Isolation,partial identification and application of diazotrophic rhizobacteria from traditional Indian rice cultivars

A diversity of N₂-fixing (diazotrophic) bacteria was isolated from two traditional rice cultivars, Sataria and Kartiki, from the rice growing area of Mithila region of North Bihar, India, where low levels of nitrogen fertilizers are applied. Nitrogen-free semisolid media NFb, JMV and LGI with different carbon sources and pH-values were used for enrichment and isolation of root-associated diazotrophs. The colonization density of roots by diazotrophs, as estimated from positive pellicle formation at highest dilution in nitrogen-free enrichment media, was 10⁶–10⁸ diazotrophic bacteria per g fresh root weight. Roots of the cultivar Kartiki were found to be more densely colonized endophytically by diazotrophs as detected after chloramine T (1%) surface disinfection. To ascertain the phylogenetic affiliation of the isolates, phylogenetic oligonucleotide probes and the Fluorescent in situ Hybridization (FISH) technique were applied. Using group-specific rRNA directed oligonucleotide probes, the majority of the isolates could be identified as alpha-, beta-, or gamma-proteobacteria. Using 16S and 23S rRNA-directed genus- or species-specific probes, Herbaspirillum seropedicae, Azospirillum amazonense, Burkholderia cepacia/vietnamiensis, Rhizobia and Pseudomonas spp. were found to be the most prominent root associated culturable diazotrophs. Diazotrophic Gluconacetobacter spp. were also demonstrated as colonizers of rice roots. Burkholderia cenocepacia, Pseudomonas sp. and three diazotrophic PGPR reference strains were used for the inoculation of axenically grown rice seedlings to determine the plant growth promoting potential. Significant increases in the shoot length (up to 60%), shoot dry weight (up to 33%) and the grain yield (up to 26%) per plant were observed in non-axenic pot and field trials. Using semisolid enrichment media after surface sterilization of field grown inoculated rice roots and oligonucleotide probing of the diazotrophic enrichment cultures, a sustainable colonization with the inoculated bacteria could be demonstrated.     

Interactions of Glomus clarum with Acetobacter diazotrophicus in infection of sweet potato (Ipomoea batatas), sugarcane (Saccharum spp.), and sweet sorghum (Sorghum vulgare)

Summary Spores of the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus clarum obtained from sweet potatoes grown in soil inoculated with this fungus and with an enrichment culture of Acetobacter diazotrophicus contained A. diazotrophicus and several other bacteria, including a diazotrophic Klebsiella sp. Inoculation of micropropagated sweet potatoes with G. clarum and A. diazotrophicus enhanced spore formation in soil compared to VAM inoculation alone. Plants inoculated with VAM spores containing the bacteria showed additional increases in the number of spores formed within roots. A. diazotrophicus infected aerial plant parts only when inoculated together with VAM or when present within VAM spores. Micropropagated sugarcane seedlings inoculated with the same VAM spores containing the diazotrophs also contained much higher numbers of A. diazotrophicus in aerial parts than seedlings inoculated in vitro with the bacteria alone. When grown in non-sterile soil, the sugarcane seedlings again showed the greatest infection of aerial parts after inoculation with VAM spores containing the diazotrophs. This treatment also increased VAM colonization and the numbers of spores formed within roots. Similar effects were observed in sweet sorghum except that the aerial plant parts were not infected by A. diazotrophicus.     

Infection and colonization of aseptically micropropagated sugarcane seedlings by nitrogen-fixing endophytic bacterium, Herbaspirillum sp. B501gfp1

In this study, we used Herbaspirillum sp. B501gfp1 (B501gfp1), an isolate from wild rice, to investigate the interaction between a non-host nitrogen-fixing endophytic bacterium and micropropagated sugarcane plants under aseptic condition. Two Japanese sugarcane plants (Saccharum sp.) cultivars (cvs) NiF8 and Ni15 were inoculated using B501gfp1 in two inoculum doses of 10⁸ and 10² bacterial-cells-per-milliliter suspension. The results showed that bacterial cells colonized both the root and stem tissues, and colonization was apparent in the intercellular spaces. Higher bacterial numbers were detected in plant tissues inoculated with the higher inoculum concentration treatment. Bacterial numbers also varied between the two cultivars, with the higher values determined in cv Ni15. This study provides evidence that Herbaspirillum sp. B501gfp1, a rice isolate, could colonize sugarcane tissues, suggesting non-specificity of host plant among endophytes.     

