Black-grass (Alopecurus myosuroides Huds.) is a common weed in Chinese wheat fields, and has become troublesome due to its evolution of herbicide resistance. One black-grass population (HN-14) collected from a wheat field where herbicides were applied was suspected to be resistant (R) to mesosulfuron-methyl. This study aims to establish a cross-resistance pattern and explore potential resistance mechanisms. The results of a whole-plant dose response assay showed that the resistant (R) population had a high of resistance to mesosulfuron-methyl (33-fold); meanwhile, no synergism of P450s activity inhibitor malathion was observed. The sequencing results revealed that ALS resistance mutation Trp-574-Leu occurred in R plants. The results of in vitro ALS enzyme activity assays also supported that the extractable ALS from R plants were 23.22-fold resistant to mesosulfuron-methyl. In the light of the “R” resistance rating system, HN-14 has evolved RRR and RR resistance to fenoxaprop-P-ethyl, clodinafop-propargyl, haloxyfop-methyl, and fluazifop-P-butyl and R? (resistance may be developing) to pinoxaden, however remains sensitive to imazethapyr, quizalofop-P-ethyl, tralkoxydim, and isoproturon. These results indicated that the mesosulfuron-methyl resistance in the black-grass population HN-14 was driven by a target-site mechanism rather than a nontarget (at least P450s-mediated) mechanism.

Amaranthus retroflexus L. is a highly competitive broadleaf weed of corn–soybean rotation in northeastern China. In recent years, the herbicide(s) resistance evolution has been threatening its effective management in crop fields. One resistant A. retroflexus (HW-01) population that survived the protoporphyrinogen oxidase (PPO) inhibitor fomesafen and acetolactate synthase (ALS) inhibitor nicosulfuron applied at their field-recommended rate was collected from a soybean field in Wudalianchi City, Heilongjiang Province. This study aimed to investigate the resistance mechanisms of fomesafen and nicosulfuron and determine the resistance profile of HW-01 to other herbicides. Whole plant dose–response bioassays revealed that HW-01 had evolved resistance to fomesafen (50.7-fold) and nicosulfuron (5.2-fold). Gene sequencing showed that the HW-01 population has a mutation in PPX2 (Arg-128-Gly) and a rare mutation in ALS (Ala-205-Val, eight/twenty mutations/total plants). In vitro enzyme activity assays showed that ALS extracted from the HW-01 plants was less sensitive to nicosulfuron (3.2-fold) than ST-1 plants. Pre-treatment with the cytochrome P450 inhibitors malathion, piperonyl butoxide (PBO), 3-amino-1,2,4-triazole (amitrole), and the GSTs inhibitor 4-chloro-7-nitrobenzofurazan (NBD-Cl) significantly increased fomesafen and nicosulfuron sensitivity in the HW-01 population compared with that of the sensitive (S) population ST-1. Moreover, the rapid fomesafen and nicosulfuron metabolism in the HW-01 plants was also confirmed via HPLC-MS/MS analysis. Furthermore, the HW-01 population showed multiple resistance (MR) to PPO, ALS, and PSII inhibitors, with resistance index (RI) values ranging from 3.8 to 9.6. This study confirmed MR to PPO-, ALS-, and PSII-inhibiting herbicides in the A. retroflexus population HW-01, as well as confirming that the cytochrome P450- and GST-based herbicide metabolic along with TSR mechanisms contribute to their multiple resistance to fomesafen and nicosulfuron.

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