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ACC deaminase producing plant growth promoting rhizobacteria enhance salinity stress tolerance in Pisum sativum.

ABSTRACT: Salinity stress is one of the most serious environmental stresses which limit plant growth, development and productivity. In this study, we screened 25 bacterial isolates based on the biochemical activity of ACC deaminase. Two potent PGPR namely Bacillus marisflavi (CHR JH 203) and Bacillus cereus (BST YS1_42) having the highest ACC deaminase (ACCD) activity were selected for further analyses such as polymerase chain reaction (PCR), salt tolerance assay, expression analysis, antioxidant assay, etc. The structural gene for ACCD activity was further confirmed by PCR showing the amplicon size ~ 800 bp. The acdS positive isolates exhibited optimum growth at 3% w/v (NaCl), indicating its ability to survive and thrive in induced saline soil. Inoculation of acdS ⁺ strain on pea plants was found to be efficient and ameliorated the induced NaCl-stress by enhancing the various parameters like plant-biomass, carbohydrates, reducing sugars, protein, chlorophylls, phenol, flavonoids content and increasing antioxidants enzymes levels in plants. Moreover, the expression of ROS scavenging genes (PsSOD, PsCAT, PsPOX, PsNOS, PsAPX, PsChla/bBP), defense genes and cell rescue genes (PsPRP, PsMAPK, PsFDH) were analyzed. Inoculated plants exhibited a higher gene expression level and salt tolerance under 1%NaCl concentration. Thus, our results indicate that CHR JH 203 and BST YS1_42 strain showed the highest plant growth-promoting attributes could be used as bio-inoculants for crops under saline stress in the field towards sustainable crop development.

Supplementary information

The online version contains supplementary material available at 10.1007/s13205-021-03047-5.

SUBMITTER: Gupta A

PROVIDER: S-EPMC8630178 | biostudies-literature | 2021 Dec

REPOSITORIES: biostudies-literature

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ACC deaminase producing plant growth promoting rhizobacteria enhance salinity stress tolerance in Pisum sativum.

Gupta Anmol A Bano Ambreen A Rai Smita S Kumar Manoj M Ali Jasarat J Sharma Swati S Pathak Neelam N

3 Biotech 20211129 12

Salinity stress is one of the most serious environmental stresses which limit plant growth, development and productivity. In this study, we screened 25 bacterial isolates based on the biochemical activity of ACC deaminase. Two potent PGPR namely Bacillus marisflavi (CHR JH 203) and Bacillus cereus (BST YS1_42) having the highest ACC deaminase (ACCD) activity were selected for further analyses such as polymerase chain reaction (PCR), salt tolerance assay, expression analysis, antiox ...[more]

PMID: 34926112

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Project description:BackgroundEndophytic bacteria reside inside healthy plant tissues and provide several benefits to their host, and help them to tolerate various stresses. Aminocyclopropane-1-carboxylate deaminase (ACCD) production is one of the mechanisms by which these bacteria help the plant to survive under ethylene stress.ObjectivesThe main focus of this study was to isolate endophytic bacteria and effectively screen them for ACCD production. The selected isolate was identified and assessed for plant growth-promoting potential under pot conditions.Materials and methodsEndophytic bacteria were isolated from root nodules of Pisum sativum plants, grown in northern India (Haryana state). ACCD activity was initially screened on DF minimal salt medium with ACC as a sole nitrogen source. To narrow down the number of the isolates, another screening method was adopted using a modified medium containing indicator dyes along with ACC. The strain producing ACCD as well as a significant amount of Indole 3 acetic acid (IAA) was identified using 16S rDNA gene sequencing and amplification of acdS gene. Its ability to promote plant growth was evaluated under pot culture conditions.ResultsTwenty-six endophytic bacteria were isolated from nodules of P. sativum plants. Sixteen isolates showed growth on DF minimal salts medium supplemented with ACC along with negative control. On the modified medium containing indicator dyes, two isolates, PJN13 and PJN17, showed zones of the color gradient. The ACC deaminase activity was further confirmed by enzymatic assay. The strains PJN13 and PJN17 produced 160 and 130 µM of α-ketobutyrate m.g-1 protein h-1, respectively. The IAA production in the strain PJN13 (79.04 ± 0.78 µg.mL -1) was significantly more than that in the strain PJN17 (38.36 ± 1.89 µg.mL-1). It could enhance pea plant growth parameters, including root and shoot length and fresh and dry weight from 1 to 4 times compare to the control (untreated pea plants) under pot conditions. The results of 16S rDNA amplification and sequencing showed that PJN13 has maximum similarity to Bacillus mojavensis, and the sequence submitted to GenBank under accession number MH298523. Also, a band about 800 bp was amplified for the acdS gene.ConclusionsThough Bacillus is known as a predominant non-rhizobial endophytic genus, however in the present study, a B. mojavensis Bacillus mojavensis PRN2 (MH298523) was reported for the first time as an endophyte from the nodules of pea plants. The isolated strain possesses ACC deaminase activity along with IAA production capability, and high potentials as PGPE (Plant growth-promoting endophyte) for plant growth, so it has potential to be used as biofertilizers in pea fields.

