Dual inoculation of pretransplant stage Oryza sativa L. plants with indigenous vesicular-arbuscular mycorrhizal fungi and fluorescent Pseudomonas spp.

Summary This study examined the response of rice (Oryza sativa L.) plants at the pretransplant/nursery stage to inoculation with vesicular-arbuscular mycorrhizal (VAM) fungi and fluorescent Pseudomonas spp., singly or in combination. The VAM fungi and fluorescent Pseudomonas spp. were isolated from the rhizosphere of rice plants. In the plants grown in soil inoculated with fluorescent Pseudomonas spp. alone, I found increases in shoot growth, and in root length and fine roots, and decreases in root growth, and P and N concentrations. In contrast, in the plants colonized by VAM fungi alone, the results were the reverse of those of the pseudomonad treatment. Dual inoculation of soil with VAM fungi and fluorescent Pseudomonas spp. yielded plants with the highest biomass and nutrient acquisition. In contrast, the plants of the control treatment had the lowest biomass and nutrient levels. The dual-inoculated plants had intermediate root and specific root lengths. The precentages of mycorrhizal colonization and colonized root lengths were significantly lower in the dual-inoculated treatment than the VAM fungal treatment. Inoculation of plants with fluorescent Pseudomonas spp. suppressed VAM fungal colonization and apparently reduced photosynthate loss to the mycorrhizal associates, which led to greater biomass and nutrient levels in dual-inoculated plants compared with plants inoculated with VAM fungi alone. Dual inoculation of seedlings with fluorescent Pseudomonas spp. and VAM fungi may be preferable to inoculation with VAM alone and may contribute to the successful establishment of these plants in the field.     

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

Summary Azolla 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.     

Yield and nitrogen response of lowland rice (Oryza sativa L.) to Sesbania rostrata and Aeschynomene afraspera green manure in different marginally productive soils in the Philippines

Organic-N fertilizers in the form of flood-tolerant, leguminous, stem-nodulating Sesbania rostrata and Aeschynomene afraspera may be useful alternatives to resource-poor rice farmers if applied as green manure. Therefore, the accumulation of N by these green manure species and their effect on the performance and yield of wetland rice (IR 64) was examined at four different sites in Luzon, Philippines. Soils deficient in N, P, and K were selected and compared with the fertile Maahas clay of the International Rice Research Institute (IRRI) at Los Baños. The green manure plants were grown under flooded conditions for 49 days in the wet season of 1987, chopped, and then ploughed in before transplanting rice seedlings. In a second experiment, the effect of S. rostrata green manure was studied under rainfed conditions. All green manure treatments were compared to an urea treatment (60 kg N ha^–1) and an untreated control. Both legumes developed well, even on the marginally productive soils. S. rostrata accumulated up to 190 kg N ha^–1 and A. afraspera even accumulated 196 kg N ha^–1 in the shoots. In all treatments, green manure increased grain yield significantly (P=0.05) over the untreated control, by 1.3–1.7 Mg ha^–1. The yields were comparable to those obtained with 60 kg N ha^–1 of urea fertilizer. S. rostrata caused the highest grain yield, of 6.5 Mg ha^–1 on the Maahas clay soil of IRRI. The apparent release of exchangeable NH ₄ ⁺ -N in the soils after green manuring and the rice grain yield response showed that both green manure species may provide sufficient available N throughout the development of IR 64 in the wet season. In the rainfed marginal soil site, green manure with S. rostrata produced even higher rice grain yields than urea. Green manure therefore seems particularly attractive for poor farmers on marginally productive soils, at least as a temporary strategy to improve yield and yield sustainability.     

Sesbania rostrata as a green manure for lowland rice: Growth,N2 fixation,Azorhizobium sp. inoculation,and effects on succeeding crop yields and nitrogen balance

Summary Root and stem nodulation, nitrogen fixation (acetylene-reducing activity), growth and N accumulation bySesbania rostrata as affected by season and inoculation were studied in a pot experiment. The effects ofS. rostrata as a green manure on succeeding wet-season and dry-season rice yields and total N balance were also studied.S. rostrata grown during the wet season showed better growth, nodulation, and greater acetylene-reducing activity than that grown during the dry season. Inoculation withAzorhizobium caulinodans ORS 571 StrSpc® (resistant to streptomycin and spectinomycin) on the stem alone or on both root and stem significantly increased N₂ fixation by the plants. Soil and seed inoculation yielded active root nodules under flooded conditions. Plants that were not inoculated on the stem did not develop stem nodules. The nitrogenase activity of the root nodules was greater than that of the stem nodules in about 50-day-oldS. rostrata. S. rostrata incorporation, irrespective of inoculation, significantly increased the grain yield and N uptake of the succeeding wet season and dry season rice crops. The inoculated treatments produced a significantly greater N gain (873 mg N pot^–1) than the noinoculation (712 mg N pot^–1) treatment. About 80% of the N gained was transferred to the succeeding rice crops and about 20% remained in the soil. The soil N in the flooded fallow-rice treatment significantly declined (–140 mg N pot^–1) but significantly increased in bothS. rostrata-rice treatments (159 and 151 mg N pot^–1 in uninoculated and inoculated treatments respectively). The N-balance data gave extrapolated values of N₂ fixed per hectare at about 303 kg N ha^–1 per two crops forS. rostrata (uninoculated)-rice and 383 forS. rostrata (inoculated)-rice.     

