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The bivoltine strain of the domestic silkworm,  Bombyx mori , exhibits a facultative diapause phenotype that is determined by maternal environmental conditions during embryonic and larval development. Although a recent study implicated a circadian clock gene  period  ( per ) in circadian rhythms and photoperiod-induced diapause, the roles of other core feedback loop genes, including  timeless  ( tim ),  Clock  ( Clk ),  cycle  ( cyc ), and  cryptochrome2  ( cry2 ), have to be clarified yet. Therefore, the aim of this study was to elucidate the roles of circadian clock genes in temperature-dependent diapause induction. To achieve this,  per ,  tim ,  Clk ,  cyc , and  cry2  knockout (KO) mutants were generated, and the percentages of diapause and non-diapause eggs were determined. The results show that  per ,  tim ,  Clk ,  cyc , and  cry2  regulated temperature-induced diapause by acting upstream of cerebral γ-aminobutyric acid (GABA)ergic and diapause hormone signaling pathways. Moreover, the temporal expression of the clock genes in wild-type ( wt ) silkworms was significantly different from that of thermosensitive transient receptor potential ankyrin 1 (TRPA1) KO mutants during embryonic development. Overall, the findings of this study provide target genes for regulating temperature-dependent diapause induction in silkworms.

Circadian Clock Genes Regulate Temperature-Dependent Diapause Induction in Silkworm Bombyx mori