Competition for nodule formation between introduced strains of <Emphasis Type="Italic">Mesorhizobium ciceri</Emphasis> and the native populations of rhizobia nodulating chickpea (<Emphasis Type="Italic">Cicer arietinum</Emphasis>) in Tunisia

Chickpea is the most cultivated grain legume in the world and it shares the first rank with faba bean in Tunisia. However, the yield remains low, mainly due to the limited availability of N and P, and to the severe bioclimatic conditions. No inoculation trials had been conducted on chickpea in the Tunisian soils. This paper reports the yield response to inoculation by two different strains of Mesorhizobium ciceri, an exogenous type strain (UPMCa7^T) and a selected local strain (CMG6). Field experiments were conducted in different sites in the north of Tunisia using three chickpea cultivars (cvs. Amdoun I, Chetoui and Kasseb). Rhizobia occupying field nodules were isolated and identified using 16S rDNA typing for both inoculated and non-inoculated plots. In contrast to the exogenous strain, the local strain gave a significant increase in nodule number and shoot dry yield in all the experimental fields for the three cultivars used. Monitoring of the nodule occupancy showed that the local strain competed well the native populations of rhizobia. The usefulness and the persistence of this strain in the different soils where it was introduced will be assessed further during the next years.     

Rhizobial strain involvement in symbiosis efficiency of chickpea–rhizobia under drought stress: plant growth,nitrogen fixation and antioxidant enzyme activities

Chickpea plants were inoculated with two strains of Mesorhizobium ciceri: local strain (C-15) and non-local strain (CP-36) in order to evaluate plant growth parameters, activities of nitrogenase and antioxidant enzymes under drought stress as well as control condition within 15 days of imposition of drought stress. Biomass production, nodulation, nitrogen fixation and antioxidant enzyme activities under drought condition were compared. Under control condition, symbiotic efficiency in symbiosis formed by C-15 was higher than that in symbiosis derived by CP-36. Although drought stress decreased shoot dry weight, root dry weight, nodule dry weight and nitrogen fixation in both symbioses, the rate of decline in plants inoculated with CP-36 was higher than that in symbiosis chickpea with C-15. Therefore, symbioses showed different tolerance level under drought condition which was essentially correlated with symbiotic performance at non-stressful conditions. Under drought stress, nodular peroxidase (POX) activity increased in both symbioses but was higher in nodules produced by C-15. Ascorbate peroxidase (APX) increased significantly in nodules of symbiosis of chickpea with C-15. Catalase (CAT) and glutation reductase (GR) declined in both symbioses which decline extent in symbiosis with C-15 was lower than that in the nodules of CP-36. These results suggested contribution of rhizobial partner in enhancing the tolerance of symbioses to drought stress, which was related with the increase of antioxidant enzyme activities (APX and POX) under drought conditions.     

Exploring Crop–Microbiome Interactions Towards Improving Symbiotic Performance of Chickpea (Cicer arietinum) Cultivars Using Cyanobacterial Inoculants

The significance of integrated nutrient management practices is well established in improving the productivity of chickpea (Cicer arietinum); however, the effects of the inoculation of cyanobacterial inoculants on nodule metabolism, microbiome and associated genes are less explored. In the present investigation, cyanobacterium Anabaena laxa (A. laxa) and biofilm developed using Anabaena torulosa, with Mesorhizobium ciceri as a partner (An-M. ciceri), were evaluated along with Mesorhizobium ciceri (M. ciceri) alone, in three chickpea cultivars. Microbial inoculation led to 40–70% enhancement in nitrogen fixation, leghaemoglobin and ureide content, and two- to threefold increment in nitrate reductase and phosphoenolpyruvate carboxylase activity of the nodules. An enhancement of 30–50% in the soil available macro- and micronutrients and plant growth attributes was also observed. A significant correlation between the soil microbiological and plant parameters was recorded, particularly in relation to the nitrogen dynamics. Increases in the leghaemoglobin content in nodules due to An-M. ciceri, A. laxa and M. ciceri ranged from 18 to 40%, particularly in chickpea cv. BG372 in which 60–80% enhancement was recorded. Whereas the nifH gene copies in the nodule tissues ranged from 5.00 × 10⁶ to 3.35 × 10⁷ g⁻¹, the application of A. laxa led to higher abundances of nifH gene copies in desi chickpea cv. BG372 and kabuli BG1053 cultivars. An-M. ciceri, followed closely by A. laxa, was the top-ranking treatment, and chickpea cv. BG372 was the best performing cultivar; An-M. ciceri—chickpea cv. BG372 proved to be the superior combination for higher plant growth and soil nutrient-related traits.

