Screening of epiphytic rhizosphere-associated bacteria in Argentinian Malbec and Cabernet-Sauvignon vineyards for potential use as biological fertilisers and pathogen-control agents

The rhizosphere-associated microbiome has diverse functions that support plant growth and health, varying among plant species, vegetation growth stages and environmental habitats. This microbiome includes a group of bacteria denominated plant growth-promoting rhizobacteria (PGPR) which can colonize plant roots. Certain PGPR isolates improve the ability of plants to adapt to a stressful environment. In this study, we collected and characterised the rhizosphere-associated bacteria, or epiphytic rhizobacteria, from Malbec and Cabernet-Sauvignon vineyards from the main wine-producing provinces of Argentina to analyse their potential use as biologic fertilisers and/or as pathogen-control agents. A total of 170 bacterial isolates were obtained, distributed into eleven different genera and classified into three phyla, Proteobacteria, Actinobacteria and Firmicutes. The in vitro analysis for plant-growth-promoting (PGP) activities demonstrated that a significant number of bacterial isolates had one or more of these traits. The Pseudomonas was the genus with the highest number of isolates and PGP activities, followed by the Arthrobacter, Serratia, Bacillus andPantoea. We observed that bacterial isolates identified as Bacillus exhibited a remarkable production of hydrolytic enzymes related to biocontrol activities. Biocontrol trials from the Bacillus collection revealed that at least five isolates were able to inhibit the fungal growth of Botrytis cinerea and Alternaria alternata. The results obtained suggest the biological potential of each isolate and the relevance of proceeding to greenhouse and field assays to obtain long-term environmentally compatible bio-products for vineyard management.

Isolation of Endophytic Salt-Tolerant Plant Growth-Promoting Rhizobacteria From Oryza sativa and Evaluation of Their Plant Growth-Promoting Traits Under Salinity Stress Condition

The application of plant growth-promoting rhizobacteria (PGPR) as vital components for plant growth promotion against biotic and abiotic stresses could be a promising strategy to improve crop production in areas vulnerable to increasing salinity. Here, we isolated Seventy-five endophytic bacteria from roots of healthy Oryza sativa grown in a saline environment of the southern coastal region of Bangladesh. The endophytes in a culture of ~108 CFU/ml showed arrays of plant growth-promoting (PGP) activities: phytohormone (Indole acetic acid) production (1.20–60.13 μg/ ml), nutrient (phosphate) solubilization (0.02–1.81 μg/ml) and nitrogen fixation (70.24–198.70 μg/ml). Four genomically diverse groups were identified namely, Enterobacter, Achromobacter, Bacillus, and Stenotrophomonas using amplified ribosomal DNA restriction analysis followed by their respective 16S rDNA sequence analyses with that of the data available in NCBI GenBank. These four specific isolates showed tolerance to NaCl ranging from 1.37 to 2.57 mol/L in the nutrient agar medium. Under a 200 mmol/L salt stress in vitro, the bacteria in a culture of 108 CFU/ml exhibited competitive exopolysaccharide (EPS) production: Stenotrophomonas (65 μg/ml) and Bacillus (28 μg/ml), when compared to the positive control, Pseudomonas spp. (23.65 μg/ml), a phenomenon ably supported by their strong biofilm-producing abilities both in a microtiter plate assay, and in soil condition; and demonstrated by images of the scanning electron microscope (SEM). Overall, the isolated endophytic microorganisms revealed potential PGP activities that could be supported by their biofilm-forming ability under salinity stress, thereby building up a sustainable solution for ensuring food security in coastal agriculture under changing climate conditions.

Amelioration of Saline Stress on Chia (Salvia hispanica L.) Seedlings Inoculated With Halotolerant Plant Growth-Promoting Bacteria Isolated From Hypersaline Environments

