Kinetics of zinc uptake by mycorrhizal (VAM) and non-mycorrhizal corn (Zea mays L.) roots

Summary The kinetics of Zn absorption were studied in mycorrhizal (Glomus macrocarpum) and non-mycorrhizal roots of corn (Zea mays L.) at pH 6.0 at Zn concentrations of 75 mol to 1.07 mol m^-3. Five concentration-dependent phases of Zn absorption were recognized; phase 0 (1.5–4.0 mmol m^-3) was linear but the other four phases (4.0 mmol to 1.07 mol m^-3) obeyed Michaelis-Menten kinetics. At low concentrations (less than 4 mmol m^-3), sigmoidal kinetics of Zn absorption were observed. The absorption of Zn by mycorrhizal maize was greater at low concentrations but decreased at higher levels. This appeared to be a result of a higher maximal uptake rate in phase 1 and lower K _m values in the subsequent phases. Kinetic models yielding continuous isotherms could not account for the observed multiphasic pattern.Research paper no. 6820 through the Director, Experiment Station, G.B. Pant University of Agriculture and Technology, Pantnagar 263 145, UP, India     

Relative significance of soil and nitrogenous fertilizer in nitrogen nutrition and growth of wetland rice (Oryza sativa L.)

Summary A pot experiment was conducted to compare the yields from five commercially cultivated varieties (Bas-198, Bas-370, Bas-Pak, Bas-385, and IR-6) of rice (Oryza sativa L.) and to establish the relative significance of soil N and fertilizer N (¹⁵N-labelled ammonium sulphate) in affecting crop performance. Another aim was to study the interaction of fertilizer N and soil N as influenced by different rice varieties. Among the five varieties tested, Bas-Pak gave the maximum dry matter and N yield. The N-use efficiency (percentage of applied N taken up by the plants) of different varieties ranged between 33.7 and 43.7%, Bas-Pak being the most efficient. Significant losses of fertilizer N occurred from the soil-plant system. The maximum N loss (52.1% of applied N) was observed with IR-6 and the minimum loss (39.2%) with Bas-Pak. A substantial increase in the uptake of soil N following the application of fertilizer and an interaction between the two N sources were observed with all varieties except Bas-385. The interaction was attributed to greater root proliferation following the application of fertilizer. It was concluded that a varietey with greater potential to use soil N is likely to give a better yield and that, of the two N sources, the availability of soil N was more important in determining the yield performance of different varieties of rice.     

Effect of pre-incubated sawdust-based cowdung on growth and nutrient uptake of Zea mays (L.) and on soil chemical properties

Summary A greenhouse soil culture experiment was set up to evaluate the effect of pre-incubation time (0, 2, 4, 6 weeks) of sawdust-based cowdung, prior to use, on the growth, dry-matter yield, and uptake of nutrients by Zea mays (L). The chemical properties of the soil (an Alfisol) were examined after the various treatments. The results indicated that incubation of cowdung for 4 weeks before planting time enhanced the growth and dry-matter yields of maize. Soil pH increased with the length of incubation while organic-C contents and cation exchange capacity remained unaffected. The length of pre-incubation enhanced the uptake of N and P but did not affect the uptake of K, Ca, Mg, and Na.     

Availability of soil and fertilizer nitrogen to wheat (Triticum aestivum L.) following rice-straw amendment

Summary A pot experiment was conducted to study the N availability to wheat and the loss of ¹⁵N-labelled fertilizer N as affected by the rate of rice-straw applied. The availability of soil N was also studied. The straw was incorporated in the soil 2 or 4 weeks before a sowing of wheat and allowed to decompose at a moisture content of 60% or 200% of the water-holding capacity. The wheat plants were harvested at maturity and the roots, straw, and grains were analysed for total N and ¹⁵N. The soil was analysed for total N and ¹⁵N after the harvest to determine the recovery of fertilizer N in the soil-plant system and assess its loss. The dry matter and N yields of wheat were significantly retarded in the soil amended with rice straw. The availability of soil N to wheat was significantly reduced due to the straw application, particularly at high moisture levels during pre-incubation, and was assumed to cause a reduction in the dry matter and N yields of wheat. A significant correlation (r=0.89) was observed between the uptake of soil N and the dry matter yield of wheat with different treatments. In unamended soil 31.44% of the fertilizer N was taken up by the wheat plants while 41.08% of fertilizer N was lost. The plant recovery of fertilizer N from the amended soil averaged 30.78% and the losses averaged 45.55%     

