Abstract

BackgroundSpinosad is important in pest management strategies of multiple insect pests. However, spinosad resistance is emerging in various pest species. Resistance has in some species been associated with alterations of the target-site receptor, but in others P450s seems to be involved. We test the possible importance of nine cytochrome P450 genes in the spinosad-resistant housefly strain 791spin and investigate the influence of spinosad on P450 expression in four other housefly strains.ResultsSignificant differences in P450 expression of the nine P450 genes in the four strains after spinosad treatment were identified in 40% of cases, most of these as induction. The highly expressed CYP4G2 was induced 6.6-fold in the insecticide susceptible WHO-SRS females, but decreased 2-fold in resistant 791spin males. CYP6G4 was constitutively higher expressed in the resistant strain compared to the susceptible strain. Furthermore, CYP6G4 gene expression was increased in susceptible WHO-SRS flies by spinosad while the expression level did not alter significantly in resistant fly strains. Expression of CYP6A1 and male CYP6D3 was constitutively higher in the resistant strain compared to the susceptible. However, in both cases male expression was higher than female expression.Conclusion CYP4G2, CYP6A1, CYP6D3 and CYP6G4 have expressions patterns approaching the expectations of a hypothesized sex specific spinosad resistance gene. CYP4G2 fit requirements of a spinosad resistance gene best, making it the most likely candidate. The overall high expression level of CYP4G2 throughout the strains also indicates importance of this gene. However, the data on 791spin are not conclusive concerning spinosad resistance and small contributions from multiple P450s with different enzymatic capabilities could be speculated to do the job in 791spin. Differential expression of P450s between sexes is more a rule than an exception. Noteworthy differences between spinosad influenced expression of P450 genes between a field population and established laboratory strains were shown.

Highlights

  • Spinosad is a mixture of two macrocyclic lactones; spinosyn A and spinosyn D isolated from the actinomycete bacteria, Saccharopolyspora spinosa and has been developed as a commercial insecticide [1]

  • Enhancement of insecticide detoxification has been associated with increased P450 expression of genes leading to resistance [11]

  • The present experimental setup was selected in order to elucidate which, if any, of the nine P450 genes associated with resistance in the housefly could be responsible or partly responsible for spinosad resistance in the 791spin strain

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Summary

Introduction

Spinosad is a mixture of two macrocyclic lactones; spinosyn A and spinosyn D isolated from the actinomycete bacteria, Saccharopolyspora spinosa and has been developed as a commercial insecticide [1]. The mechanism of spinosad resistance has been implied to involve changes of target site and metabolism [4]. Field populations of houseflies collected from livestock farms throughout Denmark showed up to 7.5-fold resistances (compared to the susceptible reference strain WHOSRS) to spinosad in feeding bioassays, which was considered to reflect the natural variation of spinosad toxicity in field populations [5]. Spinosad is important in pest management strategies of multiple insect pests. Spinosad resistance is emerging in various pest species. Resistance has in some species been associated with alterations of the target-site receptor, but in others P450s seems to be involved. We test the possible importance of nine cytochrome P450 genes in the spinosadresistant housefly strain 791spin and investigate the influence of spinosad on P450 expression in four other housefly strains

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