Yield response of various wheat genotypes to inoculation withAzospirillum brasilense

Summary Twenty different wheat genotypes representing a wide genetic variability, were tested for their yield and yield components response to inoculation withAzospirillum brasilense, at two levels of N fertilization. Only two cultivars responded by a significant yield increase of 7.4 and 8.0 per cent — both at the higher N level. The response reflected an increase in the number of grains per plant added as additional spikes. The importance of the host plant genotype for a successful wheat-bacteria association is discussed.     

Effect of inoculation ofAzospirillum spp. on nitrogen accumulation by field-grown wheat

Summary Two experiments were performed to examine the effects of inoculation of field grown wheat with various Azospirillum strains. In the first experiment the soil was sterilized with methyl bromide to reduce the Azospirillum population and¹⁵N labelled fertilizer was added to all treatments. Two strains ofAzospirillum brasilense isolated from surface sterilized wheat roots and theA. brasilense type strain Sp7 all produced similar increases in grain yield and N content. From the¹⁵N and acetylene reduction data it was apparent that these increases were not due to N₂ fixation. In the second experiment performed in the same (unsterilized) soil, twoA. brasilense strains (Sp245, Sp246) and oneA. amazonense strain (Am YTr), all isolated from wheat roots, produced responses of dry matter and N content while the response to the strain Sp7 was much smaller. These data confirm earlier results which indicate that if natural Azospirillum populations in the soil are high (the normal situation under Brazilian conditions), strains which are isolated from wheat roots are better able to produce inoculation responses than strains isolated from other sources. The inoculation of a nitrate reductase negative mutant of the strain Sp245 produced only a very small inoculation response in wheat. This suggests that the much greater inoculation response of the original strain was not due to N₂ fixation but to an increased nitrate assimilation due to the nitrate reductase activity of the bacteria in the roots. Consultant Inter-American Institute for Cooperation in Agriculture IICA/EMBRAPA World Bank Project.     

Associative effect ofAzospirilium brasilense withRhizobium japonicum on nodulation and yield of soybean (Glycine max)

Summary Azospirillum was associated with nodules of soybean. In general, seed inoculation with a broth culture ofAzospirillum brasilense alone significantly increased nodulation and grain yield of soybean grown in pots in unsterilized soil with different levels of urea ranging from 0 to 80 kg N/ha. This trend was significantly reproducible in a second experiment when a carrier based inoculant of the bacterium was used for seed inoculation.Inoculation withRhizobium japonicum andA. brasilense in combination generally increased grain yield in both the experiments, although the data were not significant.     

The response of maize (Zea mays) to Azospirillum inoculation in various types of soils in the field

Application of a peat-based powder inoculant of Azospirillum brasilense, as well as a granular inoculant (each containing 0.5–1.0×10⁷ Azospirillum/g moist peat), in the seed furrows of Zea mays resulted in significantly increased yields (11 to 14%) in light soils at low rates of N fertilization. In general, there was no effect of inoculation on plant yields in heavier soils nor when N fertilization was high. Pre-emergence application of granular inoculant and inoculation associated with irrigation were more efficient in increasing yield than inoculation post-emergence or seed coating.E. Fallik is with ARO-The Volcani Centre, Department of Postharvest Science of Fresh Produce, Bet-Dagan 50250, Israel. Y. Okon is with The Hebrew University of Jerusalem, Faculty of Agriculture, Department of Plant Pathology and Microbiology, Rehovot 76100, Israel     

Use ofAzospirillum brasilense as biofertilizer in intensive wheat cropping

Summary Three field experiments were conducted on ten cultivars of winterwheat and four cultivars of springwheat to estimate the growth promoting effect ofAzospirillum brasilense under varying nitrogen doses. Independent of cultivar selection or nitrogen dose a highly significant yield increase could be observed in winterwheat: strains S631 and SpBr14 increased the average grain yield with 9.14% and 14.82% respectively. When the yield components were studied a coinciding increase in ear density could be demonstrated of resp. 10.57% and 13.55%. Less significant results were obtained with springwheat although in one experiment strain SpBr14 significantly increased grain yield. As with winterwheat tillering of the plant was markedly affected by inoculation with both strains. In a companion greenhouse experiment it was found that inoculation with Azospirillum can cause a decrease in the root mass of wheatplants except when strain SpBr14 is used. Therefore it is suggested that the presence of a higher tillering together with an undisturbed nutrient uptake capacity can result in yield increases after inoculation withAzospirillum brasilense.     

