Aldehyde oxidase is coamplified with the World's most common Culex mosquito insecticide resistance‐associated esterases

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Abstract
The evolution and spread of insecticide resistance is an important factor in human disease prevention and crop protection. The mosquito Culex quinquefasciatus is the main vector of the disease filariasis and a member of a species complex which is a common biting nuisance worldwide. The common insecticide resistance mechanism in this species involves germline amplification of the esterases estα21 and estβ21. This amplification has arisen once and rapidly spread worldwide. Less common and more variable resistance phenotypes involve coamplification of estα3 and estβ1, or individual amplification of a single estβ1, different alleles of the same estα and estβ gene loci. Estα21 and estβ21 are on the same large fragment of amplified DNA (amplicon) 2.7 kb apart. We have now shown that this amplicon contains another full‐length gene immediately 5′ of estα21 which codes for a molybdenum‐containing hydroxylase, with highest homology to aldehyde oxidase (AO) from other organisms. The full‐length putative AO gene is not present on the estα3/estβ1 or estβ1 amplicons, but multiple truncated 5′ ends of this gene are present around the presumed estα3/estβ1 amplicon breakpoint. Polymerase chain reaction (PCR) analysis of insecticide‐susceptible genomic DNA demonstrated that a different allele of the putative AO gene in its non‐amplified form is immediately 5′ of estα. The ‘AO’ gene on the estα21/estβ21 amplicon is expressed and resistant insects have greater AO activity. This AO activity is sensitive to inhibition by an aldehyde‐containing herbicide and pesticide. This enzyme may confer a selective advantage to these insects in the presence of insecticide, as AO in mammals is believed to be important in the detoxification process of several environmental pollutants.