     

Field response of chickpea to inoculation withGlomus versiforme

The response ofCicer arietinum to inoculation withGlomus versiforme under field conditions was investigated in a phosphorus deficient sandy loam soil. Inoculation with the mycorrhizal fungusGlomus versiforme increased the rate of VAM development in chickpea. The weight of nodules and the number of nodules per plant were higher in inoculated than in uninoculated plants. The phosphorus content of the shoots and its total uptake, were increased by either the application of single super-phosphate, or by inoculation withG. versiforme. Inoculation increased shoot dry weights and grain yields by 12% and 25% respectively, as compared with the 33% and 60% increases respectively produced by P-treated plants.     

Influence of soil pH on the soybean-endomycorrhiza symbiosis

Summary Soybean (Glycine max {L.} Merr.) cultivars were inoculated withGigaspora gigantea andGlomus mosseae to determine mycorrhizal: cultivar relationships as affected by soil pH. The specific cultivarfungal response was dependent on soil pH. Overall cultivar responses in unlimed soil (pH 5.1) were greater forG. gigantea thanG. mosseae. The Bossier —G. gigantea combination was particularly responsive in unlimed soil and showed an increase of 10% in shoot length, 35% in shoot dry weight. 75% in root dry weight, and 397% in nodule dry weight over uninoculated controls. Little cultivar response was observed withG. mosseae inoculation in unlimed soil. In limed soil (pH 6.2), the larger responses were obtained withG. mosseae inoculated plants, although inoculation with eitherG. mosseae orG. gigantea appeared effective. In general, nodulation was greater on mycorrhizal roots than on control roots.     

Inoculation and production of container-grown red alder seedlings

Summary The inoculation ofAlnus rubra (red alder) withFrankia sp. can lead to a highly efficient symbiosis. Several factors contribute to the successful establishment of nitrogenfixing nodules: (1) quantity and quality ofFrankia inoculant; (2) time and method of inoculation; (3) nutritional status of the host plant.Frankia isolates were screened for their ability to nodulate and promote plant growth of container-grown red alder. Inoculations were performed on seedlings and seeds. Apparent differences in symbiotic performance could be seen when seeds or seedlings were inoculated. Plants inoculated at planting performed significantly better than those inoculated four weeks later in terms of shoot height, nodule number and shoot dry weight. If inoculation was delayed further, reduction in shoot height, nodule number and shoot dry weight resulted. The effect of fertilizer was also investigated with regard to providing optimal plant growth after inoculation. Plants receiving 1/5 Hoagland's solution minus nitrogen showed maximal plant growth with abundant nodulation. Plants receiving 1/5 Hoagland's solution with nitrogen showed excellent plant growth with significantly reduced nodulation.     

Comparative effects of Glomus mosseae and P fertilizer on foliar mineral composition of Acacia senegal seedlings inoculated with Rhizobium

Summary A factorial experiment with two controlled factors was conducted in the greenhouse with Acacia Senegal seedlings. The substrate was a degraded sandy soil (Dior soil) poor in available P (11 ppm — Olsen). The first controlled factor was soil sterilization, with two levels: (A) sterilized soil; (B) non-sterilized soil. The second factor was fertilization, with six levels: (1) uninoculated control; (2) inoculation with Rhizobium (ORS 1007); (3) inoculation with Glomus mosseae; (4) double inoculation with ORS 1007 and G. mosseae; (5) inoculation with ORS 1007 and 30 ppm phosphorus per plant; (6) inoculation with ORS 1007 and 60 ppm phosphours. The combination of the two factors and their levels led to 12 different plant treatments (A1–A6 and B1–B6). Compared to the control B1, the B5 and B6 treatments containing phosphorus increased: nodule dry weight about 7 times ; leaf dry weight about 4 times ; total N, P and Mg 4–5 times; total K and Ca 3–4 times. The mycorrhizal inoculation had the same positive effect on plant growth and mineral composition but with lower values. Plants inoculated with Rhizobium alone gave the lowest results. The A1 treatment gave lower values than B1. Foliar mineral contents varied within a narrow range (20–30%).     