Enhancement of Wheat Root Colonization and Plant Development by Azospirillum brasilense Cd. Following Temporary Depression of Rhizosphere Microflora

Inoculation of wheat with Azospirillum brasilense, combined with the application of four fungal and bacterium-inhibiting substances to which A. brasilense is resistant in the soil, decreased the rhizosphere population, while it increased wheat root colonization by A. brasilense, even in cases of poor inoculation. The inoculation significantly increased the following wheat plant parameters as well: plant dry weight, number of tillers per plant, spikelet fertility, harvest index, and grain yield. This model may provide a new approach to improve control of root colonization by beneficial bacteria.     

Growth promoting effect of Azospirillum brasilense on Casuarina cunninghamiana Miq. seedlings

The growth of Casuarina cunninghamiana seedlings was stimulated when inoculated with Azospirillum brasilense. This resulted in a higher biomass production than in uninoculated controls in the presence or absence of a non-nodulating strain of Frankia.Increase in whole plant dry weight was due to a significant increase in both shoot and root biomass, which corresponded with a higher total N content of the plants inoculated with Azospirillum. No such effects were observed under inoculation with a non-nodulating Frankia strain. These results suggest that the growth-promoting substances provided by A. brasilense may have enhanced the growth of Casuarina seedlings.     

Improvement of growth,under field conditions,of wheat inoculated with <Emphasis Type="Italic">Pseudomonas chlororaphis</Emphasis> subsp.<Emphasis Type="Italic"> aurantiaca</Emphasis> SR1

Effects of inoculation of wheat (Triticum aestivum L.) with the rhizobacterium Pseudomonas chlororaphis subsp. aurantiaca strain SR1 (termed SR1) were studied at an experimental field site in Río Cuarto, Argentina. Treatments involved SR1 inoculation with or without nitrogen/phosphorus fertilization. Inoculation produced a significant increase in plant height and root length in early growth stages. Inoculation plus fertilization with 40 kg ha⁻¹ urea/30 kg ha⁻¹ diamonic phosphate (“50% dose”) gave a yield increase of 636 kg ha⁻¹ relative to control, and an increase of 472 kg ha⁻¹ relative to fertilization with 80 kg ha⁻¹ urea/60 kg ha⁻¹ phosphate without inoculation. SR1 inoculation without fertilization, compared to control, produced increases of 6% in weight of 1,000 grains, 13% in number of spikes per plant, and 30% in number of grains per spike. Inoculation plus 50% dose fertilization also improved these parameters. Results of the study indicate that inoculation of wheat with SR1 improves various growth and yield parameters, and allows reduced dosage of nitrogen/phosphorus fertilizers in the field.     

Restoration of Growth of Durum Wheat (Triticum durum var. waha) Under Saline Conditions Due to Inoculation with the Rhizosphere Bacterium Azospirillum brasilense NH and Extracts of the Marine Alga Ulva lactuca

Inoculation with the rhizosphere bacterium Azospirillum brasilense NH, originally isolated from salt-affected soil in northern Algeria, greatly enhanced growth of durum wheat (Triticum durum var. waha) under saline soil conditions. Important plant parameters like the rate of germination, stem height, spike length, dry weight of roots and shoots, chlorophyll a and b content, proline and total sugar contents, 1000-seed weight, seed number per spike, and weight of seeds per spike were measured. At salt stress conditions (160 and 200 mM NaCl) A. brasilense NH restored almost completely vegetative growth and seed production. The combination with extracts of the marine alga Ulva lactuca resulted in even more improved salt tolerance of durum wheat. Proline and total sugar accumulation, a sign of physiological plant stress under inhibitory salt conditions, was reduced in plants inoculated with A. brasilense NH with and without addition of algal extracts. Inoculation with the salt-sensitive A. brasilense strain Sp7 could not restore salt-affected plant growth at 200 mM NaCl. Furthermore, it could be demonstrated by fluorescence in situ hybridization and confocal laser scanning microscopy that A. brasilense NH is able to colonize roots of durum wheat endophytically under salt-stressed conditions. Thus, the salt-tolerant rhizobacterium A. brasilense NH could effectively provide alone or in combination with extracts of U. lactuca a promising solution to overcome salt inhibition which is a major threat hindering productive wheat cultivation in arid saline soils.     

