A range of antibiotics was evaluated for their effect on eliminating Agrobacterium tumefaciens supervirulent strain EHA101(pEHA101) from leaf explants of `Royal Gala' apple (Malus domestica Borkh) and on regeneration. After long-term (38 days) exposure to 100-μg–ml–1 concentrations of either cefotaxime (cef), carbenicillin (carb), mefoxin (mef), or combinations of these antibiotics, only on carb or carb with mef was regeneration not inhibited. None of the above antibiotics or antibiotic combinations eliminated A. tumefaciens from leaf explants. Short-term (1-18 hours), vacuum infiltration with 500- to 1000-μg–ml–1 concentrations of either of the above antibiotics did not inhibit regeneration, but did not eliminate A. tumefaciens from leaf explants. After a 30-min vacuum infiltration with a 2000-μg–ml–1 concentration of either cef, carb, or mef, only cef reduced the number of leaf explants with A. tumefaciens.