Identification of the 2-tridecanone cis-acting element in the promoter of cytochrome P450 CYP6B7 in Helicoverpa armigera.

Abstract

The expression level of cytochrome P450 genes in insects can be induced by plant allelochemicals, which is important for insects to adapt to host plants. Cytochrome P450 CYP6B7 has been reported to be involved in pyrethroid insecticide resistance in Helicoverpa armigera, and its transcription level was induced by some inducers. Currently, the regulatory mechanism of the induced expression of CYP6B7 remains unknown, although it is very important for understanding the detoxification mechanism to allelochemicals in host plants. The objective of the present study was to investigate the cis-acting element in the promoter of CYP6B7 mediating the inducible up-regulation of CYP6B7 in H. armigera by 2-tridecanone. The promoter region of CYP6B7 was cloned by genome walking technique and analyzed by transient transfection assay. Progressive 5' deletion of the promoter region of CYP6B7 revealed that the relative luciferase activity of construct -320/+232 could be significantly induced by 2-tridecanone. Further stepwise deletion between -320 and -238 bp found that construct -292/+232 could also be significantly induced by 2-tridecanone, but the adjacent construct -256/+232 could not, suggesting the essential role of the sequence between -292 and -257 bp for 2-tridecanone induction. Nucleotide mutations between -292 and -281 bp had no influence on the induction effect by 2-tridecanone, but nucleotide mutations between -280 and -257 bp significantly decreased the induction effect. These results demonstrated that the cis-acting element for 2-tridecanone induction was between -280 and -257 bp in the promoter of CYP6B7.