Identification and expression analysis of cuticular protein genes in the diamondback moth, Plutella xylostella (Lepidoptera: Plutellidae).

Structural cuticular proteins (CPs) are major components of the insect cuticle, and they play critical roles in insect development and insecticide resistance. Here, a total of 196 CP genes were successfully annotated in the Plutella xylostella genome. On the basis of motif analysis, these CPs were classified into 10 different families, including 122 CPR, 12 CPAP1, 8 CPAP3, 9 CPLCP, 2 Tweedle, 1 CPF, 1 CPFL, 1 CPCFC, 17 CPG and 2 18 aa proteins, and the remaining 21 unclassified CPs were classed as cuticular proteins hypothetical (CPH). A phylogenetic analysis of CPs from different insects revealed species-specific clades of RR-1 and RR-2 genes, suggesting that CP gene duplication might occur independently among insect taxa, while we also found that some other CPs (such as CPAP1 and CPAP3) had a closer relationship based on their conserved domain architecture. Using available RNAseq libraries, the expression profiles of the CPs were analyzed over the four developmental stages of the insect (i.e., egg, larva, pupa, and adult), revealing stage-specific expression patterns for the CPs. In a chlorpyrifos resistant strain, 18 CP genes were found to be more than two-fold upregulated compared to the susceptible control strain, and qRT-PCR analysis showed that these CP genes were overexpressed after exposure to chlorpyrifos, suggesting a potential role in the molecular mechanism of insecticide resistance in P. xylostella. This study provides the tools and molecular basis to study the role of CPs in the post-embryonal development and the mechanisms of insecticide resistance of P. xylostella.