N2 fixation in Thai soybeans: Effect of tillage and inoculation on15N-determined N2 fixation in recommended cultivars and advanced breeding lines

The practice of seeding soybeans following paddy rice in Thailand has encountered difficulties in seedling germination, nodulation and crop establishment. This research project evaluated the choice of a non-fixing control to quantify N₂ fixation by¹⁵N isotope dilution, and the effect of tillage regime, soybean cultivar, strain ofBradyrhizobium japonicum and P fertilization on yield and N₂ fixation after paddy rice in northern and central Thailand.Japanese non-nodulating lines Tol-0 and A62-2 were the most appropriatecontrol plants for¹⁵N isotope dilution for Thai soybeans in these soils which contained indigenous rhizobia. Cereals such as maize, sorghum and barley were also appropriate controls at some sites. The choice of the appropriate non-fixing control plant for the¹⁵N isotope dilution technique remains a dilemma and no alternative exists other than to use several possible controls with each experiment. Acetylene reduction assay (ARA) proved of little value for screening varieties on their N₂ fixing capacity.The recommended Thai soybean cultivars (SJ1, 2, 4, 5) and an advanced line 16–4 differed little in their ability to support N₂ fixation or yield, possibly due to similar breeding ancestry. The ten AVRDC (ASET) lines showed considerable genotypic control in their ability to utilize their three available N sources (soil, fertilizer, atmosphere) and to translate them into yields. None of these lines were consistently superior to Thai cultivars SJ4 or SJ5 although ASET lines 129, 209 and 217 showed considerable promise.Neither recommended Thai or ASET cultivars were affected by tillage regime. Zero tillage resulted in superior N₂ fixation and yield at two sites but conventional tillage was superior at another site. Soybean cultivars grown in Thailand were well adapted to zero tillage. Levels of N₂ fixation were similar to world figures, averaging more than 100 kg N ha^–1 and supplying over 50% of the plant's N yield. However, seed yields seldom exceeded 2 t ha^–1, well below yields for temperately-grown soybeans. It is not clear why Thai soybeans support N₂ fixation, but do not translate this into higher seed yields.     

Seasonal accumulation and partitioning of nitrogen by lentil

Although wheat (Triticum aestivum L.) is the dominant crop of the semi-arid plains of Canada and the western United States, lentil (Lens culinaris Medik.) has become an important alternative crop. Sources and seasonal accumulation of N must be understood in order to identify parameters that can lead to increased N₂-fixing activity and yield. Inoculated lentil was grown in a sandy-loam soil at an irrigated site in Saskatchewan, Canada. Wheat was used as the reference crop to estimate N₂ fixation by the A-value approach. Lentil and wheat received 10 and 100 kg N ha⁻¹ of ammonium nitrate, respectively. Crops were harvested six times during the growing season and plant components analyzed. During the first 71 days after planting the wheat had a higher daily dry matter and N accumulation compared to lentil. However, during the latter part of the growing season, daily dry matter and N accumulation were greater for lentil. The maximum total N accumulation for lentil at maturity was 149 kg ha⁻¹. In contrast, wheat had a maximum N accumulation of 98 kg ha⁻¹ in the Feekes 11.1 stage, or 86 days after planting. The maximum daily rates of N accumulation were 3.82 kg N ha⁻¹ day⁻¹ for lentil and 2.21 kg N ha⁻¹ day⁻¹ for wheat. The percentage of N derived from N₂ fixation (% Ndfa) ranged from 0 at the first harvest to 92 % at final harvest. Generative plant components had higher values for % Ndfa than the vegetative components which indicates that N in the reproductive plant parts was derived largely from current N₂ fixation and lentil continued to fix N until the end of the pod fill stage. At final harvest, lentil had derived 129 kg N ha⁻¹ from N₂ fixation with maximum N₂-fixing activity (4.4 kg N ha⁻¹ day⁻¹) occurring during the early stages of pod fill. Higher maximum rates of N₂-fixing activity than net N accumulation (3.82 kg N ha⁻¹ day⁻¹) may have been caused by N losses like volatilization. In addition, lentil provided a net N contribution to the soil of 59 kg ha⁻¹ following the removal of the grain.     