Nitrogen contribution to wetland rice by green manuring with Sesbania spp. in an alkaline soil

Summary Two annual species of Sesbania, S. aculeata and Sesbania sp. PL Se-17, were field evaluated as green manure for wetland rice in an alkaline soil. The two species were raised as a catch crop during summer in a wheat-rice rotation, and added as 24.7 and 20.8 t ha^–1 of green matter, 116 and 98 kg N ha^–1, respectively, after 45 days of growth. For the optimum green manuring effect on rice grain yield and N uptake, S. aculeata required 5 days of decomposition (after turning in and before rice transplantation), whereas no decomposition period was necessary for Sesbania sp. PL Se-17. The effect on grain yield and N uptake of rice was equivalent to an application of 122 and 78 kg ha^–1 of chemical N, for the two species, respectively. There was no residual effect of the green manuring on the soil N status after rice harvest.     

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.     

Effect of phosphorus fertilization on blue-green algal inoculum production and nitrogen yield under field conditions

Summary Field experiments were conducted to assess the effects of the application of P on growth and N yield of inoculated and indigenous blue-green algae (BGA). Addition of 17.4 kg P ha^–1 in split applications led to the highest BGA biomass and N yield, 162 kg dry weight ha^–1 and 6 kg N ha^–1 per 15 days, respectively. When inoculum of Aulosira spp., Aphanothece spp., Gloeotrichia spp. were compared separately, Gloeotrichia spp. grew faster but Aulosira spp. fixed more N. The growth rate and N yield of Aulosira spp. decreased with high P applications, although growth continued until the application of 34.8 kg P ha^–1. The effects of P on inoculum production by local species compared with those collected from other states showed the superiority of the local culture. Applications of P also enhanced the growth and N yield of indigenous BGA, with Wollea spp. showing the best results.     

Inoculation withGlomus mosseae improves N2 fixation by field-grown soybeans

Summary A field study carried out in a sandy, relatively acid Senegalese soil with a low soluble P content (7 ppm) and low vesicular-arbuscular mycorrhizal (VAM) populations showed that soybean responded toGlomus mosseae inoculation when the soluble P level in the soil had been raised by the addition of 22 kg P ha^–1. In P-fertilized plots, N₂ fixation of soybean, assessed by the A value method, was 109 kg N₂ fixed hat when plants were inoculated withRhizobium alone and it reached 139 kg N₂ fixed ha^–1 when plants were dually inoculated withRhizobium andGlomus mosseae using an alginate bead inoculum. In addition to this N₂ fixation increase (+28%),Glomus mosseae inoculation significantly improved grain yield (+13%) and total N content of grains (+16%). This success was attributed mainly to the low infection potential of the native VAM populations in the experimental site. In treatments without solubleP or with rock phosphate, no effect of VAM inoculation was observed.     

Effect of urea,blue green algae and<Emphasis Type="Italic"> Azolla</Emphasis> on nitrogen fixation and chlorophyll accumulation in soil under rice

Nitrogen fixing potential in terms of acetylene reducing activity (ARA) and biomass accumulation (in terms of chlorophyll) were investigated using surface and below-surface soil cores, collected from rice fields 45 and 90 days after transplanting (DAT). Treatments included different levels of urea (30, 60, 90 and 120 kg N ha^–1) in combination with inoculation using blue green algae (BGA) and Azolla biofertilizers. Application of biofertilizers brought about a significant enhancement in chlorophyll accumulation and nitrogenase activity, when measured 45 DAT. Positive effects in below-surface soil cores, on both these parameters as a result of application of biofertilizers further emphasized their contribution to the N economy of rice fields. Plots treated with 30 and 60 kg N ha^–1 along with biofertilizers exhibited the highest percentage increase in terms of algal biomass and ARA, both in surface and below-surface soil cores at 45 DAT. A definite need to examine critically the nature and metabolic activities of below-surface microflora is highlighted through our investigation.     

