This experiments were carried out to find out the effects of different explant materials, kinds and concentration of plant growth regulators on the in vitro regeneration of Iris pseudoacorus L. The effects of growth regulators on regeneration from three explant sources (root, rhizome and leaf) were more or less same. Explants produced only callus with 2,4-D treatment and other regulators had no effects. The highest frequency of callus induction (rhizome: 84%) was on MS media supplemented with 1.0 mg/L 2,4-D. For plant regeneration, when calli were transferred to the first media supplemented with 1.0 mg/L BA and 1.0 mg/L 2,4- D. Regenerated plants were successfully acclimatized in soil containing a mixture of vermiculite : perlite : leafmold (1 : 1 : 1), and their survival was high.