Insect resistance to chemical insecticides is attributed to a combination of different mechanisms, such as metabolic resistance, knockdown resistance, and the cuticular resistance or penetration factor. The insect integument offers an efficient barrier against contact insecticides and its role as penetration factor has been previously reported; however, there is no information about its potential function in the metabolic resistance. Cytochrome P450 genes (CYP) are highly expressed in the fat body of several insects and thus play a key role in their metabolic resistance. Here, we describe new members that belong to the highly genome-wide expanded CYP3093A and CYP4EM subfamilies in the Chagas disease vectors Rhodnius prolixus and Triatoma infestans. We modeled the docking of deltamethrin in their active site and detected differences in some amino acids between both species that are critical for a correct interaction with the substrate. We also knocked down the two constitutively most expressed genes in the integument of resistant T. infestans nymphs (CYP3093A11 and CYP4EM10) in order to find clues on their participation in deltamethrin resistance. This is the first report on the role of the insect integument in detoxification events; although these two CYP genes do not fully explain the resistance observed in T. infestans.

The insect integument, formed by the cuticle and the underlying epidermis, is essential for insect fitness, regulation of lipid biosynthesis and storage, insect growth and feeding, together with development progress. Its participation in insecticide resistance has also been outlined. Triatoma infestans Klug (Hemiptera: Reduviidae) is one of the major vectors of Chagas disease in South America; however, genomic data are scarce. In this study, we performed a transcriptome analysis of the nymph integument in order to identify which genes are expressed and their putative role. Using the 454 GS-FLX sequencing platform, we obtained approximately 144,620 reads from the integument tissue. These reads were assembled into 6,495 isotigs and 8,504 singletons. Based on BLAST similarity searches, about 8,000 transcripts were annotated with known genes, conserved domains, and/or Gene Ontology terms.The most abundant transcripts corresponded to transcription factors and nucleic acid metabolism, membrane receptors, cell signaling, and proteins related to cytoskeleton, transport, and cell energy processes, among others. More than 10% of the transcripts-encoded proteins putatively involved in the metabolism of fatty acids and related components (fatty acid synthases, elongases, desaturases, fatty alcohol reductases), structural integument proteins, and the insecticide detoxification system (among them, cytochrome P450s, esterases, and glutathione transferases). Real-time qPCR assays were used to investigate their putative participation in the resistance mechanism. This preliminary study is the first transcriptome analysis of a triatomine integument, and together with prior biochemical information, will help further understandthe role of the integument in a wide array of mechanisms.

Fatty acid synthase is a multifunctional enzyme involved in the formation of fatty acids. Despite the role of fatty acids in cell signalling and energy metabolism, and as precursors to pheromones and hydrocarbons that waterproof the cuticle, the insect fatty acid synthases have been scarcely studied. Here we perform the molecular characterization of three fatty acid synthase genes (fatty acid synthase RPRC000123, RPRC000269 and RPRC002909) in the Chagas disease vector, Rhodnius prolixus. Gene expression screening by reverse transcription quantitative PCR showed that RPRC000123 and RPRC002909 are expressed almost exclusively in the integument tissue whilst RPRC000269 is mostly expressed in the fat body and also in several body organs. Phylogenetic analysis, together with gene expression results, showed that RPRC000269, RPRC002909 and RPRC000123 are orthologues of Drosophila melanogaster fatty acid synthase 1 (FASN1), FASN2 and FASN3 genes, respectively. After RNA interference‐mediated knockdown of RPRC000123, insects died immediately after moulting to the next developmental stage. However, mortality was prevented by placing the insects under saturated humidity conditions, suggesting that dehydration might play a role in the insects’ death. Lipid analyses in RPRC000123‐silenced insects showed reduced amounts of integument fatty acids and methyl‐branched hydrocarbons, compared to controls. These data support an important role for FASN3 in the biosynthesis of the precursors to hydrocarbons that waterproof the insect cuticle.