The fate of markerAzospirillum lipoferum inoculated into rice and its effect on growth,yield and N2 fixation of plants studied by acetylene reduction,15N2 feeding and15N dilution techniques

Summary A spontaneous mutant ofAzospirillum lipoferum, resistant to streptomycin and rifampicin, was inoculated into the soil immediately before and 10 days after transplanting of rice (Oryza sativa L.). Two rice varieties with high and low nitrogen-fixing supporting traits, Hua-chou-chi-mo-mor (Hua) and OS4, were used for the plant bacterial interaction study. The effect of inoculation on growth and grain and dry matter yields was evaluated in relation to nitrogen fixation, by in situ acetylene reduction assay,¹⁵N₂ feeding and¹⁵N dilution techniques. A survey of the population of marker bacteria at maximum tillering, booting and heading revealed poor effectivety. The population of nativeAzospirillum followed no definite pattern. Acetylene-reducing activity (ARA) did not differ due to inoculation at two early stages but decreased in the inoculated plants at heading. In contrast, inoculation increased tiller number, plant height of Hua and early reproductive growth of both varieties. Grain yield of both varieties significantly increased along with the dry matter. Total N also increased in inoculated plants, which was less compared with dry matter increase.¹⁵N₂ feeding of OS4 at heading showed more¹⁵N₂ incorporation in the control than in the inoculated plants. The ARA,¹⁵N and N balance studies did not provide clear evidence that the promotion of growth and nitrogen uptake was due to higher N₂ fixation.     

Nitrogen fixation by Azospirillium brasilense in soil and the rhizosphere under controlled environmental conditions

Summary Acetylene reduction activity by Azospirillum brasilense, either free-living in soils or associated with wheat roots, was determined in a sterilised root environment at controlled levels of O₂ tension and with different concentrations of mineral N. In an unplanted, inoculated soil nitrogenase activity remained low, at approximately 40 nmol C₂H₄ h^-1 per 2kg fresh soil, increasing to 300 nmol C₂H₄ h^-1 when malic acid was added as a C source via a dialyse tubing system. The N₂ fixation by A. brasilense in the rhizosphere of an actively growing plant was much less sensitive to the repressing influence of free O₂ than the free-living bacteria were. An optimum nitrogenase activity was observed at 10 kPa O₂, with a relatively high level of activity remaining even at an O₂ concentration of 20 kPa. Both NO _inf3 ^sup- and NH _inf4 ^sup+ repressed nitrogenase activity, which was less pronounced in the presence than in the absence of plants. The highest survival rates of inoculated A. brasilense and the highest rates of acetylene reduction were found in plants treated with azospirilli immediately after seedling emergence. Plants inoculated at a later stage of growth showed a lower bacterial density in the rhizosphere and, as a consequence, a lower N₂-fixing potential. Subsequent inoculations with A. brasilense during plant development did not increase root colonisation and did not stimulate the associated acetylene reduction. By using the ¹⁵N dilution method, the affect of inoculation with A. brasilense in terms of plant N was calculated as 0.067 mg N₂ fixed per plant, i.e., 3.3% of the N in the root and 1.6% in the plant shoot were of atmospheric origin. This ¹⁵N dilution was comparable to that seen in plants inoculated with non-N₂-fixing Psudomonas fluorescens.     

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.     

Pre-inoculation with arbuscular mycorrhizal fungi suppresses root knot nematode (<Emphasis Type="Italic">Meloidogyne incognita</Emphasis>) on cucumber (<Emphasis Type="Italic">Cucumis sativus</Emphasis>)

A pot experiment was conducted to evaluate the influence of pre-inoculation of cucumber plants with each of the three arbuscular mycorrhizal (AM) fungi Glomus intraradices, Glomus mosseae, and Glomus versiforme on reproduction of the root knot nematode Meloidogyne incognita. All three AM fungi tested significantly reduced the root galling index, which is the percentage of total roots forming galls. Numbers of galls per root system were significantly reduced only in the G. intraradices + M. incognita treatment. The number of eggs per root system was significantly decreased by AM fungus inoculation, no significant difference among the three AM fungal isolates. AM inoculation substantially decreased the number of females, the number of eggs g⁻¹ root and of the number of eggs per egg mass. The number of egg masses g⁻¹ root was greatly reduced by inoculation with G. mosseae or G. versiforme. By considering plant growth, nutrient uptake, and the suppression of M. incognita together, G. mosseae and G. versiforme were more effective than G. intraradices.     

