Development of insecticide resistance reduces the efficacy of controlling measures against the medical and agricultural insect pests. Cytochrome P450s are one of the major detoxification enzymes involved in insecticide metabolisms. Previously, we have reported that the P450 gene Cyp9m10 is about 260-fold overexpressed in a pyrethroid-resistant strain of Culex quinquefasciatus compared to a susceptible strain. In this study, we obtained direct evidence that the Cyp9m10 overexpression is caused by a cis-acting mutation. Additionally, a region of approximately 100 kb in length including the Cyp9m10 locus was specifically duplicated in the resistant strain. The two duplicated Cyp9m10 copies shared a completely identical sequence within the transcribed region and the flanking region up to the breakpoint located 1.1 kb upstream of the transcriptional start site. A Miniature Inverted-repeat Transposable Element (MITE)-like element was specifically inserted 0.2 kb upstream of both Cyp9m10 copies in the resistant strain. In backcross experiment, a haplotype containing the two duplicated Cyp9m10 copies was strongly associated with the pyrethroid resistance.

Copyright 2010 Elsevier Ltd. All rights reserved.

Although the importance of cis-acting mutations on detoxification enzyme genes for insecticide resistance is widely accepted, only a few of them have been determined as concrete mutations present in genomic DNA till date. The overexpression of a cytochrome P450 gene, CYP9M10, is associated with pyrethroid resistance in the southern house mosquito Culex quinquefasciatus. The haplotypes of CYP9M10 exhibiting overexpression (resistant haplotypes) belong to one specific phylogenetic lineage that shares high nucleotide sequence homology and the same insertion of a transposable element. Among the resistant haplotypes, allelic progression involving an additional cis-acting mutation and gene duplication evolved a CYP9M10 haplotype associated with extremely high transcription and strong pyrethroid resistance. Here we show that a single nucleotide substitution G-27A, which is located near the transcription start site of CYP9M10, is involved in the progression of the duplicated haplotype lineage. The deletion of a 7-bp AT-rich sequence that includes nucleotide -27 inhibited the initiation of transcription from the original transcriptional initiation site. The mutation was suspected to reside within a core promoter, TATA-box, of CYP9M10.

Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.