Influence of phenamiphos on the vesicular-arbuscular mycorrhizal symbiosis in Leucaena leucocephala

Summary Concentrations of phenamiphos ranging from 0 to 40 rg/g soil were established in a typical Oxisol (Tropeptic Eutrustox), inoculated or uninoculated with Glomus aggregatum. The effect of the nematicide on the development of vesicular-arbuscular mycorrhizal (VAM) symbiosis was evaluated in the greenhouse using Leucaena leucocephala as an indicator host plant. Treatment of soil with phenamiphos did not have a significant influence on the development of mycorrhizal activity measured in terms of subleaflet phosphorus concentrations. Similarly, the nematicide did not have an adverse effect on the level of mycorrhizal colonization or on the P content of shoots, as determined at the time of harvest. However, shoot dry weight was adversely influenced by phenamiphos when the chemical was applied to the uninoculated soil at 20 g/g soil or higher, and when it was applied to the inoculated soil at 40 g/g soil. It is concluded that phenamiphos is not likely to influence the growth of Leucaena or its symbiotic association with VAM fungi if the concentrations applied do not exceed levels known to suppress nematodes.Hawaii Institute of Tropical Agriculture and Human Resources Journal series No. 3146     

Physiological and symbiotic characteristics of Rhizobium fredii isolated from subtropical-tropical soils

Summary Physiological and symbiotic characteristics were identified in Rhizobium fredii isolated from subtropical-tropical soils. The generation times of R. fredii Taiwan isolated-SB 357 and -SB 682 were 1.7 and 2.5 h, respectively. These strains were associated with acid production in yeast-extract mannitol medium. They were able to use hexoses, pentose, sucrose, trehalose and raffinose. Strain SB 357 can resist a high concentration of kanamycin (100 g ml^–1 and penicillin (400 g ml^–1). It can tolerate up to 2.34% NaCl and 1031.3 mosmol kg^–1 (23.4 bars). The growth rate of R. fredii SB 357 under the concentration of approximately 450 mosmol kg^–1 (10.2 bars) was not affected by salinity, but responded to osmotic pressure. Both strains (SB 357 and SB 682) isolated from subtropical-tropical soils were able to form an effective N₂-fixing symbiosis with the US soybean cv Clark lanceolate leaflet.     

Molybdenum reserves of seed,and growth and N2 fixation by Phaseolus vulgaris L.

Summary Plants grown from seed with high (1.5–7.3 g Mo seed^-1) and low (0.07–1.4 g Mo seed^-1) Mo contents were grown in the presence and absence of Mo in growth media (perlite) or in a flowing-solution culture, in a controlled environment. Neither the high (1.5 g Mo seed^-1) nor the low (0.1 g Mo seed^-1) Mo content in seed from a small-seeded genotype (BAT 1297) was able to prevent Mo deficiency (reduced shoot, root and nodule dry weight, N₂ fixation and seed production) in growth media without an external supply of Mo, whereas both the high (7.3 g Mo seed^-1) and the low (0.07 g Mo seed^-1) contents in seed were able to prevent Mo deficiency in a large-seeded genotype (Canadian Wonder). Responses to Mo treatment by the Two genotypes were inconsistent between the growth media and solution culture experiments. Seed with a large Mo content (3.5 g Mo seed^-1) from the Canadian Wonder genotype was unable to prevent Mo deficiency (reduced shoot and nodule dry weight and N₂-fixation) in a solution culture without an external source of Mo, whereas both the large (1.7 g Mo seed^-1) and the small (0.13 g Mo seed^-1) contents in seed prevented a deficiency in BAT 1297. Growing plants from seed with a small Mo content, without additional Mo, reduced the seed Mo content by 83–85% and seed production by up to 38% in both genotypes. Changes in seed size and increases in shoot, root and nodule dry weight occurred, but varied with the genotype and growth conditions. These effects were also observed in some cases where plants were grown with additional Mo, demonstrating that the amount of Mo in the seed sown can influence plant nutrition irrespective of the external Mo supply. Nodule dry weight, total N content of shoots and seed production were improved by using seed with a small Mo content (1.64–3.57 g Mo seed^-1) on acid tropical soils in Northern Zambia. Plants of both the large- and small-seeded genotypes grown from seed with a small Mo content (<1.41 g Mo seed^-1) had a smaller nodule weight, accumulated less N and produced less seed. The viability of seed with a small Mo content was lower (germination up to 50% less) than that of seed with a large Mo content.     

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.     

