DIA-based proteome reveals the involvement of cuticular proteins and lipids in the wing structure construction in the silkworm.

Abstract Wing discs of Bombyx mori (B. mori) are transformed into wings during metamorphosis via dramatic morphological and structural changes. Mutations in genes related to the wings cause the adults to have altered wing shapes or abnormal wing colour. At present, there are more than 20 wing mutants recorded in the silkworm. However, the key factors that influence B. mori wing development are still unclear. Here, we used the strains +Wes/+Wes and Wes/+Wes that are typical for the normal wing and shriveled wing phenotypes, respectively, to identify differentially expressed proteins by label-free data-independent acquisition (DIA). Ten enriched GO terms and 9 KEGG pathways were identified based on the 3993 proteins in the wings. Among the identified and quantified proteins, 370 differentially expressed proteins (DEPs) were detected (P-value  0.58). Mapping of the DEPs to the reference canonical pathways in KEGG showed that the top 20% of the pathways were related to fatty acid, cutin, suberin and wax biosynthesis, protein processing in endoplasmic reticulum, protein export, etc. Of the 370 DEPs, 238 were down-regulated, and 132 were up-regulated of Wes/+Wes compared with +Wes/+Wes. Numerous cuticular proteins were down-regulated, and fatty metabolism enzymes were up-regulated, in Wes/+Wes compared with +Wes/+Wes. Significance The comparative analysis of proteomes suggested that cuticular proteins and fatty metabolism enzymes are the main abnormally expressed proteins in the pupal wings of Wes/+Wes, leading to curly and shrunken wings after moth transformation. Our results also identify the substances affecting the development of silkworm wings from the perspective of proteins. The information from this study is important for further research on the molecular mechanisms of wing development in lepidopteran insects, and these differentially expressed genes may be targets for Lepidoptera pest control.