Molecular and physiological characterization of six-way resistance in an Amaranthus tuberculatus var. rudis biotype from Missouri

BACKGROUND Previous research reported the first case of six-way herbicide resistance in a common waterhemp (Amaranthus tuberculatus var. rudis) biotype from Missouri, USA designated MO-Ren. This study investigated the mechanisms of multiple-resistance in the MO-Ren biotype to herbicides from six site-of-action (SOA) groups, i.e. synthetic auxins, 5-enolypyruvyl-shikimate-3-phosphate synthase (EPSPS)-, protoporphyrinogen oxidase (PPO)-, acetolactate synthase (ALS)-, photosystem II (PSII)-, and 4-hydroxyphenyl-pyruvate-dioxygenase (HPPD)-inhibitors. RESULTS CONCLUSION Genomic DNA sequencing confirmed the presence of known mutations associated with ALS- or PPO-inhibiting herbicide resistance: the Trp-574-Leu amino acid substitution in the ALS enzyme and the codon deletion corresponding to the Delta G210 in the PPX2 enzyme. No target-site point mutations associated with resistance to PSII- and EPSPS-inhibitors were detected. Quantitative polymerase chain reaction (qPCR) indicated that MO-Ren plants contained five-fold more copies of the EPSPS gene than susceptible plants. Malathion in combination with 2,4-D (2,4-dichlorophenoxyacetic acid), mesotrione, and chlorimuron POST enhanced the activity of these herbicides indicating that metabolism due to cytochrome P450 monooxygenase activity was involved in herbicide resistance. 4-Chloro-7-nitrobenzofurazan (NBD-Cl), a glutathione-S-transferase (GST)-inhibitor, in combination with atrazine did not reduce the biomass accumulation. Reduced absorption or translocation of 2,4-D did not contribute to resistance. However, the resistant biotype metabolized 2,4-D, seven- to nine-fold faster than the susceptible. Target-site point mutations, gene amplification, and elevated rates of metabolism contribute to six-way resistance in the MO-Ren biotype, suggesting both target site and non-target site mechanisms contribute to multiple herbicide resistance in this Amaranthus tuberculatus biotype. (c) 2018 Society of Chemical Industry