15N2还原法测定胡萝卜离体类根瘤的固氮作用

用低浓度的2,4-D处理胡萝卜离体根诱导结瘤,并接种固氮螺菌(Azospirillum.brasilense),用15N2还原法测定结果证明,胡萝卜离体类根瘤能固定15N2。电镜切片观察发现,类根瘤内有菌体存在,菌体进入细胞间隙中比进入细胞内多。     

Effect of nitrogen on the yield response of Pennisetum americanum,Triticum aestivum and Zea mays to inoculation with Azospirillum brasilense under temperate conditions

Summary A nitrate-respiring strain, a denitrifying strain, and a non-nitrogen-fixing strain of Azospirillum brasilense were compared for their effect on the growth of pearl millet (Pennisetum americanum), wheat (Triticum aestivum) and maize (Zea mays) under temperate conditions in nitrogen-limited pot cultures. Increases in yield of Z. mays shoots occurred with all three strains when inoculation coincided with the addition of low levels of combined nitrogen. The inoculation of A. brasilense did not show any effect on the yield of P. americanum and T. aestivum. Increased numbers of A. brasilense became associated with Z. mays roots following the addition of low levels of combined nitrogen. Low and very variable rates of acetylene reduction activity were observed from excised roots of inoculated Z. mays plants without preincubation. Results indicate that inoculation of cereals with A. brasilense under temperate conditions has only a limited effect on plant growth.     

Influence of arbuscular mycorrhizae and phosphorus fertilization on growth, nodulation and N2 fixation (15N) inMedicago sativa at four salinity levels

The rose of an isolate of the arbuscular mycorrhizal (AM) fungusGlomus mosseae in the protection ofMedicago sativa (+Rhizobium meliloti) against salt stress induced by the addition of increasing levels of soluble salts was studied. The interactions between soluble P in soil (four levels), mycorrhizal inoculum and degree of salinity in relation to plant growth, nutrition and infective parameters were evaluated. Salt stress was induced by sequential irrigation with saline water having four concentrations of three salts (NaCl, CaCl₂, and MgCl₂).¹⁵N-labelled ammonium sulphate was added to provide a quantitative estimate of N₂ fixation under moderate to high salinity levels. N and P concentration and nodule formation increased with the amount of plant-available P or mycorrhizal inoculum in the soil and generally declined as the salinity in the solution culture increased from a moderate to a high level. The mycorrhizal inoculation protected the plants from salt stress more efficiently than any amount of plant-available P in soil, particularly at the highest salinity level applied (43.5 dS m^–1). Mycorrhizal inoculation matched the effect on dry matter and nutrition of the addition in the soil of 150 mg P kg^–1. Nevertheless the highest saline solution assayed (43.5 dS m^–1) affected more severely plants supplemented with phosphorus than those with the addition of mycorrhizal inoculum. Such a saline-depressing effect was 1.5 (biomass), 1.4 (N) and 1.5 (P) times higher in plants supplied with soluble phosphate than with AM inoculum. Mechanisms beyond those mediated by P must be involved in the AM-protectioe effect against salinity. The¹⁵N methodology used allowed the determination of N₂ fixation as influenced by different P applications compared to mycorrhizal inoculation. A lack of correlation between nodule formation and function (N₂ fixation) was evidenced in mycorrhizal-inoculated plants. In spite of the reduced activity per nodule in mycorrhizal-inoculated In spite of the reduced activity per nodule in mycorrhizal-inoculated plants, the N contents determined indicated the highest acquisition of N occurred in plants with the symbiotic status. Moreover, N and P uptake increased while Ca and Mg decreased in AM-inoculated plants. Thus P/Ca ratios and cation/anion balance in general were altered in mycorrhizal treatments. This study therefore confirms previous findings that AM-colonized plants have optional and alternative mechanisms available to satisfy their nutritive requirements and to maintain their physiological status in stress situations and in disturbed ecosystems.     