Response of maize (Zea mays) to Azotobacter inoculation under fertilized and unfertilized conditions

Summary Inoculated seeds of maize (Zea mays) with 11 Azotobacter strains, sown in the fields receiving no fertilizer and fertilizers (N and P at the rate of 125 and 40 kg ha^–1 respectively) increased the grain yield by 19.63% and 15.89% respectively over the corresponding control. The effect was greater in unfertilized than in fertilized soil. The increase in yield due to fertilizers was 21.2% without inoculation and 37.09% with inoculation. The correlations between total yield, and N, P and K uptake were highly significant and comparable among themselves. This indicated that increase in yield due to inoculation was not due to N₂ fixation but that some other mechanisms like production of growth hormones by this bacterium may be responsible.     

Biomass and nutrient cycling in bamboo (Bambusa bambos) plantations of tropical areas

Biomass productivity and nutrient cycling in a Bambusa bambos plantation aged 4, 5, and 6 years were studied. The dry matter production of above-ground biomass increased progressively with age. Nutrient quantities in bamboo stands were in a range of 1–2 t ha^-1 for N and K, 0.5–1 t ha^-1 for Ca and Mg, and 0.1–0.2 t ha^-1 for P. Nutrient concentrations increased with the age of the plantation. About 10% year^-1 the nutrients present in the biomass of the bamboo stand are recycled to the soil by litter fall.     

Vesicular-arbuscular mycorrhizae screened for Troyer citrange

Summary Troyer citrange is an important citrus root-stock but a slow grower. Eighteen different vesicular-arbuscular mycorrhizal (VAM) fungi were screened for their symbiotic response with Troyer citrange in unsterilized soil. Of the 18 fungi,Glomus macrocarpum (Ruakura),G. caledonicum (Nedlands),G. velum (Nedlands),G. monosporum (Nedlands),Gigaspora margarita (Ruakura), gave the greatest improvements in growth and nutrition, resulting in larger leaf area, plant height, stem diameter, and plant biomass, with higher P, Zn and Cu contents. All the five fungi were statistically on a par with each other in improving the growth and nutrition of Troyer citrange. Inoculations with these fungi allowed the rootstock to reach a stem diameter of pencil thickness, i.e., ready for budding, in about 13–14 months in contrast to 18–19 months without mycorrhizal inoculations, thus saving about 5–6 months.     

Reduction of bacterial growth by a vesicular-arbuscular mycorrhizal fungus in the rhizosphere of cucumber (Cucumis sativus L.)

Summary Cucumber was grown in a partially sterilized sand-soil mixture with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus fasciculatum or left uninoculated. Fresh soil extract was places in polyvinyl chloride tubes without propagules of mycorrhizal fungi. Root tips and root segments with adhering soil, bulk soil, and soil from unplanted tubes were sampled after 4 weeks. Samples were labelled with [³H]-thymidine and bacteria in different size classes were measured after staining by acridine orange. The presence of VAM decreased the rate of bacterial DNA synthesis, decreased the bacterial biomass, and changed the spatial pattern of bacterial growth compared to non-mycorrhizal cucumbers. The [³H]-thymidine incorporation was significantly higher on root tips in the top of tubes, and on root segments and bulk soil in the center of tubes on non-mycorrhizal plants compared to mycorrhizal plants. At the bottom of the tubes, the [³H]-thymidine incorporation was significantly higher on root tips of mycorrhizal plants. Correspondingly, the bacterial biovolumes of rods with dimension 0.28–0.40×1.1–1.6 m, from the bulk soil in the center of tubes and from root segments in the center and top of tubes, and of cocci with a diameter of 0.55–0.78 m in the bulk soil in the center of tubes, were significantly reduced by VAM fungi. The extremely high bacterial biomass (1–7 mg C g^-1 dry weight soil) was significant reduced by mycorrhizal colonization on root segments and in bulk soil. The incorporation of [³H]-thymidine was around one order of magnitude lower compared to other rhizosphere measurements, probably because pseudomonads that did not incorporate [³H]-thymidine dominated the bacterial population. The VAM probably decreased the amount of plant root-derived organic matter available for bacterial growth, and increased bacterial spatial variability by competition. Thus VAM plants seem to be better adapted to compete with the saprophytic soil microflora for common nutrients, e.g., N and P, compared to non-mycorrhizal plants.     

