Spodoptera frugiperda Sf9 cells as a model system to investigate the role of detoxification gene expression in response to xenobiotics

Abstract

Spodoptera frugiperda (fall armyworm) is a highly destructive invasive pest that feeds on numerous crops including maize and rice. It has developed sophisticated mechanisms to detoxify xenobiotics such as secondary plant metabolites as well as manmade insecticides. The aim of the study was to explore the detoxification response to plant secondary metabolites and insecticides employing a S. frugiperda Sf9 cell model exposed to indole 3-carbinol (I3C) and methoprene. The cell Inhibitory Concentration 50 (IC₅₀) for these molecules was determined and IC₁₀, IC₂₀ and IC₃₀ doses were used to monitor the induction profiles of detoxification genes. Cytochrome P450 monooxygenases (P450s) of the CYP9A subfamily were the most inducible genes of the seven examined. Our results also showed the induction of the transcription factor Cap‘n'collar isoform C (CncC). Transient transformation of Sf9 cells overexpressing CncC and its partner muscle aponeurosis fibromatosis (Maf) induces overexpression of CYP4M14, CYP4M15, CYP321A9 and GSTE1 while CYP9As were not induced. Next, we determined the capacity of recombinantly expressed CYP9A30, CYP9A31 and CYP9A32 to interact with methoprene and I3C. Fluorescence-based biochemical assays revealed an interaction of methoprene with functionally expressed CYP9A30, CYP9A31 and CYP9A32 whereas almost no interaction was detected for I3C, suggesting the ability of CYP9As to metabolize methoprene. Our results showed that Sf9 cells could be a useful model to decipher detoxification pathways of S. frugiperda.