Study on the characterization of endosulfan-degrading bacterial strains isolated from contaminated rhizospheric soil.

In the present study, we have isolated endosulfan tolerant bacterial strains from the rhizosphere of plants growing in a pesticide-contaminated area. The tolerance capacities of these strains were tested up to 50,000 µg ml-1 of endosulfan. It was found that out of nineteen, four strains (EAG-EC-12, EAG-EC-13, EAG-EC-14, and EAG-EC-15) were capable of surviving up to 50,000 µg ml-1 endosulfan concentration in the media; thus, these four strains were selected for the characterization. Among four, two strains were identified as Serratia liquefaciens, while the other two strains were Bacillus sp. and Brevibacterium halotolerans. The result shows that growth of strain Serratia liquefaciens 1 and Serratia liquefaciens 2 in treated medium was statistically similar to that of control (cfu 6.8 × 107) after 24 h, while strains Bacillus sp. and Brevibacterium halotolerans have shown growth significantly less than the control. The degradation potential of these strains was analyzed against 100 to 250 µg ml-1 of endosulfan in a Minimal Broth Medium (MBM), and it was recorded that only 9, 2, 7, and 19% of endosulfan (100 µg ml-1) remain after a 72 h incubation period of Bacillus sp., Serratia liquefaciens 1, Serratia liquefaciens 2, and Brevibacterium halotolerans, respectively. This endosulfan removal potential of studied strains was decreased with an increase in concentration of endosulfan.

Silicon-Solubilizing Media and Its Implication for Characterization of Bacteria to Mitigate Biotic Stress.

Silicon (Si), the second most abundant element on earth, remains unavailable for plants' uptake due to its poor solubility. Microbial interventions to convert it in soluble forms are well documented. However, studies on discrimination of Si and P solubilizing microbes due to common estimation method and sharing of solubilization mechanism are still obscure. A defined differential media, i.e. silicon-solubilizing media (NBRISSM) is developed to screen Si solubilizers. NBRISN13 (Bacillus amyloliquefaciens), a Si solubilizer, exhibiting antagonistic property against Rhizoctonia solani, was further validated for disease resistance. The key finding of the work is that NBRISSM is a novel differential media for screening Si solubilizers, distinct from P solubilizers. Dominance of Pseudomonas and Bacillus spp. for the function of Si solubilization was observed during diversity analysis of Si solubilizers isolated from different rhizospheres. Sphingobacterium sp., a different strain has been identified for silicon solubilization other than Pseudomonas and Bacillus sp. Role of acidic phosphatase during Si solubilization has been firstly reported in our study in addition to other pH dependent phenomenon. Study also showed the combinatorial effect of feldspar and NBRISN13 on elicited immune response through (i) increased Si uptake, (ii) reduced disease severity, (iii) modulation of cell wall degrading and antioxidative enzyme activities, and (iv) induced defense responsive gene expression.

Betula utilis extract prolongs life expectancy, protects against amyloid-ß toxicity and reduces Alpha Synuclien in Caenorhabditis elegans via DAF-16 and SKN-1.

Betula utilis (BU), an important medicinal plant that grows in high altitudes of the Himalayan region, has been utilized traditionally due to it's antibacterial, hepatoprotective, and anti-tumor properties. Here, we demonstrated the longevity and amyloid-ß toxicity attenuating activity of B. utilis ethanolic extract (BUE) in Caenorhabditis elegans. Lifespan of the worms was observed under both the standard laboratory and stress (oxidative and thermal) conditions. Effect of BUE was also observed on the attenuation of age-dependent physiological parameters. Further, gene-specific mutants and green fluorescent protein (GFP)-tagged strains were used to investigate the molecular mechanism underlying the beneficial effects mediated by BUE supplementation. Our results showed that BUE (50 µg/ml) extended the mean lifespan of C. elegans by 35.99% and increased its survival under stress conditions. The BUE also reduced the levels of intracellular reactive oxygen species (ROS) by 22.47%. A delayed amyloid-ß induced paralyses was observed in CL4176 transgenic worms. Interestingly, the BUE supplementation was also able to reduce the α-synuclein aggregation in NL5901 transgenic strain. Gene-specific mutant studies suggested that the BUE-mediated lifespan extension was dependent on daf-16, hsf-1, and skn-1 but not on sir-2.1 gene. Furthermore, transgenic reporter gene expression assay showed that BUE treatment enhanced the expression of stress-protective genes such as sod-3 and gst-4. Present findings suggested that ROS scavenging activity, together with multiple longevity mechanisms, were involved in BUE-mediated lifespan extension. Thus, BUE might have potential to increase the lifespan and to attenuate neuro-related disease progression.

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.

Improvement in Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) by the inhibition of polyphenolics released during wounding of cotyledonary node explants.

Agrobacterium-mediated transformation of chickpea (Cicer arietinum L.) has been performed using cotyledonary node explants (CNs), which release phenolics upon excision that are detrimental to the viability of Agrobacterium tumefaciens and result in low transformation frequency. Twelve low molecular weight phenolic compounds and salicylic acid were identified in the exudates released upon excision during the preparation of cotyledonary nodes by reverse phase high-performance liquid chromatography (RP-HPLC). Zone inhibition assays performed with the explant exudates released at periodic intervals after excision showed the inhibition of A. tumefaciens. Agroinoculation of freshly excised cotyledonary nodes of chickpea showed 98-99 % inhibition of colony forming units (cfu). Osmium tetraoxide fixation of excised tissues showed enhanced accumulation of phenolics in the sub-epidermal regions causing enzymatic browning, affecting the viability and performance of A. tumefaciens for T-DNA delivery. The periodic analysis of exudates released from excised CNs showed enhanced levels of gallic acid (0.2945 ± 0.014 µg/g), chlorogenic acid (0.0978 ± 0.0046 µg/g), and quercetin (0.0971 ± 0.0046 µg/g) fresh weight, which were detrimental to A. tumefaciens. Quantitative assays and the elution profile showed the maximum leaching of phenolics, flavonoids, and salicylic acid immediately after the excision of explants and continued till 4 to 8 h post-excision. Pre-treatment of excised explants with inhibitors of polyphenol oxidase like L-cysteine, DTT, and sodium thiosulfate before co-cultivation showed the recovery of A. tumefaciens cfu, decreased the accumulation of phenolics, and improved transformation frequency. Our results show the hypersensitive response of excision stress for the expression of defense response-related genes and synthesis of metabolites in grain legume chickpea against pathogen infestation including Agrobacterium.