Co-inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases P use efficiency for N2 fixation in the common bean (Phaseolus vulgaris L.) under P deficiency in hydroaeroponic culture

Common bean (Phaseolus vulgaris L.) genotypes CocoT and Flamingo were inoculated with Rhizobium tropici CIAT899 and Glomus intraradices (Schenck & Smith) and grown under sufficient versus deficient phosphorus supply for comparing the effects of double inoculation on growth, nodulation, mycorrhization of the roots, phosphorus use efficiency and total nitrogen. Although the double inoculation induced a significant increase in all parameters whatever the phosphorus supply in comparison to control, significant differences were found among genotypes and treatments. Nevertheless, the highest phosphorus use efficiency and plant total nitrogen were found under P deficiency in combination with arbuscular mycorrhizal fungi. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve symbiotic nitrogen fixation even under phosphorus deficiency.     

Effect of combined inoculation ofAzospirillum brasilense and vesicular-arbuscular mycorrhiza on pearl millet (Pennisetum americanum)

Summary Growth and phosphorus uptake of pearl millet (Pennisetum americanum) on an unsterile, phosphorus-deficient soil was improved by the seed inoculation withAzospirillum brasilense or soil inoculation with the vesicular-arbuscular mycorrhizal fungi (Acaulospora,Gigaspora margarita, Glomus fasciculatum). These microorganisms acted synergistically when added simultaneously and the response was significant withAzospirillum brasilense + Gigaspora margarita andAzospirillum brasilense + Glomus fasciculatum combinations over uninoculated control as far as the dry matter content of shoots, root biomass and phosphorus uptake of the millet was concerned.     

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.     

Dual inoculation withRhizobium sp. andGlomus fasciculatum enhances nodulation,yield and nitrogen fixation in chickpea (Cicer arietinum Linn.)

Summary Seed inoculation with Rhizobium and soil inoculation withGlomus fasciculatum increased nodulation, nitrogen and phosphorus concentration in plants and yield of chickpea (Cicer arietinum) var. BG 212 in pots containing unsterilized soil especially with 50kgP₂O₅ ha⁻¹ in the form of superphosphate. Inoculation with Rhizobium orG. fasciculatum separately or in combination significantly increased the N₂ fixed in straw and grain than uninoculated controls as determined by¹⁵N atom percent excess of plants grown in soil amended with labelled ammonium sulphate (¹⁵NH₄)₂SO₄) at the rate of 20kg N ha⁻¹. These increases were most pronounced when P was applied at 50kgP₂O₅ ha⁻¹.     

Influence of phosphorus level on VA Mycorrhizal colonization and growth of cowpea cultivars

Vesicular-arbuscular mycorrhizal (VAM) associations often vary according to the abundance of available soil phosphorus (P). Therefore, understanding the response of crop plants to colonization by VAM fungi necessitates the study of the response of colonized and noncolonized plants, from a range of cultivars, to differing levels of P. Cowpea is grown throughout the world, often on impoverished soils in which it can benefit from formation of mycorrhizae. The present study was conducted to determine the response of four cultivars of cowpea (Vigna unguiculata (L.) Walp.), varying in nitrogen fixation capacity, to inoculation withGlomus fasciculatum at four levels of added P in the rooting medium. In a greenhouse experiment, four cowpea cultivars, Mississippi Silver, Brown Crowder, Six Week Browneye and MI 35, were grown with and without the mycorrhizal fungus at four levels of added P, 0, 10, 20 and 30 ppm. Root colonization (%) was negatively correlated with P content of the growth medium and shoot P concentration. Intraspecific variability was shown for shoot dry weight and leaf area in response to inoculation withG. fasciculatum at different P levels. The range of P required in the growth medium which allowed benefit fromG. fasciculatum was identified for individual cultivars using shoot dry weight and leaf area, and collectively across cultivars for other parameters.     