Identification of ergosterol in vesicular-arbuscular mycorrhizae

Summary Ergosta-5,7,22-tri-3-enol (ergosterol) was identified by gas chromatography-mass spectrometry in roots of berseem (Trifolium alexandrinum L., cv. Landsorte) and sweet corn (Zea mays L., cv. Honeycomb-F1) infected with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus intraradices. The fungal-derived compound ergosterol was determined quantitatively in root extracts using reverse-phase high performance liquid chromatography. The concentrations of ergosterol in VAM-infected roots reached 72 g^-1 dry material in berseem and 52 g^-1 in sweet corn after 80 days of growth, whereas concentrations in non-infected roots remained below 8 g^-1 dry weight. Additionally, phytosterols such as -sitosterol, campesterol, and stigmasterol were detected in both infected and non-infected roots. Ergosterol, as a characteristic fungal substance, is proposed as an indicator of fungal biomass in the early stages of VAM infection.     

External phosphorus requirement of mycorrhizal and non-mycorrhizal barley and soybean plants

An experiment was conducted under greenhouse conditions to evaluate the effects of vesicular arbuscular mycorrhizal (VAM) fungi on the external P requirements of barley and soybeans. The plants were grown in pots containing a P-deficient soil. A range of 10 P levels was obtained by adding 0, 20, 30, 40, 50, 60, 70, 110, 160, or 310 mg P kg^-1 as NaH₂PO₄ 2H₂O. Half of the pots were inoculated with the VAM fungus Glomus intraradices. The P concentration in the soil solution was determined using an adsorption isotherm and plotted against the relative yield. Barley did not respond to mycorrhizal inoculation and we concluded that P nutrition was not the limiting factor on the growth of this lowmycotrophic plant. In contrast, mycorrhizal inoculation stimulated the growth of soybeans. The external P requirements were 0.110 g ml^-1 for mycorrhizal and 0.148 g ml^-1 for non-mycorrhizal soybeans to obtain 80% of the maximum yield. In terms of P fertilization this corresponds to a saving of 222 kg P₂O₅ ha^-1. The mycorrhizal dependency of the soybean was highly correlated with the P concentration in the soil solution and it is proposed that both values should be displayed together.     

Intrinsic antibiotic resistance and competition in fast- and slow-growing soybean rhizobia on a hybrid of Asian and US cultivars

Summary Six fast-growing soybean rhizobia (Rhizobium fredii) and thirteen slow-growing soybean rhizobia (Bradyrhizobium japonicum) were examined for resistance to 10 antibiotics. Axenic studies were carried out to determine the competitiveness of dual-strain inocula consisting of fast- and slow-growing rhizobia isolated from subtropical-tropical soils for nodule occupancy on a hybrid of Asian and US soybean cultivars. Nodule occupancy was determined by intrinsic resistance to erythromycin and neomycin. The results showed wide variability in resistance to 10 antibiotics for fast- and slow-growing rhizobia. The intrinsic antibiotic resistance of fast- and slow-growing rhizobia was extremely high against nalidixic acid (400 g ml^–1) and penicillin (200 g ml^–1). The competitive ability of inoculant strains for nodule occupancy varied for different combination sets and with the plant growing media. Our results show that fast-growing rhizobia nodulate a hybrid of Asian and US soybean cultivars. Fast-growing soybean rhizobia did not completely exclude nodulation by the slow-growing strains, which formed 0–79% nodules, depending on the strain used in the inoculum.     

Effect of heavy metals on growth and survival of Macrophomina phaseolina (Tassi) Goid.

Summary The effects of Cd, Co, Ni and Zn on growth and survival of Macrophomina phaseolina were studied in vitro. Cd, Ni and Co at 500 g ml^–1 inhibited growth by 78.8%, 73.6% and 11.8%, respectively, after 4 days at 25 ± 1°C. The mycelial dry weight yield was enhanced by 2.1% at 100 g ml^–1 Zn. The population of the pathogen declined in soil amended separately with Cd, Co, Ni and Zn. Cd (4000 mg kg^–1) proved to be the most toxic, by completely inhibiting the survival of the pathogen in soil after 20 days.     