N2-fixation by ammonium-excreting Azospirillum brasilense in auxine-induced root tumours of wheat (Triticum aestivum L.)

Summary Wheat seedlings, treated with the auxine 2,4-dichlor-phenoxy acetic acid (2,4-D) during germination developed only a residual root system. Root elongation was extremely restricted and root tips were deformed to thick club-shaped tumours. When 2,4-D was added in a later stage of plant growth the plants developed additional nodule-like knots along primary roots. Root and shoot dry-matter production was slightly repressed in all 2,4-D treatments and N translocation from roots to shoots was repressed as well. When transferred to an auxine-free growth medium, the 2,4-D-affected roots were not capable of complete recovery. In plants inoculated gnotobiotically with Azospirillum brasilense, either with the wild type or with the NH ₄ ⁺ -excreting mutant strain C3, a 2,4-D addition increased rhizosphere acetylene-reduction activity at pO₂ 1.5 kPa. The O₂ sensitivity of root-associated nitrogenase activity tended to be reduced. The number of root-colonizing bacteria, at approximately 10⁸ colony-forming units (cfu) per g dry root, was similar in the 2,4-D treatments and untreated controls. Plant treatment with high concentrations of the chemical isomer 3,5-dichlor-phenoxy acetic acid (3,5-D) did not have comparable effects, either on plant development or on rhizosphere-associated nitrogenase activity. Root-tumour tissue inhabited by A. brasilense showed purple staining when subjected to a tetrazolium chloride solution, which may indicate intensive local nitrogenase activity in this tissue. Exposed to an ¹⁵N₂-enriched atmosphere, plants treated with 2,4-D and with A. brasilense incorporated significantly higher amounts of ¹⁵N than untreated controls. In all cases the highest values of ¹⁵N enrichment were found following inoculation with the NH ₄ ⁺ -excreting mutant strain C3. Present address: F. A. Janssens Laboratory of Genetics, Catholic University of Leuven, Willem de Croylan 42, B-3001 Heverlee, Belgium     

Dual inoculation with strains of Bradyrhizobium japonicum and Azospirillum brasilense to improve growth and biological nitrogen fixation of soybean (Glycine max L.)

The effects of inoculation with Bradyrhizobium japonicum and Azospirillum brasilense strains on the growth of soybean were evaluated with regard to the estimation of N₂ fixation using the ¹⁵N isotope dilution technique. Inoculation, in general, increased the dry mass of soybean as well as nitrogen content. Dual inoculation with a mixture of B. japonicum and A. brasilense strains was superior over single inoculation with B. japonicum. Nitrogen fixed (Ndfa) varied according to inoculant and soil conditions. Percentages of nitrogen derived from air (% Ndfa) using a non-nodulating isoline were 72% and 76% for B. japonicum and B. japonicum plus A. brasilense, respectively, in non-sterile soil. A similar but higher trend was recorded in sterilized soil, in which the percentages of N₂ fixed were 81% and 86% for single and dual inoculation, respectively. The correlation coefficient between N₂ fixed and N uptake (r=0.94) and dry mass (r=0.89) was significant. Application of special bacterial inoculants in agricultural systems of Egypt seems to be a promising technology and could be used for improving soybean growth as well as soil fertility, thus minimizing environmental pollution. Received: 10 January 1996     

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.     

