Suppressing the expression of glutathione S-transferase gene GSTd10 increases the sensitivity of Zeugodacus cucurbitae against β-cypermethrin

Abstract

Zeugodacus cucurbitae Coquillett (Diptera: Tephritidae) is an agriculturally and economically important pest worldwide that has developed resistance to β-cypermethrin. Glutathione S-transferases (GSTs) have been reported to be involved in the detoxification of insecticides in insects. We have found that both ZcGSTd6 and ZcGSTd10 were up-regulated by β-cypermethrin induction in our previous study, so we aimed to explore their potential relationship with β-cypermethrin tolerance in this study. The heterologous expression of ZcGSTd6 and ZcGSTd10 in Escherichia coli showed significantly high activities against 1-chloro-2,4-dinitrobenzene (CDNB). The kinetic parameters of ZcGSTd6 and ZcGSTd10 were determined by Lineweaver–Burk. The V_max and K_m of ZcGSTd6 were 0.50 μmol/min·mg and 0.3 mM, respectively. The V_max and K_m of ZcGSTd10 were 1.82 μmol/min·mg and 0.53 mM. The 3D modelling and molecular docking results revealed that β-cypermethrin exhibited a stronger bounding to the active site SER-9 of ZcGSTd10. The sensitivity to β-cypermethrin was significantly increased by 18.73% and 27.21%, respectively, after the knockdown of ZcGSTd6 and ZcGSTd10 by using RNA interference. In addition, the inhibition of CDNB at 50% (IC₅₀) and the inhibition constants (Ki) of β-cypermethrin against ZcGSTd10 were determined as 0.41 and 0.33 mM, respectively. The Ki and IC₅₀ of β-cypermethrin against ZcSGTd6 were not analysed. These results suggested that ZcGSTd10 could be an essential regulator involved in the tolerance of Z. cucurbitae to β-cypermethrin.