Uridine diphosphate (UDP)-glycosyltransferases (UGTs) catalyze the conjugation of small lipophilic endogenous and exogenous compounds with sugars to produce water-soluble glycosides, playing an important role in insect endobiotic regulation and xenobiotic detoxification. In this study, two UGT-inhibitors, sulfinpyrazone and 5-nitrouracil, significantly increased spirotetramat toxicity against third instar nymphs of resistant Aphis gossypii, whereas there were no synergistic effects in apterous adult aphids, suggesting UGT involvement in spirotetramat resistance in cotton aphids. Furthermore, the UHPLC-MS/MS was employed to determine the content of spirotetramat and its four metabolites (S-enol, S-glu, S-mono, S-keto) in the honeydew of resistant cotton aphids under spirotetramat treatment. No residual spirotetramat was detected in the honeydew, while its four metabolites were detected at a S-enol: S-glu: S-mono: S-keto ratio of 69.30: 6.54: 1.44: 1.00. Therefore, glycoxidation plays a major role in spirotetramat inactivation and excretion in resistant aphids. Compared with the susceptible strain, the transcriptional levels of UGT344M2 were significantly upregulated in nymphs and adults of the resistant strain. RNA interference of UGT344M2 dramatically increased spirotetramat toxicity in nymphs, but no such effect were found in the resistant adult aphids. Overall, UGT-mediated glycoxidation were found to be involved in spirotetramat resistance. The suppression of UGT344M2 significantly increased the sensitivity of resistant nymphs to spirotetramat, suggesting that UGT344M2 upregulation might be associated with spirotetramat detoxification. This study provides an overview of the involvement of metabolic factors, UGTs, in the development of spirotetramat resistance.
Keywords: Aphis gossypii; Insecticide resistance; Spirotetramat; UDP-glucuronosyltransferase.
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