Infectivity and acetylene reduction of diazotrophic rhizosphere bacteria in wheat (Triticum aestivum) seedlings under gnotobiotic conditions

Summary Wheat seedlings were inoculated with rhizosphere nitrogen-fixing bacteria and grown gnotobiotically for 15 days. The growth medium consisted of semisolid agar with or without plant nutrients. The bacteria, isolated from roots of field-grown wheat, were three unidentified Gram-negative rods (A1, A2, E1), one Enterobacter agglomerans (C1) and two Bacillus polymyxa (B1, B2). A strain of Azospirillum brasilense (USA 10) was included for comparison.Nitrogenase activity (acetylene reduction activity, ARA) was tested on intact plants after 8 and 15 days of growth. In semisolid agar without plant nutrients, five isolates showed ARA of 0.01–0.9 nmol C₂H₄ plant^–1 h^–1, while the two strains of B. polymyxa had higher ARA of 3.3–10.6 nmol C₂H₄ plant^–1 h^–1.Plant development was not affected by inoculation with bacteria, except that inoculation with B. polymyxa resulted in shorter shoots and lower root weight.Transmission electronmicroscopy of roots revealed different degrees of infection. A. brasilense, A1 and A2, occurred mainly in the mucilage on the root surface and between outer epidermal cells (low infectivity). B. polymyxa strains and E1 were found in and between epidermal cells (intermediate infectivity) while E. agglomerans invaded the cortex and was occasionally found within the stele (high infectivity).     

Ammonium-excreting Azospirillum sp. become intracellularly established in maize (Zea mays) para-nodules

Maize seedlings develop nodule-like tumour knots (para-nodules) along primary roots when treated with the auxin 2,4-dichlor-phenoxy-acetic acid (2,4-D). Inoculated NH ₄ ⁺ -excreting Azospirillum brasilense cells were shown to colonize these tumours, mostly intracellularly, promoting a high level of N₂ fixation when microaerophilic conditions were imposed. The nitrogenase activity inside the para-nodules was less sensitive to free O₂ than in non-para-nodulating roots. Both light and electron microscopy showed a dense bacterial population inside intact tumour cells, with the major part of the cell infection along a central tumour tissue. The bacteria colonized the cytoplasm with a close attachment to inner cell membranes. In an auxin-free growth medium, young 2,4-D-induced para-nodules grew further to become mature differentiated root organs in which introduced bacteria survived with a stable population. These results provide evidence that gramineous plants are potentially able to create a symbiosis with diazotrophic bacteria in which the NH ₄ ⁺ -excreting symbiont will colonize para-nodule tissue intracellularly, thus becoming well protected.     

Effect of Azospirillum inoculants on arbuscular mycorrhiza establishment in wheat and maize plants

Plant growth-promoting rhizobacteria and arbuscular mycorrhizal (AM) fungi represent two main groups of beneficial microorganisms of the rhizosphere. The role of different strains of Azospirillum on AM fungi development was evaluated by measuring the percentage of AM colonisation of the root system in durum wheat and maize plants, grown under both greenhouse and field conditions. The effect of wild-type Azospirillum brasilense strain Sp245 and genetically modified (GM) derivatives overproducing indole-3-acetic acid was assessed at greenhouse level in (1) three different cultivars of durum wheat, in the presence of indigenous AM fungi and (2) maize plants artificially inoculated with Glomus mosseae and Glomus macrocarpum. In addition, the establishment of natural AM fungal symbiosis was evaluated using Azospirillum lipoferum CRT1 in maize plants at field level. Despite the stimulatory effect of the different Azospirillum inocula on root growth, no significant differences in AM colonisation were found, independently of the AM fungus involved, either in wheat or in maize plants. Similarly, GM A. brasilense, which strongly stimulates root development, did not affect AM formation. Although these results were obtained in conditions in which the mycorrhization rate was moderate (15–30%), overall considered they indicate that the use of wild-type or GM Azospirillum phytostimulators does not alter mycorrhization.     