Differential responses of chickpea (Cicer arietinum L.) — Rhizobium combinations to saline soil conditions

Summary Chickpea cultivars (Cicer arietinum L.) and their symbiosis with specific strains of Rhizobium spp. were examined under salt stress. The growth of rhizobia declined with NaCl concentrations increasing from 0.01 to 2% (w : v). Two Rhizobium spp. strains (F-75 and KG 31) tolerated 1.5% NaCl. Of the 10 chickpea cultivars examined, only three (Pusa 312, Pusa 212, and Pusa 240) germinated at 1.5% NaCl. The chickpea — Rhizobium spp. symbiosis was examined in the field, with soil varying in salinity from electrical conductivity (EC) 4.5 to EC 5.2 dSm^-1, to identify combinations giving satisfactory yields. Significant interactions between strains and cultivars caused differential yields of nodules, dry matter, and grain. Four chickpea — Rhizobium spp. combinations, Pusa 240 and F-75 (660 kg ha^-1), Pusa 240 and IC 76 (440 kg ha^-1), Pusa 240 and KG 31 (390 kg ha^-1), and Pusa 312 and KG 31 (380 kg ha^-1), produced significantly higher grain yields in saline soil.     

Effects of nitrogen fertiliser and pesticide management on floodwater ecology in a wetland ricefield

We investigated the effects of N fertiliser and pesticide applications on the population dynamics of benthic molluscs in a tropical wetland rice field. Populations were monitored for two consecutive dry seasons in selected treatments during a study on the effects of agricultural practices on the floodwater ecology of tropical rice fields. The most abundant species recorded in the ricefields were the snailsMelanoides tuberculata andMelanoides granifera. Population densities and biomass values in planted plots ranged between 0 and 1530 individuals m^-2 and 0 and 1060 kg ha^-1, respectively. Snails were more abundant in unplanted than planted plots (1991: 170–2040 versus 0–1040 individuals m^-2, respectively). Populations in planted plots declined as the crop season progressed. Snail populations were significantly reduced by the broadcast application of mineral N fertiliser at 110 kg N ha^-1. There was little evidence that snails were affected by carbofuran or butachlor applications.     

Growth response of field-grown Siratro (Macroptilium atropurpureum Urb.) and Aeschynomene american L. to inoculation with selected vesicular-arbuscular mycorrhizal fungi

Summary The effect of inoculation with a selected isolate of Glomus etunicatum Becker and Gerdemann and one of G. intraradices Schenck and Smith on the growth and nutrient content of Macroptilium atropurpureum Urb. cv. Siratro and Aeschynomene americana L., at applied P levels of 10, 30, 60, and 120 kg ha^-1, was studied under field conditions. At all P levels and for all harvests, the shoot dry mass of Siratro and A. americana were greater for the plants inoculated with the vesicular-arbuscular mycorrhizal (VAM) fungi than the control plants. Differences between the VAM fungus-inoculated and the control plants were most marked between 30 and 90 kg ha^-1 of applied P and diminished at 120 kg ha^-1. At the first harvest of Siratro, the plants inoculated with G. etunicatum had a greater shoot dry mass than those inoculated with G. intraradices, for all levels of applied P. However, for subsequent harvest of Siratro and for the one harvest of A. americana the response of shoot dry mass to the two VAM fungi was equivocal. Fungal inoculation gave at least a 30% saving in the amount of P fertilizer required (40 kg ha^-1) for the maximum yield. The plants inoculated with VAM fungi had a greater tissue concentration and total content of P and N than the control plants at low and intermediate levels of applied P. The percentage of root colonized by VAM fungi for the inoculated plants of the two legumes increased linearly with P additions up to 60 kg ha^-1. The conclusion is that under amended (limed and fertilized) soil conditions, inoculation with selected VAM fungi can improve the establishement and growth of forage legumes in fields that contain ineffective populations of native VAM fungi.     

Effects of Rhizobium and vesicular-arbuscular mycorrhiza inoculations on nodulation,symbiotic nitrogen fixation and soybean yield in subtropical-tropical fields

Summary Field experiments were carried out to determine the effects of single and mixed inoculations with Rhizobium and vesicular-arbuscular mycorrhiza (VAM) on nodulation, symbiotic N₂ fixation and yield of soybeans in six Taiwan subtropical-tropical sites. Inoculation with Rhizobium alone significantly increased nodulation, nodule weight and nitrogenase activity of nodules in three out of six experimental fields, and affected soybean yields in the range –13% to + 134%. Inoculation with VAM fungi alone did not have a significant effect on nodulation and nitrogenase activity. Mycorrhiza inoculation affected soybean yields in the range –13% to + 65%, but only the yield increases at one out of six sites with N application were statistically significant. Mixed inoculation with Rhizobium and mycorrhiza affected yields in the range –8% to + 145% A synergistic effect from mixed inoculation of Rhizobium-mycorrhiza on soybean yields was found in one out of six experimental fields. The yield response to N application (40 kg N ha^–1) in these six paddy-field trials was not significant. These results suggest that single or mixed inoculation of rhizobia can greatly assist soybean grain yields and can replace N fertilizers.     

