Endophytic bacilli from Cyamopsis tetragonoloba (L.) Taub. induces plant growth and drought tolerance.

Cyamopsis tetragonoloba (L.) Taub. (guar) is a commercially important crop known for its galactomannan content in seeds. Drought stress is a significant global concern that compromises the productivity of major legumes including guar. The endophytic microbes associated with plants play a significant role in enhancing plant growth and modulating the impact of abiotic stress(s). The present study involved the isolation of 73 endophytic bacteria from the guar seeds of drought-tolerant (RGC-1002 and RGC-1066) and sensitive (Sarada and Varsha) varieties. Based on multiple PGP attributes and drought tolerance, at 50% PEG6000 w/v, 11 efficient isolates were selected and identified through 16S rRNA gene sequencing. Isolates belonging to ten different species of bacilli including Cytobacillus oceanisediminis, Mesobacillus fermenti, Peribacillus simplex from sensitive and Bacillus zanthoxyli, B. safensis, B. velezensis, B. altitudinis, B. licheniformis, B. tequilensis, and B. paralicheniformis isolated from tolerant varieties. A greenhouse experiment with a drought-sensitive guar variety demonstrated that inoculation of selected isolates showed comparatively better plant growth, higher relative water content (RWC), decreased carbon isotope discrimination ratio (Δ13C), increased chlorophyll, carotenoids, anthocyanin, and proline content, decreased malondialdehyde (MDA) and modulated defense enzymes as compared to their uninoculated controls. Tolerant variety isolates B. tequilensis NBRI14G and B. safensis NBRI10R showed the most promising results in improving plant growth and also drought stress tolerance in guar plants. This study represents for the first time that seed endophytic bacterial strains from guar can be utilized to develop the formulation for improving the productivity of guar under drought-stress conditions.

Endophytic Alkalotolerant Plant Growth-Promoting Bacteria Render Maize (Zea mays L.) Growth Under Alkaline Stress.

This study investigates the role of bacterial endophytes from extreme alkaline environments in alleviating alkaline stress and plant development. Stressful environmental factors, such as soil acidity and alkalinity/sodicity, frequently affect plant development. In the present study, alkaline-tolerant endophytic strains were isolated from three plant species Saccharum munja, Calotropis procera, and Chenopodium album, and 15 out of the total of 48 isolates were selected for further examination of their abiotic stress tolerance. Molecular analysis based on 16S rRNA gene sequencing revealed strains from Enterobacter, Acinetobacter, Stenotrophomonas, Bacillus, Lysinibacillus, and Mammaliicoccus genera. Out of 15 isolates based on their quantitative PGP traits and abiotic stress tolerance, 6 were finally selected for greenhouse experiments. Under alkaline conditions, results demonstrated that the strains from the genera Enterobacter, Bacillus, Stenotrophomonas, and Lysinibacillus had beneficial effects on maize growth. These findings suggest that using a combination of bacteria with multiple plant growth-promoting attributes could be a sustainable approach to enhance agricultural yield, even in a challenging alkaline environment. The study concludes that the application of bacterial endophytes from plants growing in extremely alkaline environments might provide other plants with similar stress-tolerance abilities. The outcome of the study provides a basis for future exploration of the mechanisms underlying endophyte-induced stress tolerance.

Arsenic stress management through arsenite and arsenate-tolerant growth-promoting bacteria in rice.

Arsenic (As) contamination is a major problem affecting soil and groundwater in India, harming agricultural crops and human health. Plant growth-promoting rhizobacteria (PGPR) have emerged as promising agents for reducing As toxicity in plants. This study aimed to isolate and characterize As-tolerant PGPR from rice field soils with varying As levels in five districts of West Bengal, India. A total of 663 bacterial isolates were obtained from the different soil samples, and 10 bacterial strains were selected based on their arsenite (As-III) and arsenate (As-V) tolerant ability and multiple PGP traits, including phosphate solubilization, production of siderophore, indole acetic acid, biofilm formation, alginate, and exopolysaccharide. These isolates were identified by 16S rRNA gene sequencing analysis as Staphylococcus sp. (4), Niallia sp. (2), Priestia sp. (1), Bacillus sp. (1), Pseudomonas sp. (1), and Citricoccus sp. (1). Among the selected bacterial strains, Priestia flexa NBRI4As1 and Pseudomonas chengduensis NBRI12As1 demonstrated significant improvement in rice growth by alleviating arsenic stress under greenhouse conditions. Both strains were also able to modulate photosynthetic pigments, soluble sugar content, proline concentration, and defense enzyme activity. Reduction in As-V accumulation inoculated with NBRI4As1 was recorded highest by 53.02% and 31.48%, while As-III by NBRI12As1 38.84% and 35.98% in the roots and shoots of rice plants, respectively. Overall, this study can lead to developing efficient As-tolerant bacterial strains-based bioinoculant application packages for arsenic stress management in rice.

