Quizalofop-resistant wheat: biochemical characterization of the AXigen™ trait and corresponding metabolism

Abstract

A new weed management tool in wheat, the CoAXium™ Wheat Production System, incorporates quizalofop-resistant wheat, a specialized formulation of quizalofop (Aggressor™), and a stewardship management program for effective management of annual grasses with otherwise limited control options. The AXigen™ trait confers resistance primarily through a single-point mutation in ACC1. The mutation causes an alanine to valine substitution at position 2004 in wheat acetyl-CoA carboxylase (ACCase) relative to the Alopecurus myosuriodes reference. Through greenhouse and biochemical studies paired with protein homology modelling and simulations, the research presented herein provides strong evidence that a conformational change imparted by the amino acid substitution results in quizalofop-resistant ACCase. Conversely, the mutation conveys negative cross-resistance to haloxyfop, a similar herbicide to quizalofop with a smaller molecular volume. The remaining research objectives focus on quizalofop metabolism in CoAXium™ wheat. Liquid chromatography-mass spectrometry measurements of quizalofop content over time from liquid demonstrate cooler temperature conditions (4.5°C) delay quizalofop metabolism by 4 times compared to warmer temperature conditions (19°C). Reduced temperatures also delay quizalofop metabolism to the same extent in the following annual grass weed species: Aegilops cylindrica, Bromus tectorum, and Secale cereale. Further, additional studies suggest herbicide metabolism mechanisms enhance overall CoAXium™ wheat quizalofop resistance. Despite similar ACCase resistance, resistant winter and spring wheat varieties convey varying degrees of whole-plant resistance. In winter wheat but not spring wheat, increased resistance corresponds to a shorter quizalofop half-life, implying faster metabolism boosts overall resistance. Treatment of resistant spring wheat varieties with cloquintocet, a metabolism-boosting safener, increases overall resistance. Follow-up differential expression analysis of cloquintocet-treated plants may support differential metabolism findings and lead to identification of putative candidate genes associated with upregulated herbicide metabolism, such as cytochrome P450 monooxygenases, glutathione-S-transferases, and glycosyltransferases.