Cicer arietinum (gram) is an important protein rich pulse crop in Indian subcontinent, the Med iterranean region, Ethiopia, and Mexico. We studied the effects of different salt concentrations on radicle growth and different markers of oxidative stress, e.g., superoxide radical, MDA, protein carbonyls, as well as antioxidant compounds. Physiological and biochemical parameters were assessed in the radicles of germinat ing gram seeds after 1 and 7 days of treatments with 15, 30, 45, and 60 mM NaCl. The results showed that salt exerted a stronger effect (17 fold) on radicle length than on their dry weight (5 fold). This growth decrease was accompanied by an excessive (3 fold) accumulation of ROS and resulting protein carbonyl and MDA formation (3-6 fold). As to the responses of antioxidant compounds to salinity of the growing medium, all the enzymatic molecules (SOD, CAT, POX, and APX) showed significant (4-6 fold) reductions in their activities. Our results suggest that under salinity substantially higher amounts of oxidative stress markers (superoxide, MDA, and protein carbonyls) in collaboration with suppression of the ROS detoxification sys tem ultimately led to gram radicle growth inhibition and severe oxidative stress.
Proline, 24-epibrassinolide and diphenylene iodonium are few of the novel antioxidant molecules, involved in growth regulation and abiotic stress tolerance of plants. However, these are scarcely explored in relation to their role in arsenic stress tolerance. Therefore, present study was designed to investigate the involvement of proline, 24-epibrassinolide and diphenylene iodonium in conferring tolerance to
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.
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