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

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

Evaluation of the availability ofAzolla-N and urea-N to rice using15N

Summary The symbiotic association of the water fernAzolla with the blue-green algaAnabaena azollae can fix 30–60 kg N ha^–1 per rice cropping season. The value of this fixed N for rice production, however, is only realized once the N is released from theAzolla biomass and taken up by the rice plants. The availability of N applied asAzolla or as urea was measured in field experiments by two¹⁵N methods. In the first,Azolla caroliniana (Willd.) was labelled with¹⁵N in nutrient solution and incorporated into the soil at a rate of 144 kg N ha^–1. The recovery ofAzolla-N in the above ground parts of rice [Oryza sativa (L) cv. Nucleoryza] was found to be 32% vs. 26% for urea applied at a rate of 100 kg N/ha; there was no significant difference in recovery. In the second, 100 kg N/ha of¹⁵N-urea was applied separately or in combination with either 250 or 330 kg N ha^–1 of unlabelledAzolla. At the higher rate, the recovery ofAzolla-N was significantly greater than that of urea. There was a significant interaction when both N sources were applied together, which resulted in a greater recovery of N from each source in comparison to that source applied separately. Increasing the combined urea andAzolla application rate from 350 kg N ha^–1 to 430 kg N ha^–1 increased the N yield but had no effect on the dry matter yield of rice plants. The additional N taken up at the higher level of N application accumulated to a greater extent in the straw compared to the panicles. Since no assumptions need to be made about the contribution of soil N in the method using¹⁵N-labelledAzolla, this method is preferable to the¹⁵N dilution technique for assessing the availability ofAzolla-N to rice. Pot trials usingAzolla stored at –20°C or following oven-drying showed that both treatments decreased the recovery of N by one third in comparison to freshAzolla.     

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.     

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.     

Field evaluation of symbiotic nitrogen fixation by rhizobial strains using15N methodology

Summary Small differences in N₂ fixation by nodulated soybeans (Glycine max. (L.) Merr.), inoculated with various strains ofRhizobium japonicum, were assessed in field experiments using¹⁵N methodology, and compared with yields of plant dry matter and total N. Percentage of plant-N derived from atmospheric N₂ and from fertilizer, and values of %¹⁵N atom excess had lower coefficients of variation than did total N and dry matter yield. Nevertheless the precision of estimates of kg N/ha fixed were sufficient to differentiate only the extremes of the range of strains tested, and there were discrepancies between ranking of strains based on % N derived from fertilizer and on total N yield.     

Effects of inoculation ofPoa pratensis andTriticum aestivum with root-associated,N2-fixing Klebsiella,Enterobacter and Azospirillum

Strains ofKlebsiella pneumoniae, Klebsiella terrigena, Enterobacter agglomerans andAzospirillum lipoferum were compared as diazotrophic inoculants in association withPoa pratensis andTriticum aestivum. Each strain colonized both plants in numbers ranging from 10⁴ to 10⁷ bacteria per root, and electron microscopy and immunofluorescence staining of inoculated roots revealed bacteria mainly on root hairs. Indirect immunofluorescence with specific antifimbriae antibodies showed that the enteric bacteria expressed their fimbria in both associations. All associations were positive in an acetylene reduction test but only in half of them was atmospheric nitrogen transferred to the plant. In the inoculated plants, variable effects in the dry matter and N yields in both hosts were observed and no correlation was found between dry matter, nitrogen content or the amount of fixed nitrogen. In infected plants, the number of root hairs and lateral roots increased and the length of the zone of elongation decreased. The changes in root morphology were more evident in associations with enteric bacteria than with Azospirillum. The results give further evidence on the importance of bacterial adhesion in associative N₂ fixation and suggest that bacteria-induced physiological changes in plant roots may be more important than the amount of nitrogen transferred to the plant.     

