Isolation and characterization of plant growth promoting endophytic diazotrophic bacteria from Korean rice cultivars

We have isolated 576 endophytic bacteria from the leaves, stems, and roots of 10 rice cultivars and identified 12 of them as diazotrophic bacteria using a specific primer set of nif gene. Through 16S rDNA sequence analysis, nifH genes were confirmed in the two species of Penibacillus, three species of Microbacterium, three Bacillus species, and four species of Klebsiella. Rice seeds treated with these plant growth-promoting bacteria (PGPB) showed improved plant growth, increased height and dry weight and antagonistic effects against fungal pathogens. In addition, auxin and siderophore producing ability, and phosphate solubilizing activity were studied for the possible mechanisms of plant growth promotion. Among 12 isolates tested, 10 strains have shown higher auxin producing activity, 6 isolates were confirmed as strains with high siderophore producing activity while 4 isolates turned out to have high phosphate-solubilizing activity. These results strongly suggest that the endophytic diazotrophic bacteria characterized in this study could be successfully used to promote plant growth and inducing fungal resistance in plants.     

Beneficial native bacteria improve survival and mycorrhization of desert truffle mycorrhizal plants in nursery conditions

Sixty-four native bacterial colonies were isolated from mycorrhizal roots of Helianthemum almeriense colonized by Terfezia claveryi, mycorrhizosphere soil, and peridium of T. claveryi to evaluate their effect on mycorrhizal plant production. Based on the phylogenetic analysis of the 16S rDNA partial sequence, 45 different strains from 17 genera were gathered. The largest genera were Pseudomonas (40.8 % of the isolated strains), Bacillus (12.2 % of isolated strains), and Varivorax (8.2 % of isolated strains). All the bacteria were characterized phenotypically and by their plant growth-promoting rhizobacteria (PGPR) traits (auxin and siderophore production, phosphate solubilization, and ACC deaminase activity). Only bacterial combinations with several PGPR traits or Pseudomonas sp. strain 5, which presents three different PGPR traits, had a positive effect on plant survival and growth. Particularly relevant were the bacterial treatments involving auxin release, which significantly increased the root-shoot ratio and mycorrhizal colonization. Moreover, Pseudomonas mandelii strain 29 was able to considerably increase mycorrhizal colonization but not plant growth, and could be considered as mycorrhiza-helper bacteria. Therefore, the mycorrhizal roots, mycorrhizosphere soil, and peridium of desert truffles are environments enriched in bacteria which may be used to increase the survival and mycorrhization in the desert truffle plant production system at a semi-industrial scale.     

Genetic diversity and plant growth promoting traits of diazotrophic bacteria isolated from two Pennisetum purpureum Schum. genotypes grown in the field

Background and aims

Some elephant grass (Pennisetum purpureum) genotypes are able to produce large amounts of biomass and accumulate N derived from BNF when growing in soil with low N levels. However, information about the diazotrophic bacteria colonizing this C4 plant is still very scarce. This study aimed to characterize the plant growth promoting traits of a fraction of culturable diazotrophs colonizing the genotypes CNPGL F06-3 and Cameroon.

Methods

A total of 204 isolates were obtained from surface sterilized leaves, stems and roots after culturing on five different N-free semisolid media. These were then analyzed by BOX-PCR, and the 16S rRNA and nifH sequences of representative isolates were obtained. The functional ability of the isolates to reduce acetylene, produce indole and to solubilize phosphate was also determined.

Results

The diazotrophic bacterial population varied from 10² up to 10⁶ bacteria g^?1 fresh tissues of both genotypes. The BOX-PCR analysis suggested a trend in the genetic diversity among the 204 diazotrophic strains colonizing the different genotypes and plant tissues. Sequencing of 16S rRNA fragments confirmed the presence of Azospirillum brasilense and Gluconacetobacter diazotrophicus and revealed for the first time the occurrence of G. liquefaciens, G. sacchari, Burkholderia silvatlantica, Klebsiella sp., Enterobacter cloacae and E. oryzae in elephant grass. Interestingly, several nifH sequences from isolates identified as G. liquefaciens and G. sacchari showed homologies with nifH sequences of Enterobacter species. The majority of the isolates (97%) produced indole compounds, 22% solubilized phosphate and 6.4% possessed both characteristics.

