Annick Matejicek

Molecular Bases for Sensitivity to Acetyl-Coenzyme A Carboxylase Inhibitors in Black-Grass

In grasses, residues homologous to residues Ile-1,781 and Ile-2,041 in the carboxyl-transferase (CT) domain of the chloroplastic acetyl-coenzyme A (CoA) carboxylase (ACCase) from the grass weed black-grass (Alopecurus myosuroides [Huds.]) are critical determinants for sensitivity to two classes of ACCase inhibitors, aryloxyphenoxypropionates (APPs) and cyclohexanediones. Using natural mutants of black-grass, we demonstrated through a molecular, biological, and biochemical approach that residues Trp-2,027, Asp-2,078, and Gly-2,096 are also involved in sensitivity to ACCase inhibitors. In addition, residues Trp-2,027 and Asp-2,078 are very likely involved in CT activity. Using three-dimensional modeling, we found that the side chains of the five residues are adjacent, located at the surface of the inside of the cavity of the CT active site, in the vicinity of the binding site for APPs. Residues 1,781 and 2,078 are involved in sensitivity to both APPs and cyclohexanediones, whereas residues 2,027, 2,041, and 2,096 are involved in sensitivity to APPs only. This suggests that the binding sites for these two classes of compounds are overlapping, although distinct. Comparison of three-dimensional models for black-grass wild-type and mutant CTs and for CTs from organisms with contrasted sensitivity to ACCase inhibitors suggested that inhibitors fitting into the cavity of the CT active site of the chloroplastic ACCase from grasses to reach their active sites may be tight. The three-dimensional shape of this cavity is thus likely of high importance for the efficacy of ACCase inhibitors.

Cross-resistance patterns to ACCase-inhibiting herbicides conferred by mutant ACCase isoforms in Alopecurus myosuroides Huds. (black-grass), re-examined at the recommended herbicide field rate.

BACKGROUND Target-site-based resistance to acetyl-CoA carboxylase (ACCase) inhibitors in Alopecurus myosuroides Huds. is essentially due to five substitutions (Isoleucine-1781-Leucine, Tryptophan-2027-Cysteine, Isoleucine-2041-Asparagine, Aspartate-2078-Glycine, Glycine-2096-Alanine). Recent studies suggested that cross-resistance patterns associated with each mutation using a seed-based bioassay may not accurately reflect field resistance. The authors aimed to connect the presence of mutant ACCase isoform(s) in A. myosuroides with resistance to five ACCase inhibitors (fenoxaprop, clodinafop, haloxyfop, cycloxydim, clethodim) sprayed at the recommended field rate. RESULTS Results from spraying experiments and from seed-based bioassays were consistent for all mutant isoforms except the most widespread, Leucine-1781. In spraying experiments, Leucine-1781 ACCase conferred resistance to clodinafop and haloxyfop. Some plants containing Leucine-1781 or Alanine-2096 ACCase, but not all, were also resistant to clethodim. CONCLUSION Leucine-1781, Cysteine-2027, Asparagine-2041 and Alanine-2096 ACCases confer resistance to fenoxaprop, clodinafop and haloxyfop at field rates. Leucine-1781 ACCase also confers resistance to cycloxydim at field rate. Glycine-2078 ACCase confers resistance to all five herbicides at field rates. Only Glycine-2078 ACCase confers clethodim resistance under optimal application conditions. It may be that Leucine-1781 and Alanine-2096 ACCases may also confer resistance to clethodim in the field if the conditions are not optimal for herbicide efficacy, or at reduced clethodim field rates.