Effects of introduced and indigenous VA mycorrhizal fungi on nodulation,growth and nutrition ofMedicago sativa in phosphate-fixing soils as affected by P fertilizers

Summary The legumeMedicago sativa was grown in two phosphate-fixing soils which received soluble or rock phosphate. The effects of the inoculation withGlomus mosseae on plant nutrition and nodulation were studied. The introduced VA fungi became successfully established and improved the degree of infection over level achieved by native endophytes. In all experimental conditions tested, plant dry weight, the total uptake of N and P and nodulation byRhizobium meliloti were increased by mycorrhizal inoculation. The size of the increase was inversely correlated with soluble P content in the soil. Mycorrhization, enhanced by introduction of suitable VA fungi, had similar effects to that of the dose of soluble phosphate tested. Indigenous and native endophytes cooperated in these effects. Results are discussed in terms of reducing the input of soluble P fertilizer to phosphate-fixing soils and the possibility of restoring the phosphate stock using a more rational supply of soluble P, that allows cooperation with VA fungi, or by the use of less soluble and expensive forms of P fertilizers.     

Response of Leucaena/Rhizobium symbiosis to mineral nutrients in Southwestern Nigeria

Pot and field experiments carried out at the International Institute of Tropical Agriculture (IITA) and at Fashola, Southwestern Nigeria, examined the effect of inoculation and N, P and micronutrients on nodulation and growth ofLeucaena leucocephala (Lam.) de Wit. In pot studies all parameters measured, except the percentage of nitrogen in shoots, were improved by inoculation, nitrogen and phosphorus. Micronutrients increased only nitrogen and allantoin contents. Interactions between inoculation and P, N and micronutrients on nodulation and growth of leucaena were observed. The effect of inoculation and fertilization with phosphorus or micronutrients was further investigated in field experiments. Establishment of uninoculated and unfertilized leucaena was poor at both locations due to low soil fertility and the presence of only a few native leucaena rhizobia. At one site, only inoculated plants were nodulated, while at the other, all plants produced nodules. Shoot dry weight, total nitrogen and phosphorus of inoculated plants were statistically equal to nitrogen-fertilized plants. Uninoculated plants were stunted. Generally, micronutrients did not influence nodulation, total nitrogen or growth of leucaena. They had only a positive effect on nitrogenase activity. Phosphorus increased total nitrogen and phosphorus uptake and plant growth. A 75% increase in shoot dry weight was obtained when 80 kg P ha^–1 was applied to inoculated leucaena with Rhizobium strain IRc 1045. Inoculated plants contained more allantoins than uninoculated ones but no significant correlation was found between these compounds and other parameters of N fixation.     

Combined inoculation with Glomus intraradices and Rhizobium tropici CIAT899 increases phosphorus use efficiency for symbiotic nitrogen fixation in common bean (Phaseolus vulgaris L.)

This study compared the response of common bean (Phaseolus vulgaris L.) to arbuscular mycorrhizal fungi (AMF) and rhizobia strain inoculation. Two common bean genotypes i.e. CocoT and Flamingo varying in their effectiveness for nitrogen fixation were inoculated with Glomus intraradices and Rhizobium tropici CIAT899, and grown for 50 days in soil–sand substrate in glasshouse conditions. Inoculation of common bean plants with the AM fungi resulted in a significant increase in nodulation compared to plants without inoculation. The combined inoculation of AM fungi and rhizobia significantly increased various plant growth parameters compared to simple inoculated plants. In addition, the combined inoculation of AM fungi and rhizobia resulted in significantly higher nitrogen and phosphorus accumulation in the shoots of common bean plants and improved phosphorus use efficiency compared with their controls, which were not dually inoculated. It is concluded that inoculation with rhizobia and arbuscular mycorrhizal fungi could improve the efficiency in phosphorus use for symbiotic nitrogen fixation especially under phosphorus deficiency.     

