The effect of carbenicillin on the dedifferentiation and the regeneration efficiency of plant tissues of horseradish(Armoracia rusticana) was evaluated, Inhibition effect for callus initiation was observed when leaf blade, root and petiole segments were grown on MS medium containing 500 mg/L to 2000 mg/L carbenicillin and 0.5 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D). The regeneration of horseradish shoots from leaf blade, root and petiole explants were decreased as the addition of carbenicillin increased from 1000 mg/L to 2000 mg/L in MS medium containing 0.5 mg/L of 6-benzylaminopurine (BAP) or kinetin. Especially, 500 mg/L carbenicillin treatment significantly inhibited shoot induction when leaf blade explants were grown on hormone-free MS medium. It was suggested that the toxic effects of combinations of carbenicillin and 2,4-D may be due to high auxin activity levels.