Biochemical and molecular characterization of neonicotinoids resistance in the tarnished plant bug, Lygus lineolaris

In the southern United States, neonicotinoids are commonly applied as foliar insecticides to control sucking insect pests, such as the tarnished plant bug (TPB, Lygus lineolaris). In this study, spraying bioassays were conducted to determine the toxicity of five neonicotinoids and sulfoxaflor to susceptible and late fall field-collected TPB adults from Mississippi Delta region. Compared to a susceptible population, the field-collected TPBs exhibited the highest resistance to imidacloprid (up to 19.5-fold), a moderate resistance to acetamiprid (9.43-fold), clothianidin (13.68-fold), thiamethoxam (7.88-fold) and the least resistance to thiacloprid (4.61-fold) and sulfoxaflor (1.82-fold), respectively. A synergist study demonstrated that piperonyl butoxide (PBO) significantly increased the toxicity of imidacloprid and thiamethoxam by 22.2- and 15.3-fold, respectively, while triphenyl phosphate (TPP) and diethyl maleate (DEM) only showed 2-3-fold synergism to both neonicotinoids. In the field-collected TPBs, activities of the three detoxification enzymes esterase, glutathione S-transferase (GST) and CYP450 monooxygenase (P450) were significantly increased by 3.43-, 1.48- and 2.70-fold, respectively, when compared to the susceptible population. Additionally, after 48 h exposure to imidacloprid or thiamethoxam, resistant TPB adults exhibited elevated esterase activities, decreased GST activities, and no significant changes in P450 activities. Further examinations revealed that the expression of certain esterase and P450 detoxification genes were significantly elevated in resistant TPBs. Overall, these results suggest that elevated esterase and P450s expression and enzyme activity are key mechanisms for metabolic resistance in TPBs to neonicotinoids. Our findings also provide valuable information for selection and adoption of neonicotinoid insecticides for resistance management of TPBs and minimizing toxic risk to foraging bees.