Nitrogen fertilization effects on dinitrogen fixation as influenced by legume species and proportion in legume-grass mixtures in Uruguay

Grasses grown in mixture with nodulated legumes often are N-limited, but N fertilization may result in reductions of N₂ fixation and legume stands. We studied N-fertilizer effects on N₂ fixation for three binary legume-grass mixtures in Uruguay. Replicated swards of white clover (Trifolium repens L.) (WC), red clover (Trifolium pratense L.) (RC), or birdsfoot trefoil (Lotus corniculatus L.) (BT), each in combination with tall fescue (Festuca arundinacea Schreb) (TF) at two legume proportions were sown in 1983 (Exp. 1) and 1984 (Exp. 2). In the fall of 1984, N treatments at 100 kg ha⁻¹ and controls were randomly assigned to subplots in Exp. 1 (established swards) and in Exp. 2 (at seeding). The soil for both experiments was a fine, montmorillonitic, mesic, Typic Argiudolls. Herbage fixed-N was estimated by ¹⁵N isotope-dilution with pure stands of TF as reference. In both experiments, N fertilization reduced the proportion of legume N derived from air (% Ndfa) and increased herbage yield only during the first 18 to 20 weeks after application. Fertilizer-N reduced annual fixed-N yield from 178 to 148 kg ha⁻¹ in Exp. 1 and from 65 to 29 kg ha⁻¹ in Exp. 2 Fixed-N yield for BT was markedly reduced by N in both experiments (33 to 53%), whereas for the clovers reduction was lesser in Exp. 1 (9 to 13%) than in Exp. 2 (46 to 64%). Negative effects of N on % Ndfa were more evident for the high legume proportion. We conclude that fertilization with 100 kg N ha⁻¹ reduced % Ndfa only for the immediate 18 to 20 weeks after application. Fertilizer-induced reduction of fixed-N yields lasted longer because of a more prolonged depression of legume proportion, especially for BT and for newly seeded swards. Journal Paper no. J.-13327 of the Iowa Agric. and Home Econ. Exp. Stn., Ames, U.S.A. (Project 2281). Supported in part by the Facultad de Agronomía, Montevideo, Uruguay; and the International Atomic Energy Agency, Vienna, Austria (Project URU/5/012). Journal Paper no. J.-13327 of the Iowa Agric. and Home Econ. Exp. Stn., Ames, U.S.A. (Project 2281). Supported in part by the Facultad de Agronomía, Montevideo, Uruguay; and the International Atomic Energy Agency, Vienna, Austria (Project URU/5/012).     

Effect of rhizobia and rock biofertilizers with <Emphasis Type="Italic">Acidithiobacillus</Emphasis> on cowpea nodulation and nutrients uptake in a tableland soil

Chemical fertilizers have been used in the cultivation of plants due to their high solubility and effect on crops yield. Biofertilizers with phosphate rock (PR) and potash rock (KR) plus sulfur inoculated with Acidithiobacillus may improve plant growth and contribute to addition of available P and K in soil. The effectiveness of biofertilizers from phosphate and potash rocks mixed with sulfur and Acidithiobacillus was studied in a Typic Fragiuldult soil of the Brazilian Northeast Tableland. Cowpea (cv. “IPA 206”) was grown with and without rhizobia inoculation. Treatments were: (a) phosphate rock (1000 kg ha⁻¹); (b) Biofertilizers-B_P (250 and 500 kg ha⁻¹); (c) triple superphosphate-TSP (250 kg ha⁻¹); (d) potash rock (1000 kg ha⁻¹); (e) biofertilizer-B_K (250; 500 and 750 kg ha⁻¹); (f) potassium chloride-KCl (250 kg K₂0 ha⁻¹); (g) control without P or K fertilization (P₀K₀). The soil was maintained under water submersion covered with black plastic (solarization process) for a period of 30 days. Biofertilizers (B_p and B_K) and soluble fertilizers increased plant growth and NPK uptake. Biofertilizers reduced soil pH, especially when applied in highest rates. Biofertilizers and TSP+KCl showed the best values of available P and K in soil. Rhizobial inoculation was effective on cowpea, but no nodules were formed by bacteria native from the soil, probably due to the effect of the solarization process. From obtained PK biofertilizers could be used as alternative for cowpea fertilization in Tableland soils.     