Short-term measurements of uptake of nitrogen fixed in the rhizospheres of sorghum (Sorghum bicolor) and millet (Pennisetum americanum)

Summary In a series of short-term experiments root systems of young sorghum and millet plants inoculated with N₂-fixing bacteria were exposed to ¹⁵N₂-enriched atmospheres for 72 h. The plants were grown in a normal atmosphere for up to 22 days after the end of the exposure to allow them to take up the fixed N₂. Environmental conditions and genotypes of sorghum and millet were selected to maximise N₂-fixation in the rhizosphere. Detectable amounts of fixed N (> 16 g/plant) were rapidly incorporated into sorghum plants grown in a sand/farmyard manure medium, but measurable fixation was found on only one occasion in plants grown in soil. N₂ fixation was detectable in some experiments with soil-grown millet plants but the amounts were small (2–4 g/plant) and represented less than 1 % of plant N accumulated over the same period. In many cases there was no detectable ¹⁵N₂ incorporation despite measurable increases in ethylene concentration found during an acetylene reduction assay.Published as ICRISAT Journal Article No. JA 740     

Influence of oxygen on denitrification and aerobic respiration in soil

Summary The influence of the partial pressure of oxygen on denitrification and aerobic respiration was investigated at defined P02 values in a mull rendzina soil. The highest denitrification and respiration rates obtained in remoistened, glucose- and nitrate-amended soil were 43 1 N₂0 h^–1g^–1 soil and 130 1 O₂ h^–1g^–1 soil, respectively. At -55 kPa matric water potential, corresponding to 40% water saturation, N₂0 was produced only below P0₂ 40 hPa. The K _m, for O₂ was 3.0 x 10^–6 M. Formation of N₂O and consumption of O₂ occurred simultaneously with half maximum rates at P0₂ 6.7–13.3 hPa. Nitrite accumulated in soil below 40 hPa and increased with decreasing pO₂. The upper threshold for N₂0 formation in amended soil was P0₂ 33–40 hPa (39-47 M O₂).     

Soil pH,rhizobia, and vesicular-arbuscular mycorrhizae inoculation effects on growth and heavy metal uptake of alfalfa (Medicago sativa L.)

Summary The interaction between soil pH and inoculation with rhizobia and vesicular-arbuscular mycorrhizae (VAM) was studied in an industrially polluted soil contaminated with high levels of Zn and Cd. A silt loam soil (pH 6.7) was amended with Ca(OH)₂ or elemental S to adjust the soil pH to 4.3, 5.3, 6.0, and 7.2. Alfalfa (Medicago sativa L.) was planted in each treated soil an subsequently inoculated with Rhizobium meliloti and/or a mixed VAM spore population. Alfalfa growing in soils at a pH of 4.3 and 5.3 failed to survive as a result of soil acidity and heavy metal toxicity. At the three higher pH values, growth and foliar N and P were significantly increased by inoculation with rhizobia or VAM. The greatest increase was observed when both VAM and rhizobia were inoculated together into the soil. With a soil pH of 6.0 and 6.7, the available heavy metal concentration in the soil was high and the VAM significantly decreased heavy metal uptake from these soils. The foliar concentration of Zn was reduced from 455 to 306 g g^–1 by inoculation with VAM (pH 6.0). At the highest soil pH (7.2), however, available heavy metal concentrations were generally lower and NAM significantly increased the heavy metal uptake. The influence of VAM on heavy metal uptake thus appears to be partly a function of the available heavy metal content in the soil.     

Nitrogen fixation and CO2 exchange in soybeans (Glycine max L.) inoculated with mixed cultures of different microorganisms

N₂ fixation, photosynthesis of whole plants and yield increases in soybeans inoculated with mixed cultures of Bradyrhizobium japonicum 110 and Pseudomonas fluorescens 20 or P. fluorescens 21 as well as Glomus mosseae were found in pot experiments in gray forest soil carried out in a growth chamber. The effects of pseudomonads and vesicular-arbuscular (VA) mycorrhizal fungus on these parameters were found to be the same. Dual inoculation of soybeans with mixed cultures of microorganisms stimulated nodulation, nitrogenase activity of nodules and enhanced the amount of biological nitrogen in plants as determined by the ¹⁵N dilution method in comparison to soybeans inoculated with nodule bacteria alone. An increased leaf area in dually infected soybeans was estimated to be the major factor increasing photosynthesis. P. fluorescens and G. mosseae stimulated plant growth, photosynthesis and nodulation probably due to the production of plant growth-promoting substances. Increasing phosphorus fertilizer rates within the range of 5–40 mg P 100 g^-1 1:1 (v/v) soil: sand in a greenhouse experiment led to a subsequent improvement in nodulation, and an enhancement of N₂ fixation and yield in soybeans dually inoculated with B. japonicum 110 and P. fluorescens 21. These indexes were considerably higher in P-treated plants inoculated with mixed bacterial culture than in plants inoculated with nodule bacteria alone.     

