Clethodim resistance was identified in 12 rigid ryegrass populations from winter cropping regions in four different states of Australia. Clethodim had failed to provide effective control of these populations in the field and resistance was suspected. Dose–response experiments confirmed resistance to clethodim and butroxydim in all populations. During 2012, the LD₅₀ of resistant populations ranged from 10.2 to 89.3 g ha⁻¹, making them 3 to 34–fold more resistant to clethodim than the susceptible population. Similarly, GR₅₀ of resistant population varied from 8 to 37.1 g ha⁻¹, which is 3 to 13.9–fold higher than the susceptible population. In 2013, clethodim-resistant populations were 7.8 to 35.3–fold more resistant to clethodim than the susceptible population. The higher resistance factor in 2013, especially in moderately resistant populations, could have been associated with lower ambient temperatures during the winter of 2013. These resistant populations had also evolved cross-resistance to butroxydim. The resistant populations required 1.3 to 6.6–fold higher butroxydim dose to achieve 50% mortality and 3 to 27–fold more butroxydim for 50% biomass reduction compared to the standard susceptible population. Sequencing of the target-site ACCase gene identified five known ACCase substitutions (isoleucine-1781-leucine, isoleucine-2041-asparagine, aspartate-2078-glycine, and cysteine-2088-arginine, and glycine-2096-alanine) in these populations. In nine populations, multiple ACCase mutations were present in different individuals. Furthermore, two alleles with different mutations were present in a single plant of rigid ryegrass in two populations.

Nomenclature: Clethodim; rigid ryegrass; Lolium rigidum Gaudin.