Abstract: Induction conditions of Crocus sativus L. callus were optimized. MS medium was optimal for shoot callus induction while B5 medium was optimal for leaf and flower callus induction. 18℃, 25 ℃ and 21℃ were the optimal temperature for callus induction of shoot, leaf and flower, respectively.Light was positive factor for leaf callus induction but negative for shoot and flower induction. Naphthaleneacetic acid (NAA, 1.5-2.0 mg.L-1) and 6-benzyladenine (6-BA, 0.25 mg.L-1) were the optimal hormones for callus induction. Under the optimized conditions, the induction rate reached 60% for shoot callus, 80% for leaf callus, and 35% for flower callus. Using eyesight and HPLC methods, one callus line which gave rise to 1 677 mg.g-1 crocin and would not brown was obtained from 229 callus lines.