I. Watanabe

Effect of fertilization on exudation, dehydrogenase activity, iron-reducing populations and Fe++ formation in the rhizosphere of rice (Oryza sativa L.) in relation to iron toxicity

SummaryTo explain the mechanism of iron toxicity, greenhouse and growth chamber (14CO2 atmosphere) experiments were carried out. In pot experiments (with a typical iron-toxic soil and a fertile clay) we studied the effect of N, P, K and Ca+Mg fertilization (alone or in combination) on dehydrogenase activity, Fe++ formation, and the populations of iron-reducing bacteria in the rhizosphere of rice IR22 and IR42. Fe uptake by the plants was measured at regular intervals. Dehydrogenase activity, the number of N2-fixing iron-reducing bacteria, and the formation and uptake of Fe++ decreased with increased supply of K, Ca, and Mg. This effect was clearer with IR22 (susceptible to iron toxicity) than with IR42 (releatively tolerant). Increased exudation and Fe uptake by IR36 at low nutrient and high Fe supply were recorded in a growth chamber experiment. Nutritional conditions, exudation rate (a measure of metabolic root leakage), the iron-reducing activity of the rhizosphere, and Fe++ uptake by wetland rice appear to be clearly related. Iron toxicity is considered a physiological disorder caused by multiple nutritional soil stress rather than by a low pH and high Fe supply per sé.

Improving nitrogen-fixing systems and integrating them into sustainable rice farming

This paper summarizes recent achievements in exploiting new biological nitrogen fixation (BNF) systems in rice fields, improving their management, and integrating them into rice farming systems. The inoculation of cyanobacteria has been long recommended, but its effect is erratic and unpredictable. Azolla has a long history of use as a green manure, but a number of biological constraints limited its use in tropical Asia. To overcome these constraints, the Azolla-Anabaena system as well as the growing methods were improved. Hybrids between A. microphylla and A. filiculoides (male) produced higher annual biomass than either parent. When Anabaena from high temperature-tolerant A. microphylla was transferred to Anabaena-free A. filiculoides, A. filiculoides became tolerant of high temperature. Azolla can have multiple purposes in addition to being a N source. An integrated Azolla-fish-rice system developed in Fujian, China, could increase farmers income, reduce expenses, and increase ecological stability. A study using Azolla labeled with 15N showed the reduction of N losses by fish uptake of N. The Azolla mat could also reduce losses of urea N by lowering floodwater-pH and storing a part of applied N in Azolla. Agronomically useful aquatic legumes have been explored within Sesbania and Aeschynomene. S. rostrata can accumulate more than 100kg N ha-1 in 45 d. Its N2 fixation by stem nodules is more tolerant of mineral N than that by root nodules, but the flowering of S. rostrata is sensitive to photoperiod. Aquatic legumes can be used in rainfed rice fields as N scavengers and N2 fixers. The general principle of integrated uses of BNF in rice-farming systems is shown.

The abundance of heterocystous blue-green algae in rice soils and inocula used for application in rice fields

SummaryAlgal populations were quantified (as colony-forming units [CFU] per square centimetre) in 102 samples of rice soils from the Philippines, India, Malaysia and Portugal, and in 22 samples of soil-based inocula from four countries. Heterocystous blue-green algae (BGA) were present in all samples. Nostoc was the dominant genus in most samples, followed by Anabaena and Calothrix. In soils, heterocystous BGA occurred at densities ranging from 1.0 × 102 to 8.0 × 106 CFU/cm2 (median 6.4 × 104) and comprised, on average, 9% of the total CFU of algae. Their abundance was positively correlated with the pH and the available P content of the soils. In soil-based inocula, heterocystous BGA occurred at densities ranging from 4.6 × 104 to 2.8 × 107 CFU/g dw (dry weight), comprising only a moderate fraction (average 13%) of the total algae. In most soils, the density of indigenous N2-fixing BGA was usually higher than that attained by applying recommended rates of soil-based inoculum. Whereas research on the practical utilization of BGA has been mostly directed towards inoculation with foreign strains, our results suggest that attention should also be given to agricultural practices that enhance the growth of indigenous strains already adapted to local environmental conditions.