Significance of timing and level of inoculation with rhizosphere bacteria on wheat plants

The importance of time of inoculation and bacterial concentration in the inoculum on the response of wheat plants was evaluated, using eight strains of rhizosphere bacteria. The optimal bacterial concentration, for all strains, was 10⁵–10⁶ colony forming units ml⁻¹. Plant response was highest when seeds had been inoculated but was less when seedlings were inoculated. Successive inoculations somewhat increased plant response. Early inoculations resulted in an increased colonization of plant roots at later stages of growth. It was concluded that time of inoculation and the concentration of bacteria in the inoculum were of significant importance in plant response to inoculation and they may govern the inconsistency found in inoculation experiments using beneficial bacteria.     

Rhizosphere and endophytic bacteria for induction of systemic resistance of banana plantlets against bunchy top virus

Bunchy top is one of the most important viral diseases affecting banana production worldwide. Several approaches have been made to control or combat the bunchy top virus disease in banana, transmitted by the banana aphid, Pentalonia nigronervosa Coq. No tactic is able to completely protect the plants against this viral infection. To ameliorate this problem, an attempt was made to use different bacterial strains, which could be inoculated to the roots of micropropagated plantlets to avoid the post transplanting problems. Virus indexed micropropagated plantlets of banana cv. Virupakshi (AAB) were subjected to root colonization followed by foliar spraying with three bacterial strains viz., Pseudomonas fluorescens strains Pf1, CHA0 and Bacillus subtilis strain EPB22 along with an un-inoculated control during primary and secondary hardening stage in the nursery and at the time of transplanting, 3rd, 5th and 7th month after planting in the field. Microbe inoculated plantlets showed improved vegetative growth, physiological attributes, PR—proteins and phenol contents besides reducing banana bunchy top disease (BBTD) incidence in the field. These results indicate that, transplant mixes amended with beneficial bacterial strains can enhance growth of banana plants, besides reducing the damage caused by banana bunchy top virus.     

Field inoculation effectiveness of native and exotic arbuscular mycorrhizal fungi in a Mediterranean agricultural soil

In sustainable agriculture, arbuscular mycorrhizal (AM) fungal inoculation in agronomical management might be very important, especially when the efficiency of native inocula is poor. Here, we assessed the effect of native and exotic selected AM fungal inocula on plant growth and nutrient uptake in a low input Trifolium alexandrinum-Zea mays crop rotation. We evaluated the effects of four exotic AM fungal isolates on T. alexandrinum physiological traits in greenhouse. Then, the field performances of T. alexandrinum inoculated with the exotic AMF, both single and mixed, were compared to those obtained with a native inoculum, using a multivariate analysis approach. Finally, we tested the residual effect of AM fungal field inoculation on maize as following crop. Multivariate analysis showed that the field AM fungal inoculation increased T. alexandrinum and Z. mays productivity and quality and that the native inoculum was as effective as, or more effective than, exotic AM fungal isolates. Moreover, the beneficial effects of AMF were persistent until the second year after inoculation. The use of native AMF, produced on farm with mycotrophic plants species, may represent a convenient alternative to commercial AM fungal inocula, and may offer economically and ecologically important advantages in sustainable or organic cropping systems.     