Carbon, nitrogen and phosphorus in volcanic soils following afforestation with native birch (Betula pubescens) and introduced larch (Larix sibirica) in Iceland

Afforestation has become an important tool for soil protection and land reclamation in Iceland. Nevertheless, the harsh climate and degraded soils are growth-limiting for trees, and little is know about changes in soil nutrients in maturing forests planted on the volcanic soils. In the present chronosequence study, changes in C, N and total P in soil (0–10 and 10–20 cm depth) and C and N in foliar tissue were investigated in stands of native Downy birch (Betula pubescens Enrh.) and the in Iceland introduced Siberian larch (Larix sibirica Ledeb.). The forest stands were between 14 and 97 years old and were established on heath land that had been treeless for centuries. Soils were Andosols derived from basaltic material and rhyolitic volcanic ash. A significant effect of tree species was only found for the N content in foliar tissue. Foliar N concentrations were significantly higher and foliar C/N ratios significantly lower in larch needles than in birch leaves. There was no effect of stand age. Changes in soil C and the soil nutrient status with time after afforestation were little significant. Soil C concentrations in 0–10 cm depth in forest stands older than 30 years were significantly higher than in heath land and forest stands younger than 30 years. This was attributed to a slow accumulation of organic matter. Soil N concentrations and soil P_tot were not affected by stand age. Nutrient pools in the two soil layers were calculated for an average weight of soil material (400 Mg soil ha⁻¹ in 0–10 cm depth and 600 Mg soil ha⁻¹ in 10–20 cm depth, respectively). Soil nutrient pools did not change significantly with time. Soil C pools were in average 23.6 Mg ha⁻¹ in the upper soil layer and 16.9 Mg ha⁻¹ in the lower soil layer. The highest annual increase in soil C under forest compared to heath land was 0.23 Mg C ha⁻¹ year⁻¹ in 0–10 cm depth calculated for the 53-year-old larch stand. Soil N pools were in average 1.0 Mg N ha⁻¹ in both soil layers and did not decrease with time despite a low N deposition and the uptake and accumulation of N in biomass of the growing trees. Soil P_tot pools were in average 220 and 320 kg P ha⁻¹ in the upper and lower soil layer, respectively. It was assumed that mycorrhizal fungi present in the stands had an influence on the availability of N and P to the trees. Responsible Editor: Hans Lambers.     

Effects of kelp (Macrocystis integrifolia) on soil chemical properties and crop response

In 1981 a two-year field plot experiment was established to assess the effects of quantities (0, 7.5, 15, 30, 60 and 120 t ha⁻¹) of fresh kelp (Macrocystis integrifolia) on crop growth and nutritional response and chemical properties of a fine-textured soil. Soil was analyzed for NO₃−N, NH₄−N, electrical conductivity, pH, Cl and exchangeable cations (K, Mg, Ca, Mn and Na). The plots were planted to beans (Phaseolus vulgaris) in the first year and peas (Pisum sativum) in the second year. Marketable bean yields increased in the first year with kelp applications up to 60 t ha⁻¹, with yields, emergence and flowering being reduced by the 120 t ha⁻¹ application. Soluble salts (EC) and Cl concentrations in the soil eight days after application increased linearly and sharply with increasing quantities of kelp. Increased K concentration and moisture content, characteristics of plants growing in a salt-stressed soil environment, were measured. A subsequent companion greenhouse experiment confirmed that the reduced bean emergence and growth with 120 t ha⁻¹ applications of kelp were primarily due to soluble salts. The only growth effects upon peas in the second year was a slight reduction in leaf plus stem yields with increasing applications of kelp.     