Conclusions

The results showed the occurrence of novel diazotrophic bacterial species colonizing different tissues of both genotypes of elephant grass. In addition, the study revealed the presence of several bacteria with growth promoting traits, and highlighted their potential to be exploited as biofertilizers.     

Identification of interior salt-tolerant bacteria from ice plant <Emphasis Type="Italic">Mesembryanthemum crystallinum</Emphasis> and evaluation of their promoting effects

Endophytic bacteria can stimulate host plant development. Insufficient information is available about NaCl-tolerant bacteria that colonize ice plants (Mesembryanthemum crystallinum) in their habitats. In this study, a culture-dependent method was used to isolate endophytic nitrogen-fixing bacteria from ice plants, and the resulting cultures were screened for salt-stress tolerance in vitro. A total of 17 salt-tolerant bacteria were obtained. The majority of the isolates grew well in 2.05 M NaCl with a maximum tolerance at 3.59 M. Most of the strains were Gram-positive bacteria with various plant growth-promoting traits. The 16S rRNA gene sequences revealed that the 17 isolates were distributed within three genera and corresponded to the bacterial species Halomonas sp., Bacillus sp., and Planococcus sp. Inoculation of cabbage (Brassica olereacea) seeds with selected strains showed that the strain MC1 promoted seed germination, and the same strain significantly increased root dry weight under saline stress by 24.5%. Our study suggests that ice plants naturally accommodate a variety of salt-tolerant endophytic bacteria and that these bacteria are able to relieve abiotic stress during plant growth.     

三株固氮类芽孢杆菌的特点及其对中国青菜的产量和土壤酶活性的影响

【目的】本研究分析三株固氮菌PGPR性状特征及其对中国青菜产量和土壤酶活的影响。【方法】氮(N)-修复(固氮)细菌被认为是一种能够促进植物生长和增产的施氮方式。在本研究中,我们用无氮培养基分离出了30株根际固氮细菌:11株来自小麦根际,16株来自中国青菜根际和3株来自莲花根际。基于16S r DNA序列分析,对小麦、中国青菜和莲花等植物根际中属于类芽孢杆菌属的主要固氮细菌进行研究。【结果】本研究从这30株固氮菌中筛选出三株属于类芽孢杆菌属(Paenibacillus)的细菌,分别命名为P-4、W-7和L-3,它们的固氮酶活性不但高于对照组(圆褐固氮菌),而且可以有效抑制两种或三种植物病原菌的生长,即核盘菌(Sclerotinia sclerotiorum)、玉蜀黍赤霉(Gibberella zeae)和棉花黄萎病菌(Verticillium dahliae)。菌株W-7还具有溶解难溶磷的能力,中国青菜在接种菌株W-7和L-3后,其鲜重显著增加,同时改变了田间土壤蔗糖酶、磷酸酶和过氧化氢酶的活性;而接种了菌株P-4对植物的生长和酶活性没有显著的影响。【结论】土壤蔗糖酶、磷酸酶和过氧化氢酶活性与中国青菜的生物量呈正相关。同时,菌株W-7和L-3具有促进植物产量和提高土壤质量的良好潜力。     

Characterization of native fluorescent Pseudomonas isolates associated with alfalfa roots in Uruguayan agroecosystems