Influence of organic fertilization on the number of culturable diazotrophic endophytic bacteria isolated from sugarcane

The numbers of culturable diazotrophic endophytic bacteria (CDEB) from roots, stems and leaves of sugarcane submitted to organic, inorganic or no fertilization were compared. In order to determine the size of the N₂ fixing populations, the Most Probable Number technique (MPN) was used. The quantification of diazotrophic bacteria by using the acetylene reduction assay (ARA) was more accurate than observing the bacterial growth in the vials; to confirm N₂ fixing capability, the detection of gene nifH was performed on a sample of 105 isolated bacteria. The production of extracellular enzymes involved in the penetration of the plants by the bacteria was also studied. The results showed that organic fertilization enhances the number of CDEB when compared with conventional fertilization used throughout the growing season. The maximum number of bacteria was detected in the roots. Roots and stems presented the greatest number of CDEB in the middle of the cropping season and in leaves numbers varied according to the treatment. Using two pairs of primers and two different methods, the nifH gene was found in 104 of the 105 tested isolates. Larger amounts of pectinase were released by isolates from sugarcane treated with conventional fertilizers (66%), whereas larger amounts of cellulase were released by strains isolated from sugarcane treated with organic fertilizers (80%).     

Response of traditional upland rice varieties to inoculation with selected diazotrophic bacteria isolated from rice cropped at the Northeast region of Brazil

The largest numbers of the Brazilian traditional upland rice varieties are found in the Maranhão state, Northeast region of Brazil. However, no information is available on the diazotrophic bacterial population associated as well as the plant growth promoting potential when these traditional genotypes are inoculated with native strains. Here, we evaluated the response of ten traditional rice varieties to inoculation with ten diazotrophic strains, previously isolated from rice soil of this region and screened for their ability to produce indole-3-acetic acid (IAA) in vitro. The procedure for selection of the best diazotrophic strain/rice variety interaction involved three steps: gnotobiotic conditions, soil pot and field experiments. The gnotobiotic experiment showed that the Azospirillum amazonense strain AR3122 increased the biomass of the traditional varieties Cana Roxa and Cana Forte (28 and 48%, respectively) while this effect was less evident for the other combination of strains/rice varieties. The soil pot experiment showed that the combination of Burkholderia vietnamiensis strain AR 1122 and traditional variety Arroz 70 was superior to the other strains/varieties and the treatment fertilized with 100 kg N ha⁻¹. The best performance of the Burkholderia vietnamiensis strain AR1122/variety Arroz 70 was confirmed in the field experiment. There was an increase of up 10 and 29% in the grain yield in comparison to both the N fertilization and Herbaspirillum seropedicae ZAE 94 strain treatments, respectively. In contrast, the response of the commercial variety Bonança to inoculation with strain AR1122 was much lower, suggesting that a biofertilizer inoculation program for traditional rice varieties should consider the genetic interaction between strain and rice variety. The diazotrophic B. vietmaniensis strain AR1122 was a good biofertilizer candidate for inoculation of traditional rice varieties and therefore should be used for further studies to confirm the strain-genotype effect envisaging a sustainable rice crop system mainly in the Northeast region of Brazil.     

四种AM真菌接种剂的田间效应及其分子检测研究

采用灭菌土壤生产了 4种AM真菌接种剂。在盆栽条件下测试了接种剂的质量 ,结果显示 ,4种接种剂促进玉米生长效果明显 ,地上部分生物量均显著高于对照 (p <0 .0 1 ) ;以MPN试验检测了接种剂的侵染能力 ,结果表明每克接种剂中真菌的繁殖体数在 95～ 1 4 0 0之间。将AM真菌的预接种技术和农业生产上的营养钵育苗技术相结合 ,进行了玉米的田间试验 ,结果显示 ,玉米根系的AM真菌感染率早期增长较快 ,然后趋于平稳 ;AM真菌接种剂A(Glomusconstrictum)、C (Glomus三种菌混合 )和D (G .intraradices)对玉米籽粒产量有显著的增产效果 (p <0 .0 5 ) ;玉米籽粒的淀粉含量和磷含量也高于对照。运用特异性分子探针和nest ed PCR技术 ,从田间接种AM真菌Glomusintraradices和G .mosseae的玉米根样中粗提DNA进行特异性扩增 ,成功地从感染根段中检测到特定的接种AM真菌。本工作从分子水平为评价高效AM真菌的应用潜力、研究AM真菌之间及其与其他微生物之间的相互关系奠定了基础。     

