ha^–1 was applied to inoculated leucaena with Rhizobium strain IRc 1045. Inoculated plants contained more allantoins than uninoculated ones but no significant correlation was found between these compounds and other parameters of N fixation.     

Estimation of symbiotically fixed nitrogen in field grown soybeans: An application of natural15N/14N abundance and a low level15N-tracer technique

Summary Plants from agricultural and natural upland ecosystem were investigated for¹⁵N content to evaluate the role of symbiotic N₂-fixation in the nitrogen nutrition of soybean. Increased yields and lower δ¹⁵N values of nodulating soybeansvs, non-nodulating isolines gave semi-quantitative estimates of N₂ fixation. A fairly large discrepancy was found between estimations by δ¹⁵N and by N yield at 0 kg N/ha of fertilizer. More precise estimates were made by following changes in plant δ¹⁵N when fertilizer δ¹⁵N was varied near¹⁵N natural abundance level. Clearcut linear relationships between δ¹⁵N values of whole plants and of fertilizer were obtained at 30 kg N/ha of fertilizer for three kinds of soils. In experimental field plots, nodulating soybeans obtained 13±1% of their nitrogen from fertilizer, 66±8% from N₂ fixation and 21±10% from soil nitrogen in Andosol brown soil; 30%, 16% and 54% in Andosol black soil; 7%, 77% and 16% in Alluvial soil, respectively. These values for N₂ fixation coincided with each corresponding estimation by N yield method. Other results include: 1)¹⁵N content in upland soils and plants was variable, and may reflect differences in the mode of mineralization of soil organics, and 2) nitrogen isotopic discrimination during fertilizer uptake (δ¹⁵N of plant minus fertilizer) ranged from −2.2 to +4.9‰ at 0–30 kg N/ha of fertilizer, depending on soil type and plant species. The proposed method can accurately and relatively simply establish the importance of symbiotic nitrogen fixation for soybeans growing in agricultural settings.