Greenhouse and laboratory experiments were conducted to investigate differences in glyphosate susceptibility among three species of the genus Conyza introduced as weeds in Spain: tall fleabane (Conyza sumatrensis), hairy fleabane (Conyza bonariensis), and horseweed (Conyza canadensis). Plant material was obtained from seeds collected in weed populations growing in olive groves and citrus orchards in southern Spain, with no previous history of glyphosate application. Dose-response curves displayed ED(50) values of 2.9, 15.7, and 34.9 g ai ha(-1), respectively, for C. sumatrensis, C. bonariensis, and C. canadensis plants at the rosette stage (6-8 leaves). Significant differences were found among the three species in the glyphosate retention on leaves as well as the leaf contact angle. The species order according to glyphosate retention was C. sumatrensis > C. bonariensis > C. canadensis, while the mean contact angles of glyphosate droplets were 59.2, 65.5, and 72.9 degrees , respectively. There were no significant differences among species in the absorption of [(14)C]glyphosate (ranged from 37.4% for C. canadensis to 52.4% for C. sumatrensis), but the order among species was the same as glyphosate retention. The amount of radioactivity translocated from treated leaves was lower in C. canadensis as compared to the other two species (C. sumatrensis > C. bonariensis > C. canadensis). Combined, all of the studied parameters identified differential susceptibility to glyphosate among the Conyza species. Each species accumulated shikimate in leaf tissues following application of glyphosate at 200 g ai ha(-1). However, C. canadensis exhibited lower shikimate levels than the other two species at 168 h after herbicide application. For hairy fleabane, a greenhouse study explored its susceptibility to glyphosate at three developmental stages: rosette, bolting (stem height, 10-15 cm), and flowering. The ED(50) was lower at the rosette stage (15.7 g ai ha(-1)) as compared to bolting (86.6 g ai ha(-1)), with the highest ED(50) values occurring at flowering (117.5 g ai ha(-1)); plants at the earlier developmental stage retained more glyphosate. These results agree with field observations that plants at early developmental stages are more sensitive to glyphosate.