Selection and evaluation ofAzospirillum brasilense strains growing at a sub-optimum temperature in rhizocoenosis with wheat

FourteenAzospirillum brasilense strains growing at a sub-optimum temperature were selected based on their ability to grow and carry out plant growth promoting activities at 22°C. The strains were tested for their response to inoculation in wheat (two popular cultivars, HD2285 and WH547, under sterile conditions) crop using sterile and nonsterile rooting medium. Significant increase in plant growth parameters was observed: the overall response to inoculation was better in cultivar HD 2285. Based on their performance under sterile conditions, 4 strains were selected and compared under nonsterile conditions with strain sensitive to a sub-optimum temperature in pots using wheat variety HD2285. The strains capable of growing at the sub-optimum temperature can colonize the wheat endorhizosphere efficiently and improve the plant growth and yield as compared to sensitive strain; a 25–27% increase in grain yield was found on inoculating two selected strains compared to NO ₃ ^- control.     

Seed inoculation withAzospirillum brasilense andAzotobacter chroococcum and the root biomass of rice (Oryza sativa L.)

Summary The root biomass of rice seedlings increased due to inoculation withAzospirillum brasilense andAzotobacter chroococcum singly or in combination. In general, the increase in the biomass of roots was better in unsterilized soil than in sterilized one with or without inorganic nitrogen in the form of urea.     

Field inoculation of sorghum and rice withAzospirillum spp. andHerbaspirillum seropedicae

Two field experiments were carried out at the UAPNPBS experimental station, Seropédica, with two sorghum and one rice cultivars. The establishment, and inoculation effects, ofAzospirillum spp. andHerbaspirillum strains marked with antibiotic resistance were investigated. One grain sorghum (BR 300) and one sugar sorghum (Br 505) cultivar were used.Azospirillum lipoferum strain S82 (isolated from surface sterilized roots of sorghum) established in both cultivars and comprised 40 to 80% of theAzospirillum spp. population in roots and stems 60 days after plant emergence (DAE).Azospirillum amazonense strain AmS91 (isolated from surface-sterilized roots of sorghum) reached only 50%. At 90 DAE, S82 almost disappeared (less than 30% of establishment) while the establishment of AmS91 remained constant in roots and stems. No establishment ofH. seropedicae strain H25 (isolated from surface-sterilized roots of sorghum) orA. lipoferum strain S65 (isolated from the root surface of sorghum) could be observed on inoculated roots. Inoculation with S82, AmS91 or S65 but not withH. seropedicae H25, increased plant dry weight of both cultivars and total N in grain of the grain sorghum. In rice,A. lipoferum Al 121 andA. brasilense Sp 245 (isolated from surface sterilized rice and wheat roots respectively) established in the roots but there was no increase inAzospirillum spp. numbers due to inoculation. None of the strains affected plant growth or rice grain yield.Azospirillum amazonense, A82 andH. seropedicae Z95, which did not establish in roots, significantly enhanced seed germination.     

Soybean biofertilization in sandy soils of Egypt using15N tracer technique

The effect of inoculating soybean plants withBradyrhizobium japonicum andAzospirillum brasilense either solely or in mixture, and of using different N fertilizer levels was studied in pot experiments. The nodulation of soybean grown in a sandy soil was enhanced by the inoculation while the highest nodule numbers and fresh mass, recorded at a N-fertilizer dose of 20 kg N/hm², decreased when the fertilizer quantity increased to 40 kg N/hm². By contrast, the dry mass of above-ground parts and the N uptake was increased with increasing N fertilizer level. A similar effect was observed for inoculation as compared with the uninoculated variant. Although the nodulating and non-nodulating soybeans has nearly the same dry mass, the nodulating isoline accumulated more N than the non-nodulating. The percentage of nitrogen derived from air (%, Ndfa) and estimated by isotope dilution (ID) or N difference method (DM) dropped with increasing N level from 10 to 40 kg N/hm². Dual inoculation resulted in a high per cent of fixed N₂ (42.5%) at 10 kg N/hm². Correlation between the ID and DM methods was found to be dependent on inoculation treatments. The amount of nitrogen utilized by nodulating soybean (FUE %) was enhanced as a function of inoculation withB. japonicum.     