Effects of inoculation with ectomycorrhizal fungi on microbial biomass and bacterial functional diversity in the rhizosphere of Pinus tabulaeformis seedlings

This study investigated the effects of inoculation with three individual ectomycorrhizal (ECM) fungal species on soil microbial biomass carbon and indigenous bacterial community functional diversity in the rhizosphere of Chinese pine (Pinus tabulaeformis Carr.) seedlings under field experimental conditions. The results showed that ECM fungal inoculation significantly increased the ectomycorrhizal colonization compared with non-inoculated seedlings. ECM fungal inoculations have higher soil microbial biomass carbon than that of control, ranging from 49.6 μg C g^?1 dry soil in control to 134.02 μg C g^?1 dry soil in treatment inoculated with Boletus luridus Schaeff ex Fr. Multivariate analyses (PCA) of BIOLOG data revealed that the application of ECM fungi significantly influenced bacterial functional diversity in the rhizosphere of P. tabulaeformis seedlings. The highest average well-color development (AWCD) and functional diversity indices were also observed in treatment inoculated with B. luridus. A wider range of sole carbon sources were utilized by the bacterial community in the rhizosphere of inoculated seedlings. The data gathered from this study provides important information for utilization of ECM fungi in forest restoration project in the Northwestern China. The present study will also significantly broaden our understanding of practical importance in the application of ECM fungal inoculum to promote soil microbial community diversity of soil.     

接种密度对植物生长及烃降解的影响

The combined use of plants and bacteria is a promising approach for the remediation of soil contaminated with organic pollutants. Different biotic and abiotic factors can affect the survival and activity of the applied bacteria and consequently plant growth and phytoremediation efficiency. The effect of inoculum density on the abundance and expression of alkane-degrading genes in the rhizosphere of plant vegetated in hydrocarbon-contaminated soil has been rarely observed. In this study, an alkane-degrading bacterium (Pantoea sp. strain BTRH79), at different inoculum densities (10⁵ to 10⁸ cells cm^-3 soil), was inoculated to ryegrass (Lolium perenne) vegetated in diesel-contaminated soil to find the optimum inoculum density needed for its efficient colonization and hydrocarbon degradation activity. Bacterial inoculation improved plant growth and hydrocarbon degradation. Maximum plant growth and hydrocarbon degradation wwereobserved with the inoculum having the highest cell density (10⁸ cells cm^-3 soil). Moreover, the inoculum with higher cell density exhibited more abundance and expression of alkane hydroxylase gene, CYP153. This study suggests that the inoculum density is one of the main factors that can affect bacterial colonization and activity during phytoremediation.     

Endophytic nitrogen-fixing Klebsiella variicola strain DX120E promotes sugarcane growth

Klebsiella variicola bacteria are found in association with plants. Little is known about their colonization patterns, roles, and mechanisms during association with the plant hosts. Here, we identified a nitrogen-fixing bacterium, DX120E, which was isolated from surface-sterilized roots of the ROC22 sugarcane cultivar, as K. variicola by phylogenetic analyses of its 16S rRNA gene, RNA polymerase β-subunit gene, and DNA gyrase subunit A gene sequences. gfp-tagged DX120E was found to colonize at the roots and aerial parts of micropropagated sugarcane plantlets by fluorescence microscopy and confocal microscopy. DX120E was able to survive in soils and colonize in root epidermal cells, intercellular spaces in root cortices, and leaf mesophyll and vascular tissues. DX120E preferentially colonized at root maturation and elongation zones and entered roots via cracks at the emergence site of lateral roots and at disrupted epidermis. DX120E may penetrate root epidermal cells with the aid of their cellulose-degrading enzymes. ¹⁵N isotope dilution assay demonstrated that DX120E was able to fix N₂ in association with ROC22 sugarcane plants under gnotobiotic condition. DX120E was also able to promote GT21 cultivar growth and plant uptake of N, P, and K under greenhouse condition. Together, this study for the first time shows that a K. variicola strain is able to colonize in its sugarcane plant hosts, to fix N₂ in association with plants, and to promote plant growth.     

