Recurrent Selection of Echinochloa crus-galli with a Herbicide Mixture Reduces Progeny Sensitivity.

Herbicide mixtures are used to increase the spectrum of weed control and to manage weeds with target-site resistance to some herbicides. However, the effect of mixtures on the evolution of herbicide resistance caused by enhanced metabolism is unknown. This study evaluated the effect of a fenoxaprop-p-ethyl and imazethapyr mixture on the evolution of herbicide resistance in Echinochloa crus-galli using recurrent selection at sublethal doses. The progeny from second generations selected with the mixture had lower control than parental plants or the unselected progeny. GR50 increased 1.6- and 2.6-fold after two selection cycles with the mixture in susceptible (POP1-S) and imazethapyr-resistant (POP2-IR) biotypes, respectively. There was evidence that recurrent selection with this sublethal mixture had the potential to evolve cross-resistance to diclofop, cyhalofop, sethoxydim, and quinclorac. Mixture selection did not cause increased relative expression for a set of analyzed genes (CYP71AK2, CYP72A122, CYP72A258, CYP81A12, CYP81A14, CYP81A21, CYP81A22, and GST1). Fenoxaprop, rather than imazethapyr, is the main contributor to the decreased control in the progenies after recurrent selection with the mixture in low doses. This is the first study reporting the effect of a herbicide mixture at low doses on herbicide resistance evolution. The lack of control using the mixture may result in decreased herbicide sensitivity of the weed progenies. Using mixtures may select important detoxifying genes that have the potential to metabolize herbicides in patterns that cannot currently be predicted. The use of fully recommended herbicide rates in herbicide mixtures is recommended to reduce the risk of this type of resistance evolution.

Mechanisms of evolved herbicide resistance.

The widely successful use of synthetic herbicides over the past 70 years has imposed strong and widespread selection pressure, leading to the evolution of herbicide resistance in hundreds of weed species. Both target-site resistance (TSR) and nontarget-site resistance (NTSR) mechanisms have evolved to most herbicide classes. TSR often involves mutations in genes encoding the protein targets of herbicides, affecting the binding of the herbicide either at or near catalytic domains or in regions affecting access to them. Most of these mutations are nonsynonymous SNPs, but polymorphisms in more than one codon or entire codon deletions have also evolved. Some herbicides bind multiple proteins, making the evolution of TSR mechanisms more difficult. Increased amounts of protein target, by increased gene expression or by gene duplication, are an important, albeit less common, TSR mechanism. NTSR mechanisms include reduced absorption or translocation and increased sequestration or metabolic degradation. The mechanisms that can contribute to NTSR are complex and often involve genes that are members of large gene families. For example, enzymes involved in herbicide metabolism-based resistances include cytochromes P450, GSH S-transferases, glucosyl and other transferases, aryl acylamidase, and others. Both TSR and NTSR mechanisms can combine at the individual level to produce higher resistance levels. The vast array of herbicide-resistance mechanisms for generalist (NTSR) and specialist (TSR and some NTSR) adaptations that have evolved over a few decades illustrate the evolutionary resilience of weed populations to extreme selection pressures. These evolutionary processes drive herbicide and herbicide-resistant crop development and resistance management strategies.