Ammonia volatilization from nitrogen fertilizers surface-applied to corn (Zea mays) and grass pasture (Dactylis glomerata)

Summary The major agronomic concern with NH₃ loss from urea-containing fertilizers is the effect of these losses on crop yields and N fertilizer efficiency. In this 2-year study, NH₃ volatilization from surface-applied N fertilizers was measured in the field, and the effects of the NH₃ losses detected on corn (Zea mays L.) and orchardgrass (Dactylis glomerata L.) yield and N uptake were determined. For corn, NH₄NO₃ (AN), a urea-AN solution (UAN), or urea, were surface-broadcast at rates of 0, 56 and 112 kg N ha^–1 on a Plano silt loam (Typic Argiudoll) and on a Fayette silt loam (Typic Hapludalf). Urea and AN (0 and 67 kg N ha^–1) were surface-applied to grass pasture on the Fayette silt loam. Significant NH₃ losses from urea-containing N sources were detected in one of four corn experiments (12%–16% of applied N) and in both experiments with grass pasture (9%–19% of applied N). When these losses occurred, corn grain yields with UAN and urea were 1.0 and 1.5 Mg ha^–1, respectively, lower than yields with AN, and orchardgrass dry matter yields with urea were 0.27 to 0.74 Mg ha^–1 lower than with AN. Significant differences in crop N uptake between N sources were detected, but apparent NH₃ loss based on N uptake differences was not equal to field measurements of NH₃ loss. Rainfall following N application markedly influenced NH₃ volatilization. In corn experiments, NH₃ loss was low and yields with all N sources were similar when at least 2.5 mm of rainfall occurred within 4 days after N application. Rainfall within 3 days after N application did not prevent significant yield reductions due to NH₃ loss from urea in grass pasture experiments.     

Biological nitrogen fixation in maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.) by <Superscript>15</Superscript>N isotope-dilution and identification of associated culturable diazotrophs

The nitrogen-fixing capacity of a range of commercial cultivars of maize (Zea mays L.) was evaluated by the ¹⁵N isotope-dilution method. Biological nitrogen fixation (BNF) expressed as percent nitrogen derived from air (Ndfa) ranged from 12 to 33 regardless of nitrogen fertilization. BNF was not affected by mineral nitrogen fertilization except on cultivar Topacio and PAU-871 cultivars. Subsequently, culturable bacterial diazotrophs were isolated from endophytic tissue of maize: seed, root, stem, and leaf. All isolates were able to grow on N-free semisolid medium. Eleven bacteria isolates showed nitrogen-fixing capacity by the reduction of acetylene to ethylene and confirmed by PCR the presence of nifH gene in their genome. Identification of the 11 isolates was performed by bacteriological methods, 16S rRNA gene sequences, and phylogenetic analysis, which indicated that the bacteria isolated were closely related to Pantoea, Pseudomonas, Rhanella, Herbaspirillum, Azospirillum, Rhizobium (Agrobacterium), and Brevundimonas. This study demonstrated that maize cultivars obtain significant nitrogen from BNF, varying by maize cultivar and nitrogen fertilization level. The endophytic diazotrophic bacteria isolated from root, stem, and leaf tissues of maize cultivars may contribute to BNF in these plants.     

Effects of 15N-labelled ammonium nitrate and urea on soil nitrogen during growth of wheat (Triticum aestivum L.) under field conditions

We studied the effects of ¹⁵N-labelled ammonium nitrate and urea on the yield and uptake of labelled and unlabelled N by wheat (Triticum aestivum L., cv. Mexi-Pak-65) in a field experiment. The dry matter and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The wheat crop used 33.6–51.5 and 30.5–40.9% of the N from ammonium nitrate and urea, respectively. Splitting the fertilizer N application had a significant effect on the uptake of fertilizer N by the wheat. The fertilizer N uptake showed that ammonium nitrate was a more available source of N for wheat than urea. The effective use of fertilizer N (ratio of fertilizer N in grain to fertilizer N in whole plant) was statistically similar for the two N fertilizers. The application of fertilizer N increased the uptake of unlabelled soil N by wheat, a result attributed to a positive added N interaction, which varied according to the fertilizer N split; six split applications gave the highest added N interaction compared to a single application or two split applications for both fertilizers. Ammonium nitrate gave 90.5, 33.5, and 48.5% more added N interaction than urea with one, two, and six split N applications. A values were not significantly correlated with the added N interaction (r=0.557). The observed added N interaction may have been the result of pool substitution, whereby added labelled fertilizer N replaced unlabelled soil N.     