Ochrobactrum sp. NBRISH6 Inoculation Enhances Zea mays Productivity, Mitigating Soil Alkalinity and Plant Immune Response.

Intensifying sodic land characterized by high alkaline pH is an incipient environmental hazard-limiting agricultural potential. In this study, we investigated the effects of plant growth-promoting bacteria Ochrobactrum sp. strain NBRISH6 on the growth and physiology of maize (Zea mays L.) grown under alkaline stress at two soil pH levels. Additionally, we also studied the effects of NBRISH6 on soil fertility parameters. A greenhouse experiment was designed using two live soils (pH 8.2 and 10.2) in earthen pots using maize as a host. Results revealed a significant increase in plant growth and a decrease in defense enzymes in both soil types due to NBRISH6 inoculation as compared to non-treated control. Furthermore, activities of all soil enzymes along with bacterial diversity increased in NBRISH6 treatment under normal as well as stressed conditions. In addition, field evaluation of NBRISH6 inoculation using maize was carried out under normal and alkaline conditions, which resulted in significant enhancement of all vegetative parameters as compared to respective controls. Therefore, the study suggested that Ochrobactrum sp. NBRISH6 can be used to develop a bioinoculant formulation to ameliorate abiotic stresses and enhanced crop productivity.

Bacillus Consortia Modulate Transcriptional and Metabolic Machinery of Arabidopsis Plants for Salt Tolerance.

Rhizobacteria that are helpful to plants can lessen the impacts of salt stress, and they may hold promise for the development of sustainable agriculture in the future. The present study was intended to explicate consortia of salt-tolerant plant-beneficial rhizobacteria for the amelioration of salinity stress in Arabidopsis plants. Inoculation with both the consortia positively influenced the growth of plants as indicated by total chlorophyll content, MDA content, and antioxidant enzyme activities under stressful conditions. Both the multi-trait consortia altered the expression profiles of stress-related genes including CSD1, CAT1, Wrky, Ein, Etr, and ACO. Furthermore, the metabolomic analysis indicated that inoculated plants modulated the metabolic profiles to stimulate physiological and biochemical responses in Arabidopsis plants to mitigate salt stress. Our study affirms that the consortia of salt-tolerant bacterial strains modulate the transcriptional as well as metabolic machinery of plants to protect them from salinity stress. Nevertheless, the findings of this study revealed that consortia are composed of salt-tolerant bacterial strains viz. Bacillus safensis NBRI 12M, B. subtilis NBRI 28B, and B. subtilis NBRI 33N demonstrated significant improvement in Arabidopsis plants under saline stress conditions.

Revealing the complexity of protein abundance in chickpea root under drought-stress using a comparative proteomics approach.

Global warming has reached an alarming situation, which led to a dangerous climatic condition. The irregular rainfalls and land degradation are the significant consequences of these climatic changes causing a decrease in crop productivity. The effect of drought and its tolerance mechanism, a comparative roots proteomic analysis of chickpea seedlings grown under hydroponic conditions for three weeks, performed at different time points using 2-Dimensional gel electrophoresis (2-DE). After PD-Quest analysis, 110 differentially expressed spots subjected to MALDI-TOF/TOF and 75 spots identified with a significant score. These identified proteins classified into eight categories based on their functional annotation. Proteins involved in carbon and energy metabolism comprised 23% of total identified proteins include mainly glyceraldehyde-3-phosphate dehydrogenase, malate dehydrogenase, transaldolase, and isocitrate dehydrogenase. Proteins related to stress response (heat-shock protein, CS domain protein, and chitinase 2-like) contributed 16% of total protein spots followed by 13% involved in protein metabolism (adenosine kinase 2, and protein disulfide isomerase). ROS metabolism contributed 13% (glutathione S-transferase, ascorbate peroxidase, and thioredoxin), and 9% for signal transduction (actin-101, and 14-3-3-like protein B). Five percent protein identified for secondary metabolism (cinnamoyl-CoA reductase-1 and chalcone-flavononeisomerase 2) and 7% for nitrogen (N) and amino acid metabolism (glutamine synthetase and homocysteine methyltransferase). The abundance of some proteins validated by using Western blotting and Real-Time-PCR. The detailed information for drought-responsive root protein(s) through comparative proteomics analysis can be utilized in the future for genetic improvement programs to develop drought-tolerant chickpea lines.