The role of the earthworm Lumbricus terrestris in the transport of bacterial inocula through soil

Two laboratory experiments were used to investigate the effect of the earthworm Lumbricus terrestris on transport of genetically marked Pseudomonas fluorescens inocula through soil microcosms. The microcosms comprised cylindrical cores of repacked soil with or without earthworms. Late log-phase cells of P. fluorescens, chromosomally marked with lux genes encoding bioluminescence, were applied to the surface of soil cores as inoculated filter paper discs. In one experiment, 5 and 10 days after inoculation, cores were destructively harvested to determine concentrations of marked pseudomonads with depth relative to the initial inoculum applied. Transport of the bacteria occurred only in the presence of earthworms. In a second experiment cores were subjected to simulated rainfall events 18 h after inoculation with lux-marked bacteria at 3-day intervals over a 24-day period. Resulting leachates were analysed for the appearance of the marked bacteria, and after 28 days cores were destructively harvested. Although some marked cells (less than 0.1% of the inoculum applied) were leached through soil in percolating water, particularly in the presence of earthworms, the most important effect of earthworms on cell transport was through burial of inoculated litter rather than an increase in bypass flow due to earthworm channels.     

Long-term agronomic practices alter the composition of asymbiotic diazotrophic bacterial community and their nitrogen fixation genes in an acidic red soil

The asymbiotic diazotrophic bacteria are important for nitrogen (N) input to soil. Here, we investigated asymbiotic diazotrophic bacteria in an acidic red soil from functional, phylogenetic, and ecological perspectives. We firstly confirmed that phosphorus (P) availability determines the overall asymbiotic N fixation potential in the red soil. Then, we analyzed the soil bacterial community and N fixing (nifH) gene composition. Long-term different fertilizations significantly affected the composition of soil bacterial community. In addition, long-term organic cultivations increased most of the asymbiotic diazotrophic bacteria and the corresponding nifH gene abundances. Few asymbiotic diazotrophic bacteria, belonging to Chloroflexaceae, Methylocystaceae, Enterobacteriaceae, and Pseudomonadaceae, and their corresponding nifH genes were more abundant in N and P co-limited than in not co-limited soils, suggesting that some bacterial taxa from these families might be activated under nutrient limited conditions. Our findings provided new information for the distribution of asymbiotic diazotrophic bacteria in red soil and gave insights into the ecology of diazotrophic bacteria.     

Azospirillum inoculation and nitrogen fertilization effect on grain yield and on the diversity of endophytic bacteria in the phyllosphere of rice rainfed crop

We assessed the Azospirillum inoculation and N-fertilization effect on grain yield and on the phyllosphere endophytic diversity of nitrogen-fixing bacteria in a rice rainfed crop. We used cultivation-based techniques and cultivation-independent methods involving PCR-16S rRNA and denaturing gradient gel electrophoresis (DGGE). In general, we observed that grain yield was improved when inoculated with Azospirillum (depending on the genotype) and/or fertilized with urea. A similar behavior was observed in total N-content in grain and the MPN determination, as the highest values occurred when seeds were inoculated with A. brasilense REC3 (S1) than with A brasilense 13-2C (S2). A positive nitrogenase activity and PCR-nifH amplification suggests that the bacteria associated to inner tissues of rice phyllosphere could have contributed to the different N-contents detected. The bacterial diversity, observed in the number and intensity of DGGE profiles, showed a higher number of bands when total DNA was obtained using only CTAB than with CTAB + PVP. The DGGE profiles revealed great stability in the dominating bands, which presumably represent numerically dominant species. Application of A. brasilense strains as inoculants did not influence the dominant members of the endophytic microbial communities in the phyllosphere, but improved N-content and production of rainfed rice crop.     

