Imperative roles of salicylic acid and nitric oxide in improving salinity tolerance in Pisum sativum L.

This study was undertaken to scrutinize efficacy of salicylic acid (SA) and/or sodium nitroprusside [SNP, source of nitric oxide (NO)] to mitigate injury symptoms of saline stress in Pisum sativum L. Exposure to sodium chloride (NaCl) was found to be injurious to germinating P. sativum L. (var. Shubhra IM-9101) and a direct correlation between severity of toxicity and NaCl-concentrations could be discernible. Both SA and NO serves as signal molecules in plant stress responses, and play crucial roles in key regulatory pathways of growth, development and metabolism. The limiting effects of salinity on radicle length and biomass accumulation were considerably released by SA and/or SNP and among which their combined application was found to be the most promising. Supplemented SA and/or SNP, particularly their cocktail, resulted in a substantial decline in reactive oxygen species accumulation, which later caused reduced accumulations of malondialdehyde, 4-hydroxy-2-nonenal and protein carbonyl, in NaCl subjected germinating P. sativum L. seeds. SA and/or SNP had significant inducing effects on activities of superoxide dismutase, catalase, guaiacol peroxidase and ascorbate peroxidase. Additionally, exogenous SA and/or SNP led to the higher proline, sugar and glycinebetaine contents, than that of the control. On the basis of accumulated results, it could be concluded that the cocktail of SA and SNP may be efficiently used to overcome the adverse signatures of salinity stress.

Efficient extraction of proteins from recalcitrant plant tissue for subsequent analysis by two-dimensional gel electrophoresis.

Protein extraction for two-dimensional electrophoresis from tissues of recalcitrant species is quite problematic and challenging due to the low protein content and high abundance of contaminants. Proteomics in Shorea robusta is scarcely conducted due to the lack of a suitable protein preparation procedure. To establish an effective protein extraction protocol suitable for two-dimensional electrophoresis in Shorea robusta, four procedures (borate buffer/trichloroacetic acid extraction, organic solvent/trichloroacetic acid precipitation, sucrose/Tris/phenol, and organic solvent/phenol/sodium dodecyl sulfate) were evaluated. Following these, proteins were isolated from mature leaves and were analyzed for proteomics, and also for potential contaminants, widely reported to hinder proteomics. The borate buffer/trichloroacetic acid extraction had the lowest protein yield and did not result in any banding even in one-dimensional electrophoresis. In contrast, organic solvent/phenol/sodium dodecyl sulfate extraction allowed the highest protein yield. Moreover, during proteomics, organic solvent/phenol/sodium dodecyl sulfate extracted protein resolved the maximum number (144) of spots. Further, when proteins were evaluated for contaminants, significant (77-95%) reductions in the nucleic acids, phenol, and sugars were discernible with refinement in extraction procedure. Accumulated data suggested that the organic solvent/phenol/sodium dodecyl sulfate extraction was the most effective protocol for protein isolation for proteomics of Shorea robusta and can be used for plants that have a similar set of contaminants.

Carbon dot induces tolerance to arsenic by regulating arsenic uptake, reactive oxygen species detoxification and defense-related gene expression in Cicer arietinum L.

The scientific and technological applications of one of the nanomaterials viz.; carbon dot (C-dots), having extraordinary properties, is becoming an emerging and ongoing research area in recent times. In the present study, we have evaluated the effectiveness of C-dots in reducing arsenic (As) toxicity by analyzing physiological, biochemical and molecular parameters in Cicer arietinum L. The results revealed that As decreased the germination rate, growth, biomass, and membrane stability of the cell to a significant extent. Further, As was taken up by the growing seeds which eventually caused cell death. Levels of reactive oxygen species (ROS), stress markers (malondialdehyde), activities of defensive enzymes (glutathione-S-transferase and pyrroline-5-carboxylate synthetase) and non-enzymatic antioxidant contents (proline and glutathione) were increased under As stress. Moreover, As treatment resulted in the up-regulation of expressions of NADPH oxidase and defense-related genes in Cicer arietinum L. However, application of C-dots along with As improved the germination and growth of Cicer arietinum L. Exogenous application of C-dots, enhanced the expressions of defense-related genes and, contents of proline and glutathione, thereby causing considerable reductions in ROS, and malondialdehyde levels. Overall, this study suggests the possible involvement of C-dots in lowering the toxic effects of As on biomass by reducing As uptake and, inducing the activities/gene expressions and contents of enzymatic and non-enzymatic antioxidants.

Silver nanoparticle modulates gene expressions, glyoxalase system and oxidative stress markers in fluoride stressed Cajanus cajan L.

Application of engineered nanomaterials has increased these days due to their beneficial impacts on several sectors of the economy, including agriculture. Silver nanoparticles (AgNP) are commonly used to improve rate of seed germination, and growth and development of plants. The present study was aimed to monitor the role of engineered AgNP (non-dialysed) in the amelioration of fluoride (F)-induced oxidative injuries in Cajanus cajan L. Experimental results revealed that F-exposure inhibited growth and membrane stability index, while were enhanced with the augmentation of AgNP. The results also demonstrated that F treatment enhanced the accumulations of reactive oxygen species, malondialdehyde and oxidized glutathione, gene expression of NADPH oxidase, and activity of lipoxygenase, but were decreased by the addition of AgNP. The results indicated that exogenous application of AgNP provided tolerance against F-toxicity via enhancing the levels of proline, total and reduced glutathione, glyoxalase I and II activities, and expression of pyrroline-5-carboxylate synthetase gene. Conducted study uniquely suggested potential role of AgNP in the remediation of F-toxicity, at least in the Cajanus cajan L. radicles. Further research would be intended to unravel the molecular mechanism(s) involved precisely in the AgNP mediated alleviation of F-toxicity.

Arsenic-induced genotoxic responses and their amelioration by diphenylene iodonium, 24-epibrassinolide and proline in Glycine max L.

Presence of the toxic metalloid, "arsenic (As)" is ubiquitous in the environment especially in the soil and water. Its excess availability in the soil retards growth and metabolism of plants via (a) slowing down the cell division/elongation, (b) overproduction of reactive oxygen species (ROS), (c) modulation of antioxidant enzymes, and (d) alteration of DNA profile/genomic template stability (GTS). In the current study, diphenylene iodonium (DPI), 24-epibrassinolide (EBL) and proline (Pro) were used to analyze their roles in eliminating the adverse effects of As. Glycine max L. (variety JS 335) seeds were subjected to As (75 µM, Sodium arsenite was used as source of As), and in combination with DPI (10 µM), EBL (0.5 µM) or Pro (10 mM), for five consecutive days, and effects of these treatment combinations were analyzed on germination percentage, biomass, membrane stability, GTS and expressions of defensive genes. In addition, the levels of As, ROS, malondialdehyde, DNA content, oxidation, fragmentation, polymorphism, DNase activity, endogenous Pro and pyrroline-5-carboxylate synthetase activity were evaluated. The results indicated that the treatments of DPI, EBL or Pro are capable to alleviate detrimental effects of As, gauged from above variables, but with different magnitudes. Apropos As-stress mitigation, Pro was found to be the most effective under the confines of the study protocol. This study certainly provides new ideas for intensifying studies to unravel elusive central mechanism of amelioration involving use of DPI, EBL or Pro in plants with confirmed As-toxicity.