The response of field grownPhaseolus vulgaris to Rhizobium inoculation and the quantification of N2 fixation using15N

Summary A field experiment was performed to assess the effects of Rhizobium inoculation and nitrogen fertilizer (100 kg N ha^–1) on four cultivars of Phaseolus beans; Carioca, Negro Argel, Venezuela 350 and Rio Tibagi. In the inoculated treatment 2.5 kg N ha^–1 of¹⁵N labelled fertilizer was added in order to apply the isotope dilution technique to quantify the contribution of N₂ fixation to the nutrition of these cultivars.Nodulation of all cultivars in the uninoculated treatments was poor, but the cultivars Carioca and Negro Argel were well nodulated when inoculated. Even when inoculated, nodulation of the cultivars Venezuela 350 and Rio Tibagi was poor and these cultivars showed little response to inoculation in terms of nitrogen accumulation or grain yield. The estimates of the contribution of N₂ fixation estimated using the isotope dilution technique, for the Carioca and Negro Argel cultivars, amounted to 31.7 and 18.4 kg N ha^–1 respectively. These two cultivars produced 991 and 883 kg ha^–1 of grain, respectively, when inoculated and 663 and 620 kg ha^–1 with the addition of 100 kg N ha^–1 of N fertilizer. The response to nitrogen was particularly poor due to high leaching losses in the very sandy soil at the experimental site.The Venezuela 350 and Rio Tibagi cultivars only responded to N fertilizer and not to inoculation with Rhizobium which stresses the great importance of selecting plant cultivars for nitrogen fixation in the field.     

Application of 15N and xylem ureide methods for assessing N2 fixation of three shrub legumes periodically pruned for forage

The apparently diminished capacity for N₂ fixation by the shrub legume Calliandra calothyrsus (Calliandra) relative to other woody perennial legumes was investigated in a field experiment in northern Queensland, Australia. In this trial, (i) the proportion of plant nitrogen (N) derived from symbiotic N₂ fixation (%Pfix) and the amounts of N₂ fixed were compared in Calliandra, Gliricidia sepium (Gliricidia) and Codariocalyx gyroides (Codariocalyx), (ii) variations in N₂ fixation due to season or tree age were determined, (iii) estimates of Pfix derived with the ¹⁵N natural abundance technique were compared with values obtained from ¹⁵N enrichment or xylem sap ureide procedures to determine whether the previous conclusions about Calliandra's ability to fix N had resulted from specific problems with the natural abundance methodology used in the earlier studies.Inoculated seedlings of each of the three shrub legume species were planted in dense stands (1.5 m rows, 0.5 m between trees) in two randomised blocks. The northern block was used solely for natural abundance measurements, while ¹⁵N-enriched KNO₃ (10 atom % ¹⁵N excess) was applied four times over a 52 week period to plots in the southern block. The non-nodulating tree legume Senna spectabilis (formally Cassia spectabilis) was used as a non-N₂-fixing reference for the ¹⁵N-based procedures, with Guinea grass (Panicum maximum) included as an additional non-fixing check. Growth by the trees above 75 cm was first cut and removed after 22 weeks and regrowth was subsequently pruned periodically for another 95 weeks. Sampling for dry matter production, N yield and estimates of Pfix were restricted to the central four of the 32 plants which constituted each replicate plot. Information generated during the 117 week study indicated that estimates of Pfix by ¹⁵N natural abundance were closely similar to values derived with ¹⁵N-enrichment or sap ureides. The data indicated that Calliandra had a reduced reliance upon N₂ fixation relative to Gliricidia and Codariocalyx for the first 65 weeks after establishment. This appeared to be due to more prolifc root growth by Calliandra than either of the other N₂-fixing species and an ability to extract a greater proportion of its N requirements from soil mineral N. However, after week 65 and for the remainder of the experiment, estimates of Pfix for Calliandra were similar to the other shrub legumes. Over 117 weeks, prunings from Calliandra and Gliricidia had removed 52–58 t dry matter ha^-1, and between 1471 and 1678 kg N ha^-1, of which 1026–1063 kg N ha^-1 was estimated to have been derived from N₂ fixation. At the time of final harvest, 65–73% of the fixed N was present in shoot regrowth of the N₂ fixing shrubs, 9–18% in the roots, 15% in the trunk, and 2–6% in fallen leaves.     

Remobilization of labelled nitrogen from vegetative tissues to pods of mungbean

Summary Remobilization of¹⁵N from vegetative tissue of mungbean (Vigna radiata (L.) Wilczek) into pods was measured during the reproductive phase of growth. Plant tissue was labelled with¹⁵N during vegetative development. Experiments were conducted in the field at two sites. At one site the soil provided cowpeas with most of their N but at the other site N fixation provided most of the N. Remobilized N from vegetative tissue to pods occurred soon after they began to develop. The quantity of the labelled N ultimately remobilized to the pods amounted to 50% for one cultivar (Tx33) at the high soil N site and 70% at the low N site. For the other cultivar (Tx13) the values were 25% and 30%, respectively. The two cultivars performed very differently with respect to partitioning of N into pods and the rate of N fixation. Even though more N was accumulated in the shoots of the high N fixing cultivar (Tx13) less total N was contained in the pods.     