Arbuscular mycorrhizal root colonization and soil P availability are positively related to agrodiversity in Mexican maize polycultures

In Los Tuxtlas, Mexico, the local Popoluca people maintain the traditional management of their maize agroecosystems. However, it is not known whether the loss of agrodiversity over recent decades has affected mycorrhizal populations, nutrient availability, and crop productivity. This study utilized linear mixed effect models to analyze the relationship between agrodiversity (three, six, and greater than or equal to eight cultivated species) and (a) arbuscular mycorrhizal fungi (AMF) inoculum potential, measured as the most probable number (MPN) of propagules and colonization level, (b) nutrient availability, and (c) aboveground maize productivity. We also investigated the relationship between soil nutrient content and inoculum potential. Soil samples were taken before planting, and during flowering, in the 2009 maize cycle. We found that AMF colonization level of maize roots and P availability increased with planted species richness, but that this effect only occurred at the flowering sampling date. Plots with a higher MPN of propagules presented increased C and NO ₃ ^? contents and lower C/N ratio than those with lower MPN of propagules, regardless of agrodiversity. Soils that produced the highest maize root colonization level also featured high P availability and N content. We conclude that decreased agrodiversity in these traditional systems does not significantly affect the soil MPN of propagules, but may have a negative impact on the ability of the mycorrhizal community to colonize maize roots, as well as reducing the availability of P, which is often the most limiting nutrient in tropical soils.     

Tolerance of potentially diazotrophic bacteria to adverse environmental conditions and plant growth-promotion in sugarcane

Sugarcane is one of the crops responsible for the high consumption of fertilizers in Brazil. To minimize this demand a sustainable alternative is to increase the studies to evaluate the beneficial effects of the relationship between plants/microorganisms, mainly plant growth-promoting bacteria. The objectives of this study were: a) to evaluate potentially diazotrophic bacteria isolates from sugarcane in adverse environmental conditions; b) inoculate these bacteria in sugarcane to evaluate their ability in plant growth-promotion. The study was carried in the Northeast of Brazil. Bacterial genera Burkholderia sp., Pantoea sp., Stenotrophomonas sp. and Enterobacter sp. were evaluated in different culture medium and later the bacterial isolates were inoculated in sugarcane evaluating the plant growth-promotion. Stenotrophomonas sp. and Pantoea sp. were tolerant to salinity and to different sources of carbon in acid medium and too tolerated high concentrations of pesticides and produced quorum sensing molecules (QS), but did not increase the dry matter production of sugarcane. Burkholderia sp. and Enterobacter sp. were more sensitive to salinity and pest control, but were more effective in plant growth-promotion. The tolerance of the bacteria to the adverse environmental conditions interfered negatively in the ability to plant growth-promotion.     

Überleben inokulierter Rhizosphärenbakterien und deren Einfluß auf native Bakterienpopulationen im Wurzelraum von Luzerne

Survival of inoculated rhizosphere bacteria and their influence on native bacterial populations in the rhizosphere of alfalfa The survival of inoculated bacteria and their influence on native bacterial populations in the rhizosphere of alfalfa were investigated in a greenhouse experiment. The plant growth promoting strains Rhizobium meliloti me18 and Pseudomonas fluorescens PsIA12 were reisolated from the rhizosphere about 7 weeks after single and mixed strain inoculation. They did not induce lasting changes in the diversity of the native bacterial communities of the rhizosphere. Only within the first week after inoculation was an increase in total bacterial abundance observed. In general, the diversity of bacterial communities increased with plant age and with proximity to the root tip.     