Establishment of alfalfa crops is continuously threatened by seedling diseases caused by soilborne pathogens. The use of plant beneficial bacteria as inoculants is a feasible and environmentally friendly means to control soil pathogens. Identifying effective plant growth-promoting strains to use on local crops under local environmental conditions requires the screening of large collections of native isolates. A collection of 738 rhizospheric fluorescent Pseudomonas isolates was obtained from alfalfa plants from three agroecological regions representative of Uruguayan agricultural systems. The isolates were evaluated for in vitro pathogen inhibition, biosurfactant production, phosphate solubilization and the presence of genes involved in antibiotic synthesis. Isolates with strong in vitro antagonistic activity toward Pythium debaryanum were more abundant in alfalfa plants established in a previously natural ecosystem while biosurfactant producers were less abundant in that location. A subset of isolates was selected for genotypic characterization by rep-PCR using BOX primers. Twenty-four genotypes were defined, sixteen from a single geographical origin and eight composed of isolates from multiple origins. Genotypic profiles correlated well with phenotypic traits. A subset of isolates was assayed to determine their ability to protect alfalfa against P. debaryanum damping-off and to promote plant growth. Five native Pseudomonas isolates showed significant effects on alfalfa by increasing plant biomass and/or protecting from pathogen infection. Plant growth promoting isolates from each location were genotypically similar. Our work contributes to the knowledge of the phenotypic and genotypic diversity of rhizospheric fluorescent pseudomonads of forage legumes and the frequency of plant growth promoting traits associated with this group of bacteria in different agricultural systems.     

Diversity of sporadic symbionts and nonsymbiotic endophytic bacteria isolated from nodules of woody, shrub, and food legumes in Ethiopia

Fifty-five bacterial isolates were obtained from surface-sterilized nodules of woody and shrub legumes growing in Ethiopia: Crotalaria spp., Indigofera spp., and Erythrina brucei, and the food legumes soybean and common bean. Based on partial 16S rRNA gene sequence analysis, the majority of the isolates were identified as Gram-negative bacteria belonging to the genera Achromobacter, Agrobacterium, Burkholderia, Cronobacter, Enterobacter, Mesorhizobium, Novosphingobium, Pantoea, Pseudomonas, Rahnella, Rhizobium, Serratia, and Variovorax. Seven isolates were Gram-positive bacteria belonging to the genera Bacillus, Paenibacillus, Planomicrobium, and Rhodococcus. Amplified fragment length polymorphism (AFLP) fingerprinting showed that each strain was genetically distinct. According to phylogenetic analysis of recA, glnII, rpoB, and 16S rRNA gene sequences, Rhizobium, Mesorhizobium, and Agrobacterium were further classified into six different genospecies: Agrobacterium spp., Agrobacterium radiobacter, Rhizobium sp., Rhizobium phaseoli, Mesorhizobium sp., and putative new Rhizobium species. The strains from R. phaseoli, Rhizobium sp. IAR30, and Mesorhizobium sp. ERR6 induced nodules on their host plants. The other strains did not form nodules on their original host. Nine endophytic bacterial strains representing seven genera, Agrobacterium, Burkholderia, Paenibacillus, Pantoea, Pseudomonas, Rhizobium, and Serratia, were found to colonize nodules of Crotalaria incana and common bean on co-inoculation with symbiotic rhizobia. Four endophytic Rhizobium and two Agrobacterium strains had identical nifH gene sequences with symbiotic Rhizobium strains, suggesting horizontal gene transfer. Most symbiotic and nonsymbiotic endophytic bacteria showed plant growth-promoting properties in vitro, which indicate their potential role in the promotion of plant growth when colonizing plant roots and the rhizosphere.     

Molecular diversity of nitrogen-fixing bacteria associated with Chrysopogon zizanioides (L.) Roberty (vetiver), an essential oil producer plant

The essential oil produced by vetiver can vary in amount and composition depending on the bacterial community associated with its roots. Some of these bacteria could also promote plant growth by fixing nitrogen. This study aimed to analyze the diversity of diazotrophic bacteria tightly associated with roots of different vetiver genotypes. nifH-based PCR-denaturing gradient gel electrophoresis (DGGE) and clone libraries were used. DGGE profiles obtained from bulk and rhizosphere soils and root DNA amplified with nifH primers showed that samples from rhizosphere soil and root were separated at 68% similarity. Twelve bands were excised from the DGGE and sequenced. High similarity with nifH sequences of Bradyrhizobium sp., Pseudacidovorax sp. and Xanthobacter sp. was observed. Moreover, three nifH clone libraries were generated using polF/polR-primers from root DNA samples obtained from vetiver genotypes UFS-VET001, UFS-VET003 and UFS-VET004. In UFS-VET001, 24.2% of 95 clones were affiliated with sequences of Mesorhizobium loti while in UFS-VET003 41.5% of 89 clones were affiliated with Sphingomonas azotifigens, and in UFS-VET004 36.4% of 85 clones were affiliated with Klebsiella pneumoniae. The data obtained can be used to guide the isolation of diazotrophic bacteria, which may contribute to plant growth promotion and improvement of the production of essential oil in vetiver.     

