sHsp22.6, an intronless small heat shock protein gene, is involved in stress defence and development in Apis cerana cerana

Abstract

Small heat shock proteins (sHSPs) play an important role in protecting against stress-induced cell damage and fundamental physiological processes. In this study, we identified an intronless sHsp gene from Apis cerana cerana (AccsHsp22.6). The open reading frame of AccsHsp22.6 was 585 bp and encoded a 194 amino acid protein. Furthermore, a 2064 bp 5'-flanking region was isolated, and potential transcription factor binding sites associated with development and stress response were identified. Quantitative PCR and western blot analyses demonstrated that AccsHsp22.6 was detected at higher levels in the midgut than in other tissues tested, and it is highly expressed during the shift to different development stages. Moreover, AccsHsp22.6 was significantly up-regulated by abiotic and biotic stresses, such as 4 °C, 16 °C, 42 °C, cyhalothrin, pyridaben, H2O2, UV, CdCl2, 20-hydroxyecdysone and Ascosphaera apis treatments. However, AccsHsp22.6 was slightly repressed by other stresses, including 25 °C, phoxim, paraquat and HgCl2 treatments. The recombinant AccsHSP22.6 also exhibited significant temperature tolerance, antioxidation and molecular chaperone activity. In addition, we found that knockdown of AccsHsp22.6 by RNA interference remarkably reduced temperature tolerance in A. cerana cerana. Taken together, these results suggest that AccsHsp22.6 plays an essential role in the development stages and defence against cellular stress.

Keywords: Apis cerana cerana; Expression analysis; RNA interference; Stresses response; sHsp.