Eastern black nightshade is one of the problematic weeds in vegetables and soybeans in the Midwest. It is representative of a rapidly growing complex of broadleaf weeds where herbicide resistance would be expected to occur. Eastern black nightshade calli lines that are resistant and susceptible to acifluorfen were maintained on the medium without the herbicide. After two years, these lines were tested for tolerance to acifluorfen and paraquat. Tolerance to acifluorfen was maintained in the previously selected lines. The lines were also cross tolerant to paraquat. Plants were regenerated from these calli lines and grown in the greenhouse. 14C-acifluorfen and 14C-paraquat uptake, translocation, and metabolism were studied.