Interactions of arbuscular-mycorrhizal fungi and Bacillus strains and their effects on plant growth,microbial rhizosphere activity (thymidine and leucine incorporation) and fungal biomass (ergosterol and chitin)

The effects of two Bacillus strains (Bacillus pumillus and B. licheniformis) on Medicago sativa plants were determined in single or dual inoculation with three arbuscular-mycorrhizal (AM) fungi and compared to P-fertilization. Shoot and root plant biomass, values of thymidine and leucine incorporation as well as ergosterol and chitin in rhizosphere soil were evaluated to estimate metabolic activity and fungal biomass, respectively, according to inoculation treatments. For most of the plant parameters determined, the effectiveness of AM fungal species was influenced by the bacterial strain associated. Dual inoculation of Bacillus spp. and AM fungi did not always significantly increase shoot biomass compared to single AM-colonized plants. The most efficient treatment in terms of dry matter production was the dual Glomus deserticola plus B. pumillus inoculation, which produced similar shoot biomass and longer roots than P-fertilization and a 715% (shoot) and 190% (root length) increase over uninoculated control. The mycorrhizas were more important for N use-efficiency than for P use-efficiency, which suggests a direct mycorrhizal effect on N nutrition not mediated by P uptake. Both chemical and biological treatments affected thymidine and leucine incorporation in the rhizosphere soil differently. Thymidine was greater in inoculated than in control rhizospheres and B. licheniformis was more effective than B. pumillus in increasing thymidine. Non-inoculated rhizospheres showed the lowest thymidine and leucine values, which shows that indigenous rhizosphere bacteria increased with introduced inocula. The highest thymidine and leucine values found in P-fertilized soils indicate that AM plants are better adapted to compete with saprophytic soil bacteria for nutrients than P-amended plants. Chitin was only increased by coinoculation of B. licheniformis and G. intraradices. B. pumillus increased ergosterol (indicative of active saprophyte fungal populations) in the rhizosphere of AM plants and particularly when colonized by G. mosseae. The different AM fungi have different effects on bacterial and/or fungal saprophytic populations and for each AM fungus, this effect was specifically stimulated or reduced by the same bacterium. This is an indication of ecological compatibilities between microorganisms. Particular Glomus–bacterium interactions (in terms of effect on plant growth responses or rhizosphere population) do not seem to be related to the percentage of AM colonization. The effect on plant growth and stimulation of rhizosphere populations, as a consequence of selected microbial groups, may be decisive for the plant establishment under limiting soil conditions.     

Diversity and plant growth promoting evaluation abilities of bacteria isolated from sugarcane cultivated in the South of Brazil

Plant growth promoting (PGP) bacteria associated with sugarcane are a promising alternative for the expansion of this crop in Southern Brazil. In this study bacterial strains from different sugarcane fields were isolated to estimate their diversity, to evaluate some of their PGP activities and to use them as inoculant strains in field experiment. Samples of rhizospheric soil, roots, and stems of sugarcane were collected in six Rio Grande do Sul localities. The isolation of bacteria was made in three different N-free media. DNA from each isolate was subjected to nifH or 16S rDNA PCR-RFLP, and to the 16S rDNA partial sequencing. Five hundred and sixteen strains were isolated and several PGP characteristics were analyzed. Shannon index was used to evaluate the bacterial diversity. Indexes varying from 0.94 to 2.46 were obtained. Soil pH and clay were the characteristics most closely related to bacterial diversity. Achromobacter, Agrobacterium, Burkholderia, Gluconacetobacter, and Stenotrophomonas were the most abundant genera. Concerning the PGP activities, indolic compounds production was detected in 368 isolates; 138 isolates were able to solubilize phosphate; and 390 were siderophores producers. The inoculation of sugarcane with Gluconacetobacter diazotrophicus VI27 strain showed a significant increase in the number of sets germinated, in the amount of soluble solids, and in the yield of sugarcane juice compared with the control. As a conclusion, a diverse population of PGP bacteria was found in the sugarcane samples. These bacteria, especially G. diazotrophicus strain VI27, could be used as biofertilizers of sugarcane as well as other cereal crops under controlled conditions to avoid or reduce the use of standard N fertilizers.