Mesorhizobium ciceri LMS-1 expressing an exogenous 1-aminocyclopropane-1-carboxylate (ACC) deaminase increases its nodulation abilities and chickpea plant resistance to soil constraints

Aims: Our goal was to understand the symbiotic behaviour of a Mesorhizobium strain expressing an exogenous 1‐aminocyclopropane‐1‐carboxylate (ACC) deaminase, which was used as an inoculant of chickpea (Cicer arietinum) plants growing in soil. Methods and Results: Mesorhizobium ciceri LMS‐1 (pRKACC) was tested for its plant growth promotion abilities on two chickpea cultivars (ELMO and CHK3226) growing in nonsterilized soil that displayed biotic and abiotic constraints to plant growth. When compared to its wild‐type form, the M. ciceri LMS‐1 (pRKACC) strain showed an increased nodulation performance of c. 125 and 180% and increased nodule weight of c. 45 and 147% in chickpea cultivars ELMO and CHK3226, respectively. Mesorhizobium ciceri LMS‐1 (pRKACC) was also able to augment the total biomass of both chickpea plant cultivars by c. 45% and to reduce chickpea root rot disease susceptibility. Conclusions: The results obtained indicate that the production of ACC deaminase under free living conditions by Mesorhizobium strains increases the nodulation, plant growth abilities and biocontrol potential of these strains. Significance and Impact of the Study: This is the first study regarding the use of a transformed rhizobial strain expressing an exogenous ACC deaminase in different plant cultivars growing in soil. Hence, obtaining Mesorhizobium strains with high ACC deaminase activity is a matter of extreme importance for the development of inoculants for field applications.     

Field response of tomato (Lycopersicon esculentum Mill ‘Pusa Ruby’) to inoculation with a VA mycorrhizal fungusGlomus fasiculatum and withAzotobacter vinelandii

Summary The response of tomato (Lycopersicon esculentum Mill) to inoculation with the vasicular arbuscular mycorrhizal (VAM) fungusGlomus fasiculatum andAzotobacter vinelandii singly and in combination was tested in the field. It was found thatG. fasiculatum as well asA. vinelandii significantly increased leaf area, shoot dry weight, nitrogen content phosphorus content and yield in respect to uninoculated control. While, VAM fungal treatment alone could bring about substantial increase in growth, nitrogen content, phosphorus content and yield, its combination withA. vinelandii produced additional effects on leaf area, shoot dry weight, phosphorus content and yield. Contribution No. 304/83 of Indian Institute of Horticultural Research, Bangalore-89.     

Associative effect of Rhizobium and phosphate-solubilizing bacteria on the yield and nutrient uptake of chickpea

Combined inoculation of Rhizobium and ‘Phosphate-solubilizing’Pseudomonas striata orBacillus polymyxa with and without added chemical fertilizer on chickpea yield and nutrient content was studied under greenhouse conditions. While the single inoculation of Rhizobium increased the nodulation and nitrogenase activity, the ‘phosphate-solubilizers’ increased the available phosphorus content of the soil. Combined inoculation of Rhizobium andP. striata orB. polymyxa increased the above parameters and also the dry matter content, the grain yield and nitrogen and phosphorus uptake significantly over the uninoculated control. The inoculation effects were more pronounced in the presence of added fertilizers. The possibilities of saving half the dose of N and replacing superphosphate with rockphosphate and inoculation with ‘phosphate-solubilizers’ are discussed.     

Rhizobium leguminosarum inoculation decreases damage to faba bean (Vicia faba) caused by broad bean mottle bromovirus and bean yellow mosaic potyvirus

Faba bean (Vicia faba) plants were inoculated with rhizobia and then their sap was infected with broad bean mottle bromovirus (BBMV) or bean yellow mosaic potyvirus (BYMV) in a field experiment. Both viral infections significantly decreased shoot and root dry weight, number of nodules, nodule dry weight, numbers of flowers and pods/plant, total plant N, grain yield and N₂ fixation. However, inoculation withRhizobium leguminosarum significantly increased all these parameters, both in healthy and virus-infected plants. Although BYMV was more destructive than BBMV, inoculation with rhizobia could be used, with other control measures, to limit damage by both viruses.The authors are with the Department of Biochemistry and Soil Science, Faculty of Agriculture, Shambat, Sudan.     

