The survival of species is negatively impacted by the increased frequency and intensity of high‐temperature episodes caused by increased global warming. Thermal tolerance can influence population size and the geographic distribution of species, which translates into species‐wide responses to climate and affects interactions between communities of species. The silkworm, Bombyx mori (L.) (Lepidoptera: Bombycidae), is an ectotherm and an internationally recognized model insect for studying sensitivity to the environment. In the present study, we investigated the effects of different diets on the thermal tolerance of silkworm. As such, it was identified that silkworms reared on an artificial diet in instars I–III and mulberry leaves in instar IV (referred as the AM group) exhibited faster weight gain and higher antioxidant enzyme activity at 35 °C, whereas silkworms reared only on mulberry leaves (referred as the ML group) exhibited upregulated transcript levels of key genes regulating antioxidant enzyme signaling pathways including Akt, CncC, and keap1, significantly increased reactive oxygen species (ROS) levels, and severely impaired midgut activity at 35 °C. Thus, the AM group presented stronger thermal tolerance. These findings suggest that diet can affect the thermal tolerance of silkworm. This research improved our understanding of the molecular, biochemical, and physiological changes caused by thermal stress and laid the groundwork for future research regarding the mechanism underlying thermal tolerance. Furthermore, this study provides an important reference for the conservation of biodiversity in the context of global warming.