Estimating N2 fixation by Sesbania rostrata and S. cannabina (syn. S. aculeata) in lowland rice soil by the 15N dilution method

SummaryA field experiment in concrete-based plots was conducted to estimate the contribution of N derived from air (Ndfa) or biological N2 fixation in Sesbania rostrata and S. cannabina (syn. S. aculeata), using various references, by the 15N dilution method. The two Sesbania species as N2-fixing reference plants and four aquatic weed species as non-N2-fixing references were grown for 65 days after sowing in two consecutive crops, in the dry and the wet seasons, under flooded conditions. Soil previously labeled with 15N at 0.26 atom % 15N excess in mineralizable N was further labeled by ammonium sulfate with 3 and 6 atom % 15N excess. The results showed that 15N enrichment of soil NH4+-N dropped exponentially in the first crop to half the original level in 50 days while in the second crop, it declined gradually to half the level in 130 days. The decline in 15N enrichment, in both N2-fixing and non-fixing species, was also steeper in the first crop than in the second crop. Variations in 15N enrichment among non-fixing species were smaller in the second crop. The ratio of the uptake of soil N to that of fertilizer N in N2-fixing and non-fixing species was estimated by the technique of varying the 15N level. In the second crop, this ratio in non-fixing species was higher than that in N2-fixing species. Comparable estimates of % Ndfa were obtained by using 15N enrichment of various non-fixing species. There was also good agreement between the estimates obtained by using 15N enrichment of non-fixing species and those by using soil NH4+-N, particularly in the second crop. By 25 days after sowing, the first crop of both Sesbania spp. had obtained 50% of total N from the atmosphere and the second crop had obtained 75%. The contribution from air increased with the age of the plant and ranged from 70% to 95% in 45–55 days. S. rostrata fixed substantially higher amounts of N2 due to its higher biomass production compared with S. cannabina. Mathematical considerations in applying the 15N dilution method are discussed with reference to these results.

Green manure production of Azolla microphylla and Sesbania rostrata and their long-term effects on rice yields and soil fertility

SummaryAzolla spp. and Sesbania spp. can be used as green manure crops for wetland rice. A long-term experiment was started in 1985 to determine the effects of organic and urea fertilizers on wetland rice yields and soil fertility. Results of 10 rice croppings are reported. Azolla sp. was grown for 1 month and then incorporated before transplanting the rice and 3–4 weeks after transplanting the rice. Sesbania rostrata was grown for 7–9 weeks and incorporated only before transplanting the rice. Sesbania sp. grew more poorly before dry season rice than before wet season rice. Aeschynomene afraspera, which was used in one dry season rice trial, produced a larger biomass than the Sesbania sp. The quantity of N produced by the Azolla sp. ranged from 70 to 110 kg N ha-1. The Sesbania sp. produced 55–90 kg N ha-1 in 46–62 days. Rice grain yield increases in response to the green manure were 1.8–3.9 t ha-1, similar to or higher than that obtained in response to the application of 60 kg N ha-1 as urea. Grain production per unit weight of absorbed N was lower in the green manure treatments than in the urea treatment. Without N fertilizer, N uptake by rice decreased as the number of rice crops increased. For similar N recoveries, Sesbania sp. required a lower N concentration than the Azolla sp. did. Continuous application of the green manure increased the organic N content in soil on a dry weight basis, but not on a area basis, because the application of green manure decreased soil bulk density. Residual effects in the grain yield and N uptake of rice after nine rice crops were found with a continuous application of green manure but not urea.

