Abstract:Explants of Orychophragmus violaceus (L.) O. E. Schulz were acquired from young leaves which lower epidermis was stripped, Differentiation of calli in high frequency is in the case of that calli grown on B5 culture medium supplemented with 1 mg/l 2, 4-D should be transferred onto MS culture medium supplemented with 0.2 mg/l NAA. Effect of basic culture medium on the differentiation was discussed. In addition, protoplasts derived from calli of Orychophragmus violaceus were cultured, and small calli consisted of more than hundred cells had been obtained.