Chinese sprangletop (Leptochloa chinensis (L.) Nees) is one of main grass weeds invading Chinese rice fields. The target-site resistance (TSR) of cyhalofop-butyl have been widely reported in L. chinensis populations, but the non-target-site resistance (NTSR) mechanisms have not yet been well-characterized. This study aims to investigate the likely NTSR in a cyhalofop-butyl-resistant L. chinensis population (YZ-R), which was collected from Yangzhou city, Jiangsu Province, China. Dose-response assays showed the YZ-R population exhibited 191.6-fold resistance to cyhalofop-butyl, compared to the susceptible population (YZ-S). This resistance is not target-site based, because no mutations in the two ACCase genes were detected in the YZ-R plants compared to the YZ-S plants, and the ACCase genes expression levels were similar in YZ-S and YZ-R plants. In addition, the cytochrome P450 inhibitor malathion and piperonyl butoxide (PBO), and glutathione S-transferase (GST) inhibitor 4-chloro-7-nitrobenzoxadiazole (NBD-Cl) did not significantly reverse cyhalofop-butyl resistance in the YZ-R population. However, liquid chromatography-mass spectrometry (LC-MS) analysis indicated that the metabolic rates of cyhalofop acid in YZ-R plants was significantly faster (5 to 10- fold) than in YZ-S plants. Furthermore, the YZ-R population showed no cross-resistance to other ACCase-inhibiting herbicides. These results indicated that cyhalofop-butyl resistance in the YZ-R population is due to non-target-site based enhanced herbicide metabolism. Resistance in this population is likely involved in a specific detoxification enzyme, with possible high catalytic efficiency and strong substrate specificity, therefore leading to high-level and single resistance to cyhalofop-butyl.

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