Root turnover in a beech and a spruce stand of the Belgian Ardennes

The theoretical basis of fine root turnover estimation in forest soils is discussed, in relation to appropriate experimental techniques of measurement. After sequential coring, the correct expression is the sum of significant positive increments of live and dead roots of the various diameter categories, to which the transfer of dead roots to organic matter derived from roots, OMDR, has to be added. This should not be confounded with dead root mineralization. The transfer rates should first be estimated in root dimensions and not in weight of dry matter. The measurements were carried out in a 120 year old beech (Fagus sylvatica L.) stand and a 35 year old Norway spruce (Picea abies Karst) stand, in the Eastern Ardennes, Belgium. The turnover rate of fine roots (diam. <5 mm) was 4393 kg ha⁻¹ year⁻¹ (root dry weight), including 711.2 kg ha⁻¹ year⁻¹ for dead root transfer to OMDR, for beech. For spruce, turnover rate was 7011 kg ha⁻¹ year⁻¹ (root dry weight), including 1498 kg ha⁻¹ year⁻¹ for dead root transfer to OMDR. Under beech, there was a slight root density increase in spring. No seasonal fluctuations were observed under spruce, but a strong irreversible drop in live root growth was found in the later season 1980–1981, corresponding to a decrease of tree height growth and trunk radius increment. Turnover rates were further expressed in dry weight and in amounts of elements (kg ha⁻¹ year⁻¹) (Ca, Mg, K, Na, Al, N, P, S). Correlative relations between root dimensions and dry weight and element concentrations show that the derived values, and in particular root specific density (dry weight volume⁻¹) vary according to species, root category, and seasonal sampling. Various schemes of seasonal variations of root growth, described in Europe, show that the major dependance on general climate is obscured by environmental factors (soil, exposure, species). It is suggested that root density fluctuation approach the steady state on an annual basis under mild Atlantic conditions.     

Nitrogen fixation (15N dilution) with soybeans under Thai field conditions

Controlled environment and field studies were conducted to determine relationships between various measurements of N₂ fixation using soybeans and to use these measures to evaluate a number ofBradyrhizobium japonicum strains for effectiveness in N₂ fixation in Thai soils.¹⁵N dilution measurements of N₂ fixation showed levels of fixation ranging from 32 to 161 kg N ha⁻¹ depending on bacterial strain, host cultivar and location. Midseason measures of N₂ fixation were correlated with each other, but not related measures taken at maturity. Ranking ofB. japonicum strains based on performance under controlled conditions in N-free media were highly correlated with rankings based on soybean seed yields and N₂ fixation under field conditions. This study showed that inoculation of soybeans with effectiveB. japonicum strains can result in significant increases in yield and uptake of N through fixation. The most effective strains tested for use in Thai conditions were those isolated from Thai soils; however, effective strains from other locations were also of benefit.     

Dual inoculation withRhizobium sp. andGlomus fasciculatum enhances nodulation,yield and nitrogen fixation in chickpea (Cicer arietinum Linn.)

Summary Seed inoculation with Rhizobium and soil inoculation withGlomus fasciculatum increased nodulation, nitrogen and phosphorus concentration in plants and yield of chickpea (Cicer arietinum) var. BG 212 in pots containing unsterilized soil especially with 50kgP₂O₅ ha⁻¹ in the form of superphosphate. Inoculation with Rhizobium orG. fasciculatum separately or in combination significantly increased the N₂ fixed in straw and grain than uninoculated controls as determined by¹⁵N atom percent excess of plants grown in soil amended with labelled ammonium sulphate (¹⁵NH₄)₂SO₄) at the rate of 20kg N ha⁻¹. These increases were most pronounced when P was applied at 50kgP₂O₅ ha⁻¹.     

The influence of long-term tillage systems on symbiotic N2 fixation of pea (Pisum sativum L.) and red clover (Trifolium pratense L.)