Potential role of rhizobia to enhance chickpea-growth and yield in low fertility-soils of Tunisia

Plant growth-promoting rhizobacteria are bacteria that improve plant growth and reduce plant pathogen damages. In this study, 100 nodule bacteria were isolated from chickpea, screened for their plant growth-promoting (PGP) traits and then characterised by PCR-RFLP of 16 S rDNA. Results showed that most of the slow-growing isolates fixed nitrogen but those exhibiting fast-growth did not. Fourteen isolates solubilized inorganic phosphorus, 16 strains produced siderophores, and 17 strains produced indole acetic acid. Co-culture experiments identified three strains having an inhibitory effect against Fusarium oxysporum, the primary pathogenic fungus for chickpea in Tunisia. Rhizobia with PGP traits were assigned to Mesorhizobium ciceri, Mesorhizobium mediterraneum, Sinorhizobium meliloti and Agrobacterium tumefaciens. We noted that PGP activities were differentially distributed between M. ciceri and M. mediterraneum. The region of Mateur in northern Tunisia, with clay–silty soil, was the origin of 53% of PGP isolates. Interestingly, we found that S. meliloti and A. tumefaciens strains did not behave as parasitic nodule-bacteria but as PGP rhizobacteria useful for chickpea nutrition and health. In fact, S. meliloti strains could solubilize phosphorus, produce siderophore and auxin. The A. tumefaciens strains could perform the previous PGP traits and inhibit pathogen growth also. Finally, one candidate strain of M. ciceri (LL10)—selected for its highest symbiotic nitrogen fixation and phosphorus solubilization—was used for field experiment. The LL10 inoculation increased grain yield more than three-fold. These finding showed the potential role of rhizobia to be used as biofertilizers and biopesticides, representing low-cost and environment-friendly inputs for sustainable agriculture.

     

Mycorrhizal inoculation effect on nodulation and N accumulation in Cajanus cajan at soil P concentrations sufficient or inadequate for mycorrhiza-free growth

The interaction of Cajanus cajan with Rhizobium and vesicular-arbuscular mycorrhizal fungi (VAMF) was investigated in a greenhouse experiment. C. cajan was planted in soil that had been inoculated with Glomus aggregatum or treated with benlate to suppress VAMF activity. Initial soil solution P concentrations of 0.06, 0.2, 0.4, and 0.8 mg l^-1 were established to test the interaction at external P levels that ranged from inadequate to nonlimiting for the host plant. At 0.06 and 0.2 mg P l^-1, mycorrhizal inoculation significantly increased plant P concentrations as well as nodule numbers and shoot dry weight. Mycorrhizal inoculation also significantly increased nodule dry weight at a soil P concentration of 0.4 mg l^-1 but did not significantly influence any of the other variables. The mycorrhizal inoculation effect observed at this soil solution P concentration could not be explained by any of the measures of plant P status. At 0.8 mg P l^-1, none of the measured variables were affected significantly by mycorrhizal inoculation. The results indicate that the enhanced nodulation associated with mycorrhizal inoculation at soil P concentrations lower than 0.4 mg l^-1 was explainable by mycorrhizal-mediated P uptake. The small but significant increase in nodule mass due to VAMF inoculation at 0.4 mg P l^-1 suggests that factors not related to plant P nutrition may be involved. On the other hand, the lack of a VAMF inoculation effect at 0.8 mg P l^-1 despite VAMF colonization at a level comparable to that observed at the former P concentration appear to discount this hypothesis. This observation is also supported by the lack of response of plant N status and nodule number to VAMF inoculation at this soil P concentration.Contribution from the Hawaii Institute of Tropical Agriculture and Human Resources Journal Series No.4066     

Field nursery inoculation of Hevea brasiliensis Muell. Arg. seedling rootstock with vesicular-arbuscular mycorrhizal (VAM) fungi

