Rice yields as influenced by Azolla N2 fixation and urea N-fertilization

Application of 0, 30, 60, 90 and 120 kg N ha^–1 of urea (U) in split doses with (and without)Azolla pinnata, R. Brown was studied for three consecutive seasons under planted field condition. Fresh weight (FW), acetylene reduction activity (ARA) and N yield of Azolla were found to be maximum 14 days after inoculation (DAI). Among the different treatments, maximum Azolla growth was recorded in no N control. The FW, ARA and N yield of Azolla were inhibited increasingly with the increase in N levels. Irrespective of season, FW and N yield of Azolla were inhibited only a small extent with 90 kg N ha^–1 U, beyond which the inhibition was pronounced. ARA was inhibited only slightly up to 60 kg N ha^–1 of U. Grain yield and crop N uptake of rice increased significantly up to 90 kg N ha^–1 of U (alone or in combination with Azolla) in the dry seasons (variety IR 36) and up to 60 kg N ha^–1 U in the wet season (variety CR 1018).     

Availability of nitrogen from 15N-labelled Azolla pinnata and urea to flooded rice

In view of the recently generated interest in Azolla and the high cost of N fertilizers, this field study was aimed at measuring the availability of Azolla-N applied in two split application in comparison to urea-N. Azolla was cultivated and labelled with ¹⁵N isotope in the field. A total of about 60 kg N ha^-1 was applied as Azolla, urea or Azolla and urea in combination, in two equal splits at transplanting and at maximum tillering, i.e. 30 days after transplanting (30 DAT).The recovery by the crop of Azolla-N applied at 30 DAT was significantly higher than that applied at transplanting, viz. 30.2% and 20.2%, respectively. The recoveries of urea-N applied at the same stages were similarly low, viz. 22.5% at transplanting and 38.6% at 30 DAT. Total recoveries of fertilizer N at the time of harvest were 26.8% from Azolla, 30.7% from urea applied in the same two splits and 49.1% from urea applied in locally recommended three splits. Recoveries of labelled Azolla-N in succeeding rice crop were twice higher than those of labelled urea-N. The recoveries ranged from 1.9 to 2.1% from urea-N and 4.0 to 4.9% from Azolla-N. There were no differences in residual ¹⁵N recovery in the succeeding crop between Azolla and urea either applied at transplanting or at 30 DAT.     

Studies on the availability of Azolla N and urea N for rice growth using 15N

Field experiments (20 m² plots) were conducted to compare Azolla and urea as N sources for rice (Oryza sativa L.) in both the wet and dry seasons. Parallel microplot (1 m²) experiments were conducted using ¹⁵N. A total of approximately 60 kg N ha^-1 was applied as urea, Azolla, or urea plus Azolla. Urea or Azolla applied with equal applications of 30 kg N ha^-1 at transplanting (T) and at maximum tillering (MT) were equally effective for increasing rice grain yields in both seasons. Urea at 30 kg N ha^-1 at T and Azolla 30 kg N ha^-1 at MT was also equally effective. Urea applied by the locally recommended best split (40 kg at T and 20 kg at MT) gave a higher yield in the wet season, but an equal yield in the dry season. The average yield increase was 23% in the wet season, and 95% in the dry season. The proportion of the N taken up by the rice plants which was derived from urea (%NdfU) or Azolla (%NdfAz) was essentially identical for the treatments receiving the same N split. Recovery of ¹⁵N in the grain plus straw was also very similar. Positive yield responses to residual N were observed in the succeeding rice crop following both the wet and dry seasons, but the increases were not always statistically significant. Recovery of residual ¹⁵N ranged from 5.5 to 8.9% for both crops in succeeding seasons. Residual recovery from the urea applications was significantly higher than from Azolla in the crop succeeding the dry season crop. Azolla was equally effective as urea as an N source for rice production on a per kg N basis.     