Pea as a grain legume and red clover as a forage legume in the seeding year were cultivated in two long-term differentiated tillage systems on a loess soil in Germany. A continuous conventional tillage system (plow; CT) and a continuous minimum tillage system (rotary harrow; MT) were established in 1970. With pea and red clover dry matter accumulation and N parameters (N accumulation, Ndfa, N-harvest-index, N balance) were investigated in 1998 and 1999. Differences in the N₂ fixation of pea due to the tillage system could clearly be shown whereas grain yields and total N accumulation were equal in both tillage systems and years. In both years a significantly (P < 0.05) higher Ndfa in the MT system was found at least in the final harvest (maturity of pea): 1998/1999, 0.42/0.54 in CT, 0.62/0.75 in MT. The differences in N₂ fixation of pea may be explained by the delayed soil N supply in MT at the beginning of the vegetative period. Simplified N balances of pea were -18 and –25 kg N ha^–1 in CT and –5 and +1 kg N ha^–1 in MT for 1998 and 1999, respectively. Red clover showed no significant differences in the DM and N accumulation between both tillage systems but a year dependent effect caused by different stubble and root yields between the years was apparent. With red clover slightly, but also significantly (P < 0.05) increased Ndfa values were found in the MT system compared to the CT system with 0.55/0.62 in CT (1998/1999) and 0.64/0.71 in MT. However, the difference in Ndfa between the tillage systems (9 percentage points) was much smaller with red clover than with pea (20 and 21 percentage points in 1998 and 1999, respectively). Soil N uptake of red clover using the longer growing season reflected the more adjusted N supply in both long-term differentiated tillage systems, whereas pea in using only a short-term vegetative period reacted stronger to the lower N mineralization in the MT system in springtime.     

Methane Budget of a Black Forest Spruce Ecosystem Considering Soil Pattern

Study included seven soils, an adjacent spring and brook and was conducted to estimate CH₄ source and sink strengths of forest soils along a wetness gradient, i.e. their exchange with atmosphere (direct emission), and hydrosphere (indirect emission). Soils are represented by anaerobic Histosol, oxic Cambisols, Histosol with degraded peatlayers and Gleysols having intermediate redox state. They could be separated into three emission groups: CH₄ emitting (248–318 kg C ha⁻¹ a⁻¹), CH₄ uptake (−0.1 to −5 kg C ha⁻¹ a⁻¹), and soils on the edge of CH₄ uptake and release (−0.2–20 kg C ha⁻¹ a⁻¹). Although soils with CH₄ uptake were dominant (75%), the soil specific CH₄ budget identified the study field (6.53 ha) as CH₄ source (40.9 kg C ha⁻¹ a⁻¹). Not only CH₄ emissions, but also dissolved CH₄ in soil solution varied regularly with soil type. Individual soil solutions contained 0.008–151 μmol CH₄ l⁻¹. CH₄ vanished to negligible loads, when dissolved CH₄ passed an oxidative downslope soil zone, but promoted CH₄ uptake was measured at this soil. In turn, CH₄ was discharged to the atmosphere, when the soil solution left the pedosphere across an anaerobic soil zone. These measured indirect emissions were low (34 g C a⁻¹), but the values of individual soil solution indicate possible higher discharges (3.9 kg a⁻¹) at a different soil pattern. The results suggest that CH₄ uptake rates of temperate forests are overestimated.     

Nodulation,nitrogen fixation and nitrogen uptake in pigeonpea (Cajanus cajan (L.) Millsp) of different maturity groups

Summary The seasonal patterns of nodulation, acetylene reduction, nitrogen uptake and nitrogen fixation were studies for 11 pigeonpea cultivars belonging to different maturity groups grown on an Alfisol at ICRISAT Center, Patancheru, India. In all cultivars the nodule number and mass increased to a maximum around 60–80 days after sowing and then declined. The nodule number and mass of medium- and late-maturing cultivars was greater than that of early-maturing cultivars. The nitrogenase activity per plant increased to 60 days after sowing and declined thereafter, with little activity at 100 days when the crop was flowering. At later stages of plant growth nodules formed down to 90 cm below the soil surface but those at greater depth appeared less active than those near the surface. All the 11 cultivars continued to accumulate dry matter until 140 days, with most biomass production by the late-maturing cultivars (up to 11 t ha⁻¹) and least by the early-maturing determinate cultivars (4 t ha⁻¹). Total nitrogen uptake ranged from 69 to 134 kg ha⁻¹. Nitrogen fixation by pigeonpea was estimated as the difference in total nitrogen uptake between pigeonpea and sorghum and could amount to 69 kg N ha⁻¹ per season, or half the total nitrogen uptake. Fixation by pigeonpea increased with crop duration, but there were differences within each maturity group. The limitations of the methods used for estimating N₂ fixation by pigeonpea are discussed. Submitted as J.A. No. 552 by the International Crops Research Institute for the Semi-Arid Tropics (ICRISAT).     

