Resistance to ALS inhibitors conferred by non-target-site resistance mechanisms in Myosoton aquaticum L

Myosoton aquaticum L. is a competitive broadleaf weed commonly found in wheat fields in China and has become challenging due to its evolving herbicide resistance. In this study, one subpopulation, RF1 (derived from the tribenuron-methyl-resistant population HN10), with none of the known acetolactate synthase (ALS) resistance mutations was confirmed to exhibit resistance to tribenuron-methyl (SU), pyrithiobac‑sodium (PTB), florasulam (TP), flucarbazone-Na (SCT), and diflufenican (PDS). In vitro ALS activity assays showed that the total ALS activity of RF1 was lower than that of the susceptible (S) population. However, there was no difference in ALS gene expression induced by tribenuron-methyl between the two populations. The combination of the cytochrome P450 monooxygenase (P450) inhibitor malathion and tribenuron-methyl resulted in the RF1 population behaving like the S population. The rapid P450-mediated tribenuron-methyl metabolism in RF1 plants was also confirmed by liquid chromatography tandem mass spectrometry (LC–MS/MS) analysis. In addition, approximately equal glutathione S-transferase (GST) activity was observed in RF1 and S plants of untreated and tribenuron-methyl treated groups. This study reported one M. aquaticum L. population without ALS resistance mutations exhibiting resistance to ALS inhibitors and the PDS inhibitor diflufenican, and the non-target-site resistance mechanism played a vital role in herbicide resistance. [Display omitted] •A Myosoton aquaticum L. subpopulation RF1 without known ALS resistance mutations exhibits resistance to tribenuron-methyl.•No difference occurred in ALS gene expression and enzyme sensitivity to herbicide between both populations.•Although GSTs responses to tribenuron-methyl, it seems not likely contribute to herbicide resistance in RF1 population.•The enhanced tribenuron-methyl metabolism mediated by P450s acts major role in herbicide resistance in RF1 population.•RF1 population evolved resistance to ALS inhibitors and PDS inhibitor diflufenican.