Abstract

YSK2 type dehydrin from Sorghum bicolor (SbDhn1) showed a high level of transcript accumulation when subjected to high temperature and osmotic stress. The high transcript level occurring in such stress situation might lead to a protective effect; though the exact mechanism by which this is achieved remains poorly understood. Nevertheless, our results provide compelling evidence to prove that transgenic tobacco lines overexpressing SbDhn1 gene showed improve stress tolerance as assessed by reduced membrane damage and low MDA content. Furthermore, we demonstrate here SbDhn1 expressing lines were only able to recover after stress treatment. In this study, we have provided direct evidence for the protection rendered by SbDHN1 protein to a temperature-sensitive enzyme under both high temperature and osmotic stress. We extended this analysis to the whole plant proteome where the addition of SbDHN1 protein helped in retaining the solubility of the protein was demonstrated. Interestingly, in vitro experiments carried out with lactate dehydrogenase (LDH), showed aggregate formation upon subjecting it to high temperature. However, in presence of SbDHN1 protein very few aggregates were observed. Aggregation assay showed a high level of aggregates in wild-type or empty vector transformed plants as compared to SbDhn1 transgenic lines. Confocal microscopy images in leaf peel sections of wild-type plants showed high amounts of aggregates as compared with transgenic lines. This study provides evidence for the protection rendered by SbDHN1 protein under high temperature by inhibiting the aggregate formation and provide the rational for the mechanism how these proteins ameliorate the adverse stress conditions.

Highlights

  • Plants face several environmental challenges during their life cycle

  • Our results showed that SbDHN1 protein could protect the lactate dehydrogenase (LDH) activity better than glycerol and BSA when subjected to high temperature or osmotic stress

  • A condition of high temperature and osmotic stress is intimately associated with drought condition

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Summary

Introduction

Plants face several environmental challenges during their life cycle. A complex mechanism exists in plants which help them to sense, react, and adapt . The LEA proteins comprise about 4% of the total cellular proteins (Roberts et al, 1993) The expression of these proteins is coupled with the acquisition of osmotic stress tolerance in orthodox seeds, pollens, and anhydrobiotic plants. The LEA proteins (PFAM: LEA_4; LEA_5 and dehydrins) are endowed with protective mechanisms to circumvent the structural changes in the membranes and biological macromolecules associated with osmotic stress tolerance. Prevention of conformational changes in the biological macromolecules by these LEA proteins might aid in achieving osmotic stress tolerance in plants. Recombinant forms of AavLEA1, a group 3 LEA protein from Aphelenchus avenae; and Em, a group 1 LEA protein from wheat, can independently protect citrate synthase from aggregation due to osmotic and freezing stress. Dehydrins from several plant species were revealed to possess cryoprotective activity (Hara et al, 2001)

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