Abstract: Leaf calli of Silybum marianum Gaertn. subcultured for one year were used for protoplast isolation and culture. First division was observed three days after culture on medium M12, and the highest division frequency was 35.4%. One to three months later, small ralli were seen with naked eyes, and grew up gradually. Upon transferring them onto D6 differentiation medium, the green bud apices were observed two months later. However, no shoot differentiation was obtained. Hypocotyl calli were induced on MS+NAA 0.8mg/1, 6-BA 0.5mg/1. Two months after transferring calli onto D6 medium, shoots were regenerated from the surface of the calli. The freqency of shoot differentiation was 75%. On a MS rooting medium containing NAA 0.5 mg/1, IBA 0.1 mg/1, whole plants with healthy roots were obtained.

Key words: Silybum marianum Gaertn., Protoplast culture, Tissue culture, Plant regeneration