Plant tissue culture is an efficient technique for conserving endemic plant species. A reproducible in vitro regeneration protocol was developed for the endemic Iris sari and Iris schachtii in the present study. The highest number of shoots per explant was obtained on Murashige and Skoog (MS) medium supplemented with 0.5 mg/L thidiazuron (TDZ) plus 0.5 mg/L alpha-naphthaleneacetic acid (NAA) and 1 mg/L TDZ plus 0.5 mg/L NAA, whereby 96.88% and 100% shoot induction with 9.55 and 11.34 shoots per explant of Iris sari and Iris schachtii were recorded, respectively. Regenerated shoots were successfully rooted on MS medium with either 1 mg/L indole-3-butyric acid (IBA) or 1 mg/L IBA plus 0.2 mg/L NAA. Rooted shoots were transferred to pots containing either a peat-soil-sand (1:1:1) mixture or a hydroponic culture containing Hoagland solution to acclimatize the regenerated plants to the greenhouse chamber. Approximately 90% of plants were transferred ex vitro successfully.