Nitrogen gas (N2+N2O) flux from urea applied to lowland rice as affected by green manure

A field study was conducted on a clay soil (Andaqueptic Haplaquoll) in the Philippines to directly measure the evolution of (N₂+N₂O)−¹⁵N from 98 atom %¹⁵N-labeled urea broadcast at 29 kg N ha⁻¹ into 0.05-m-deep floodwater at 15 days after transplanting (DT) rice. The flux of (N₂+N₂O)−¹⁵N during the 19 days following urea application never exceeded 28 g N ha⁻¹ day⁻¹. The total recovery of (N₂+N₂O)−¹⁵N evolved from the field was only 0.51% of the applied N, whereas total gaseous¹⁵N loss estimated from unrecovered¹⁵N in the¹⁵N balance was 41% of the applied N. Floodwater (nitrate+nitrite)−N in the 5 days following urea application never exceeded 0.14 g N m⁻³ or 0.3% of the applied N. Prior cropping of cowpea [Vigna unguiculata (L.) Walp.] to flowering with subsequent incorporation of the green manure (dry matter=2.5 Mg ha⁻¹, C/N=15) at 15 days before rice transplanting had no effect on fate of urea applied to rice at 15 DT. The recovery of (N₂+N₂O)−¹⁵N and total¹⁵N loss during the 19 days following urea application were 0.46 and 40%, respectively. Direct recovery of evolved (N₂+N₂O)−¹⁵N and total¹⁵N loss from 27 kg applied nitrate-N ha⁻¹ were 20% and 53% during the same 19-day period. The failure of directly-recovered (N₂+N₂O)−¹⁵N to match total¹⁵N loss from added nitrate-¹⁵N might be due to entrapment of denitrification end products in soil or transport of gaseous end products to the atmosphere through rice plants. The rapid conversion of added nitrate-N to (N₂+N₂O)−N, the apparently sufficient water soluble soil organic C for denitrification (101 μg C g⁻¹ in the top 0.15-m soil layer), and the low floodwater nitrate following urea application suggested that denitrification loss from urea was controlled by supply of nitrate rather than by availability of organic C.     

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.     

Optimising biological N2 fixation by legumes in farming systems

Whether grown as pulses for grain, as green manure, as pastures or as the tree components of agro-forestry systems, the value of leguminous crops lies in their ability to fix atmospheric N₂, so reducing the use of expensive fertiliser-N and enhancing soil fertility. N₂ fixing legumes provide the basis for developing sustainable farming systems that incorporate integrated nutrient management. By exploiting the stable nitrogen isotope ¹⁵N, it has been possible to reliably measure rates of N₂ fixation in a wide range of agro-ecological field situations involving many leguminous species. The accumulated data demonstrate that there is a wealth of genetic diversity among legumes and their Rhizobium symbionts which can be used to enhance N₂ fixation. Practical agronomic and microbiological means to maximise N inputs by legumes have also been identified.     

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.     

Variability in nitrogen fixation of common bean (Phaseolus vulgaris L.) intercropped with maize

Thirty one selected bean lines were evaluated in the field for ability to support N₂ fixation when intercropped with maize which received 0, 30 and 60 kg N ha^–1 as ammonium sulphate. The amount of fixed N₂ was estimated using the natural variation of ¹⁵N and wheat as the standard non-fixing crop. Nitrogen as low as 15 kg N ha^–1 at sowing suppressed nodule weight and activity (acetylene reduction activity) but not nodule number, suggesting that the main effect of mineral N was on nodule development and function. ¹⁵N data revealed a high potential of the bean genotypes to fix N₂, with the most promising ones averaging between 50–60% of seed N coming from fixation. Bean lines CNF-480, Puebla-152, Mexico-309, Negro Argel, CNF-178, Venezuela-350 and WBR22-3, WBR22-50 and WBR22-55 were ranked as good fixers.     

Natural 15N abundances of maize and soil amended with urea and composted pig manure

To investigate the effect of inorganic fertilizer and composted manure amendments on the N isotope composition (delta ¹⁵N) of crop and soil, maize (Zea mays L.) was cultivated under greenhouse conditions for 30, 40, 50, 60, and 70 days. Composted pig manure (delta ¹⁵N= +13.9) and urea (-2.3) were applied at 0 and 0 kg N ha^–1 (C0U0), 0 and 150 kg N ha^–1 (C0U2), 150 and 0 kg N ha^–1 (C2U0), and 75 and 75 kg N ha^–1 (C1U1), respectively. The delta ¹⁵N of total soil-N was not affected by both amendments, but delta ¹⁵N of NH⁺ ₄ and NO^– ₃ provided some information on the N isotope fractionation in soil. During the early growth stage, significant differences (P < 0.05) in delta ¹⁵N among maize subjected to different treatments were observed. After 30 days of growth, the delta ¹⁵N values of maize were +6.6 for C0U0, +1.1 for C0U2, +7.7 for C2U0, and +4.5 for C1U1. However, effects of urea and composted manure application on maize delta ¹⁵N progressively decreased with increasing growth period, probably due to isotope fractionation accompanying N losses and increased uptake of soil-derived N by maize. After 70 days of growth, delta ¹⁵N of leaves and grains of maize amended with composted pig manure were significantly (P < 0.05) higher than those with urea. The temporal variations in delta ¹⁵N of maize amended with urea and composted manure indicate that plant delta ¹⁵N is generally not a good tracer for N sources applied to field. Our data can be used in validation of delta ¹⁵N fractionation models in relation to N source inputs.     