Nitrogen nutrition of rice plants under salinity

Two rice (Oryza sativa L.) cultivars, Koshihikari and Pokkali, were grown in solution culture at three concentrations of NaCl or Na₂SO₄ [0 (S0), 50 (S1), and 100 (S2) mmol dm^–3] and three N contents [0.7 (N1), 7 (N2) and 14 (N3) mmol dm^–3]. Salinity significantly decreased dry matter of both cultivars. Pokkali had better growth than Koshihikari under both saline and non-saline conditions. Applications of N enhanced development of shoot dry mass under S0 and S1 treatments up to N2. Under S2, N application had no effect on shoot dry mass of both cultivars. Root dry mass of both cultivars decreased with increasing N application at S1 and S2. Shoot and root NO₃-N content in both rice cultivars increased with increasing N concentration in the nutrient solutions. The absorption of NO₃-N was less in Koshihikari than Pokkali plants, and also was much less in Cl^– than SO₄ ^2– salinity suggesting the antagonism between Cl^– and NO₃ ^–. In addition a significant negative correlation between concentrations of NO₃-N and Cl^– in the shoots or roots was observed in both cultivars     

Vegetative growth and sporulation ofBipolaris sorokiniana on sequentially older infected leaves ofPoa pratensis exposed to postemergence herbicides

Poa pratensis was exposed to the postemergence herbicides 2,4-D, MCPP, and dicamba at concentrations of 10^–6 M applied to the soil surface. Sequentially older leaf blades were inoculated withBipolaris sorokiniana and detached to determine the effect of the herbicides on saprophytic hyphal growth and sporulation on the tissue of each older leaf. 2,4-D and dicamba stimulated hyphal growth in vitro from leaf tissue of all ages; MCPP had no effect on hyphal growth. All herbicides increased sporulation byB. sorokiniana on leaf tissue of all ages. Both hyphal growth and sporulation increased progressively on tissue from sequentially older leaves ofP. pratensis exposed to dicamba.Journal Paper No. J-14985 of the Iowa Agriculture and Home Economics Experiment Station, Ames, Iowa, USA, Project No. 2616.     

Measurements of nitrogen fixation in fababean at different N fertilizer rates using the 15N isotope dilution and ‘A-value’ methods

The ¹⁵N isotope dilution and A-value methods were used to measure biological nitrogen (N₂) fixation in field grown fababean (Vicia faba L.), over a 2-year period. Four N rates, 20, 100, 200 and 400 kg N ha^–1 were examined. The two isotope methods gave similar values of % N derived from the atmosphere (%Ndfa). With 20 kg N ha^–1, %Ndfa in fababean was about 85% in both years. Increasing the N rate to 100 kg N ha^–1 decreased N₂ fixation slightly to 75%. Further reductions in N₂ fixed to 60 and 43% occurred where 200 and 400 kg N ha^–1 were applied, respectively. Thus even higher rates of N than normally applied in farming practice could not completely suppress N₂ fixation in fababean.We also devised one equation for both the isotope dilution and A-value approaches, thereby (i) avoiding the need for different calculations for the ¹⁵N isotope methods, and (ii) showing once again that the isotope dilution and A-value methods are mathematically and conceptually identical.     

Estimation of residual N effect of faba bean and pea on two succeeding cereals using15N methodology