Survival and competitiveness of Bradyrhizobium sp. in the rhizosphere of pigeonpea (Cajanus cajan)

Summary The survival of Bradyrhizobium Cajanus strain CC1021 in the soil and rhizosphere of pigeonpea [Cajanus cajan (L.) Millsp] was determined by most probable number and fluorescent antibody techniques. The survival of strain CC1021 was poor under field compared with pot-culture conditions. Although the rhizosphere of pigeonpea promoted the growth of native pigeonpea rhizobia, although no increase in the number of rhizobia was observed with the inoculant strain. Under similar conditions of rhizosphere colonization the competitiveness of strain CC1021 with cv UPAS 120 and ICPL 312 was 10% and 66016 during the first year and 8.4% and 33.3%, respectively, during the second year.     

Comparative growth and symbiotic performance of four Acacia mangium provenances from Papua New Guinea in response to the supply of phosphorus at various concentrations

Plant growth performance, the P content in root and nodule tissues, and nodulation and N₂-fixing ability were studied in four provenances of Acacia mangium from Papua New Guinea following different levels of P fertilizer application. A. mangium did not seem to need high levels of P for growth and N₂ fixation. The response by this leguminous tree to the P supply varied significantly according to provenance and to P concentrations in the culture solution. The provenances of A. mangium were classified into three types according to their P response: (1) Growth performance, nodulation, and N₂ fixation of plants were stimulated as concentrations of P increased (provenance PH 482); (2) the maximal effect of P on plant growth was found only at P concentrations higher than 500 M (provenance PH 484); and (3) the plant response to P fertilization was low, even with nutrient solutions containing P concentratins higher than 500 M (provenances PH 483 and PH 485). Provenance PH 483 was distinguished by its low nodulating ability. However, this provenance grew well, probably because of its high N₂ fixation efficiency as expressed by specific acetylene reduction activity and its high P content in nodule tissues. Therefore, in certain cases, these two parameters may be useful criteria in selecting leguminous plants for field use. Statistical analyses of the study results showed that the effect of the factor P supply on N₂ fixation efficiency and nodule development was only significant at P concentrations lower than 250 M whereas the effect of the factor plant provenance was significant regardless of the P concentration used. This observation emphasizes the value of provenance screening in the identification of plants for use in a wide range of soil types.     

Uptake and toxicity of copper and zinc for the African earthworm,Eudrilus eugeniae (Oligochaeta)

Growth rate change in earthworms is considered to be a suitable endpoint when determining sublethal effects. In this study we evaluated growth and maturation in the vermicomposting earthworm speciesEudrilus eugeniae as marker of sublethal toxicity of copper and zinc. We also compared routes of uptake. Apart from exposing worms experimentally for 73 days to contaminated food, a series of contact filter paper tests was also performed to determine LD₅₀ for copper and zinc. Both copper and zinc at sublethal concentrations affected growth and maturation in worms exposed to contaminated food. These worms had a copper content of 34.5 g g^–1 after 73 days and a zinc content of 184.9 g g^–1, showing a differential uptake. Copper was more toxic than zinc. Also in the contact test worms did take up more zinc than copper and the LD₅₀ (48 h) for copper was 0.011 mg cm^–2 and for zinc 0.066 mg cm^–2, which translated to body burdens of 6 g g^–1 for copper and 131 g g^–1 for zinc. Indications were that a regulatory mechanism existed for both metals. Both metals were taken up through the body wall at a relatively fast rate. This study indicated that the skin was the major route of metal uptake. This study also showed a poor relation between the two types of tests for purposes of evaluating lethality of zinc and copper.     

Effect of increasing oxygen concentration on total denitrification and nitrous oxide release from soil by different bacteria

Summary The effect of increasing oxygen concentrations (0, 5, 10 and 20 Vol% O₂) on total denitrification and N₂0 release was studied in model experiments using a neutral pH loamy soil relatively rich in easily decomposable organic matter and supplied with nitrate (300 g nitrate N/g dry soil). The sterilized soil was inoculated with three different denitrifying bacteria (Bacillus licheniformis,Aeromonas denitrificans andAzospirillum lipoferum) and incubated (80% WHC, 30°C). The gas volume was analysed for O₂, CO₂, N₂O, NO and N₂ by gas chromatography and the soil investigated for changes in ammonium, nitrite, nitrate, pH, total N and C as well as water-extractable C. WithB. licheniformis andAeromonas denitrificans total denitrification increased remarkably with increasing pO₂ as the result of intensified mineralization.Azospirillum lipoferum, however, showed the highest activity at 5 vol% O₂. WithB. licheniformis N₂O was released only in anaerobic conditions and at 5 Vol% O₂ (maximum) or 10 Vol% 02, but not at 20 Vol%, whereasAeromonas denitrificans produced N₂O only in the presence of He gas (maximum) or at 5 Vol% O₂. In contrast to these bacteria, N₂O production withAzospirillum lipoferum was restricted to 10 Vol% O₂ (maximum) and to 20 Vol% 02, with some traces at 5 vol% O₂. With a certain set of conditions, total denitrification and N₂O formation seem to be governed by the mineralization rate of the organisms in question. The increased demand for electron acceptors by a high turnover rate rather than the presence of anaerobic conditions seems to have determined the rate of denitrification.     