The rhizosphere and microbiome of halotolerant plants could be crucial for alleviating salinity stress during plant growth. The aims of this work were (1) to isolate bacteria from rhizosphere and bulk soil samples from the Salar del Hombre Muerto (Catamarca, Argentina), (2) to characterize different plant growth-promoting (PGP) activities produced by these bacterial isolates, and (3) to evaluate their effect on the initial growth of chia (Salvia hispanica L.) under saline stress. A total of 667 microorganisms were isolated, using different culture media with NaCl, and their abilities for nitrogen fixation, phosphate solubilization, siderophores production, and indole-3-acetic acid production were evaluated. Thirteen strains were selected for showing all the tested PGP activities; they belonged to the genera Kushneria, Halomonass, Pseudomonas, Planomicrobium, and Pseudarthrobacter. The strains Kushneria sp. and Halomonas sp. showed the highest salinity tolerance (from 50 to 2,000 mM NaCl) and biomass and biofilm production. Chia seeds were treated with six of the first 13 selected strains to evaluate their plant growth-promoting effect under saline stress (without and with 50 and 100 mM NaCl). Halomonas sp. 3R.12 and Kushneria sp. T3.7 produced heavier seedlings with a balanced shoot/root length ratio, while Pseudomonas sp. AN23 showed the best effect upon chia seedlings, with a morphological response similar to non-stressed seedlings. On the other hand, seedlings displayed no responses when inoculated with Planomicrobium sp. 3S.31 and Pseudarthrobacter sp. ER25. This study contributes to the knowledge on microorganisms from hypersaline environments as plant growth promoters for their use in the production of salt-sensitive crops, among other potential uses.

Characterization of Bradyrhizobium spp. Nodulating Lupinus cosentinii and L. luteus Microsymbionts in Morocco

In this work, we analyzed the diversity of the nodule-forming bacteria associated with Lupinus luteus and Lupinus cosentinii grown in the Maamora Cork oak forest acidic soils in Morocco. The phenotypic analysis showed the high diversity of the strains nodulating the two lupine's species. The strains were not tolerant to acidity or high alkalinity. They do not tolerate salinity or high temperatures either. The strains isolated from L. luteus were more tolerant to antibiotics and salinity than those isolated from L. cosentinii. The plant growth promoting (PGP) activities of our strains are modest, as among the 28 tested isolates, only six produced auxins, six produced siderophores, whereas three solubilized phosphates. Only two strains possess the three activities. The rrs gene sequences from eight representative strains selected following ARDRA and REP-PCR results revealed that they were members of the genus Bradyrhizobium. Six strains were then retained for further molecular analysis. The glnII, recA, gyrB, dnaK, and rpoB housekeeping gene sequence phylogeny showed that some strains were close to B. lupini LMG28514T whereas others may constitute new genospecies in the genus Bradyrhizobium. The strains were unable to nodulate Glycine max and Phaseolus vulgaris and effectively nodulated L. luteus, L. cosentinii, L. angustifolius, Chamaecytisus albidus, and Retama monosperma. The nodC and nodA symbiotic gene phylogenies showed that the strains are members of the genistearum symbiovar.

Phenetic and Molecular Diversity of Nitrogen Fixating Plant Growth Promoting Azotobacter Isolated from Semiarid Regions of India

In the present study, 24 Azotobacter strains were isolated from soils of different areas of southern Rajasthan and characterized at biochemical, functional, and molecular levels. The isolated Azotobacter strains were gram negative and cyst forming when viewed under the microscope. These strains were also screened for their plant growth promoting activities and the ability of these isolates to survive under abiotic stress conditions viz. salt, pH, temperature, and drought stress. All the isolates showed IAA, siderophore, HCN, and ammonia production, whereas seven Azotobacter strains showed phosphate solubilization. Amplified Ribosomal DNA Restriction Analysis (ARDRA) revealed significant diversity among Azotobacter strains and the dendrogram obtained differentiated twenty-four of the strains into two major clusters at a similarity coefficient of 0.64. Qualitative and quantitative N2 fixation abilities of these strains were also detrained, and the amounts of acetylene reduced by Azotobacter strains were in the range of 1.31 to 846.56 nmol C2H4 mg protein−1 h−1. The strains showing high nitrogen fixation ability with multiple PGP activities were selected for further pot studies, and these Azotobacter strains significantly increased the various plant growth parameters of maize plantlets. Furthermore, the best Azotobacter isolates were subjected to 16S rRNA sequencing and confirmed their identities as Azotobacter sp. The indigenous Azotobacter strains with multiple PGP activities could be further used for commercial production.