Dinitrogen fixation in white clover grown in pure stand and mixture with ryegrass estimated by the immobilized 15N isotope dilution method

Dinitrogen fixation in white clover (Trifolium repens L.) grown in pure stand and mixture with perennial ryegrass (Lolium perenne L.) was determined in the field using ¹⁵N isotope dilution and harvest of the shoots. The apparent transfer of clover N to perennial ryegrass was simultaneously assessed. The soil was labelled either by immobilizing ¹⁵N in organic matter prior to establishment of the sward or by using the conventional labelling procedure in which ¹⁵N fertilizer is added after sward establishment. Immobilization of ¹⁵N in the soil organic matter has not previously been used in studies of N₂ fixation in grass/clover pastures. However, this approach was a successful means of labelling, since the ¹⁵N enrichment only declined at a very slow rate during the experiment. After the second production year only 10–16% of the applied ¹⁵N was recovered in the harvested herbage. The two labelling methods gave, nonetheless, a similar estimate of the percentage of clover N derived from N₂ fixation. In pure stand clover, 75–94% of the N was derived from N₂ fixation and in the mixture 85–97%. The dry matter yield of the clover in mixture as percentage of total dry matter yield was relatively high and increased from 59% in the first to 65% in the second production year. The average daily N₂ fixation rate in the mixture-grown clover varied from less than 0.5 kg N ha⁻¹ day⁻¹ in autumn to more than 2.6 kg N ha⁻¹ day⁻¹ in June. For clover in pure stand the average N₂ fixation rate was greater and varied between 0.5 and 3.3 kg N ha⁻¹ day⁻¹, but with the same seasonal pattern as for clover in mixture. The amount of N fixed in the mixture was 23, 187 and 177 kg N ha⁻¹ in the seeding, first and second production year, respectively, whereas pure stand clover fixed 28, 262 and 211 kg N ha⁻¹ in the three years. The apparent transfer of clover N to grass was negligible in the seeding year, but clover N deposited in the rhizosphere or released by turnover of stolons, roots and nodules, contributed 19 and 28 kg N ha⁻¹ to the grass in the first and second production year, respectively. This revised version was published online in June 2006 with corrections to the Cover Date.     

15N-Determined effect of inoculation with N2 fixing bacteria on nitrogen assimilation in Western Canadian wheats

Summary A two-year field study was undertaken using¹⁵N isotope techniques to differentiate between stimulation of N uptake and N₂ fixation in Western Canadian cultivars of spring wheat (Triticum aestivum L. emend Thell) and durum (T. turgidum L. emend Bowden) in response to inoculation with N₂-fixing bacteria. Bacterial inoculation either had no effect or lowered the % N derived from the fertilizer and the fertilizer use efficiency. Despite the depression of fertilizer uptake, inoculants did not alter the relative uptake from soil and fertilizer-N pools indicating that bacterial inoculation did not alter rooting patterns. Nitrogen-15 isotope dilution indicated that N₂ fixation did occur. In 1984, % plant N derived from the atmosphere (% Ndfa) due to inoculation with Bacillus C-11-25 averaged 23.9% while that withAzospirillum brasilense ATCC 29729 (Cd) averaged 15.5%. In 1985, higher soil N levels reduced these values by approximately one-half. Cultivar x inoculant interactions, while significant, were not consistent across years. However, these interactions did not affect cultivars ‘Cadet’ and ‘Rescue’. In agreement with previous results, ‘Cadet’ performed well with all inoculants in both years while ‘Rescue’ performed poorly. Among 1984 treatments, the N increament in inoculated plants was positively correlated with % Ndfa but no such correlation existed in 1985. N₂ fixation averaged over all cultivars and strains was 17.9 and 6.7 kg N fixed ha⁻¹ in 1984 and 1985, respectively. Highest rates of N₂ fixation were estimated at 52.4 kg N ha⁻¹ for ‘Cadet’ in 1984 and 31.3 kg N ha⁻¹ for ‘Owens’ in 1985, both inoculated with Bacillus C-11-25, an isolate from southern Alberta soils. Inoculation with either ofAzospirillum brasilense strain Cd (ATCC29729) or 245 did not result in as consistent or as high N₂ fixation, suggesting that these wheats had not evolved genetic compatability with this exogenous microorganism. These agronomically significant amounts of N₂ fixation occurred under optimally controlled experimental conditions in the field. It is yet to be determined if N₂ fixation would occur in response to bacterial inoculation under dryland conditions commonly occurring in Western Canada. Contribution from Agriculture Canada Research Station, Lethbridge, Alberta, Canada.     