Impact of seed-applied fungicide and insecticide on Azospirillum brasilense survival and wheat growth-promoting ability

The use of Azospirillum brasilense as a crop inoculant has increased in recent years. Thus, the compatibility of the inoculation technology with seed treatments using pesticides needs to be evaluated. In this study, we evaluated the effect of an insecticide and fungicide formulation on A. brasilense strain FP2 population by culturing and culture-independent approaches. In addition, we evaluated the impact of these pesticides on the ability of A. brasilense to promote plant growth by monitoring biometric traits (root and shoot dry mass and length) of wheat grown in Greenhouse conditions. Seed pesticide dressings, mainly fungicide, led to a significant mortality of A. brasilense over time. The ability of A. brasilense to promote wheat growth also decreased due to pesticide treatments combined with sowing delay. Considering that pesticides confer fitness advantages to the wheat in field condition, our results suggest that sowing within the first 4 h after inoculation maintain the beneficial effects of A. brasilense on wheat growth promotion. Furthermore, we conclude that inoculation and treatment of seeds with pesticides may be compatible techniques when carried out immediately before sowing.     

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.     

Biofertilizers and/or some micronutrients role on wheat plants grown on newly reclaimed soil

Pot experiment was conducted to study the effect of biofertilizers (inoculation with different bacterial isolates), foliar spraying with some micronutrients (Mn, Zn, Fe and Mn+Zn+Fe) and their interaction on growth, physiological parameters and nutrients content of wheat plants grown on reclaimed soil. Pot experiment was conducted in the greenhouse of National Research center, The experimental design was split plot with four replicates. Four biofertilizer treatments (un‐inoculated, Bacillus polymyxa, Azotobacter chroococcum or Azosprillium barasilense) were used and randomly distributed in the main pots. The foliar treatments with micronutrients were randomly distributed in the sub plots. The growth parameters (plant height, leaf area, roots, shoots and whole plant dry weights and number of tillers & leaves per plant); some physiological parameters (soluble sugar %, protein %, polysaccharide %, chl. A+b μg cm^?1 leaf per plant, carotenoids μg g^?1, IAA mg kg^?1 and psll mol DCPIP reduced per mg chl. per h) and nutrient contents (N, P, K, Mg, Mn, Zn and Cu) of wheat plants were significantly increased by inoculating wheat grains with different bacteria as compared with un‐inoculated plants (control). The highest values of all the mentioned parameters were obtained by using Azospirillum brasilense followed by Azotobacter chroococcum and Bacillus polymyxa in decreasing order. Foliar spraying treatments significantly increased the growth parameters, physiological parameters as well as nutrients content of wheat plants as compared with control. Highest values were obtained by using (Mn+Fe+Zn) treatment followed by Zn, Fe and Mn in decreasing order. Micronutrients in wheat plants differed as the foliar treatments were differed, so application of any micronutrient individually significantly increased its content and enhanced the content of other micronutrients in wheat. Interaction between the used biofertilizers and foliar spraying with micronutrients significantly affected all the studied parameters of wheat plants, the highest were obtained by inoculating wheat grains with Azospirillum brasilense and spraying the plants with (Mn+Fe+Zn) treatment, while the lowest values were attained by un‐inoculated grains (control) and spraying the wheat plants with tap water (control). Effective microorganisms in combination with micronutrients could be recommended to farmers to lead higher wheat yield.     