Quorum Sensing System Affects the Plant Growth Promotion Traits of Serratia fonticola GS2

Quorum sensing (QS) enables bacteria to organize gene expression programs, thereby coordinating collective behaviors. It involves the production, release, and population-wide detection of extracellular signaling molecules. The cellular processes regulated by QS in bacteria are diverse and may be used in mutualistic coordination or in response to changing environmental conditions. Here, we focused on the influence of the QS-dependent genes of our model bacterial strain Serratia fonticola GS2 on potential plant growth promoting (PGP) activities including indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and biofilm formation. Based on genomic and phenotypic experimental data we identified and investigated the function of QS genes in the genome of the model strain. Our gene deletion study confirmed the biological functionality of the QS auto-inducer (gloI) and receptor (gloR) on potential PGP activities of GS2. A transcriptomic approach was also undertaken to understand the role of QS genes in regulation of genes primarily involved in PGP activities (IAA, ACC deaminase activity, and biofilm formation). Both transcriptomic and phenotypic data revealed that the QS-deletion mutants had considerably less PGP activities, as compared to the wild type. In addition, in vivo plant experiments showed that plants treated with GS2 had significantly higher growth rates than plants treated with the QS-deletion mutants. Overall, our results showed how QS-dependent genes regulate the potential PGP activities of GS2. This information may be helpful in understanding the relationship between QS-dependent genes and the PGP activity of bacteria, which aid in the production of practical bio-fertilizers for plant growth promotion.

Evaluation of plant growth promotion properties and induction of antioxidative defense mechanism by tea rhizobacteria of Darjeeling, India

A total of 120 rhizobacteria were isolated from seven different tea estates of Darjeeling, West Bengal, India. Based on a functional screening of in vitro plant growth-promoting (PGP) activities, thirty potential rhizobacterial isolates were selected for in-planta evaluation of PGP activities in rice and maize crops. All the thirty rhizobacterial isolates were identified using partial 16S rRNA gene sequencing. Out of thirty rhizobacteria, sixteen (53.3%) isolates belong to genus Bacillus, five (16.6%) represent genus Staphylococcus, three (10%) represent genus Ochrobactrum, and one (3.3%) isolate each belongs to genera Pseudomonas, Lysinibacillus, Micrococcus, Leifsonia, Exiguobacterium, and Arthrobacter. Treatment of rice and maize seedlings with these thirty rhizobacterial isolates resulted in growth promotion. Besides, rhizobacterial treatment in rice triggered enzymatic [ascorbate peroxidase (APX), catalase (CAT), chitinase, and phenylalanine ammonia-lyase (PAL)], and non-enzymatic [proline and polyphenolics] antioxidative defense reactions indicating their possible role in the reduction of reactive oxygen species (ROS) burden and thereby priming of plants towards stress mitigation. To understand such a possibility, we tested the effect of rhizobacterial consortia on biotic stress tolerance of rice against necrotrophic fungi, Rhizoctonia solani AG1-IA. Our results indicated that the pretreatment with rhizobacterial consortia increased resistance of the rice plants towards the common foliar pathogen like R. solani AG1-IA. This study supports the idea of the application of plant growth-promoting rhizobacterial consortia in sustainable crop practice through the management of biotic stress under field conditions.

Metagenomic Insights and Genomic Analysis of Phosphogypsum and Its Associated Plant Endophytic Microbiomes Reveals Valuable Actors for Waste Bioremediation

The phosphogypsum (PG) endogenous bacterial community and endophytic bacterial communities of four plants growing in phosphogypsum-contaminated sites, Suaeda fruticosa (SF), Suaeda mollis (SM), Mesembryanthmum nodiflorum (MN) and Arthrocnemum indicum (AI) were investigated by amplicon sequencing. Results highlight a more diverse community of phosphogypsum than plants associated endophytic communities. Additionally, the bacterial culturable communities of phosphogypsum and associated plant endophytes were isolated and their plant-growth promotion capabilities, bioremediation potential and stress tolerance studied. Most of plant endophytes were endowed with plant growth-promoting (PGP) activities and phosphogypsum communities and associated plants endophytes proved highly resistant to salt, metal and antibiotic stress. They also proved very active in bioremediation of phosphogypsum and other organic and inorganic environmental pollutants. Genome sequencing of five members of the phosphogypsum endogenous community showed that they belong to the recently described species Bacillus albus (BA). Genome mining of BA allowed the description of pollutant degradation and stress tolerance mechanisms. Prevalence of this tool box in the core, accessory and unique genome allowed to conclude that accessory and unique genomes are critical for the dynamics of strain acquisition of bioremediation abilities. Additionally, secondary metabolites (SM) active in bioremediation such as petrobactin have been characterized. Taken together, our results reveal hidden untapped valuable bacterial actors for waste remediation.