Project description:Microbes enhance crop resilience to abiotic stresses, aiding agricultural sustainability amid rising global land salinity. While microbes have proven effective via seed priming, soil amendments, and foliar sprays in diverse crops, their mechanisms remain less explored. This study explores the utilization of ACC deaminase-producing Nocardioides sp. to enhance wheat growth in saline environments and the molecular mechanisms underlying Nocardioides sp.-mediated salinity tolerance in wheat. The Nocardioides sp. inoculated seeds were grown under four salinity regimes viz., 0 dS m-1, 5 dS m-1, 10 dS m-1, and 15 dS m-1, and vegetative growth parameters including shoot-root length, germination percentage, seedling vigor index, total biomass, and shoot-root ratio were recorded. The Nocardioides inoculated wheat plants performed well under saline conditions compared to uninoculated plants and exhibited lower shoot:root (S:R) ratio (1.52 ± 0.14 for treated plants against 1.84 ± 0.08 for untreated plants) at salinity level of 15 dS m-1 and also showed improved biomass at 5 dS m-1 and 10 dS m-1. Furthermore, the inoculated plants also exhibited higher protein content viz., 22.13 mg g-1, 22.10 mg g-1, 22.63 mg g-1, and 23.62 mg g-1 fresh weight, respectively, at 0 dS m-1, 5 dS m-1, 10 dS m-1, and 15 dS m-1. The mechanisms were studied in terms of catalase, peroxidase, superoxide dismutase, and ascorbate peroxidase activity, free radical scavenging potential, in-situ localization of H2O2 and superoxide ions, and DNA damage. The inoculated seedlings maintained higher enzymatic and non-enzymatic antioxidant potential, which corroborated with reduced H2O2 and superoxide localization within the tissue. The gene expression profiles of 18 stress-related genes involving abscisic acid signaling, salt overly sensitive (SOS response), ion transporters, stress-related transcription factors, and antioxidant enzymes were also analyzed. Higher levels of stress-responsive gene transcripts, for instance, TaABARE (~+7- and +10-fold at 10 dS m-1 and 15 dS m-1); TaHAk1 and hkt1 (~+4- and +8-fold at 15 dS m-1); antioxidant enzymes CAT, MnSOD, POD, APX, GPX, and GR (~+4, +3, +5, +4, +9, and +8 folds and), indicated actively elevated combat mechanisms in inoculated seedlings. Our findings emphasize Nocardioides sp.-mediated wheat salinity tolerance via ABA-dependent cascade and salt-responsive ion transport system. This urges additional study of methylotrophic microbes to enhance crop abiotic stress resilience.

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ACC deaminase producing plant growth promoting rhizobacteria enhance salinity stress tolerance in Pisum sativum.

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ACC deaminase producing plant growth promoting rhizobacteria enhance salinity stress tolerance in <i>Pisum sativum</i>.

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