Enhancement of nitrogen fixation in lentil,faba bean,and soybean by dual inoculation with Rhizobia and mycorrhizae

The combined effect of Vesicular Arbuscular Mycorrhizae (VAM) and Rhizobium on the cold season legumes, lentil and faba bean, as well as on summer legume, soybean, were studied in soils with low indeginous VA mycorrhizal spores. Inoculation of the plant with VA mycorrhizal fungi increased the level of mycorrhizal root infection of lentil, faba bean and soybean. The inoculation with Rhizobium had no significant effect on VA mycorrhizal infection percent, but VA mycorrhizal inoculation increased nodulation of the three legumes. The inoculation with Rhizobium alone significantly increased plant dry weight and N content of lentil and faba bean as well as seed yield of soybean. VA mycorrhizal inoculation also significantly increased plant dry weight and phosphorus content of the plants as did fertilization with superphosphate. Rock phosphate fertilization, however, had no significant effect on plant growth or phosphorus uptake. The addition of rock phosphate in combination with VA mycorrhizal inoculation significantly increased plant dry weight and P uptake of the plants. The dual inoculation with both rhizobia and mycorrhizae induced more significant increases in plant dry weight, N and P content of lentil and faba bean as well as seed yield of soybean than inoculation with either VA mycorrhizae or Rhizobium alone.     

VA-mycorrhiza mediated P effect on growth and yield of sunflower (Helianthus annuus L.) at different P levels

A field trial was conducted to study the response of sunflower (Helianthus annuus L.) to different phosphorus levels (16, 24 or 32 kg P ha^–1) and inoculation with vesicular-arbuscular mycorrhizal fungus, Glomus fasciculatum on vertisol during summer 1993. At the vegetative stage of sunflower, percent mycorrhizal root colonization, spore count, dry biomass and P uptake did not differ significantly between inoculated and uninoculated control plants. However, at later stages (flowering and maturity) percent root colonization, spore count, total dry biomass and total P uptake were significantly higher in inoculated plants than in uninoculated control plants. The total dry biomass, P content and seed yield increased with increasing P level in uninoculated plants, whereas no significant difference was observed between 16 and 32 kg P ha^–1 in inoculated plants. The positive effect of mycorrhizal inoculation decreased with increasing P level above 16 kg P ha^–1, due to decreased percent root colonization and spore count at higher P levels.     

Symbiotic effectiveness of Hup+ Rhizobium,VAM fungi and phosphorus levels in relation to nitrogen fixation and plant growth ofCajanus cajan

Effect of hydrogen uptake positive (Hup⁺) strain ofRhizobium sp. (pigeon pea) and VAM fungus (Glomus fasciculatum) was studied on the symbiotic parameters of pigeon pea (Cajanus cajan) cv. AL-15 at various levels of phosphorus. The Hup⁺ Rhizobium strain showed more nodulation, plant biomass and plant nitrogen content than its Hup⁻ counterpart. VAM infection in pigeon pea roots helped in translocating phosphorus from the soil and improved nitrogen fixation. Similarly, addition of phosphorus was found to play a positive role in enhancing all these parameters. Dual inoculation of Hup⁺ Rhizobium strain and VAM significantly increased nodulation, nitrogenase activity, plant nitrogen and phosphorus content and plant biomass compared to single inoculation of either organism and dual inoculation with Hup⁻ and VAM fungus.     

Interactions of an arbuscular mycorrhizal fungus with free or co-encapsulated cells of Rhizobium trifoli and Yarowia lipolytica inoculated into a soil-plant system

Four times higher nodule number was observed when Glomus deserticola (an arbuscular mycorrhizal fungus) was introduced into a soil-plant system as compared to the control inoculated only with Rhizobium trifoli. This symbiotic parameter was further enhanced by Yarowia lipolytica together with an increase in root mycorrhizal infection of about 14%. Soil inoculation with co-encapsulated R. trifoli and Y. lipolytica provoked a 10-fold increase in root nodulation and led to 55% mycorrhization of the test plant.     

