Biochemical and structural characterization of quizalofop-resistant wheat acetyl-CoA carboxylase

Raven Bough,Franck E Dayan

doi:10.1038/s41598-021-04280-x

Abstract

A novel nucleotide mutation in ACC1 resulting in an alanine to valine amino acid substitution in acetyl-CoA carboxylase (ACCase) at position 2004 of the Alopecurus myosuroides reference sequence (A2004V) imparts quizalofop resistance in wheat. Genotypes endowed with the homozygous mutation in one or two ACC1 homoeologs are seven- and 68-fold more resistant to quizalofop than a wildtype winter wheat in greenhouse experiments, respectively. In vitro ACCase activities in soluble protein extracts from these varieties are 3.8- and 39.4-fold more resistant to quizalofop with the homozygous mutation in either one or two genomes, relative to the wildtype. The A2004V mutation does not alter the specific activity of wheat ACCase, suggesting that this resistance trait does not affect the catalytic functions of ACCase. Modeling of wildtype and quizalofop-resistant wheat ACCase demonstrates that the A2004V amino acid substitution causes a reduction in the volume of the binding pocket that hinders quizalofop’s interaction with ACCase. Docking studies confirm that the mutation reduces the binding affinity of quizalofop. Interestingly, the models suggest that the A2004V mutation does not affect haloxyfop binding. Follow up in vivo and in vitro experiments reveal that the mutation, in fact, imparts negative cross-resistance to haloxyfop, with quizalofop-resistant varieties exhibiting higher sensitivity to haloxyfop than the wildtype winter wheat line.

Highlights

A novel nucleotide mutation in ACC1 resulting in an alanine to valine amino acid substitution in acetyl-CoA carboxylase (ACCase) at position 2004 of the Alopecurus myosuroides reference sequence (A2004V) imparts quizalofop resistance in wheat
Group 1 herbicides target the eukaryotic form of ACCase by binding to the carboxyl transferase domain and inhibiting the second catalytic s tep[5], whereas these herbicides have little activity on the multi-subunit chloroplastic form
The recently commercialized CoAXium® winter wheat production system consists of a specialized quizalofop formulation under the tradename Aggressor® for use on wheat varieties with the AXigen® quizalofop resistance trait coupled with a stewardship program aiming to delay resistance in weeds[7]

Summary

Introduction

A novel nucleotide mutation in ACC1 resulting in an alanine to valine amino acid substitution in acetyl-CoA carboxylase (ACCase) at position 2004 of the Alopecurus myosuroides reference sequence (A2004V) imparts quizalofop resistance in wheat. Group 1 herbicides target the eukaryotic form of ACCase by binding to the carboxyl transferase domain and inhibiting the second catalytic s tep[5], whereas these herbicides have little activity on the multi-subunit chloroplastic form. The nucleotide substitution in ACC1 imparting quizalofop resistance in CoAXium® wheat corresponds to position 2004 in the ACCase amino acid sequence from the Alopecurus myosuroides reference sequence[14]. This mutation causes an amino acid substitution of an alanine for a slightly larger valine residue in ACCase (A2004V). We further quantified fold-differences in whole-plant and enzymatic resistance between wheat with wildtype ACCase and wheat with one or two mutant ACCase homoeologs

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