A field experiment was conducted using¹⁵N methodology to study the effect of cultivation of faba bean (Vicia faba L.), pea (Pisum sativum L.) and barley (Hordeum vulgare L.) on the N status of soil and their residual N effect on two succeeding cereals (sorghum (Sorghum vulgare) followed by barley). Faba bean, pea and barley took up 29.6, 34.5 and 53.0 kg N ha^–1 from the soil, but returned to soil through roots only 11.3, 10.8 and 5.7 kg N ha^–1, respectively. Hence, removal of faba bean, pea and barley straw resulted in a N-balance of about –18, –24, and –47 kg ha^–1 respectively. A soil nitrogen conserving effect was observed following the cultivation of faba bean and pea compared to barley which was of the order of 23 and 18 kg N ha^–1, respectively. Cultivation of legumes resulted in a significantly higher A_N value of the soil compared to barley. However, the A_N of the soil following fallow was significantly higher than following legumes, implying that the cultivation of the legumes had depleted the soil less than barley but had not added to the soil N compared to the fallow. The beneficial effect of legume cropping also was reflected in the N yield and dry matter production of the succeeding crops. Cultivation of legumes led to a greater exploitation of soil N by the succeeding crops. Hence, appreciable yield increases observed in the succeeding crops following legumes compared to cereal were due to a N-conserving effect, carry-over of N from the legume residue and to greater uptake of soil N by the succeeding crops when previously cropped to legumes.     

Nitrate deposition in northern hardwood forests and the nitrogen metabolism of Acer saccharum marsh

It is generally assumed that plant assimilation constitutes the major sink for anthropogenic Nitrate NO ₃ ^– deposited in temperate forests because plant growth is usually limited by nitrogen (N) availability. Nevertheless, plants are known to vary widely in their capacity for NO ₃ ^– uptake and assimilation, and few studies have directly measured these parameters for overstory trees. Using a combination of field and greenhouse experiments, we studied the N nutrition of Acer saccharum Marsh. in four northern hardwood forests receiving experimental NO ₃ ^– additions equivalent to 30 kg N ha^–1 year^–1. We measured leaf and fine-root nitrate reductase activity (NRA) of overstory trees using an in vivo assay and used ¹⁵N to determine the kinetic parameters of NO ₃ ^– uptake by excised fine roots. In two greenhouse experiments, we measured leaf and root NRA in A. saccharum seedlings fertilized with 0–3.5 g NO ₃ ^– –N m^–2 and determined the kinetic parameters of NO ₃ ^– and NH ₄ ⁺ uptake in excised roots of seedlings. In both overstory trees and seedlings, rates of leaf and fine root NRA were substantially lower than previously reported rates for most woody plants and showed no response to NO ₃ ^– fertilization (range = non-detectable to 33 nmol NO ₂ ^– g^–1 h^–1). Maximal rates of NO ₃ ^– uptake in overstory trees also were low, ranging from 0.2 to 1.0 mol g^–1 h^–1. In seedlings, the mean V _max for NO ₃ ^– uptake in fine roots (1 mol g^–1 h^–1) was approximately 30 times lower than the V _max for NH ₄ ⁺ uptake (33 mol g^–1 h^–1). Our results suggest that A. saccharum satisfies its N demand through rapid NH ₄ ⁺ uptake and may have a limited capacity to serve as a direct sink for atmospheric additions of NO ₃ ^– .     

The role of Azolla cover in improving the nitrogen use efficiency of lowland rice

The development of management techniques to improve the poor N use efficiency by lowland rice (Oryza sativa L.) and reduce the high N losses has been an important focus of agronomic research. The potential of an Azolla cover in combination with urea was assessed under field conditions in Laguna, Philippines. Two on-station field experiments were established in the 1998–1999 dry season and eight on-farm experiments per season were carried out in the 2000–2001 wet and dry seasons. Treatment combinations consisting of N levels applied alone or combined with Azolla were evaluated with respect to floodwater chemistry, ¹⁵N recovery, crop growth, and grain yield. A full Azolla cover on the floodwater surface at the time of urea application prevented the rapid and large increase in floodwater pH and floodwater temperature. As a consequence, the partial pressure of ammonia (NH₃), which is an indicator of potential NH₃ volatilization, was significantly depressed. ¹⁵N recovery was higher in plots covered with Azolla where the total ¹⁵N recovery ranged between 77 and 99%, and the aboveground (grain and straw) recovery by rice ranged between 32 and 61%. The tiller count in Azolla-covered plots was significantly increased by 50% more than the uncovered plots at all urea levels. Consequently, the grain yield was likewise improved. Grain yields from the 16 on-farm trials increased by as much as 40% at lower N rates (40 and 50 kg N ha^–1) and by as much as 29% at higher N rates (80 and 100 kg N ha^–1). In addition, response of rice to treatments with lower N rates with an Azolla cover was comparable to that obtained with the higher N rates without a cover. Thus, using Azolla as a surface cover in combination with urea can be an alternative management practice worth considering as a means to reduce NH₃ volatilization losses and improve N use efficiency.     