ACC deaminase-producing rhizosphere competent Bacillus spp. mitigate salt stress and promote Zea mays growth by modulating ethylene metabolism.

Plant growth-promoting rhizobacteria (PGPR) are known for growth promotion and mitigating environmental stresses. Here, we examined the propitiousness of three indigenous salt-tolerant PGPR, i.e., Bacillus subtilis (NBRI 28B), B. subtilis (NBRI 33 N), and B. safensis (NBRI 12 M) for plant growth promotion and salt stress amelioration in Zea mays. Results of the in vitro plant growth-promoting attribute revealed NBRI 12 M demonstrated the highest values at 1 M salt (NaCl) concentration. Furthermore, the greenhouse experiment using three Bacillus strains confirmed plant growth-promoting and salt stress-ameliorating ability, through colonizing successfully and mitigating the adverse effects of ethylene by modulating 1-aminocyclopropane-1-carboxylic acid (ACC) accumulation, ACC-oxidase (ACO), and ACC-synthase (ACS) activities under salt stress. Bacillus sp. inoculation has also induced plant response for defense enzymes, chlorophyll, proline and soluble sugar under salt stress. Among three Bacillus strains, NBRI 12 M not only demonstrated higher values for plant growth-promoting (PGP) attributes but also the same was observed in the greenhouse experiment. Thus, the outcomes of this comparative study represent for the first time that salt-tolerant Bacillus strains exhibiting multiple PGP attributes under salt stress along with high rhizosphere competence can alleviate salt stress by reducing the stress ethylene level in the host plant.

Characterization of plant growth-promoting alkalotolerant Alcaligenes and Bacillus strains for mitigating the alkaline stress in Zea mays.

Intensification of sodic soil due to increasing pH is an emerging environmental issue. The present study aimed to isolate and characterise alkaline stress-tolerant and plant growth-promoting bacterial strains from moderately alkaline soil (pH 8-9), strongly alkaline soil (pH 9-10), and very strongly alkaline soil (> 10). Total 68 bacteria were isolated, and screened for multiple plant growth promoting (PGP) attributes. Out of total, 42 isolates demonstrating at least three plant growth promoting PGP traits selected for further assays. Then out of 42, 15 bacterial isolates were selected based on enhanced maize plant growth under greenhouse experiment, and 16S rRNA gene sequencing revealed Bacillus spp. as a dominant genus. Furthermore, based on improved seed germination percentage and biomass of maize (Zea mays L.) under alkaline stress conditions Alcaligenes sp. NBRI NB2.5, Bacillus sp. NBRI YE1.3, and Bacillus sp. NBRI YN4.4 bacterial strains were selected, and evaluated for growth-promotion and alkaline stress amelioration under greenhouse condition. Amongst the selected 3 plant growth promoting rhizobacterial (PGPR) strains, Bacillus sp. NBRI YN4.4 significantly improved the photosynthetic pigments and soluble sugar content, and decreased proline level in inoculated maize plants as compared to uninoculated control under stress conditions. Moreover, significantly enhanced soil enzymes such as dehydrogenase, alkaline phosphatase and betaglucosidase due to inoculation of Bacillus sp. NBRI YN4.4 in maize plants grown in alkaline soil attributes to its role in improving the soil health. Therefore, alkaline stress-tolerant PGPR NBRI YN4.4 can be useful for developing strategies for the reclamation of saline/sodic soils and improving the plant growth and soil health in sustainable manner.

Drought tolerant Ochrobactrum sp. inoculation performs multiple roles in maintaining the homeostasis in Zea mays L. subjected to deficit water stress.

