Abstract
This study describes the first detailed molecular characterization of the heat shock protein 70 (Hsp70) gene from Sorghum bicolor, MN1618 designated as SbHsp70-1. The full-length cDNA of SbHsp70-1 consists of 2524 bp with a 1950 bp open reading frame, which encodes a protein of 649 amino acids. SbHsp70-1 is a cytoplasmic protein with high homology to other plant Hsp70s, especially grain crops. Recombinant SbHsp70-1 was able to bind and hydrolyse ATP in a dose-dependent manner, suggesting that SbHsp70-1 functions as an ATPase. Immunoblot assays showed that the expression of SbHsp70-1 is induced at temperatures of 37, 45, and 4 °C but reduced at 42 °C. In addition, the SbHsp70-1 mRNA transcript is constitutively expressed in both leaves and stem but is significantly increased upon heat shock at 42 °C. Upon cold shock at 4 °C, SbHsp70-1 mRNA transcript level increased in the leaf, but no significant change was observed in the stem. In addition, expression of the pET28a-SbHsp70-1 construct in Escherichia coli cells under heat stress resulted in their survival even at higher temperature (65 °C). Our results suggest that SbHsp70-1 is a heat-inducible protein that confer thermal tolerance to bacterial cells and can be claimed as a promising target to study stress tolerance in crops.Electronic supplementary materialThe online version of this article (doi:10.1007/s12192-015-0591-2) contains supplementary material, which is available to authorized users.
Highlights
Plants are constantly exposed to abiotic and biotic stresses, with abiotic stress being the major limiting factor of crop growth and productivity worldwide (Mahajan and Tuteja 2005)
heat shock proteins (Hsps) are normally classified according to their molecular weight as Hsp90, heat shock protein 70 (Hsp70), Hsp60, and small heat shock protein families
The recombinant clone was confirmed by Polymerase chain reaction (PCR) and double digestion of the pET28a-SbHsp70-1 construct with BamHI and XhoI (Fig. 1b)
Summary
Plants are constantly exposed to abiotic and biotic stresses, with abiotic stress being the major limiting factor of crop growth and productivity worldwide (Mahajan and Tuteja 2005). Stressed plants undergo various physiological and metabolic changes. They have evolved a wide range of mechanisms such as the induction of heat shock proteins (Hsps) to adapt to stressful conditions (Boston et al 1996; Waters et al 1996). Hsps are a class of ubiquitous and highly conserved proteins which are expressed in response to temperature or other environmental stresses (de Maio 1999). They are constitutively present in normal or stressed cells. Hsps are normally classified according to their molecular weight as Hsp, Hsp, Hsp, and small heat shock protein families. Hsp genes are encoded by a major Hsp multi-gene family made up of the cytosolic Hsp70s including the inducible Hsp and the cognate Hsc, glucose-regulated protein 78 (Grp78), and the
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