Salt-tolerant and plant growth-promoting bacteria isolated from Zn/Cd contaminated soil: identification and effect on rice under saline conditions

The bacteria of PDMCd0501, PDMCd2007, and PDMZnCd2003 were isolated from a Zn/Cd contaminated soil. They were classified as salt-tolerant bacteria in this experiment. The bacteria had indole-3-acetic acids (IAA) production, nitrogen fixation, and phosphate solubilization, under 8% (w/v) NaCl condition. Biochemical test (API 20E) and 16S rDNA sequencing identified PDMCd2007 and PDMCd0501 as Serratia sp. and PDMZnCd2003 was Pseudomonas sp. The effect of Pseudomonas sp. PDMZnCd2003 on the germination and seedlings of Oryza sativa L.cv. RD6 was determined under a salinity of 0–16 dS/m. The salinity levels of 4–16 dS/m affected to decrease germination and seedlings of rice. Comparison between uninoculated and inoculated system, however, Pseudomonas sp. PDMZnCd2003 had a negative impact on the rice growth. This unexpected effect was a case that should be concerned and studied further before application as a plant growth-promoting bacteria (PGPB).     

The bacterial microflora of witloof chicory (Cichorium intybus L. var.foliosum Hegi) leaves

The bacterial flora on the heads of four different witloof chicory varieties was examined. The 590 isolates were characterized by their SDS-PAGE protein profiles; they revealed 149 different protein fingerprint types. The fluorescentPseudomonas fingerprint type CH001 was abundantly found on all heads examined. Fourteen other fingerprint types occurred in high densities more than twice. Among these, the following were identified: fluorescentPseudomonas, nonfluorescentPseudomonas sp.,Erwinia herbicola, Erwinia sp., andFlavobacterium sp. The majority of the fingerprint types (90%) was found only once. It was also our objective to isolate bacteria applicable in the biological control of chicory phytopathogens. Isolates of all fingerprint types were tested for in vitro antagonistic activity and for possible deleterious effect on plant growth. FluorescentPseudomonas andSerratia liquefaciens isolates were antagonistic against fungi. Among the 161 fluorescentPseudomonas strains, five were able to produce disease symptoms on chicory leaves upon inoculation. Comparison of the results of this study with those obtained in two previous analyses revealed that the leaf microflora showed some similarities with the bacterial flora of chicory roots. The chicory seed microflora differed from that of both leaves and roots.     

The Nitrogen-Fixation Island Insertion Site Is Conserved in Diazotrophic Pseudomonas stutzeri and Pseudomonas sp. Isolated from Distal and Close Geographical Regions