The response ofAzolla pinnata R. Brown to the split application of phosphorus and the transfer of assimilated phosphorus to flooded rice plants

A field experiment was conducted at the Bangladesh Rice Research Institute, Joydebpur, Dhaka during the late wet season. Basal application of P at both 5 and 10 kg ha⁻¹ significantly increased total biomass production and nitrogen fixation byAzolla pinnata R. Brown (local strain). Addition of both 5 and 10 kg P ha⁻¹ in equal splits at inoculation and at six day intervals thereafter during growth periods of 12, 24 and 36 days increased biomass production and nitrogen fixation by Azolla over that attained with the basal application. Biomass and nitrogen fixation using a split application of 5 kg P ha⁻¹ exceeded that attained with basal application of 10 kg P ha⁻¹ and split application of 10 kg P ha⁻¹ resulted in 0.58, 11.2, and 18.3 t ha⁻¹ more biomass, and 0.47, 18.9, and 18.3 more kg fixed N ha⁻¹ at 12, 24 and 36 days, respectively, than the same amount applied as a basal application. Analyses indicated that the critical level of dry weight P in Azolla for sustained growth was in the range of 0.15–0.17%. Compared with the control, where no P was added, and additional 30 and 36 kg N ha⁻¹ were fixed after 24 and 36 days, respectively, when P was provided at 10 kg ha⁻¹ using a split application. A separate field study showed that flooded rice plants received P from incorporated Azolla with about 28% of the P present in the supplied Azolla being incorporated into the rice plants.     

Dinitrogen-fixation by three neotropical agroforestry tree species under semi-controlled field conditions

Cultivating dinitrogen-fixing legume trees with crops in agroforestry is a relatively common N management practice in the Neotropics. The objective of this study was to assess the N₂ fixation potential of three important Neotropical agroforestry tree species, Erythrina poeppigiana, Erythrina fusca, and Inga edulis, under semi-controlled field conditions. The study was conducted in the humid tropical climate of the Caribbean coastal plain of Costa Rica. In 2002, seedlings of I. edulis and Vochysia guatemalensis were planted in one-meter-deep open-ended plastic cylinders buried in soil within hedgerows of the same species. Overall tree spacing was 1 × 4 m to simulate a typical alley-cropping design. The ¹⁵N was applied as (NH₄)₂SO₄ at 10% ¹⁵N atom excess 15 days after planting at the rate of 20 kg [N] ha⁻¹. In 2003, seedlings of E. poeppigiana, E. fusca, and V. guatemalensis were planted in the same field using the existing cylinders. The ¹⁵N application was repeated at the rate of 20 kg [N] ha⁻¹ 15 days after planting and 10 kg [N] ha⁻¹ was added three months after planting. Trees were harvested 9 months after planting in both years. The ¹⁵N content of leaves, branches, stems, and roots was determined by mass spectrometry. The percentage of atmospheric N fixed out of total N (%N_f) was calculated based on ¹⁵N atom excess in leaves or total biomass. The difference between the two calculation methods was insignificant for all species. Sixty percent of I. edulis trees fixed N₂; %N_f was 57% for the N₂-fixing trees. Biomass production and N yield were similar in N₂-fixing and non-N₂-fixing I. edulis. No obvious cause was found for why not all I. edulis trees fixed N₂. All E. poeppigiana and E. fusca trees fixed N₂; %N_f was ca. 59% and 64%, respectively. These data were extrapolated to typical agroforestry systems using published data on N recycling by the studied species. Inga edulis may recycle ca. 100 kg ha⁻¹ a⁻¹ of N fixed from atmosphere to soil if only 60% of trees fix N₂, E. poeppigiana 60–160 kg ha⁻¹ a⁻¹, and E. fusca ca. 80 kg ha⁻¹ a⁻¹.     