Competition between inoculated and indigenous Rhizobium/Bradyrhizobium spp. strains for nodulation of grain and fodder legumes in Pakistan

Summary The competitive ability of inoculated and indigenous Rhizobium/Bradyrhizobium spp. to nodulate and fix N₂ in grain legumes (Glycine max, Vigna unguiculata, Phaseolus vulgaris) and fodder legumes (Vicia sativa, Medicago sativa, and Trifolium subterraneum) was studied in pots with two local soils collected from two different fields on the basis of cropping history. The native population was estimated by a most-probable-number plant infectivity test in growth pouches and culture tubes. The indigenous rhizobial/bradyrhizobial population ranged from 3 to 2×10⁴ and 0 to 4.4×10³ cells g^-1 in the two soils (the first with, the second without a history of legume cropping). Inoculated G. max, P. vulgaris, and T. subterraneum plants had significantly more nodules with a greater nodule mass than uninoculated plants, but N₂ fixation was increased only in G. max and P. vulgaris. A significant response to inoculation was observed in the grain legume P. vulgaris in the soil not previously used to grow legumes, even in the presence of higher indigenous population (>10³ cells g^-1 soil of Rhizobium leguminosarum bv phaseoli). No difference in yield was observed with the fodder legumes in response to inoculation, even with the indigenous Rhizobium sp. as low as <14 cells g^-1 soil and although the number and weight of nodules were significantly increased by the inoculation in T. subterraneum. Overall recovery of the inoculated strains was 38–100%, as determined by a fluorescent antibody technique. In general, the inoculation increased N₂ fixation only in 3 out of 12 legume species-soil combinations in the presence of an indigenous population of rhizobial/bradyrhizobial strains.     

Peculiar effect of Azospirillum inoculation on growth and nitrogen balance of winter wheat (Triticum aestivum)

Summary Wheat plants (Triticum aestivum) grown in pots and in the field under the Mediterranean climate of the south of France were inoculated with a strain of Azospirillum brasilense. Comparisons with non-inoculated plants grown under the same conditions showed significant responses to inoculation with an increase in the number of fertile tillers, shoot and root dry weight, and root to shoot biomass ratio. The roots of inoculated plants attracted relatively more assimilates than those of the control plants until a late stage of growth (heading stage) but the rhizosphere respiration expressed per unit of root growth was not increased by inoculation. Nitrogen yield, both total and in grains, was also enhanced; however, N percentages of all aerial parts of the plants grown in pots were always statistically lower after inoculation than in the control. At maturity, the N % in seeds was 1.81 and 2.45, respectively. The possible mechanisms of this effect of inoculation under the experimental conditions of this study are discussed.     

Inoculation of field-grown wheat (Triticum aestivum) with Azospirillum spp. in Brazil

Summary Three field experiments with wheat were conducted in 1983, 1984, and 1985 in Terra Roxa soil in Paraná, the major Brazilian wheat-growing region, to study inoculation effects of various strains of Azospirillum brasilense and A. amazonense. In all three experiments inoculation with A. brasilense Sp 245 isolated from surface-sterilized wheat roots in Paraná produced the highest plant dry weights and highest N% in plant tops and grain. Grain yield increases with this strain were up to 31 % but were not significant. The application of 60 or 100 kg N ha^–1 to the controls increased N accumulation and produced yields less than inoculation with this strain. Another A. brasilense strain from surface-sterilized wheat roots (Sp 107st) also produced increased N assimilation at the lower N fertilizer level but reduced dry weights at the high N level, while strain Sp 7 + Cd reduced dry weights and N% in the straw at both N levels. The A. amazonense strain isolated from washed roots and a nitrate reductase negative mutant of strain Sp 245 were ineffective. Strains Sp 245 and Sp 107st showed the best establishment within roots while strain Cd established only in the soil.     

Yield response of wheat and barley to inoculation of plant growth promoting rhizobacteria at various levels of nitrogen fertilization

The yield response of a wheat (Kirik) and a barley (Tokak 157/37) cultivar to inoculation with Azospirillum brasilense Sp246 and Bacillus sp. OSU‐142 was studied in relation to three levels of N fertilization (0, 40, and 80 kg ha^–1) under field conditions in Erzurum, Turkey, in 1999 and 2000. Seed inoculation with A. brasilense Sp246 significantly affected yield and yield components, both in wheat and barley. On average of years and N doses, inoculation with A. brasilense Sp246 increased spike number per m², grain number per spike, grain yield, and crude protein content by 7.2, 5.9, 14.7, and 4.1 % in wheat and by 6.6, 8.1, 17.5, and 5.1 % in barley, respectively, as compared to control. Inoculation with Bacillus sp. OSU‐142 significantly increased kernel number per spike in wheat, but no significant effect was determined in the other characteristics. Grain yields and yield components were also higher at all levels of nitrogen fertilizer in the inoculated plots as compared to the control. However, these increases diminished at high fertilizer levels. These results suggest that application of the growth promoting bacteria A. brasilense Sp246 may have the potential to be used as a biofertilizer for spring wheat and barley cultivation in organic and low‐N input agriculture.     

