Studies were conducted from 2001 through 2003 to determine the extent of resistance to acetolactate synthase (ALS) inhibitors and glyphosate in Ohio horseweed biotypes. The response of 66 horseweed biotypes to cloransulam-methyl and glyphosate was determined in the greenhouse. Application of 0.07 kg ai cloransulam/ha reduced plant biomass by less than 60% for 38 of the 66 biotypes. Application of 3.4 kg ae glyphosate/ha reduced biomass by at least 80% for the 51 biotypes collected in 2001, but biomass was similar to that of nontreated plants for 11 of the 15 populations collected in 2002. A dose–response study was conducted with selected biotypes, and a nonlinear, logistic dose–response curve was fit to the data to calculate the herbicide dose required to reduce fresh weight 50% (GR50). On the basis of GR50 values, the resistance ratio (R/S) for two ALS-resistant biotypes was 34 and 943 for chlorimuron-ethyl and 32 and 168 for cloransulam, respectively. The R/S ratio for two glyphosate-resistant biotypes was 33 and 39. Results of these studies indicate that, in 2002, ALS-resistant horseweed was widespread throughout Ohio, whereas resistance to glyphosate occurred primarily in several counties in southwestern Ohio.