Inter-row crop competition for band-injected ammonium nitrate

Inter-species competition for applied nutrients may be managed by weed control in cultivated communities. However, intra-crop competition may still occur. Thus, regarding a fertilizer band as a finite nutrient source, individual rows of a cereal crop may compete for the applied nitrogen. The aim was to explore the partition of applied N between crop rows affected by the displacement of the fertilizer band relative to the rows. In a micro-plot experiment, a ¹⁵N-ammonium–¹⁵N-nitrate solution was injected in four positions parallel to two rows of spring wheat (Triticum aestivum ssp. vulgare). The course of ¹⁵N crop uptake in each row was estimated by a sigmoid growth function using eight samplings during the elongation phase. The estimated parameters were used for calculating the asymptotic value (a), the time until a/2 was reached (T_0.5a), and the uptake rate at this time (MaxRate), and related to the distance between the crop row and fertilizer band. The a-value decreased by 5.1%-point recovery cm^–1, T_0.5a increased by 0.5 day cm^–1 and the MaxRate decreased by 0.3%-point recovery day^–1 cm^–1. Placement of the fertilizer band centred between two crop rows caused an even access to the applied nitrogen, but a parallel displacement from the centre by a few centimetres significantly affected the parameters. The proliferation of the roots in the soil and thus the time until the roots reached the fertilizer zone is assumed to cause the inter-row competition for the band-injected N source. The access to the fertilizer band was clearly reflected in the dry matter production causing an uneven crop stand. The consequences of inter-row crop competition for a band-injected N source with respect to grain yield and grain quality were not explored, but attention on these parameters are required, particularly in the production of high quality wheat grains.     

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.     

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.     

Effect ofAzospirillum inoculation on nitrogen fixation and growth of several winter legumes

Summary Inoculation of naturally nodulatedPisum sativum L. (garden pea) withAzospirillum in the greenhouse caused a significant increase in nodule numbers above controls. Field inoculation of garden peas in the winter 1981–1982 andCicer arietinum L. (chick pea), in winter 1982–1983, withAzospirillum one week after plant emergence, produced a significant increase in seed yield, but did not affect plant dry matter yield. ForVicia sativa L. (vetch) grown in soil in the greenhouse and in the field for forage, winter 1980–1981, inoculation significantly increased dry matter yield, %N, N-content, and acetylene reduction (nitrogen fixation) activity. InHedysarum coronarium L. (sulla clover), winter 1981–1982, inoculated with both its specificRhizobium (by the slurry method) andAzospirillum, 7 days after emergence, there was an increase in acetylene reduction above controls inoculated withRhizobium alone. These results suggest that it is possible, under conditions tested in this work, to increase nodulation, nitrogen fixation, and crop yields of winter legumes by inoculation withAzospirillum.     

Rootless Aquatic Plant Aldrovanda Vesiculosa: Physiological Polarity, Mineral Nutrition, and Importance of Carnivory

Various ecophysiological investigations are presented in Aldrovanda vesiculosa, a rootless aquatic carnivorous plant. A distinct polarity of N, P, and Ca tissue content per dry mass (DM) unit was found along Aldrovanda shoots. Due to effective re-utilization, relatively small proportions of N (10 – 13 %) and P (33 – 43 %) are probably lost with senescent leaf whorls, while there is complete loss of all Ca, K, and Mg. The total content of starch and free sugars was 26 – 47 % DM along adult shoots, with the maximum in the 7^th – 10^th whorls. About 30 % of the total maximum sugar content was probably lost with dead whorls. The plant was found to take up 5 – 7 times more NH₄ ⁺ to NO₃ ^– from a mineral medium. Under nearly-natural conditions in an outdoor cultivation container, catching of prey led to significantly more rapid growth than in unfed plants. DM of the fed controls was 48 % higher than in the unfed plants. The controls produced 0.69 branches per plant, while the unfed plants did not produced any. However, the N and P content per DM unit increased by 6 – 25 % in the apices and the first 6 whorls in the unfed variant, as compared to the fed controls. It may be suggested that carnivory is very important for Aldrovanda.