The presence of nitrogen fixers within the genus Pseudomonas has been established and so far most isolated strains are phylogenetically affiliated to Pseudomonas stutzeri. A gene ortholog neighborhood analysis of the nitrogen fixation island (NFI) in four diazotrophic P. stutzeri strains and Pseudomonas azotifigens revealed that all are flanked by genes coding for cobalamin synthase (cobS) and glutathione peroxidise (gshP). The putative NFIs lack all the features characterizing a mobilizable genomic island. Nevertheless, bioinformatic analysis P. stutzeri DSM 4166 NFI demonstrated the presence of short inverted and/or direct repeats within both flanking regions. The other P. stutzeri strains carry only one set of repeats. The genetic diversity of eleven diazotrophic Pseudomonas isolates was also investigated. Multilocus sequence typing grouped nine isolates along with P. stutzeri and two isolates are grouped in a separate clade. A Rep-PCR fingerprinting analysis grouped the eleven isolates into four distinct genotypes. We also provided evidence that the putative NFI in our diazotrophic Pseudomonas isolates is flanked by cobS and gshP genes. Furthermore, we demonstrated that the putative NFI of Pseudomonas sp. Gr65 is flanked by inverted repeats identical to those found in P. stutzeri DSM 4166 and while the other P. stutzeri isolates harbor the repeats located in the intergenic region between cobS and glutaredoxin genes as in the case of P. stutzeri A1501. Taken together these data suggest that all putative NFIs of diazotrophic Pseudomonas isolates are anchored in an intergenic region between cobS and gshP genes and their flanking regions are designated by distinct repeats patterns. Moreover, the presence of almost identical NFIs in diazotrophic Pseudomonas strains isolated from distal geographical locations around the world suggested that this horizontal gene transfer event may have taken place early in the evolution.     

Structural and functional diversity of rhizobacteria associated with Rauwolfia spp. across the Western Ghat regions of Karnataka, India

The present study carried out with denaturing gradient gel electrophoresis of DNA extracted from rhizosphere soils of Rauwolfia spp. collected from Western Ghat (WG) regions of Karnataka indicated that Pseudomonas sp. was prevalently found followed by Methylobacterium sp., Bacillus sp. and uncultured bacteria. A total of 200 rhizobacteria were isolated from 58 rhizosphere soil samples comprising of 15 different bacterial genera. The Shannon Weaver diversity index (H′) and Simpson’s diversity index (D) were found to be 2.57 and 0.91 for cultivable bacteria, respectively. The total species richness of cultivable rhizobacteria was high in Coorg district comprising 15 bacterial genera while in Mysore district, four bacterial genera were recorded. Rarefaction curve analysis also indicated the presence of higher species richness in samples of Shimoga and Coorg. All the rhizobacteria were screened for their multiple plant growth promotion and disease suppression traits. The results revealed that 70 % of the isolates colonized tomato roots, 42 % produced indole acetic acid, 55 % solubilized phosphorus, while 43, 22, 27, 19, 40, 15 and 44 % produced siderophore, salicylic acid, hydrogen cyanide, chitinase, phytase, cellulase and protease, respectively. Rhizobacterial isolates showing antagonistic activity against Fusarium oxysporum and Aspergillus flavus were 53 and 33 %, respectively. Plant growth promotion studies revealed that most of the isolates increased percent germination with significantly higher vigour index as compared to untreated control. Most predominant rhizobacteria found in the rhizospheres of Rauwolfia spp. of WG regions are potential PGPR which can serve as biofertilizers and biopesticides.     

Biological Control of Chickpea Collar Rot by Co-inoculation of Antagonistic Bacteria and Compatible Rhizobia

Two hundred and seven bacteria were isolated from composts and macrofauna and screened for plant growth promoting and antagonistic traits. Seven of the 207 isolates showed antagonistic activity against Sclerotium rolfsii in plate culture. Inhibition of S. rolfsii by the bacterial isolates ranged between 61 and 84%. Two of the seven isolates were Bacillus sp. and rest belonged to Pseudomonas sp. Two isolates, Pseudomonas sp. CDB 35 and Pseudomonas sp. BWB 21 was compatible with chickpea Rhizobium sp. IC 59 and IC 76 in plate culture conditions. Increase in plant biomass (dry weight) ranged between 18 and 30% on application of these bacteria by seed coating and seed priming methods. However, by seed-priming there was an increase in plant biomass by 5–7% compared to seed coating. Number of nodules and the nodule weight was similar by both seed coating and seed priming methods. Disease incidence was reduced up to 47% in treatments where captan (fungicide) or antagonistic Pseudomonas sp. CDB 35 was applied. Increase in shoot weight was 36% by seed coating with Rhizobium sp. IC 59 and Pseudomonas sp. CDB 35 when compared to captan application. Whereas by seed priming with IC 59 and CDB 35 increased shoot weight by 3 and 39% increase in nodulation was observed.     