The effect of aluminium on phosphate uptake by three isolated ectomycorrhizal fungi

Three wheat cultivars with different tolerances against free aluminium were grown monoxenically in association with Azospirillum brasilense. In situ nitrogen fixation, measured with the acetylene reduction assay, was higher by the aluminium-tolerant cultivars than by the sensitive cultivar. The transfer of fixed nitrogen to the host plant, determined by the ¹⁵N dilution technique, was also significantly higher in the aluminium-resistant wheat plants. The total accumulation of fixed nitrogen in the host plants due to an A. brasilense inoculation varied from approximately 13% to 17% of the total nitrogen in the root and 2.9% to 3.9% of the nitrogen in the shoot.The quantity and quality of exudates released in liquid nutrient solution were analysed separately for two of the wheat cultivars, one aluminium-tolerant and one aluminium-sensitive. After 29 days of growth the aluminium-tolerant plants exudated significantly higher total amounts of carbon than aluminium-sensitive plants. No differences between the two cultivars existed in the carbon exudation rate per gram dry root.Much higher concentrations of low molecular dicarboxylic acids i.e. succinic, malic and oxalic acid, were found in the exudates of aluminium-tolerant plants. Dicarboxylic acids are potential chelating compounds for positively charged metals such as aluminium and they may play an important role in protecting the plant against aluminium incorporation. They are also very suitable substrates for Azospirillum spp. It is therefore suggested that these factors may be causing the higher associative nitrogen fixation rates which was found in the aluminium-tolerant wheat cultivars.     

Soil and litter respiration rates in different microhabitats of a mixed oak-conifer forest and their control by edaphic conditions and substrate quality

Summary Seven isolates belonged toA. brasilense and 3 belonged toA. lipoferum. Isolates having more denitrifying capacity fixed less nitrogen in nitrogen free semi-solid malate medium. One strain ofA. lipoferum having high nitrogen fixing capacity with negative test for denitrification was tested as inoculant to supplement the nitrogen need of a wheat crop in field condition with different doses of N with and without the inoculant. While control without nitrogen yielded 1260 kg/ha the yield in inoculated treatment was 2070 kg/ha resulting in significant increase. In a treatment receiving 40 kg N/ha the grain yield was 2370 kg/ha as against yield of 3110 kg/ha in a similar treatment receiving fertiliser plus inoculant. Thus increase in yield was about 30%. Further the treatment receiving 80 kg N/ha yielded 2970 kg/ha as against yield of 4150 kg/ha in a treatment receiving inoculant alongwith the above dose of the fertiliser. Thus increase in yield due to application of inoculant was about 36%. Similarly, the uptake of N in different treatments was augmented due to inoculation of seeds with the culture.     

Nutrition of sorghum plants fertilized with nitrogen or inoculated withAzospirillum brasilense

Summary Sorghum plants were inoculated withAzospirillum brasilense or received an N-amended nutrient solution. Azospirillum inoculation increased plant dry weight and nitrogen assimilation by 25%. Most plant growth responses to Azospirillum were comparable to application of 2.0 mM N. Increased scavenging of nutrients, altered root permeability or nitrogen fixation are possible explanations for these effects.This work was supported by the United States Department of Agriculture, Agricultural Research Service (CRIS No. 5102-20170-001) in collaboration with the University of California, Berkeley. Requests for reqrints to G. J. Bethlenfalvay.     

Response ofSetaria italica to inoculation withAzospirillum brasilense as compared toAzotobacter chroococcum

Summary Under controlled conditions in pots filled with sand, vermiculite and field soil, inoculation withA. brasilense-Cd ATCC 29729 or withAzotobacter chroococcum caused increases above controls in the weight and N content of panicles ofSetaria italica. In no case, however, did N increases in test plants exceed the initial total N content. High acetylene reduction activities (1,000–2,000 nmole/h/pot) could be found only in plants inoculated withAzospirillum. Inoculation withAzospirillum (strain-Cd) in the field caused a significant increase above noninoculated controls of 18.5% in shoot dry weight, ofSetaria italica. Azotobacter caused a non significant increase of 8%. No significant differences were found between yields ofSetaria italica grown in soil inoculated withA. chroococcum, and those of plants grown in the presence ofA. vinelandii. A. brasilense-Cd was more effective in the field thanA. brasilense Sp-7 ATCC 29145. The results suggest that Azospirillum may increase yields ofS. italica more efficiently than Azotobacter under local field conditions.