The growth response of Hevea brasiliensis to vesicular-arbuscular mycorrhizal (VAM) fungi inoculation was assessed in two field nursery sites containing indigenous mycorrhizal fungi (IMF). Seedling rootstocks were inoculated with mixed VAM-fungal species in a factorial combination with phosphorus (P) fertilizer application, and planted in randomised blocks on sandy (site 1) and clayey (site 2) soils. Plants were harvested after 26 weeks for measurements of shoot dry weight (DW), stem diameter, height, mycorrhizal root colonization and leaf nutrient contents. At site 1, VAM increased shoot DW, stem diameter and plant height only in treatments without P applied. Increases in shoot DW due to VAM were 70% greater than the uninoculated controls although this was reduced to 5% when P was applied. At site 2, VAM inoculation also increased shoot DW and stem diameter but the magnitude of the increases was smaller. Shoot DW response due to VAM was only 29%. At this second site, applying phosphate to uninoculated plants did not increase shoot yields further. Leaf concentrations of all nutrients were unaffected by VAM at both sites, except for copper (Cu) which was increased by VAM in treatments where P was not applied. However, leaf contents of P, potassium (K), magnesium (Mg) and Cu were increased by VAM at site 1, and of leaf nitrogen (N) and K at site 2. These experiments demonstrate that VAM-fungi could be introduced into field nursery sites to improve growth and P uptake by H. brasiliensis. The relevance of VAM-fungi to H. brasiliensis seedling rootstock development and the influence of IMF in determining field responses is discussed.     

Growth increment ofAlnus glutinosa upon dual inoculation with effective and ineffectiveFrankia strains

Investigations on the ecological function of ineffectiveFrankia strains and their behaviour in competition with effectiveFrankia strains indicated an enhanced plant growth upon dual inoculation with increasing amounts of effective (i.e. N₂-fixing)Frankia strains and simultaneous inoculation with a constant amount of an ineffectiveFrankia strain. Enhanced plant growth was measured as increase in plant height and total dry weight at constant shoot/root ratio. The stimulating effect of the ineffective strain was independent of the plant clone and was obtained with bothAlnus glutinosa clones W I and B II, which were resistant and susceptable, respectively, to the ineffective strain. Stimulation was also independent of the nodulation conditions. Short-term studies (7 weeks) under axenic conditions and greenhouse experiments during 3 months showed comparable results, not only in plant growth but also in nodule formation. Increment in plant growth was not necessarily correlated to higher nodule formation with the effectiveFrankia strains.     

Response of field-grownCasuarina equisetifolia to inoculation withFrankia strain ORS 021001 entrapped in alginate beads

A large scale field experiment (ca 1 ha) was carried out in Senegal, to evaluate the response ofCasuarina equisetifolia to inoculation withFrankia strain ORS 021001 entrapped in alginate beads. Biomasses (expressed as dry weight or total nitrogen) of assimilatory branchlets, wood and roots, and nodules were measured in uninoculated and inoculated trees, randomly sampled 1,2 and 3 years after transplantation in the field. When biomasses were expressed as dry weight, increases due to inoculation were similar at the three sampling dates, 45, 36 and 40%, respectively. When biomasses were expressed as total nitrogen, the response to inoculation with time was much higher in the 2nd year than in the 1st and 3rd year. N₂ fixation, estimated using the difference method reached 2.48, 12.25 and 13.44 g N₂ fixed annually per tree. Correspondingly, nodule dry weights, expressed in g per tree, were 2.5, 12.18 and 22.75 at the end of the 1 st, 2nd and 3rd year, respectively. In spite of the positive response of field-grownCasuarina equisetifolia to inoculation, the decrease of N₂ fixation observed in the third year was probably due to unfavorable climatic conditions coupled with insect attacks at the beginning of the third year.     

Genotypic alteration and competitive nodulation of Mesorhizobium muleiense against exotic chickpea rhizobia in alkaline soils

Mesorhizobium muleiense, Mesorhizobium mediterraneum and Mesorhizobium ciceri are chickpea (Cicer arietinum L.) rhizobia that share a high similarity of the symbiotic genes nodC and nifH, but they have different geographic distributions. M. muleiense has been isolated and found only in alkaline soils of Xinjiang, China, whereas the other two strains have been found in the Mediterranean and India. To investigate the species stability of M. muleiense during natural evolution and its capability of competitive nodulation against the other two exotic species, re-sampling of nodules in the field and competition experiments between the three species were conducted. The results showed that the predominant microsymbiont associated with chickpea grown in Xinjiang was still M. muleiense, but the predominant genotypes of M. muleiense had changed significantly during the four years since a previous survey. The data also showed that M. mediterraneum and M. ciceri were more competitive than the residential strain of M. muleiense CCBAU 83963^T in sterilized vermiculite or soils from Xinjiang. However, in non-sterilized soils, M. muleiense was the predominant nodule occupier. These results indicated that natural or adapting evolution of M. muleiense was occurring in fields subjected to changing environmental factors. In addition, the biogeography and symbiotic associations of rhizobia with their host legumes were also influenced by biological factors in the soil, such as indigenous rhizobia and other organisms.     