Seasonal variation in the nitrogenase activity of aPanicum maximum var.trichoglume pasture and identification of associated bacteria

Summary The influence of seasonal variation on nitrogenase (N₂-ase) activity of undisturbed soil-plant cores ofPanicum maximum var.trichoglume was measured using the C₂H₂ reduction assay. The largest N₂-ase activity in the field, 14.7 g N ha⁻¹ day⁻¹, occurred in spring when soil moisture was high, soil temperature was low and nitrogenous fertiliser influence was at a minimum. The potential N₂-ase activity of the cores, measured under controlled conditions, reached a maximum of 27.2 g N ha⁻¹ day⁻¹ and averaged 26.3 g N ha⁻¹ day⁻¹ over the 14 month sampling period. N₂-ase activity was positively correlated (P=0.05) with field soil moisture and negatively correlated with field soil temperature (r=0.59 and −0.78 respectively). Multiple regression showed that 69% of the variation of N₂-ase activity in the field was associated with the combined effects of soil moisture and soil temperature. Nitrogen fixing bacteria were isolated from the roots ofP. maximum and based upon morphology, biochemical tests and fluorescent antibody reaction, were found to be closely related toAzospirillum lipoferum.     

Symbiotic N2 fixation in pea and field bean estimated by15N fertilizer dilution in field experiments with barley as a reference crop

Summary The total amount of nitrogen derived from symbiotic nitrogen fixation in two pea and one field bean cultivar, supplied with 50 kg N ha⁻¹ at sowing (‘starter’-N), was estimated to 165, 136, and 186 kg N ha⁻¹, respectively (three-year means). However, estimates varied considerably between the three years. At the full bloom/flat pod growth stage from 30 to 59 per cent of total N₂ fixation had taken place. The proportion of total N derived from N₂ fixation at maturity was higher in seeds than in vegetative plant parts and amounted to 59.5, 51.3 and 66.3 per cent of total above-ground plant N in the two pea cultivars and field bean, respectively (three-year means). The recovery of fertilizer N was 62.2, 70.2, 52.1, and 69.5 per cent in the two pea cultivars, field bean and barley, respectively. Growth analysis indicated that barley did not meet the claims for an ideal reference crop in the¹⁵N fertilizer dilution technique for estimating N₂ fixation in pea and field bean. ‘Starter’-N neither increased the seed yield nor the N content of the grain legumes.     

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.     

Turfgrass (Cynodon dactylon L.) sod production on sandy soils: I. Effects of irrigation and fertiliser regimes on growth and quality

The effects on growth, quality and N uptake by turfgrass (Cynodon dactylon L.) during sod production of four fertiliser types applied at three application rates (100, 200 or 300 kg N ha⁻¹ per ‘crop’) under two irrigation treatments (70% and 140% daily replacement of pan evaporation) were investigated. The fertiliser types were: water-soluble (predominately NH₄NO₃), control-release, pelletised poultry manure, and pelletised biosolids; and the experiment was conducted on a sandy soil in a Mediterranean-type climate. Plots were established from rhizomes, with the turfgrass harvested as sod every 16–28 weeks depending upon the time of the year. Four crops were produced during the study. Applying water-soluble and control-release fertilisers doubled shoot growth and improved turfgrass greenness by up to 10% in comparison with plots receiving pelletised poultry manure and pelletised biosolids. Nitrogen uptake into the shoots after four crops (averaged across irrigation treatments and N rates) was 497 kg N ha⁻¹ for the water-soluble fertiliser, 402 kg N ha⁻¹ for the control-release, 188 kg N ha⁻¹ for the pelletised poultry manure and 237 kg N ha⁻¹ for the pelletised biosolids. Consequently, the agronomic nitrogen-use efficiency (NAE, kg DM kg⁻¹ N applied) of the inorganic fertilisers was approximately twice that of the organic fertilisers. Increasing irrigation from 70% to 140% replacement of pan evaporation was detrimental to turfgrass growth and N uptake for the first crop when supplied with the water-soluble fertiliser. Under the low irrigation treatment, inorganic N fertilisers applied at 200–300 kg N ha⁻¹ were adequate for production of turfgrass sod. Section Editor: P. J. Randall