Functional characterization of the Pinellia ternata cytoplasmic class II small heat shock protein gene PtsHSP17.2 via promoter analysis and overexpression in tobacco

Abstract

High temperature is one of the main abiotic factors limiting agricultural production, particularly for heat-sensitive plant species. Small heat-shock proteins contribute substantially to alleviating damage to plants caused by heat stress. In the present study, the heat shock protein gene PtsHSP17.2 from Pinellia ternata was functionally characterized through promoter analysis and its overexpression in tobacco. Respectively, relative expression using real-time RT-PCR and ex situ promoter activity assay indicated that PtsHSP17.2 is strongly inducible under heat stress, and in silico promoter analysis discovered multiple stress-related cis elements including heat shock element. When overexpressing PtsHSP17.2 in tobacco, the thermotolerance of transgenic plants was markedly enhanced. Furthermore, the transgenic tobacco plants exhibited less variation in chlorophyll content, relative electrolyte leakage, and malondialdehyde content under heat stress compared with wild-type (WT) plants. The activities of antioxidant enzymes and content of proline were significantly enhanced under heat stress in transgenic plants relative to WT plants. Transgenic plants also had enhanced water retention and increased antioxidative capacity. Further, the expression levels of genes encoding antioxidant enzymes were more highly induced by heat stress in transgenic lines than WT. These results enrich the current understanding of thermal adaptation of heat-sensitive plant species and encourage further genetic improvement.