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Background: Soybean pod borer (Leguminivora glycinivorella) is an important soybean pest in north-eastern Asia, whose mature larvae overwinter in a diapause state. Disruption of winter diapause may be a valuable tool in pest management. However, the molecular mechanisms regulating diapause in this species have not yet been elucidated., Results: We compared the transcriptomes and proteomes between diapause and mature larvae and between mature and newly developed pupae to identify the genes and proteins associated with diapause. Thirty-seven differentially expressed genes and their proteins changed synchronously between diapause and mature larvae and 82 changed synchronously between diapause larvae and newly developed pupae. Among these, genes involved in fatty acid biosynthesis and the longevity regulating pathway were up-regulated in diapause larvae and down-regulated in newly developed pupae, suggesting that they may regulate diapause. One fatty acid synthase (FAS) gene and two small heat shock genes (HSP19.8 and HSP18.9) were chosen for further functional analysis. After RNA interference (RNAi)-mediated knockdown of FAS, the survival of mature larvae was significantly lower than that of control larvae, but the mean developmental time from first-instar larva to adult remained unchanged. RNAi-mediated knockdown of HSP19.8 and HSP18.9 severely shortened the mean developmental time, causing approximately 50% larvae to develop directly into pupae., Conclusion: FAS and the small heat shock gene play roles in diapause regulation and larvae survival. This study provides important information that may assist in understanding the molecular regulatory mechanisms of overwintering diapause of this important agricultural insect pest. © 2020 Society of Chemical Industry., (© 2020 Society of Chemical Industry.)