Effect of urea placement on leaching losses of nitrogen from flooded rice soils

In an effort to provide an explanation for the reported variability in fertilizer N efficiency from deepplaced urea on flooded rice, a set of controlled experiments was conducted to evaluate the effect of water percolation on fertilizer loss and plant uptake from¹⁵N labeled urea supergranules. Three soils of different texture (silt loam-clay) were subjected to various percolation rates (0–20 mm/day) while planted to rice which was harvested after approximately 40 days.The results indicate that moderate to high percolation through silt loam soil will lead to significant fertilizer N losses and drastically decrease the fertilizer uptake by plants. The permeability of the clay soil was too low for any leaching to take place. It is therefore concluded that deep placement of urea supergranules not be recommended in soils where percolation rates may exceed 5 mm/day, particularly if the cation exchange capacity of the soil is low. This experiment points to the need of evaluating and reporting the percolation rates in soils where experiments with supergranular urea are conducted.Contribution from the Agro-Economic Division of the International Fertilizer Development Center (IFDC), Muscle Shoals, Alabama 35660.     

Nitrogen-15 balances for urea and neem coated urea applied to lowland rice following two cowpea cropping systems

Little is known about whether the high N losses from inorganic N fertilizers applied to lowland rice (Oryza sativa L.) are affected by the combined use of either legume green manure or residue with N fertilizers. Field experiments were conducted in 1986 and 1987 on an Andaqueptic Haplaquoll in the Philippines to determine the effect of cowpea [Vigna unguiculata (L.) Walp.] cropping systems before rice on the fate and use efficiency of¹⁵N-labeled, urea and neem cake (Azadirachta indica Juss.) coated urea (NCU) applied to the subsequent transplanted lowland rice crop. The pre-rice cropping systems were fallow, cowpea incorporated at the flowering stage as a green manure, and cowpea grown to maturity with subsequent incorporation of residue remaining after grain and pod removal. The incorporated green manure contained 70 and 67 kg N ha⁻¹ in 1986 and 1987, respectively. The incorporated residue contained 54 and 49 kg N ha⁻¹ in 1986 and 1987, respectively. The unrecovered¹⁵N in the¹⁵N balances for 58 kg N ha⁻¹ applied as urea or NCU ranged from 23 to 34% but was not affected by pre-rice cropping system. The partial pressure of ammoniapNH₃, and floodwater (nitrate + nitrite)-N following application of 29 kg N ha⁻¹ as urea or NCU to 0.05-m-deep floodwater at 14 days after transplanting was not affected by pre-rice cropping system. In plots not fertilized with urea or NCU, green manure contributed an extra 12 and 26 kg N ha⁻¹, to mature rice plants in 1986 and 1987, respectively. The corresponding contributions from residue were 19 and 23 kg N ha⁻¹, respectively. Coating urea with 0.2g neem cake per g urea had no effect on loss of urea-N in either year; however, it significantly increased grain yield (0.4 Mg ha⁻¹) and total plant N (11 kg ha⁻¹) in 1987 but not in 1986.     

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.     

Nitrogen fixation and N transfer from peanut to rice cultivated in aerobic soil in an intercropping system and its effect on soil N fertility