Effect of high temperature on plasmid curing of Rhizobium spp. in relation to nodulation of pigeonpea [Cajanus cajan (L.) Millsp.]

Summary Fifty-six isolates of Rhizobium and Bradyrhizobium spp. (Cajanus) were studied for their plasmid profile and N₂-fixation efficacy. One to three plasmids were reproducibly detected in all the Rhizobium spp. strains but no plasmid was detected in the Bradyrhizobium spp. strains. Rhizobium sp. strain P-1 was mutagenized by Tn5 and three nod^– and six nod⁺fix^– were screened for symbiotic parameters. Neomycin-sensitive mutants were isolated by elevated temperatrue (40°C) from tranconjugants carrying Tn5 insertions. The high temperature cured these mutants from the single large plasmid present in the parent strain P-1. All these cured mutants were nod^–, indicating that the genes for nodulation were present on this plasmid, which is readily cured at a high temperature (40°C). The high temperature in the semi-arid zones of Haryana could be responsible for the low nodulation of pigeonpea because the plasmid carrying the nodulation genes is cured at 40°–45°C giving rise to non-nodulating mutants.     

Crop residue and Collembola interact to determine the growth of mycorrhizal pea plants

The effects of collembolan grazing on arbuscular mycorrhizal (AM) fungi and plant growth were studied in a controlled experiment utilizing a mix of AM fungi and the dominant collembolan species (Isotoma sp.) indigenous to the experimental soil. Collembolan (+/– Col) effects were examined in the presence and absence of crop residue (+/– Litter) incorporated into the experimental soil. Significant interactions between collembolans and crop residue occurred for mycorrhizal colonization of roots and plant growth. In the absence of crop residue, collembolans reduced root length colonized by AM fungi, total plant dry mass and seed pod yield. However, in the presence of crop residue, collembolans had no effect on root colonization by AM fungi, and increased total plant mass and pod yield. Crop residue increased root colonization by AM fungi, numbers of bacteria and saprophytic fungi (colony forming units), small- (<5 m) and large- (>5 m) diameter hyphal lengths in soil, and the final population of collembolans in soil. Collembolans reduced both small- and large-diameter hyphae in soil and the number of saprophytic fungi (colony forming units, p =0.052). Feeding preference experiments conducted in vitro showed that Isotoma sp. preferred to graze on mycorrhizal roots over nonmycorrhizal roots when given no other food choice. However, when crop residue was added as a food choice, Isotoma sp. showed a clear feeding preference for crop residue. We conclude that collembolan grazing on mycorrhizae can be detrimental to plant growth when other fungal food sources are limited, but grazing on mycorrhizal fungi does not occur when ample organic matter and associated saprophytic fungi are present in soils.     

Effect of encapsulated calcium carbide on dinitrogen,nitrous oxide,methane, and carbon dioxide emissions from flooded rice

Summary The efficiency of N use in flooded rice is usually low, chiefly due to gaseous losses. Emission of CH₄, a gas implicated in global warming, can also be substantial in flooded rice. In a greenhouse study, the nitrification inhibitor encapsulated calcium carbide (a slow-release source of acetylene) was added with 75, 150, and 225 mg of 75 atom % ¹⁵N urea-N to flooded pots containing 18-day-old rice (Oryza sativa L.) plants. Urea treatments without calcium carbide were included as controls. After the application of encapsulated calcium carbide, 3.6 g N₂, 12.4 g N₂O-N, and 3.6 mg CH₄ were emitted per pot in 30 days. Without calcium carbide, 3.0 mg N₂, 22.8 g N₂O-N, and 39.0 mg CH₄ per pot were emitted during the same period. The rate of N added had a positive effect on N₂ and N₂O emissions, but the effect on CH₄ emissions varied with time. Carbon dioxide emissions were lower with encapsulated calcium carbide than without. The use of encapsulated calcium carbide appears effective in eliminating N₂ losses, and in minimizing emissions of the greenhouse gases N₂O and CH₄ in flooded rice.     

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.