Eighteen Lolium spp. (ryegrass) accessions collected in 1998 from several locations in Arkansas were tested for resistance (R) to diclofop in both seed and whole-plant response bioassays. Eleven accessions were L. temulentum and eight were L. perenne. Fourteen of eighteen accessions were confirmed resistant to diclofop in whole-plant assay. Three of the susceptible (S) accessions were L. temulentum. The GR50 (diclofop concentration that reduced shoot or root length by 50%) R/S ratios based on whole-plant response were greater than those of the seed bioassay in all test populations, indicating that the whole-plant bioassay was more sensitive than the seed bioassay for determining diclofop resistance in Lolium spp. The most resistant (#18) and most susceptible (#3) accessions of L. temulentum were used for multiple resistance and enzyme assay experiments. Based on whole-plant bioassay, accession #18 was 411 times more resistant to diclofop than the susceptible accession #3. Accession #18 exhibited cross-resistance to fenoxaprop and multiple resistance to chlorsulfuron applied preemergence or postemergence. Resistance to other herbicide families was not observed. Resistance to chlorsulfuron was not detected in the seed bioassay. Acetyl-CoA carboxylase (ACCase) from accession #18 was 833 times more resistant to diclofop and 10 times more resistant to sethoxydim than ACCase from accession #3. Cross-resistance to sethoxydim was not observed at the whole-plant level. Resistance to diclofop among Lolium spp. from Arkansas may be due to an alteration in the target enzyme, ACCase.