The effect of Azospirillum brasilense Sp7 and Azotobacter chroococcum on nitrogen balance in soil under cropping with oats (Avena sativa L.)

Summary Pot experiments with oats were carried out to study the effect of Azospirillum brasilense Sp 7 and Azotobacter chroococcum 94K on the yield of plants, the N content of soil and the ¹⁴N balance. The plants were grown on gray forest soil under irrigation with deionized water and application of ¹⁵N-labelled fertilizer at a rate of 4 mg N 100 g^-1 soil. Inoculation of plants with Azospirillum spp. and Azotobacter spp. failed to increase the plant yield. However, the increase in total N in the soil at the end of the experiment and the positive ¹⁴N balance in the soil-plant system due to increased nitrogenase activity in the rhizosphere were statistically significant. The amount of N accumulated in the soil was comparable with the rate of N applied as fertilizer.     

Kinetics of 15N-labelled nitrate uptake by maize (Zea mays L.) root segments

Abstract

Isotherms and kinetic constants of nitrate uptake by excised root segments from the apical root zone of 6-d-old maize seedlings pretreated with nitrate were investigated using ¹⁵N-labelled nitrate. The isotherms were resolved into two systems namely a multiphasic saturable system at substrate concentrations lower than 2 mol m^?-3 and a linear system at higher concentrations. The detailed analysis of the multiphasic saturable system suggested the existence of at least three phases, which followed the Michaelis-Menten kinetics. The I _max and K _m of each phase increase from the lower phase to the upper phase. The distance from the root tip and the presence of stele affected considerably the linear system but only slightly the saturable system.     

Interaction of 15N-labelled ammonium nitrate,urea and ammonium sulphate with soil N during growth of wheat (Triticum aestivum L.) under field conditions

The effects of ¹⁵N-labelled ammonium nitrate, urea and ammonium sulphate on yield and uptake of labelled and unlabelled N by wheat (Triticum aestivum L. cv. Mexi-Pak-65) were studied in a field experiment. The dry matter and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The wheat crop used 64.0–74.8%, 61.5–64.7% and 61.7–63.4% of the N from ammonium nitrate, urea and ammonium sulphate, respectively. The fertilizer N uptake showed that ammonium nitrate was a more available source of N for wheat than urea and ammonium sulphate. The effective use of fertilizer N (ratio of fertilizer N in grain to fertilizer N in whole plant) was statistically similar for the three N fertilizers. The application of fertilizer N increased the uptake of unlabelled soil N by wheat, a result attributed to a positive added N interaction, which varied with the method of application of fertilizer N. Ammonium nitrate, urea and ammonium sulphate gave 59.3%, 42.8% and 26.3% more added N interaction, respectively, when applied by the broadcast/worked-in method than with band placement. A highly significant correlation between soil N and grain yield, dry matter and added N interaction showed that soil N was more important than fertilizer N in wheat production. A values were not significantly correlated with added N interaction (r=0.719). The observed added N interaction may have been the result of pool substitution, whereby added labelled fertilizer N stood proxy for unlabelled soil N.     

Shoot,root, soil and microbial nitrogen dynamics in two contrasting soils cropped to barley (Hordeum vulgare L.)