The novel cultivation of paddy rice in aerobic soil reveals the great potential not only for water-saving agriculture, but also for rice intercropping with legumes and both are important for the development of sustainable agriculture. A two-year field experiment was carried out to investigate the yield advantage of intercropping peanut (Arachis hypogaea L., Zhenyuanza 9102) and rice (Oryza sativa L., Wuyujing 99-15) in aerobic soil, and its effect on soil nitrogen (N) fertility. A pot experiment was also conducted to examine the N₂-fixation by peanut and N transfer from peanut to rice at three N fertilizer application rates, i.e., 15, 75 and 150 kg N ha^–1 using a ¹⁵N isotope dilution method. The results showed that the relative advantage of intercropping, expressed as land equivalent ratio (LER), was 1.41 in 2001 and 1.36 in 2002. Both area-adjusted yield and N content of rice were significantly increased in the intercropping system while those of peanut were not significantly different between intercropping and monocropping systems. The yields of rice grain and peanut, for example, were increased by 29–37% and 4–7% in the intercropping system when compared to the crop grown in the monocropping system. The intercropping advantage was mainly due to the sparing effect of soil inorganic N contributed by the peanut. This result was proved by the higher soil mineral N concentration under peanut monocropping and intercropping than under the rice monocropping system.%Ndfa (nitrogen derived from atmosphere) by peanut was 72.8, 56.5 and 35.4% under monocropping and 76.1, 53.3 and 50.7% under the intercropping system at N fertilizer application rates of 15, 75 and 150 kg ha^–1, respectively. The ¹⁵N-based estimates of N transfer from peanut (%NTFL) was 12.2, 9.2 and 6.2% at the three N fertilizer application rates. N transferred from peanut accounted for 11.9, 6.4 and 5.5% of the total N accumulated in the rice plants in intercropping at the same three N fertilizer application rates, suggesting that the transferred N from peanut in the intercropping system made a contribution to the N nutrition of rice, especially in low-N soil.     

Nitrogen uptake and recovery from urea and green manure in lowland rice measured by15N and non-isotope techniques

In the recent past considerable attention is paid to minimize dependence on purchased inputs such as inorganic nitrogen fertilizer. Green manure in the form of flood-tolerant, stem-nodulatingSesbania rostrata andAeschynomene afraspera is an alternative N source for rice, which may also increase N use efficiency. Therefore research was conducted to determine the fate of N applied to lowland rice (Oryza sativa L.) in the form ofSesbania rostrata andAeschynomene afraspera green manure and urea in two field experiments using¹⁵N labeled materials.¹⁵N in the soil and rice plant was determined, and¹⁵N balances established. Apparent N recoveries were determined by non-tracer method. ¹⁵N recoveries averaged 90 and 65% of N applied for green manure and urea treatments, respectively. High partial pressures of NH₃ in the floodwater, and high pH probably resulted from urea application and favoured losses of N from the urea treatment. Results show that green manure N can supply a substantial proportion of the N requirements of lowland rice. Nitrogen released fromSesbania rostrata andAeschynomene afraspera green manure was in synchrony with the demand of the rice plant. The effect of combined application of green manure and urea on N losses from urea fertilizer were also investigated. Green manure reduced the N losses from¹⁵N labeled urea possibly due to a reduction in pH of the floodwater. Positive added N interactions (ANIs) were observed. At harvest, an average of 45 and 25% of N applied remained in the soil for green manure and urea, respectively.Contribution from IRRI, Los Baños, Philippines and Justus-Liebig-University, Giessen, GermanyContribution from IRRI, Los Baños, Philippines and Justus-Liebig-University, Giessen, Germany     

Estimation of symbiotic N2 fixation in an Amazon floodplain forest

Sustainable management for existing Amazonian forests requires an extensive knowledge about the limits of ecosystem nutrient cycles. Therefore, symbiotic nitrogen (N₂) fixation of legumes was investigated in a periodically flooded forest of the central Amazon floodplain (Várzea) over two hydrological cycles (20 months) using the ¹⁵N natural abundance method. No seasonal variation in ¹⁵N abundance (δ ¹⁵N values) in trees which would suggest differences in N₂ fixation rates between the terrestrial and the aquatic phase was found. Estimations of the percentage of N derived from atmosphere (%Ndfa) for the nodulated legumes with Neptunia oleracea on the one side and Teramnus volubilis on the other resulted in mean %Ndfa values between 9 and 66%, respectively. More than half of the nodulated legume species had %Ndfa values above 45%. These relatively high N gains are important for the nodulated legumes during the whole hydrological cycle. With a %Ndfa of 4–5% for the entire Várzea forest, N₂ fixation is important for the ecosystem and therefore, has to be taken into consideration for new sustainable land-use strategies in this area.     

