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Isolation and characterization of plant growth-promoting rhizobacteria from wheat rhizosphere and their effect on plant growth promotion.

ABSTRACT: The present study was conducted to characterize the native plant growth promoting (PGP) bacteria from wheat rhizosphere and root-endosphere in the Himalayan region of Rawalakot, Azad Jammu and Kashmir (AJK), Pakistan. Nine bacterial isolates were purified, screened in vitro for PGP characteristics and evaluated for their beneficial effects on the early growth of wheat (Triticum aestivum L.). Among nine bacterial isolates, seven were able to produce indole-3- acetic acid in tryptophan-supplemented medium; seven were nitrogen fixer, and four were able to solubilize inorganic phosphate in vitro. Four different morphotypes were genotypically identified based on IGS-RFLP fingerprinting and representative of each morphotype was identified by 16S rRNA gene sequencing analysis except Gram-positive putative Bacillus sp. Based on 16S rRNA gene sequence analysis, bacterial isolates AJK-3 and AJK-9 showing multiple PGP-traits were identified as Stenotrophomonas spp. while AJK-7 showed equal homologies to Acetobacter pasteurianus and Stenotrophomonas specie. Plant inoculation studies indicated that these Plant growth-promoting rhizobacteria (PGPR) strains provided a significant increase in shoot and root length, and shoot and root biomass. A significant increase in shoot N contents (up to 76%) and root N contents (up to 32%) was observed over the un-inoculated control. The study indicates the potential of these PGPR for inoculums production or biofertilizers for enhancing growth and nutrient content of wheat and other crops under field conditions. The study is the first report of wheat associated bacterial diversity in the Himalayan region of Rawalakot, AJK.

SUBMITTER: Majeed A

PROVIDER: S-EPMC4362341 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Project description:Plant growth-promoting rhizobacteria (PGPR) are a functionally diverse group of microbes having immense potential as biostimulants and biopesticides. We isolated four PGPR (designated n, L, K, and Y) that confer growth-promoting effects on Arabidopsis thaliana. The present study describes the detailed polyphasic characterization of these PGPR. Classical methods of bacterial identification and biochemical test kits (API20E, API20NE, API ZYM, and API 50CH) revealed their metabolic versatility. All rhizobacterial isolates were positive for 1-aminocyclopropane-1-carboxylate (ACC) deaminase (ACCD) and indole acetic acid production and phosphorous solubilization. PCR analysis confirmed the presence of the nifH gene in strains n, L, and Y, showing their N2-fixation potential. In vitro dual culture methods and bacterial infestation in planta demonstrated that strains n and L exerted antagonistic effects on Pseudomonas syringae pv. tomato DC3000 and Botrytis cinerea 191 and provided protection to Arabidopsis plants against both phytopathogens. Short- or long-term bacterial treatment revealed significant changes in transcript levels of genes annotated to stress response and hormone metabolism in A. thaliana. In particular, the expression of stress-responsive genes in A. thaliana showed an upregulation under salinity stress. MAP kinase 6 (MPK6) was involved in the growth promotion induced by the four bacterial strains. Furthermore, these strains caused a significant increase in root dry weight of maize seedlings under gnotobiotic conditions. We conclude that the four rhizobacteria are good candidates as biofertilizers for enhancing growth of maize, among which strains n and L showed marked plant growth-promoting attributes and the potential to be exploited as functional biostimulants and biopesticides for sustainable agriculture. IMPORTANCE There are pressing needs to reduce the use of agrochemicals, and PGPR are receiving increasing interest in plant growth promotion and disease protection. This study follows up our previous report that the four newly isolated rhizobacteria promote the growth of Arabidopsis thaliana. We test the hypothesis that they have multiple PGP traits and that they can be used as biofertilizers and biopesticides. In vitro assays indicated that these four strains have various PGP properties related to nutrient availability, stress resistance, and/or pest organism antagonism. They significantly influenced the transcript levels of genes involved in stress response and hormone metabolism in A. thaliana. MPK6 is indispensable to the growth stimulation effects. Strains n and L protected A. thaliana seedlings against phytopathogens. Three strains significantly increased maize growth in vitro. In summary, introducing these four strains onto plant roots provides a benefit to the plants. This is the first study regarding the potential mechanism(s) applied by Mucilaginibacter sp. as biostimulants.