Summary Dynamics of barley N, mineral N, and organic N were compared at Ellerslie (Black Chernozem, Typic Cryoboroll) and Breton (Gray Luvisol, Typic Cryoboralf) in central Alberta, using ¹⁵N-urea. On average, shoot N and shoot ¹⁵N recoveries at Ellerslie (14.1 g m^–2, 36%) were greater than at Breton (4.5 g m^–2, 17%). Root N (g m^–2) did not significantly differ between sites (0–30 cm) but root ¹⁵N recovery was greater at Breton (3.4%) than Ellerslie (1.8%). Low levels of shoot N and shoot ¹⁵N at Breton were partly due to very wet soil conditions in July, which resulted in premature shoot senescence and low plant N uptake. Although the total ¹⁵N recoveries from the system (to 30 cm depth) at Ellerslie (63%) and Breton (56%) were similar, soil ¹⁵N was greater at Breton (35%) than at Ellerslie (26%). There were no differences in mineral N between sites but the average ¹⁵N recovery in the mineral-N pool was significantly greater at Ellerslie (3.3%) than at Breton (1.6%). There was no difference in ¹⁵N recovery in the microbial biomass (3%) between sites, although non-microbial organic ¹⁵N was greater at Breton (31 %) than at Ellerslie (20%). The two soils showed differences in the relative size of kinetically active N pools and in relative mineralization rates. Microbial N (0–30 cm) was greater at Ellerslie (13.3 g m^–2) than at Breton (9.9 g m^–2), but total microbial N made up a larger proportion of total soil N at Breton (1.6%) than at Ellerslie (0.9%). In the 0–10 cm interval, microbial N was 1.7-fold greater and non-microbial active N was 3-fold greater at Breton compared to Ellerslie, when expressed as a proportion of total soil N. Net N mineralization in a 10-day laboratory incubation was 1.4-fold greater in the Black Chernozem (0–10 cm interval) from Ellerslie, compared to the Gray Luvisol from Breton, when expressed per gram of soil. Net N mineralization in the soil from Breton was double that of the soil from Ellerslie, when expressed as a proportion of soil N. Although soil N (g m^–2) was 2.5-fold greater at Ellerslie compared to Breton, it was cycled more rapidly at Breton.     

Natural N abundances of inorganic nitrogen in soil treated with fertilizer and compost under changing soil moisture regimes

This study was conducted to examine whether the applications of N-inputs (compost and fertilizer) having different N isotopic compositions (δ¹⁵N) produce isotopically different inorganic-N and to investigate the effect of soil moisture regimes on the temporal variations in the δ¹⁵N of inorganic-N in soils. To do so, the temporal variations in the concentrations and the δ¹⁵N of NH₄⁺ and NO₃⁻ in soils treated with two levels (0 and 150 mg N kg⁻¹) of ammonium sulfate (δ¹⁵N=−2.3‰) and compost (+13.9‰) during a 10-week incubation were compared by changing soil moisture regime after 6 weeks either from saturated to unsaturated conditions or vice versa. Another incubation study using ¹⁵N-labeled ammonium sulfate (3.05 ¹⁵N atom%) was conducted to estimate the rates of nitrification and denitrification with a numerical model FLUAZ. The δ¹⁵N values of NH₄⁺ and NO₃⁻ were greatly affected by the availability of substrate for each of the nitrification and denitrification processes and the soil moisture status that affects the relative predominance between the two processes. Under saturated conditions for 6 weeks, the δ¹⁵N of NH₄⁺ in soils treated with fertilizer progressively increased from +2.9‰ at 0.5 week to +18.9‰ at 6 weeks due to nitrification. During the same period, NO₃⁻ concentrations were consistently low and the corresponding δ¹⁵N increased from +16.3 to +39.2‰ through denitrification. Under subsequent water-unsaturated conditions, the NO₃⁻ concentrations increased through nitrification, which resulted in the decrease in the δ¹⁵N of NO₃⁻. In soils, which were unsaturated for the first 6-weeks incubation, the δ¹⁵N of NH₄⁺ increased sharply at 0.5 week due to fast nitrification. On the other hand, the δ¹⁵N of NO₃⁻ showed the lowest value at 0.5 week due to incomplete nitrification, but after a subsequence increase, they remained stable while nitrification and denitrification were negligible between 1 and 6 weeks. Changing to saturated conditions after the initial 6-weeks incubation, however, increased the δ¹⁵N of NO₃⁻ progressively with a concurrent decrease in NO₃⁻ concentration through denitrification. The differences in δ¹⁵N of NO⁻₃ between compost and fertilizer treatments were consistent throughout the incubation period. The δ¹⁵N of NO₃⁻ increased with the addition of compost (range: +13.0 to +35.4‰), but decreased with the addition of fertilizer (−10.8 to +11.4‰), thus resulting in intermediate values in soils receiving both fertilizer and compost (−3.5 to +20.3‰). Therefore, such differences in δ¹⁵N of NO₃⁻ observed in this study suggest a possibility that the δ¹⁵N of upland-grown plants receiving compost would be higher than those treated with fertilizer because NO₃⁻ is the most abundant N for plant uptake in upland soils.     

Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules

Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH ₄ ⁺ -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N₂ fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O₂ than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH ₄ ⁺ -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.     

Diversity of diazotroph populations in the rhizosphere of maize (Zea mays L.) growing on different French soils

Summary We studied the dominant diazotrophs associated with maize roots and rhizosphere soil originating from three different locations in France. An aseptically grown maize plantlet, the spermosphere model, was used to isolate N₂-fixing (acetylene-reducing) bacteria. Bacillus circulans was the dominant N₂-fixing bacterium in the rhizosphere of maize-growing soils from Ramonville and Trogny, but was not found in maize-growing sandy soil from Pissos. In the latter soil, Enterobacter cloacae, Klebsiella terrigena, and Pseudomonas sp. were the most abundant diazotrophs. Azospirillum sp., which has been frequently reported as an important diazotroph accociated with the maize rhizosphere, was not isolated from any of these soils. The strains were compared for their acetylene-reducing activity in the spermosphere model. The Bacillus circulans strains, which were more frequently isolated, also exhibited significantly greater acetylene-reducing activity (3100 nmol ethylene day^-1 plant^-1) than the Enterobacteriaceae strains (180 nmol ethylene day^-1 plant^-1). This work indicates for the first time that Bacillus circulans is an important maizerhizosphere-associated bacterium and a potential plant growth-promoting rhizobacterium.     

Effects of Selected Surfactants on Nutrient Uptake in Corn (Zea mays L.)

Surfactants in herbicide formulations eventually enter soil and may disrupt various processes. Research examined effects on nutrient uptake in corn caused by surfactants, herbicides, and surfactant-herbicide combinations applied to silt loam and silty clay loam soils in the greenhouse. Surfactants evaluated were Activator 90, Agri-Dex, and Thrust; herbicides were glyphosate, atrazine, and bentazon. Corn was planted in fertilized soils with moisture content maintained for optimum growth. Foliage (V8 growth stage) was collected for elemental analyses. Nutrient uptake differed with soil texture. Nutrient uptake from silty clay loam was more affected by surfactants and/or herbicides than in silt loam. Potassium uptake was significantly (P = 0.05) decreased in silt loam only by Thrust but uptake of phosphorus (P), potassium (K) calcium (Ca), sulfur (S), copper (Cu), and zinc (Zn) decreased by ≤30% in silty clay loam treated with surfactants. Surfactants and/or herbicides may interact with soil texture to affect nutrient uptake. Long-term field studies to validate changes in nutrient uptake and grain yields after annual applications of surfactants plus herbicides are needed.     

Grain yield response of Zea mays (hybrid AE 703) to Azotobacter chroococcum H23

Summary Field experiments were conducted during 1985 and 1986 to examine the effect of Azotobacter chroococcum on the grain yield of maize. Application of 40 kg N ha^–1 plus A. chroococcum caused a significant increase in maize yield. Azotobacter inoculation was more efficient at lower doses (40 kg N ha^–1) than at high doses (80 kg N ha^–1) of urea.     

Field evaluation of N2 fixation and N partitioning in climbing bean (Phaseolus vulgaris L.) using 15N

Summary The common bean (Phaseolus vulgaris L.) is generally regarded as a poor N₂ fixer. This study assessed the sources of N (fertilizer, soil, and fixed N), N partitioning and mobilization, and soil N balance under field conditions in an indeterminate-type climbing bean (P. vulgaris L. cv. Cipro) at the vegetative, early pod-filling, and physiological maturity stages, using the A-value approach. This involved the application of 10 and 100 kg N ha^-1 of ¹⁵N-labelled ammonium sulphate to the climbing bean and a reference crop, maize (Zea mays L.). At the late pod-filling stage (75 days after planting) the climbing bean had accumulated 119 kg N ha^-1, 84% being derived from fixation, 16% from soil, and only 0.2% from the ¹⁵N fertilizer. N₂ fixation was generally high at all stages of plant growth, but the maximum fixation (74% of the total N₂ fixed) occurred during the interval between early (55 days after planting) and late podfilling. The N₂ fixed between 55 and 75 days after planting bas a major source (88%) of the N demand of the developing pod, and only about 11% was contributed from the soil. There was essentially no mobilization of N from the shoots or roots for pod development. The cultivation of common bean cultivars that maintain a high N₂-fixing capacity especially during pod filling, satisfying almost all the N needs of the developing pod and thus requiring little or no mobilization of N from the shoots for pod development, may lead to a net positive soil N balance.     

