Overuse of acetolactate synthase (ALS)–inhibiting herbicides in rice has led to the evolution of halosulfuron-resistant rice flatsedge in Arkansas and Mississippi. Resistant accessions were cross-resistant to labeled field rates of ALS-inhibiting herbicides from four different families, in comparison to a susceptible (SUS) biotype. Resistance index of Arkansas and Mississippi accessions based on an R/S ratio of the lethal dose required for 50% plant mortality (LD₅₀) to bispyribac-sodium, halosulfuron, imazamox, and penoxsulam was ≥ 21-fold. Control of Arkansas, Mississippi, and SUS accessions with labeled field rates of 2,4-D, bentazon, and propanil was ≥ 93%. An enzyme assay revealed that an R/S ratio for 50% inhibition (I₅₀) of ALS for halosulfuron was 2,600 and 200 in Arkansas and Mississippi, respectively. Malathion studies did not reveal enhanced herbicide metabolism in resistant plants. The ALS enzyme assay and cross-resistance studies point toward altered a target site as the potential mechanism of resistance. Trp₅₇₄–Leu amino acid substitution within the ALS gene was found in both Arkansas and Mississippi rice flatsedge accessions using the Illumina HiSeq platform, which corresponds to the mechanism of resistance found in many weed species. Field-rate applications of 2,4-D, bentazon, and propanil can be used to control these ALS-resistant rice flatsedge accessions.

Nomenclature: 2,4-D; acetolactate synthase; bentazon; bispyribac-sodium; halosulfuron; imazamox; malathion; propanil; rice flatsedge, Cyperus iria L; rice, Oryza sativa L.