Bacterial Biosynthesis of 1-Aminocyclopropane-1-Carboxylate (ACC) Deaminase and Indole-3-Acetic Acid (IAA) as Endophytic Preferential Selection Traits by Rice Plant Seedlings

In this study, bacteria were isolated from the rhizosphere and inside the roots and nodules of berseem clover plants grown in the field in Iran. Two hundred isolates were obtained from the rhizosphere (120 isolates), interior roots (57 isolates), and nodules (23 isolates) of clover plants grown in rotation with rice plants. Production of chitinase, pectinase, cellulase, siderophore, salicylic acid, hydrogen cyanide, indole acetic acid (IAA), 1-aminocyclopropane-1-carboxylate (ACC) deaminase, solubilization of phosphate, antifungal activity against various rice plant pathogen fungi, N₂ fixation, and colonization assay on rice seedlings by these strains was evaluated and compared (endophytic isolates vs. rhizosphere bacteria). The results showed both the number and the ability of plant growth-promoting (PGP) traits were different between endophytic and rhizosphere isolates. A higher percentage of endophytic isolates were positive for production of IAA, ACC deaminase, and siderophore than rhizosphere isolates. Therefore, it is suggested that clover plant may shape its own associated microbial community and act as filters for endophyte communities, and rhizosphere isolates with different (PGP) traits. We also studied the PGP effect of the most promising endophytic and rhizosphere isolates on rice seedlings. A significant relationship among IAA and ACC deaminase production, the size of root colonization, and plant growth (root elongation) in comparison with siderophore production and phosphate solubilization for the isolates was observed. The best bacterial isolates (one endophytic isolate and one rhizosphere isolate), based on their ability to promote rice growth and colonize rice roots, were identified. Based on 16S rDNA sequence analysis, the endophytic isolate CEN7 and the rhizosphere isolate CEN8 were closely related to Pseudomonas putida and Pseudomonas fluorescens, respectively. It seems that PGP trait production (such as IAA, ACC deaminase) may be required for endophytic and rhizosphere competence as compared to other PGP traits in rice seedlings under constant flooded conditions. The study also shows that the presence of diverse rhizobacteria with effective growth-promoting traits associated with clover plants may be used for sustainable crop management under field conditions.     

The phenotypic,phylogenetic and symbiotic characterization of rhizobia nodulating Lotus sp. in Tunisian arid soils

Fifteen bacterial isolates, representatives of different 16S rRNA-RFLP genomogroups which were isolated from root nodules of Lotus creticus and L. pusillus growing in the arid areas of Tunisia were characterized by phenotypic features and 16S rDNA sequences. Phenotypically, all isolates are fast growers with the ability to grow at a pH between 5.5 and 9. Most of the tested isolates tolerate NaCl concentrations from 1.39 to 3.48 %. Phylogenetically, the studied isolates are affiliated into the genera: Sinorhizobium (5 strains), Rhizobium (2 strains), and Mesorhizobium (4 strains). The 16S rDNA sequences of Tunisian Lotus sp. nodule isolates: LAC7511, LAC733, and Mesorhizobium alhagi (Alhagi sparsifolia symbiont) shared 100 % identical nucleotides similar to the 16S rDNA sequences of LAC831, LAC814 and Mesorhizobium temperatum CCNWSX0012-2 (Astragalus adsurgens symbiont). Non-nodulating bacteria, considered as endophytes of Lotus sp. nodules, were also found in our studies and they were classified into the genera: Phyllobacterium (2 strains), Starkeya (1 strain) and Pseudomonas (1 strain). Except for these four endophytic Lotus sp. bacteria, all other strains under investigation induce nodules on Lotus sp., but they differ in the number of induced root nodules and the effectiveness of atmospheric nitrogen fixation. The Sinorhizobium sp., Mesohizobium sp. and Lotus sp. nodule isolates, forming the most effective symbiosis with the plant host, are potential candidates for inoculants in revegetation programs.     

