Overexpression of cytochrome P450 CYP71AF43 contributing resistance to fenoxaprop-P-ethyl in Alopecurus myosuroides from China

Abstract

Black-grass (Alopecurus myosuroides), one of the most economically destructive herbicide-resistant weeds in Europe, is rapidly expanding in winter wheat regions of China. In recent years, the recommended application rate of fenoxaprop-P-ethyl in the field has failed to effectively control Alopecurus myosuroides populations, thereby threatening wheat yields at risk. In this study, we collected a suspected herbicide-resistant population (R-HB) of Alopecurus myosuroides from a wheat field in Hebei Province and confirmed its resistance to fenoxaprop-P-ethyl, with a resistance index of 26.73-fold. Sensitivity analyses of other ACCase-inhibiting herbicides revealed cross-resistance in the R-HB population to clethodim and pinoxaden. Molecular analysis indicated that the resistance phenotype in this population was not due to alterations in the target site. Pretreatment with the cytochrome P450 (P450) inhibitor malathion partially reversed fenoxaprop-P-ethyl resistance in the R-HB population. RNA-seq and RT-qPCR validation revealed the constitutive overexpression of the P450 gene CYP71AF43 in the R-HB population. Molecular docking predictions suggest that the CYP71AF43 protein may have metabolic activity toward fenoxaprop-P-ethyl. In genetically modified yeast, overexpression of AmCYP71AF43 was found to enhance tolerance to fenoxaprop-P-ethyl, but not to clethodim and pinoxaden. Additionally, rice calli overexpressing the AmCYP71AF43 gene exhibited resistance to fenoxaprop-P-ethyl, but not to clethodim or pinoxaden. Collectively, the increased expression of CYP71AF43 may enhance P450-mediated metabolism, conferring resistance to fenoxaprop-P-ethyl in the R-HB population. This is the first report of this mechanism in Alopecurus myosuroides. This discovery provides a novel perspective for the in-depth analysis of resistance mechanisms in weeds against the ACCase-inhibiting herbicide fenoxaprop-P-ethyl.