Bradyrhizobium technology: a promising substitute for chemical nitrogen fertilizer in Bangladesh agriculture

Nitrogen is the most limiting element in Bangladesh soils and urea is the fertilizer commonly used for supplying it. Bradyrhizobium/Rhizobium inoculant was tried as a source of N nutrition for grain legumes in a number of field experiments. The inoculants markedly increased nodule number, nodule mass, shoot weight and yield of the crops compared to uninoculated control and urea-N treatments. For soybean (Glycine max), inoculation increased yield 113 percent over the control and 49 percent over the urea treament. For groundnut (Arachis hypogaea), the increases were 36 and 11 percent; for lentil (Lens culinaris), 30 and 13 percent; and for mungbean (Vigna radiata), 47 and 7 percent. The local inoculant strains were suitable for dependable inoculant production. The inoculant technology can be used as a promising and cheap substitute of urea for growing food legume crops in Bangladesh.     

Biodiversity of populations of phosphate solubilizing rhizobia that nodulates chickpea in different Spanish soils

Within rhizobia, two species nodulating chickpea, Mesorhizobium ciceri and Mesorhizobium mediterraneum, are known as good phosphate solubilizers. For this reason, we have analysed the ability to solubilize phosphate of a wide number of strains isolated from Cicer arietinum growing in several soils in Spain. The aim of this work was to analyse microbial populations nodulating chickpea, that are able to solubilize phosphates, using molecular techniques. In the present work we analyzed 19 strains isolated from effective nodules of C. arietinum growing in three soils from the North of Spain. Nineteen strains showed ability to solubilize phosphate in YED-P medium. These strains were separated into 4 groups according to the results obtained by 879F-RAPD fingerprinting. The 16S rDNA sequencing of a representative strain from each group allowed the identification of strains as belonging to the genus Mesorhizobium. Strains from groups I and II showed a 99.4% and 99.2% similarity with M. mediterraneum UPM-CA142^T, respectively. The strains from group III were related to M. tianshanense USDA 3592^T at a 99.4% similarity level. Finally, the strain from group IV was related to M. ciceri USDA 3383^T with a 99.3% similarity. The LMW RNA profiles confirmed these results. Strains from groups I and II showed an identical LMW RNA profile to that of M. mediterraneum UPM-CA142^T; the profile of strains from group III was identical to that of M.␣tianshanense USDA 3592^T and the profile of strains from group IV was identical to that of M. ciceri USDA 3383^T. Different 879F-RAPD patterns were obtained for strains of the group I, group II and the M.␣mediterraneum type strain (UPM-CA142^T). The 879-RAPD patterns obtained for group III also differed from the pattern shown by M. tianshanense USDA 3592^T. Finally, the patterns between group IV and M. ciceri USDA 3383^T were also different. These results suggest that groups I and II may be subspecies of M. mediterraneum, group III a subspecies of M. tianshanense and group IV a subspecies of M. ciceri. Nevertheless, more studies are needed to establish the taxonomic status of strains isolated in this study.     

Effect of mycorrhizal inoculation and soluble phosphorus fertilizer on growth and phosphorus uptake of pearl millet

Summary Six mycorrhizal fungi were tested as inoculants for pearl millet (Pennisetum americanum Leeke) grown in pots maintained in a greenhouse. VAM fungi varied in their ability to stimulate plant growth and phosphorus uptake. Inoculation withGigaspora margarita, G. calospora andGlomus fasciculatum increased shoot drymatter 1.3 fold over uninoculated control. In another pot trial, inoculation withGigaspora calospora andGlomus fasciculatum resulted in dry matter and phosphorus uptake equivalent to that produced by adding phosphorus at 8 kg/ha.The influence of inoculatingGigaspora calospora on pearl millet at different levels of phosphorus fertilizer (0 to 60 kg P/ha) as triple superphosphate in sterile and unsterile alfisol soil was also studied. In sterile soil, mycorrhizal inoculation increased dry matter and phosphorus uptake at levels less than 20 kg/ha. At higher P levels the mycorrhizal effect was decreased. These studies performed in sterilized soil suggest that inoculation of pearl millet with efficient VAM fungi could be extremely useful in P deficient soils. However, its practical utility depends on screening and isolation of fungal strains which perform efficiently in natural (unsterilized) field conditions.