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Multiple resistance mechanisms to ALS inhibitors and auxin mimics in two Papaver rhoeas populations were investigated in wheat fields from Portugal. Dose-response trials, also with malathion (a cytochrome P450 inhibitor), cross-resistance patterns for ALS inhibitors and auxin mimics, alternative herbicides tests, 2,4-D and tribenuron-methyl absorption, translocation and metabolism experiments, together with ALS activity, gene sequencing and enzyme modelling and ligand docking were carried out. Results revealed two different resistant profiles: one population (R1) multiple resistant to tribenuron-methyl and 2,4-D, the second (R2) only resistant to 2,4-D. In R1, several target-site mutations in Pro197 and enhanced metabolism (cytochrome P450-mediated) were responsible of tribenuron-methyl resistance. For 2,4-D, reduced transport was observed in both populations, while cytochrome P450-mediated metabolism was also present in R1 population. Moreover, this is the first P. rhoeas population with enhanced tribenuron-methyl metabolism. This study reports the first case for P. rhoeas of the amino acid substitution Pro197Phe due to a double nucleotide change. This double mutation could cause reduced enzyme sensitivity to most ALS inhibitors according to protein modelling and ligand docking. In addition, this study reports a P. rhoeas population resistant to 2,4-D, apparently, with reduced transport as the sole resistance mechanism. [Display omitted] • Tribenuron P450-mediated metabolism is first reported in Papaver rhoeas. • The double mutation Pro197Phe is found for the first time in P. rhoeas , second world case. • Ligand docking predicts cross resistance to ALS inhibitors for the Pro197Phe. • Impaired transport alone could confer cross-resistance to all auxin mimics. • Various resistance mechanisms evolve in P. rhoeas to ALS inhibitors and auxin mimics. [ABSTRACT FROM AUTHOR]

The evolution of herbicide resistance in weeds has emerged as one of the most serious threats to sustainable food production systems, which necessitates the evaluation of herbicides to determine their efficacy. The first herbicide resistance case in the Iberian Peninsula was reported about 50 years ago, wherein Panicum dichotomiflorum was found to be resistant (R) to atrazine in Spanish maize fields. Since then, herbicide resistance has evolved in 33 weed species, representing a total of 77 single-herbicide-resistance cases in this geographic area: 66 in Spain and 11 in Portugal. Changes in agricultural practices, namely the adoption of non-tillage systems and the increased use of herbicides, led to the selection of weed biotypes resistant to a wide range of herbicides. Nowadays the most important crops in Spain and Portugal (maize, winter cereals, rice, citrus, fruits, and olive orchards) are affected, with biotypes resistant to several mechanisms of action (MoAs), namely: ALS inhibitors (20 species), ACCase inhibitors (8 species), PS II inhibitors (18 species), and synthetic auxin herbicides (3 species). More recently, the fast increase in cases of resistance to the EPSPS-inhibiting herbicide glyphosate has been remarkable, with 11 species already having evolved resistance in the last 10 years in the Iberian Peninsula. The diversity of resistance mechanisms, both target-site and non-target-site, are responsible for the resistance to different MoAs, involving point mutations in the target site and enhanced rates of herbicide detoxification, respectively. More serious are the 13 cases reported with multiple-herbicide resistance, with three cases of resistance to three–four MoAs, and one case of resistance to five MoAs. Future research perspectives should further study the relationship between management strategies and the occurrence of TSR and NTSR resistance, to improve their design, develop monitoring and diagnostic tools for herbicide resistance, and deepen the study of NTSR resistance. [ABSTRACT FROM AUTHOR]