Uptake and use patterns of nitrogen from urea,oxamide, and isobutylidene diurea by rice plants

Summary Two¹⁵N-labelled slow-release nitrogen (N) sources, oxamide and isobutylidene diurea (IBDU), each at two particle sizes, and¹⁵N-labelled urea were compared at two rates as sources of N for rice (Oryza sativa) under two watering regimes which simulated a transplant (continuous flood, CF) and a direct-seeded (A/F) system of paddy rice culture. Highest grain yields were obtained from −8+10-mesh oxamide particles applied at the rate of 2,000 mg of N/5 kg of soil, CF series; this yield was slightly higher than that obtained from −3+4-mesh oxamide, A/F series. Incubating the N fertilizers in moist (22% water) soil for 21 days immediately before flooding and transplanting rice greatly reduced N supply because of nitrification during the preflood period, followed by denitrification after flooding. This resulted in less plant uptake of N and less grain yield from urea, fine oxamide and IBDU, A/F series. For coarse oxamide, N release during the preflood period resulted in higher N uptake and grain yield in the A/F rather than in the corresponding CF series. The pattern of fertilizer N uptake by rice plants was affected by kind of fertilizer, particle size of oxamide and IBDU, and watering regime. Uptake of fertilizer N generally paralleled uptake of soil N throughout the growth period. Plant tops continued to accumulate some N during the period of grain filling, but much of the N in plant tops was translocated to the grain after heading. There was a large decrease in dry weight, N content, and¹⁵N content of tops after heading. Root weight and N content increased rapidly at first, and then at a diminishing rate until maturity. Unexplained N deficits occurred in the CF series (14–23% of the N applied, depending on N rate and source), and in the A/F series for IBDU (37–43% of the N applied).     

Interactions between nitrate uptake and N2 fixation in white clover

Summary White clover (Trifolium repens L.) plants grown in pots and supplied with the same concentration x days of¹⁵N labelled nitrate, but in contrasting patterns and doses had similar N concentrations but differed in the proportions devived from N₂ fixation and nitrate. N₂-fixation and nodule dry weight responded rapidly (2–3 days) to changes in nitrate availability. Plants exposed frequently to small doses of nitrate took up more nitrate (and hence relied less on N₂-fixation) and had greater dry weights and shoot: root ratios than those exposed to larger doses less often. In mixed ryegrass (Lolium perenne L.)/clover communities clover's ability to either successfully compete for nitrate or fix N₂ gave it consistently higher N concentrations than grass whether they were given high or low nitrate nutrient. This higher N concentration was accompanied by greater dry weights than grass in the low nitrate swards but not where high levels of nitrate were applied.     

Effects of deep placement and surface application of urea on the yield of wetland rice in pot trials

Summary The effects of deep placement and surface application of urea fertilizer on the yield of rice grown in pots of alluvial clay soil covered with 5 cm water was studied under controlled conditions. Application of two levels of urea supergranules and prills (2 g and 4 g urea/0.1 m²) on the surface of submerged soil increased the vegetative growth and enhanced the grain yield as much as 85%. However, no difference in yield was found between urea prills applied in three split doses and one application of urea supergranules.Deep placement of two levels of urea supergranules in the soil at four different depths (2.5, 5.0, 10.0 and 15.0 cm) resulted in the highest yields. The fertilizer was most efficient when the highest concentration was placed in the soil at a depth of 5.0 cm. This application method increased the grain yield by 20% as compared with the soil surface application.     

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

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

Amino acid content in xylem sap of regrowing alfalfa (Medicago sativa L.): Relations with N uptake,N2 fixation and N remobilization

During vegetative regrowth of Medicago sativa L., soil N, symbiotically fixed N₂ and N reserves meet the nitrogen requirements for shoot regrowth. Experiments with nodulated or non-nodulated plants were carried out to investigate the changes in N flows originating from the different N sources and in xylem transport of amino acids during regrowth. Exogenous N uptake, N₂ fixation and endogenous N remobilization were estimated by ¹⁵N labelling and amino acids in xylem sap were analysed. Removal of shoots resulted in great declines of exogenous N flows derived either from N₂ or from NH₄NO₃ during the first week of regrowth, thereafter recovery increased linearly. Mineral N uptake as well as N₂ fixation occurred mainly between the 10th and 18th day after removal of shoots while exogenous N assimilation in intact plants remained at a steady level. Nitrogen remobilization rates in defoliated plants increased by at least three to five-fold, especially during the first 10 days following shoot removal. Compared to control plants, contents of amino acids in xylem sap, during the first 10 days of regrowth, were reduced by about 72% and 82% in NH₄NO₃ grown and in N₂ fixing plants, respectively. Asparagine was the main amino acid transported in xylem sap of both treated plants. Its relative contents during this period significantly decreased from 75% to 59% and from 67% to 36% respectively in non-nodulated plants and in nodulated ones. This decline was accompanied by compensatory increase in the relative contents of aspartate and glutamine.