Improved growth of white clover in hill grasslands by mycorrhizal inoculation

Field trials in a hill grassland area of mid-Wales showed that growth of white clover could be improved by inoculation with selected vesicular-arbuscular (VA) mycorrhizal fungi. After one growing season mycorrhizal inoculation had approximately doubled shoot dry weight, runner number and nodulation at two sites previously improved by adding lime and fertiliser and by replacing the native vegetation mainly with rye-grass and white clover. At a previously unimproved site, limed at the time of planting, mycorrhizal inoculation increased shoot dry weight by up to two and a half times, almost doubled runner formation, and increased nodulation up to five-fold where 90 kg P/ha as basic slag had been applied. Responses to mycorrhiza at this site were smaller with 22-5 kg P/ha as basic slag and were absent where no P was added. After two growing seasons inoculation responses were even more pronounced at this previously unimproved site; where clovers were being overgrown by adjacent native grasses only those given both 90 kg P/ha and mycorrhizal inoculation survived well.     

Mycorrhizal inoculation in neem (Azadirachta indica) enhances azadirachtin content in seed kernels

In view of the high mycorrhizal dependency of neem trees (Azadirachta indica), an experiment was conducted to study if Arbuscular Mycorrhizal (AM) inoculation can enhance the azadirachtin content in seed kernels of trees grown in the field. Azadirachtin is an important active ingredient in neem seed kernels based on which a large biopesticide industry has emerged in India and few countries in Europe and the USA. Inoculation of neem seedlings in the nursery with Glomus fasciculatum and Glomus mosseae resulted in increased height, dry weight, root colonization and phosphorus (P) content. In a separate experiment, field-grown neem plants inoculated in the nursery and during transplantation with Glomus fasciculatum were evaluated after 5 years. No significant differences were found in the tree height, girth at breast height (GBH) and fruit yield but oil percentage, total triterpenoids and azadirachtin content in kernels increased significantly as a result of AM inoculation. A similar enhancement in azadirachtin was noted with P application. These results open up possibilities of producing quality neem seed with high bioactive ingredients through AM inoculation.     

Mycorrhizal colonization decreases respiration of common bean nodulated root in hydroaeroponic culture

The bean-rhizobia symbiosis allows atmospheric nitrogen fixation through nodule formation. Nevertheless, nodule establishment in Mediterranean areas is subjected to various biotic and abiotic constraints such as phosphorus soils deficiency. This study compares plant-growth response to moderate (75 μmol KH₂PO₄ plant^?1 week^?1) versus severe phosphorus deficiency (30 μmol KH₂PO₄ plant^?1 week^?1) after inoculation with Rhizobium tropici CIAT 899 and Glomus intraradices of four Phaseolus vulgaris lines contrasting in P use efficiency (PUE) for their symbiotic nitrogen fixation (SNF) in hydroaeroponic culture. After 5 weeks of growth under glasshouse conditions, the oxygen consumption related to nitrogen fixation was measured on intact nodulated roots. The obtained results revealed that mycorrhizal colonization decreased the nodulated-roots O₂ consumption of P. vulgaris under both P deficiencies although it increased the growth of all plant organs and the nodulation with a large genotypic variability. Moreover, mycorrhizal colonization was higher under severe P deficiency than under moderate one. In conclusion, the tripartite inoculation improved growth parameters under severe P-deficiency with a decrease in nodulated root O₂ consumption.     

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.     

Influence of Glomus fasciculatum on Meloidogyne hapla Infecting Allium cepa

The impact of Glomus fasciculatum on Meloidogyne hapla associated with Allium cepa was evaluated in two experiments. Nematode density was not different in mycorrhizal and nonmycorrhizal plants 10 weeks after the joint inoculation of M. hapla and G. fasciculatum. Differences in the age structure of M. hapla populations reared on mycorrhizal and nonmycorrhizat plants were noted. G. fasciculatum enhanced leaf and bulb growth of A. cepa in the absence of M. hapla, but did not affect plant weight when nematodes were present. Survival and reproduction of M. hapla were not affected by G. fasciculatum or phosphorus (P). The estimated time required for inoculated second-stage juveniles (J2) to mature to the adult stage was 1,000 degree hours (base = 9 C) greater in mycorrhizal than in nonmycorrhizal plants supplemented with P. Although the infectivity of J2 was not measured directly, colonization of A. cepa by G. fasciculatum appeared to alter the ability of M. hapla to penetrate roots.     