Effects of nitrogen rate and nitrogen form on glycine uptake by pakchoi (Brassica chinensis L.) under sterile culture

It is well known that plants are capable of taking up intact amino acids. However, how the nitrogen (N) rates and N forms affect amino acid uptake and amino acid nutritional contribution for plant are still uncertain. Effects of the different proportions of nitrate (NO₃^?), ammonium (NH₄⁺) and ¹⁵N-labeled glycine on pakchoi seedlings glycine uptake were investigated for 21 days hydroponics under the aseptic media. Our results showed that plant biomass and glycine uptake was positively related to glycine rate. NO₃^? and NH₄⁺, the two antagonistic N forms, both significantly inhibited plant glycine uptake. Their interactions with glycine were also negatively related to glycine uptake and glycine nutritional contribution. Glycine nutritional contribution in the treatments with high glycine rate (13.4%–35.8%) was significantly higher than that with low glycine rate (2.2%–13.2%). The high nutritional contribution indicated amino acids can serve as an important N source for plant growth under the high organic and low inorganic N input ecosystem.     

Priming effects of 15N-labelled ammonium nitrate on uptake of soil N by wheat (Triticum aestivum L.) under field conditions

Summary The uptake of labelled and unlabelled N by wheat was measured in a field experiment using ¹⁵N-labelled ammonium nitrate fertilizer. The dry matter yield and N yields were significantly increased with fertilizer N application compared to those from unfertilized soil. The uptake of applied N by wheat ranged between 25 and 34%. Fertilizer N application increased the uptake of unlabelled soil N which was attributed to a positive priming effect or added N interaction. The added N interaction observed by applying 20, 60, and 120 kg fertilizer N was 11.4, 19.1, and 27.9 kg, corresponding to 26, 44 and 64%, respectively of the N taken up from unfertilized soil. The A values did not alter with the increase in fertilizer N application. The observed added N interaction may have been the result of pool substitution whereby added labelled fertilizer N stood proxy for unlabelled soil N. A significant correlation coefficient (r=0.996^**) between the uptake of soil N and the dry matter yield showed that soil N was more important than fertilizer N in wheat production.     

Fate of Azolla spp. and urea nitrogen applied to wetland rice (Oryza sativa L.)

Summary Using ¹⁵N, the fate of N applied to wetland rice either as Azolla or urea was studied in a field at the International Rice Research Institute (IRRI). In bigger plots nearby, yield response and N uptake were also determined with unlabelled N sources. Azolla microphylla was labelled by repeated application of labelled ammonium sulfate. Labelled and unlabelled N were used alternately in applications of Azolla or urea 0 and 42 days after transplanting, in order to determine the effect of the time of application on the availability of Azolla N. The quantities of Azolla N incorporated were 23% more than those of urea N (30 kg N ha^–1) in the isotope plots or 7% less in the yield response plots. Grain yield and total N uptake by the rice plants in the yield-response plots were higher in the urea-treated plots than in the Azolla-treated plots, but the physiological effect of Azolla N (grain yield response/increase in N uptake) was higher than that of rea. The labelled N balance was studied after the first and second crops of rice. Losses of labelled N after the first crop were higher from urea (30%–32%) than from Azolla (0%–11 %). Losses in N applied as a side dressing 42 days after transplanting were less than those of N applied basally. No further losses of ¹⁵N occurred after the first crop. The recovery of Azolla ¹⁵N in the first crop of rice was 39% from the basal application and 63% from the side dressing. The recovery of urea ¹⁵N was 27% from the basal application and 48% from the side dressing. Recoveries of residual N from both Azolla and urea during the second rice crop were similar. Laboratory incubation of the Azolla used and the changes in labelled exchangeable N in the soil showed that at least 65% of Azolla N (4.7% N content) was mineralized within 10 days.