Effect of inoculation with different strains of Azospirillum brasilense on cotton (Gossipium barbadense)

Summary The response of the cotton plant to inoculation with six strains of Azospirillum brasilense was investigated under subtropical conditions in Egypt. Azospirilla populations and activities were increased as a result of root inoculation with liquid inoculum of Azospirillum sp. Highest C₂H₂ — reduction activities on roots were obtained with strains S631 and Sp Br 14 (means of 216.85 and 209.50 nmol C₂H₄g^–1 root h^–1 respectively) while strain M4 gave the lowest activity (mean of 100.8 nmol C₂H₄g^–1 root h^–1). Statistical analysis showed that Azospirillum strains 5631, Sp Br 14, E15 and SC22 significantly increased the plant dry weight and nitrogen uptake while inoculation with strains M4 and SE had no significant effect in that respect.     

Effect of plant growth-promoting bacteria and soil compaction on barley seedling growth, nutrient uptake, soil properties and rhizosphere microflora

Inoculants are of great importance in sustainable and/or organic agriculture. In the present study, plant growth of barley (Hordeum vulgare) has been studied in sterile soil inoculated with four plant growth-promoting bacteria and mineral fertilizers at three different soil bulk densities and in three harvests of plants. Three bacterial species were isolated from the rhizosphere of barley and wheat. These bacteria fixed N₂, dissolved P and significantly increased growth of barley seedlings. Available phosphate in soil was significantly increased by seed inoculation of Bacillus M-13 and Bacillus RC01. Total culturable bacteria, fungi and P-solubilizing bacteria count increased with time. Data suggest that seed inoculation of barley with Bacillus RC01, Bacillus RC02, Bacillus RC03 and Bacillus M-13 increased root weight by 16.7, 12.5, 8.9 and 12.5% as compared to the control (without bacteria inoculation and mineral fertilizers) and shoot weight by 34.7, 34.7, 28.6 and 32.7%, respectively. Bacterial inoculation gave increases of 20.3–25.7% over the control as compared with 18.9 and 35.1% total biomass weight increases by P and NP application. The concentration of N and P in soil was decreased by increasing soil compaction. In contrast to macronutrients, the concentration of Fe, Cu and Mn was lower in plants grown in the loosest soil. Soil compaction induced a limitation in root and shoot growth that was reflected by a decrease in the microbial population and activity. Our results show that bacterial population was stimulated by the decrease in soil bulk density. The results suggest that the N₂-fixing and P-solubilizing bacterial strains tested have a potential on plant growth activity of barley.     

Interactions between azospirillum and va mycorrhiza and their effects on growth and nutrition of maize and ryegrass

Interactions between the N₂-fixing bacterium Azospirillum brasilense and the mycorrhizal fungus Glomus mosseae were studied in relation to their effects on the growth and nutrition of Zea mays (C₄) and Lolium perenne (C₃) plants. Although roots from plants inoculated with Azospirillum exhibited C₂H₂ reduction activity no significant effect of inoculation on N concentration in the plant shoots was found. With non-mycorrhizal plants, inoculation with Azospirillum resulted in increased dry matter production at the first harvest compared to the effect achieved by supplying N as fertilizer, but this trend was reversed at the last harvest. However, with mycorrhizal maize plants, Azospirillum, which stimulated the development of VA mycorrhiza, was still effective in improving plant growth and nutrient uptake at the last harvest. Azospirillum and N behaved similarly in enhancing the growth and nutrition of mycorrhizal maize. The dual inoculation of maize by Azospirillum and Glomus produced plants of a similar size, N content, and a higher P content, than those supplied with N and P.