Project description:BackgroundPlant growth-promoting rhizobacteria (PGPR) have a specific symbiotic relationship with plants and rhizosphere soil. The purpose of this study was to evaluate the effects of PGPR on blueberry plant growth, rhizospheric soil nutrients and the microbial community.MethodsIn this study, nine PGPR strains, belonging to the genera Pseudomonas and Buttiauxella, were selected and added into the soil in which the blueberry cuttings were planted. All the physiological indexes of the cuttings and all rhizospheric soil element contents were determined on day 6 after the quartic root irrigation experiments were completed. The microbial diversity in the soil was determined using high-throughput amplicon sequencing technology. The correlations between phosphorus solubilization, the auxin production of PGPR strains, and the physiological indexes of blueberry plants, and the correlation between rhizospheric microbial diversity and soil element contents were determined using the Pearson's correlation, Kendall's tau correlation and Spearman's rank correlation analysis methods.ResultsThe branch number, leaf number, chlorophyllcontentand plant height of the treated blueberry group were significantly higher than those of the control group. The rhizospheric soil element contents also increased after PGPR root irrigation. The rhizospheric microbial community structure changed significantly under the PGPR of root irrigation. The dominant phyla, except Actinomycetota, in the soil samples had the greatest correlation with phosphorus solubilization and the auxin production of PGPR strains. The branch number, leaf number, and chlorophyllcontent had a positive correlation with the phosphorus solubilization and auxin production of PGPR strains and soil element contents. In conclusion, plant growth could be promoted by the root irrigation of PGPR to improve rhizospheric soil nutrients and the microenvironment, with modification of the rhizospheric soil microbial community.DiscussionPlant growth could be promoted by the root irrigation of PGPR to improve rhizospheric soil nutrients and the microenvironment, with the modification of the rhizospheric soil microbial community. These data may help us to better understand the positive effects of PGPR on blueberry growth and the rhizosphere soil microenvironment, as well as provide a research basis for the subsequent development of a rhizosphere-promoting microbial fertilizer.

Project description:A diverse group of rhizobacteria persists in the rhizospheric soil, on the surface of roots, or in association with rice plants. These bacteria colonize plant root systems, enhance plant growth and crop yield. Indigenous rhizobacteria are known to promote soil health, grain production quality and serve as sustainable bioinoculant. The present study was aimed to isolate, identify and characterize indigenous plant growth promoting (PGP) diazotrophic bacteria associated with the rhizosphere of rice fields from different areas of Jammu and Kashmir, India. A total of 15 bacteria were isolated and evaluated for various PGP traits, antagonistic activity against phytopathogens, production of hydrolytic enzymes and biofilm formation under in-vitro conditions. The majority of the isolated bacteria were Gram-negative. Out of 15 bacterial isolates, nine isolates produced IAA (12.24 ± 2.86 to 250.3 ± 1.15 μg/ml), 6 isolates exhibited phosphate solubilization activity (36.69 ± 1.63 to 312.4 ± 1.15 μg/ml), 7 isolates exhibited rock phosphate solubilization while 5 isolates solubilized zinc (10-18 mm), 7 isolates showed siderophore production, 8 isolates exhibited HCN production, 6 isolates exhibited aminocyclopropane-1-carboxylate (ACC) deaminase activity, 13 isolates exhibited cellulase activity, nine isolates exhibited amylase and lipase activity and six isolates exhibited chitinase activity. In addition, 5 isolates showed amplification with the nifH gene and showed a significant amount of nitrogenase activity in a range of 0.127-4.39 μmol C2H4/mg protein/h. Five isolates viz., IHK-1, IHK-3, IHK-13, IHK-15 and IHK-25 exhibited most PGP attributes and successfully limited the mycelial growth of Rhizoctonia solani and Fusarium oxysporum in-vitro. All the five bacterial isolates were identified based on morphological, biochemical and 16S rDNA gene sequencing study, as Stenotrophomonas maltophilia, Enterobacter sp., Bacillus sp., Ochrobactrum haematophilum and Pseudomonas aeruginosa. Rice plants developed from seeds inoculated with these PGP strains individually had considerably higher germination percentage, seed vigor index and total dry biomass when compared to control. These findings strongly imply that the PGP diazotrophic bacteria identified in this work could be employed as plant growth stimulators in rice.

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Isolation and characterization of plant growth-promoting rhizobacteria from wheat rhizosphere and their effect on plant growth promotion.

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