The physiological and molecular basis of apparent resistance to glyphosate in horseweed (Conyza canadensis L. Cronq.) plants that had survived being sprayed with the herbicide at Prague-Bubny railway station in the Czech Republic was investigated. For the sake of comparison, plants expected to be susceptible were collected in areas where no herbicides had been used. Plants of both sets were treated, at the rosette stage (10-25 leaves, diameter of 3-5 cm), with herbicide at the rate recommended for use in the Czech Republic to control horseweed (960 g of glyphosate-IPA/ha; Roundup Klasik, Monsanto, 480 g of glyphosate-IPA ae L(-1)). Phytotoxic symptoms of the treated plants varied substantially, both between and within these sets of plants. Leaves of susceptible (S) plants wilted and turned yellow, and the plants subsequently died; leaves of plants with reduced susceptibility (RS) remained green, or new leaves were created in the center of their rosettes a few weeks after glyphosate application. There were no significant differences in the accumulation of shikimate between S and RS plants 3 days after treatment (DAT). However, the time course of changes in shikimic acid contents differed between the two biotypes; from 3 to 10 DAT, they decreased more than 4-fold in RS plants, while in S plants, they increased (3-fold, on average) from 3 to 7 DAT. A conserved region of the epsps gene, in which mutations are known to confer resistance in several plant species, was amplified from samples of both S and RS plants and sequenced, but no changes in the encoded amino acid sequence were found, indicating that mutations at another epsps site were responsible for the observed resistance, or that the mechanism may be at least partially non-target-based. Our results suggest that the reduced susceptibility to glyphosate may be due to impaired herbicide translocation, as previously found in studies of horseweed in the United States.