Plant growth-promoting rhizobacteria (PGPR) improve plant health under various biotic and abiotic stresses. However, the underlying mechanisms of the protective effects of PGPR in deficit water stress (WS) remain less explored. This study aimed to characterize the role of Ochrobactrum sp. NBRISH6 inoculation on maize (Zea mays "Maharaja") under WS conditions using multiple approaches such as physiological, anatomical, metabolic, and molecular. The effect of NBRISH6 inoculation using maize as a host plant was characterized under greenhouse conditions in deficit water stress. Results from this study demonstrated that NBRISH6 significantly lowered the expression of genes involved in the abscisic acid cycle, deficit water stress-response, osmotic stress, and antioxidant enzyme activity (superoxide dismutase, catalase, ascorbate peroxidase, guaiacol peroxidase, and polyphenol oxidase). Phytohormones, i.e. indole acetic acid (IAA) and salicylic acid (SA) levels, intercellular CO2 concentration, metabolites such as simple sugars, amino acids, aliphatic hydrocarbons, and the number of shrunken pith cells modulated in maize roots inoculated with NBRISH6. The NBRISH6 inoculation also improved the plant vegetative properties (root length, 33.80%; shoot length, 20.68%; root dry weight, 39.21%; shoot dry weight, 61.95%), shoot nutrients, xylem cells, root hairs, vapor pressure deficit (75%), intrinsic water-use efficiency (41.67%), photosynthesis rate (83.33%), and total chlorophyll (16.15%) as compared to the respective stress controls. This study provides valuable insights into mechanistic functions of PGPR in WS amelioration and promoting plant physiological response.

Exploitation of agro-climatic environment for selection of 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase producing salt tolerant indigenous plant growth promoting rhizobacteria.

A comprehensive survey for 09 agro-climatic zones of Uttar Pradesh, India was conducted to isolate and characterize salt tolerant 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase possessing plant growth promoting (PGP) rhizobacteria for salt stress amelioration in rice. Here, we have isolated 1125 bacteria having the ability to tolerate 1M NaCl and out of those, 560 were screened for utilizing ACC as sole nitrogen source. 560 isolates were subjected for bacteria coated seed germination assay under 100mM salt (NaCl) stress resulting to 77 isolates which were further evaluated for seed germination assay, PGP and abiotic stress tolerance ability in vitro. This evaluation revealed 15 potent rhizobacteria representing each agro-climatic zone and salt stress mitigation in vitro. In particular, the biomass obtained for bacteria coated rice seedlings were corroborated with the performance of isolates exhibiting maximum average indole acetic acid (IAA) production respective to the agro-climatic zone. Surprisingly based on 16S rRNA, much of the propitious isolates belonged to same specific epithet exhibited variedly in their characteristics. Overall, Bacillus spp. was explored as dominant genera in toto with highest distribution in Western Plain zone followed by Central zone. Therefore, this study provides a counter-intuitive perspective of selection of native microflora for their multifarious PGP and abiotic stress tolerance abilities based on the agro-climatic zones to empower the establishment and development of more suitable inoculants for their application in agriculture under local stress environments.

Characterisation of Pseudomonas spp. and Ochrobactrum sp. isolated from volcanic soil.

Soil bacteria may have properties of plant growth promotion but not be sufficiently beneficial for plants under stress conditions. This challenge has led researchers to extend their searches into extreme environments for potential soil bacteria with multiple plant beneficial traits as well as abiotic stress tolerance abilities. In the current study, an attempt was made to evaluate soil bacteria from an extreme environment, volcano soils, based on plant growth promoting and abiotic stress mitigating characteristics. The screening led to the isolation of eight (NBRISH4, NBRISH6, NBRISH10, NBRISH11, NBRISH13, NBRISH14, NBRISH16 and NBRISH26) bacterial isolates capable of withstanding stresses, namely temperature (up to 45 °C), salt (up to 2 M NaCl) and drought (up to 60% Poly Ethylene Glycol 6000) in vitro. Further, the selected isolates were notable for their in vitro temporal performance with regards to survival (in terms of colony count), phosphate solubilisation, biofilm formation, auxin, alginate and exo-polysaccharide production abilities under abiotic stresses i.e. 40 °C temperature; 500 mM NaCl salt and drought (PEG) conditions. In vivo seed treatments of individual selected bacteria to maize plants resulted into significant enhancement in root and shoot length, root and shoot fresh and dry weight and number of leaves per plant. Overall, the plant growth promoting and abiotic stress tolerance ability was most evident for bacterial isolate NBRISH6 which was identified as an Ochrobactrum sp. using 16S rRNA based phylogenetic analysis.