Root bacteria from nematicidal plants and their biocontrol potential against trichodorid nematodes in potato

Trichodorid nematodes (Nematoda: Trichodoridae) are vectors of tobacco rattle virus (TRV), one of the causal agents of spraing disease in potato. Root bacteria from nematicidal plants and their control potential against Trichodoridae were the focus of this study. Bacteria isolated from the roots of 12 nematicidal plants and potato were characterized for their production of hydrolytic enzymes, hydrogen cyanide, phenol oxidation ability and antifungal activity towards the potato pathogen Rhizoctonia solani. Based on these functional traits, bacteria isolates were selected and tested in greenhouse conditions on potato (cv. Saturna) for their effect on plant growth, and screened for nematicidal activity against Paratrichodorus pachydermus and Trichodorus primitivus in naturally infested soil. Sixteen bacteria isolates out of 44 reduced nematode densities by 50–100%. Nine selected isolated were further tested by bacterizing potato tubers (cv. King Edward) which were planted in a trichodorid and TRV-infested soil. Four bacterial isolates consistently reduced nematode densities (by 56.7–74.4%) with no visual negative effect on plant growth. These isolates were tentatively identified, partly by fatty acid methyl ester (FAME) analysis as: Stenotrophomonas maltophilia, Bacillus mycoides, Pseudomonas sp., and one unidentified bacterium. The isolates originated from potato, Plantago major, Thymus vulgaris and Asparagus officinalis, respectively. Two Pseudomonas isolates obtained from Zinnia elegans and selected for their strong nematicidal activity in soil screening tests, did not reduce the nematode population when tested on potato. It is concluded that plants releasing nematicidal compounds may harbour nematode-antagonistic bacteria as well.     

Plant growth-promoting activities of fluorescent pseudomonads,isolated from the Iranian soils

Fluorescent pseudomonads are among the most influencing plant growth-promoting rhizobacteria in plants rhizosphere. In this research work the plant growth-promoting activities of 40 different strains of Pseudomonas fluorescens and Pseudomonas putida, previously isolated from the rhizosphere of wheat (Triticum aestivum L.) and canola (Brassica napus L.) and maintained in the microbial collection of Soil and Water Research Institute, Tehran, Iran, were evaluated. The ability of bacteria to produce auxin and siderophores and utilizing P sources with little solubility was determined. Four strains of Wp1 (P. putida), Cfp10 (Pseudomonas sp.), Wp150 (P. putida), and Wp159 (P. putida) were able to grow in the DF medium with ACC. Thirty percent of bacterial isolates from canola rhizosphere and 33% of bacterial isolates from wheat rhizosphere were able to produce HCN. The results indicate that most of the bacteria, tested in the experiment, have plant growth-promoting activities. This is the first time that such PGPR species are isolated from the Iranian soils. With respect to their great biological capacities they can be used for wheat and canola inoculation in different parts of the world, which is of very important agricultural implications.     

Improved germination efficiency of Salicornia ramosissima seeds inoculated with Bacillus aryabhattai SP1016‐20

Saline agriculture and the crop cultivation of halophytes represent an alternative for the reclamation of salinized soils and for the management of irrigation water. Halotolerant plant growth promoting bacteria with biocontrol effect, as an alternative to commercial fungicides, may contribute to improve crop productivity while mitigating saline stress effects. The objective of this work was to isolate autochthonous rhizobacteria with biocontrol features, to be used as germination enhancers and plant‐growth promoting agents in the crop cultivation of Salicornia ramosissima. A set of isolates obtained from the rhizosphere of S. ramosissima was characterised in terms of Gram, motility, salt tolerance and biocontrol traits (hydrogen cyanide production, antifungal activity and production of extracellular lipases and proteases). One Gram‐positive motile isolate that tested positive for all biocontrol traits was identified by 16S rRNA gene sequencing as Bacillus aryabhattai. The inoculation of S. ramosissima seeds with B. aryabhattai SP1016‐20 reduced the negative effect of salinity on the germination efficiency. At the highest tested salinity (30 g/L NaCl) the final germination efficiency of inoculated seeds doubled in relation to non‐inoculated controls. Although the mechanisms involved in the biocontrol effect were not defined in the current work, the results highlight the potential of Bacillus aryabhattai SP1016‐20 as a plant‐growth promoting agent for the crop cultivation of Salicornia and contribute to the strengthening of the scientific basis of biosaline agriculture and plant growth promoting rhizobacteria‐assisted crop cultivation of halophytes in saline soils and estuarine sediments.     