Composition of Azospirillum species associated with wetland rice plant grown in different soils

Total aerobic heterotrophs and N2-fixing putative azospirilla associated with rice plant grown in long-term unfertilized wetland rice field at 5 sites in the Philippines were enumerated. Several azospirilla isolates were identified based on cellular morphology, biochemical tests and reaction to immunodiffusion. Azospirilla constitute about one percent of the total aerobic heterotrophs. Most (85%) of the Azospirillum isolates belong toA. lipoferum indicating its preferential colonization to the rice plant.

Nitrogen fixation by non-legumes in tropical agriculture with special reference to wetland rice

SummaryOf the 143 million hectares of cultivated rice land in the world, 75% are planted to wetland rice. Wet or flooded conditions favour biological nitrogen fixation by providing (1) photic-oxic floodwater and surface soil for phototrophic, free-living or symbiotic blue-green algae (BGA), and (2) aphotic-anoxic soil for anaerobic or microaerobic, heterotrophic bacteria. TheAzolla-Anabaena symbiosis can accumulate as much as 200 kg N ha−1 in biomass. In tropical flooded fields, biomass production from a singleAzolla crop is about 15 t fresh weight ha−1 or 35 kg N ha−1. Low tolerance for high temperature, insect damage, phosphorus requirement, and maintenance of inoculum, limit application in the tropics. Basic work on taxonomy, sporulation, and breeding ofAzolla is needed. Although there are many reports of the positive effect of BGA inoculation on rice yield, the mechanisms of yield increase are not known. Efficient ways to increase N2-fixation by field-grown BGA are not well exploited. Studies on the ecology of floodwater communities are needed to understand the principles of manipulating BGA. Bacteria associated with rice roots and the basal portion of the shoot also fix nitrogen. The system is known as a rhizocoenosis. N2-fixation in rhizocoenosis in wetland rice is lower than that ofAzolla or BGA. Ways of manipulating this process are not known. Screening rice varieties that greatly stimulate N2-fixation may be the most efficient way of manipulating the rhizocoenosis. Stimulation of N2-fixation by bacterial inoculation needs to be quantified.

Effects of N-fertilizers, straw, and dry fallow on the nitrogen balance of a flooded soil planted with rice

SummaryNitrogen balance studies were made on rice (Oryza sativa) grown in flooded soil in pots. A low rate of fertilizer (5.64 mg N. kg−1 soil) did not depress the N gain, but a high rate (99.72 mg N. kg−1 soil) elminated the N gain. Soil N loss was negligible since15N applied as ammonium sulfate and thoroughly mixed with the soil was recovered from the soil-plant system after 3 crops. The observed N gain, therefore, was caused by N2-fixation, not by a reduction of soil N loss.Straw enhanced N gain at the rate of 2–4 mg per g straw. However, this gain was not observed when soil N availability was high. Dry fallow between rice crops decreased the N gain.

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

SummaryUsing 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 Azolla15N 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.

The mechanism of excessive iron‐uptake (iron toxicity) of wetland rice

Abstract The hypothesis that iron toxicity is the result of a multiple nutritional stress (deficiencies in P, K, Ca and Mg) was studied in the greenhouse (pot experiments) using a typical iron toxic soil and the rice varieties IR 22 (susceptible to iron toxicity) and IR 42 (relatively tolerant). During the growth period the effect of N, P, K, Ca + Mg‐fertillzation on Fe(II)‐formation, the total rhizosflora (reflected by the dehydrogenase activity), iron reducing microorganisms (total number and N2‐fixing), Fe‐reducing bacteria) as well as on the sulfate reducing bacteria was investigated by regular sampling. Plant and root samples were analysed for total Fe and N, P, K, Ca and Mg. Though the total number of iron reducing bacteria increased with fertilization, Fe(II)‐formation and dehydrogenase activity was much more intensive in the rhizosphere of the unfertilized treatment transplanted to IR 22. Population densities of the strictly anaerobic N2‐fixing Fe‐ or sulfate reducers in the unfertilized pots were...