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BACKGROUNDChlorantraniliprole (CAP) is widely used in agriculture and forestry to prevent and control pests. The effects of environmental CAP residue on non‐target insect metamorphosis have not been reported. Our research aimed to investigate the sublethal effect of CAP on larva–pupa transformation in silkworm, and explore the mechanism of sublethal CAP exposure‐mediated pupation metamorphosis defects.RESULTSublethal CAP exposure affected the growth and development of silkworm larvae and caused defects in pupation metamorphosis. After CAP exposure, formation the of prepupa procuticle, ecdysial membrane and new epidermis was inhibited. Also, the level of 20‐hydroxyecdysone (20E) and mRNA levels of the 20E signaling pathway‐related genes EcR, USP, E74, E75 and Ftz‐f1 were significantly reduced. Moreover, genes involved in chitin synthesis, such as ChsA, CDA1 and CDA2, were downregulated. Injection of 20E led to the upregulation of chitin synthesis‐related genes and increased formation of new epidermis in CAP‐treated silkworm. However, injection of 20E failed to prevent downregulation of Ftz‐f1 and the defects in pupation metamorphosis.CONCLUSIONOur results suggested that 20E is a target hormone of CAP exposure‐mediated epidermis formation phenotype. Ftz‐f1 was silenced by CAP and might be a direct target gene of sublethal CAP exposure. Our study provided new evidence of the effects of sublethal CAP exposure on insect development and metamorphosis. © 2020 Society of Chemical Industry