Biochemical characterization of acetylcholinesterase, cytochrome P450 and cross-resistance in an omethoate-resistant strain of Aphis gossypii Glover

Abstract

The degree of insecticide resistance, synergism effects, acetylcholinesterase (AChE) activity kinetics, specific activities of detoxification enzymes and cross-resistance were investigated in omethoate resistant and relatively susceptible strains of Aphis gossypii Glover. The resistant cotton aphid strain (ORR) exhibited 231.3-fold resistance to omethoate compared to the susceptible strain (OSS). Synergist piperonyl butoxide (PBO) dramatically increased the toxicity of omethoate in the resistant strain, while triphenyl phosphate (TPP) and diethyl maleate (DEM) did not exhibit synergism effects. The calculated AChE activity, V max and K m ratios of ORR to OSS were 0.1, 0.2 and 0.4, respectively. Based on analysis of IC 50 indices, enzyme inhibition experiments showed that AChE from the ORR strain was 10.6-, 3.2-, 6.2-, 10.5- and 4.4-fold more insensitive to inhibition by eserine, omethoate, paraoxon, paraoxon-methyl and malaoxon, respectively, than that from the OSS strain. The cytochrome P450-mediated O-demethylation activity (3.7-fold) and ethoxycoumarin-O-deethylase activity (2.6-fold) in the ORR strain were significantly higher than in the OSS strain. Specific activity of carboxylesterase (CarE) and glutathione S-transferase (GST) were not significantly different in both the ORR and OSS strains. Bioassay results indicated the ORR strain had developed high levels of cross-resistance to chlorpyrifos (24.2-fold), malathion (21.1-fold), acephate (10.2-fold), esfenvalerate (30.6-fold), methomyl (22.4-fold), carbofuran (33.2-fold), but had negative cross-resistance to bifenthrin (0.4-fold). Overall, these results demonstrate that reduced AChE sensitivity, combined with increased cytochrome P450 monooxygenase detoxification, plays an important role in the high levels of omethoate resistance and can cause cross-resistance to other insecticides in the ORR strain.