The contribution of nitrogen fixation to sugarcane (Saccharum officinarum L.), and the identification and characterization of part of the associated diazotrophic bacterial community

Background and aims

Rhizospheric, epiphytic and endophytic bacteria are associated with several non-legumes, colonizing their surface and inner tissues. Many of these bacteria are beneficial to their hosts, and are collectively termed plant growth-promoting rhizobacteria (PGPR). Recent interest has focused particularly upon PGPR that are endophytic (i.e. PGPE), and which have been reported to be associated with important crops such as rice, wheat and sugarcane. Different mechanisms are involved in bacteria-induced plant growth promotion (PGP), including biological nitrogen fixation (BNF), mineral solubilization, production of phytohormones and pathogen biocontrol. In Uruguay, sugarcane (Saccharum officinarum L.) is considered a strategic multipurpose crop, used for bioenergy, feed, sugar and bioethanol production. The aim of this work was to estimate the BNF contribution to Uruguayan sugarcane cultivars, as well as to identify and characterize the (culturable) putatively endophytic diazotrophic bacteria associated with these varieties.

Methods and results

Results using the ¹⁵N-dilution technique have shown that these sugarcane varieties obtain significant inputs of N from BNF (34.8–58.8% Ndfa). In parallel, a collection of 598 isolates of potentially endophytic diazotrophs was obtained from surface-sterilized stems using standard isolation techniques, and nifH ⁺ isolates from these were the subject of further studies. The bacteria were shown to belong to several genera, including Pseudomonas, Stenotrophomonas, Xanthomonas, Acinetobacter, Rhanella, Enterobacter, Pantoea, Shinella, Agrobacterium and Achromobacter. Additionally, some PGP features were studied in 35 selected isolates. The data obtained in this study represent the initial steps in a program aimed at determining the mechanisms of PGP of non-legume crops in Uruguay (such as sugarcane) with potentially beneficial plant-associated bacteria.     

Deciphering Diversity of Salt-Tolerant Bacilli from Saline Soils of Eastern Indo-gangetic Plains of India

The intensive use of chemical fertilizers, monoculture and irrigation with surface saline water has resulted in the deterioration of soil health by enhancing the level of salinity in the Eastern Indo-Gangetic Plains of India. Therefore, diversity of halotolerant bacteria adapted to that environment and possessed the ability to produce plant growth hormones was explored, that could be used for salt stress amelioration. The 16S rRNA gene sequencing and fatty acid methyl ester (FAME) were used for diversity analysis of salt-tolerant bacilli. Among the 95 isolates, 55 strains showed plant growth promotion traits, production of industrially important enzymes (amylase, protease and cellulase) and tolerance to more than 4% NaCl. Using partial 16S rRNA sequences and FAME comparisons, 21 different species of Bacillus and Bacillus-derived genera were identified, viz. Bacillus megaterium, B. subtilis, B. licheniformis, B. firmus, B. horikoshii, B. pumilus, Bacillus sp., B. safensis, B. thuringiensis, B. simplex, B. agri, B. flexus, B. oceanisediminis, B. cereus, B. arsenicus, Paenibacillus dendritiformis, Lysinibacillus sp., L. sphaericus, B. marisflavi, Terribacillus sp., and B. mycoides. These isolates possess the ability to tolerate high salt, form endospores, withstand harsh environments, and also have the potential for plant growth promotion, which could be useful in formulation of new inoculants to enhance the availability of nutrients for crop growth under saline conditions.