Preparation of gel-entrapped mycorrhizal inoculum in the presence or absence of Yarowia lipolytica

Two arbuscular mycorrhizal fungi (Glomus deserticola and Glomus fasciculatum) were entrapped in calcium alginate, alone or in combination with a phosphate-solubilizing yeast (Yarowia lipolytica) and, after storage for 60 days, were inoculated into soil microcosms with tomato as the test plant. The average extent of root colonization by gel-entrapped G. deserticola and G. fasciculatum were 32 ± 5.6 and 24 ± 12.1%, respectively. Improved infective potential and colonization efficiency were observed when Y. lipolytica was co-entrapped with the mycorrhizal fungi. The best value, 49%, of mycorrhizal colonization was in roots of plants inoculated with G. deserticola co-entrapped with Y. lipolytica.     

The effect of VA mycorrhizal fungi,phosphorus and drought stress on the growth of Acacia nilotica and Leucaena leucocephala seedlings

The effect of vesicular-arbuscular mycorrhizal (VAM) fungi on growth and drought resistance of Acacia nilotica and Leucaena leucocephala seedlings was studied in a glasshouse experiment. The experimental design was a 2·2·2 factorial: ± mycorrhizal inoculation, ± application of phosphorus fertilizer and ± repeated drought treatment. The growth promoting effect of VAM fungi equalled the effect of phosphorus fertilization after 12 weeks. The drought treatment reduced seedling biomass and nodulation. Differences between the plant species were found with respect to growth improvements due to VAM inoculation and/or phosphorus fertilization under drought stress conditions. The results are discussed in relation to plant drought resistance and reforestation in the subhumid to arid tropics.     

The interaction of rockphosphate,Bradyrhizobium, vesicular-arbuscular mycorrhizae and phosphate-solubilizing microbes on soybean grown in a sub-Himalayan mollisol

A factorial design 2³ × 4 with two levels of Mussorie rockphosphate (RP) with or without vesicular-arbuscular mycorrhizal (VAM) fungi and Bradyrhizobium japonicum, and four treatments of phosphate-solubilizing microbes (PSM) Pseudomonas striata, Bacillus polymyxa, Aspergillus awamori was employed using Patharchatta sandy loam soil (Typic Hapludoll). The observations included mycorrhization, nodulation, grain and straw yield, N and P uptake, available soil P and the PSM population in the soil after crop harvest. Inoculation with endophytes alone caused about 70% root colonization. Addition of rockphosphate or inoculation with PSM, except B. polymyxa, stimulated root infection of native as well as introduced VAM endophytes. Application of RP or inoculation with Bradyrhizobium japonicum, mycorrhizal fungi or phosphate-solubilizing microorganisms significantly increased nodulation, N uptake, available soil P and the PSM population in the soil after the crop harvest. The grain and straw yields did not increase following RP addition or mycorrhizal inoculation but increased significantly after inoculation wit Bradyrhizobium or PSM. In general, the application of RP, Bradyrhizobium, VAM and PSM in combinations of any two or three resulted in significant increases in nodulation, plant growth, grain yield and uptake of N and P. Among the four factor interactions, rockphosphate, Bradyrhizobium and P. striata in the absence of VAM resulted in maximal nodulation, grain and straw yields and N uptake by soybean. The highest P uptake by soybean grain was recorded with Bradyrhizobium and A. awamori in the absence of rockphosphate and VAM. Generally, available soil P and PSM population after crop harvest were not significantly increased by the treatment combinations giving the maximal uptake of nutrients. However, they increased significantly in response to PSM, which produced no significant increase in total uptake of nutrients.Research paper no. 7498