Enhanced heat tolerance in transgenic silkworm via overexpression of Pyrococcus furiosus superoxide reductase

Abstract

Heat shock causes a serious harm to organisms by accelerating the production of reactive oxygen species (ROS). Pyrococcus furiosus superoxide reductase (PfSOR) is an enzyme that efficiently detoxifies ROS. In order to generate a silkworm strain with high heat tolerance for sericulture, we synthesized an artificial DNA sequence encoding PfSOR based on the codon bias of Bombyx mori. PfSOR was successfully overexpressed in transgenic silkworm (named A4SOR) and BmE cells, as determined by RT-PCR and western blot analyses. An SOR activity assay confirmed that the expressed enzyme was functional in A4SOR. After exposure to a temperature of 35 °C for 44 h, the mortality rate was about 30% lower in transgenic A4SOR than in non-transgenic silkworms. Moreover, transgene expression had no apparent effect on economic characteristics of silkworms. The heat tolerance of silkworm was thus enhanced by expressing an archaeal SOR; this can be useful for sericulture in regions where the average temperature exceeds the optimal environmental temperature for B. mori of 25 °C.

Keywords: Heat tolerance; Overexpression; Pyrococcus furiosus superoxide reductase (PfSOR); Reactive oxygen species (ROS); Silkworm; Transgenic.