Buprofezin Is Metabolized by CYP353D1v2, a Cytochrome P450 Associated with Imidacloprid Resistance in Laodelphax striatellus

Abstract

CYP353D1v2 is a cytochrome P450 related to imidacloprid resistance in Laodelphax striatellus. This work was conducted to examine the ability of CYP353D1v2 to metabolize other insecticides. Carbon monoxide difference spectra analysis indicates that CYP353D1v2 was successfully expressed in insect cell Sf9. The catalytic activity of CYP353D1v2 relating to degrading buprofezin, chlorpyrifos, and deltamethrin was tested by measuring substrate depletion and analyzing the formation of metabolites. The results showed the nicotinamide–adenine dinucleotide phosphate (NADPH)-dependent depletion of buprofezin (eluting at 8.7 min) and parallel formation of an unknown metabolite (eluting 9.5 min). However, CYP353D1v2 is unable to metabolize deltamethrin and chlorpyrifos. The recombinant CYP353D1v2 protein efficiently catalyzed the model substrate p-nitroanisole with a maximum velocity of 9.24 nmol/min/mg of protein and a Michaelis constant of Km = 6.21 µM. In addition, imidacloprid was metabolized in vitro by the recombinant CYP353D1v2 microsomes (catalytic constant Kcat) 0.064 pmol/min/pmol P450, Km = 6.41 µM. The mass spectrum of UPLC-MS analysis shows that the metabolite was a product of buprofezin, which was buprofezin sulfone. This result provided direct evidence that L. striatellus cytochrome P450 CYP353D1v2 is capable of metabolizing imidacloprid and buprofezin.

Deltamethrin is metabolized by CYP6FU1, a cytochrome P450 associated with pyrethroid resistance, in Laodelphax striatellus

“…CYP6AY3v2 could hydrolyze only imidacloprid, 16 and CYP353D1v2 could metabolize both imidacloprid 17 and buprofezin. 18 None of the tested CYP genes could degrade chlorpyrifos. However, the over-expressed esterase identified together with these five CYP genes was found to function in resistance to different insecticides, including chlorpyrifos.…”

“…Among the tested insecticides, CYP439A1v3 (Miah MA, unpublished) and CYP6FU1 were demonstrated to metabolize only deltamethrin. CYP6AY3v2 could hydrolyze only imidacloprid, and CYP353D1v2 could metabolize both imidacloprid and buprofezin . None of the tested CYP genes could degrade chlorpyrifos.…”

“…Previous work found that five CYP genes were over‐expressed in a deltamethrin‐resistant strain of L. striatellus, and experiments with in vitro expressed enzymes and transformed fruit flies have confirmed that CYP6AY3v2 can hydrolyze imidacloprid, CYP353D1v2 can metabolize both imidacloprid and buprofezin, and CYP439A1v3 can metabolize deltamethrin (Miah MA, unpublished). In this study, we report findings on another CYP enzyme, CYP6FU1, which was functionally expressed in Sf9 cells to examine its ability to metabolize deltamethrin, chlorpyrifos, imidacloprid and traditional CYP probing substrates.…”

Functionally characterized arthropod pest and pollinator cytochrome P450s associated with xenobiotic metabolism

“…S. litura CYP9A40 is implicated in resistance/tolerance of both insecticides (deltamethrin (pyrethroids), methoxyfenozide (diacylhydrazines)) and natural compounds (cinnamic acid, quercetin (flavonoid)) (Wang et al, 2015b). (Homoptera, Hemiptera, Diptera, Coleoptera, Orthoptera) L. striatellus CYP353D1v2 is able to degrade the chemically unrelated insecticides buprofezin and imidacloprid (neonicotinoid) (Elzaki et al, 2017), while CYP417A2v2, CYP439A1v3, CYP4C71 have been found to metabolize chemical substrates: imidacloprid (neonicotinoid), deltamethrin (pyrethroid) and imidacloprid (neonicotinoid), respectively (Miah et al, 2019(Miah et al, , 2017Xiao et al, 2020). CYP6CW1 has been associated with buprofezin and pymetrozine (pyridine azomethine derivative) resistance, CYP4DE1 and CYP6W3v2 with ethiprole (phenyl pyrazole) resistance (Elzaki et al, 2015;Zhang et al, 2015).…”

An overexpressed cytochrome P450 CYP439A1v3 confers deltamethrin resistance in Laodelphax striatellus Fallén (Hemiptera: Delphacidae)

“…The first three P450s have been functionally expressed in vitro and tested for insecticide degradation. Among these three P450s, CYP353D1v2 was found capable of catalyzing the degradation of imidacloprid (Elzaki, Miah, Wu, et al, ) and buprofezin (Elzaki, Miah, & Han, ), whereas CYP6AY3v2 catalyzed imidacloprid degradation (R. Wang et al, ). Only CYP6FU1 was found capable of deltamethrin degradation (Elzaki, Miah, Peng, et al, ).…”