Effect of soil treatment with cereal straw and method of crop establishment on field pea (Pisum sativum L.) N2 fixation

The effects of soil incorporation with cereal straw (nil, 2.5, 5 and 10 t straw ha^–1) and direct drilling on the proportion and amount of pea N derived from biological N fixation were investigated in three field experiments. Fixed N was determined by¹⁵N dilution using barley as a reference plant. The three sites were on acidic, red clay-loams in the cropping zone of southeastern Australia. Seasonal plant available soil N, as determined by the N accumulated in barley, was 31, 56 and 158 kg N ha^–1, for the three sites. Incorporated straw reduced soil nitrate at sowing by 10–50 kg N ha^–1 (0–30 cm), and 5 or 10 t straw ha^–1 reduced barley uptake of N by 10–38 kg N ha^–1. However, reducing plant available soil N was generally ineffective for increasing the N fixed by pea. Fixed N increased only at the site with the least plant-available N, and only one-third of the increase could be attributed to lower soil N uptake by pea. There was no evidence that direct drilling pea increased fixed N by decreasing crop uptake of soil N. It is proposed that a lower requirement for soil N by pea as compared to barley, and availability of mineral N beneath the soil layer treated with straw, minimise the effectiveness of straw incorporation for increasing the N fixed by pea.     

Effects of inoculation with N2-fixing cyanobacteria on the nitrogenase activity in soil and rhizosphere of wetland rice (Oryza sativa L.)

Summary A greenhouse experiment was conducted with wetland rice (Oryza sativa cv. IR-50) in a clay-loam soil (Fluventic Eutrochrept) to study the effect of cyanobacterial inoculation a mixed culture of Aulosira fertilissima, Nostoc muscorum, Nostoc spp., and Anabaena spp., applied at the rate of 0.15 g (dry weight pot^-1 or 43 kg ha^-1) on acetylene reduction activity in soil and the root system (excised root), and the grain and straw yield. The effects of applying P (40 kg ha^-1), N (60 kg ha^-1), and P+N to the soil were also evaluated. Cyanbacterial inoculation significantly increased (more than 200% on average) photo-dependent acetylene reduction activity in soils, particularly where the indigenous activity was considerably low, i.e. under unfertilized and N-fertilized conditions. The effect of inoculation was prominent at the maximum tillering and grain formation stages of the crop. This inoculation benefit was, however, marginal in P-applied soils (P and P+N), where the indigenous activity was stimulated more than threefold. The inoculation led to a remarkable increase in root-associative acetylene reduction activity after the maximum tillering stage of the crop, particularly with applied N but for other treatments this inoculation effect was not significant. Cyanobacterial inoculation also increased the grain and straw yield of the crop when N was not applied. The grain and straw yield was significantly correlated with the acetylene reduction activity in flooded soils and in the root system during the tillering and maximum tillering stages of rice growth, respectively.     

Amelioration of subsoil acidity in an Oxisol of the humid tropics

Summary This work investigated the effectsof amendments of fertilizer N and lime on subsoil acidity and maize rooting depth in an acid soil of the central Amazon basin. A split-plot designed field experiment was conducted on a clayey Oxisol (Typic Acrudox) during a 16-month period. Main plots received 0 or 4 Mt ha^-1 of lime. Subplots were four crop sequences: (1) Maize-green manure (Canavalia ensiformes); (2) maize-green manure (Mucuna aterrima); (3) maize-bare fallow, with the maize receiving 300 kg ha^-1 of urea-N; and (4) bare fallow, with an application of 300 kg ha^-1 of urea-N at the same time as sequence 3. Plots were periodically sampled to 1.2 m. The experimental site received 4265 mm of precipitation during 16 months; approximately 60%–90% of this rain percolated through the profile. Substantial amounts of Ca were leached from the 0–30 cm horizon during the experimental period, but only limited amounts accumulated in the subsoil. Base saturation below 45 cm was less than 50% at the end of the experiment regardless of lime treatment. Roots of maize were concentrated in the 0–30 cm layers in limed plots and the 0–20 cm layers in unlimed plots. In all treatments less than 5% of the roots was found below 50 cm. An acidity balance indicated that considerable acidity was leached below the plow layer and out of the profile.