Critically endangered (CR) endemic plant Centaurea tchihatcheffii Fisch. Et. Mey. grows at Golbasi district of Turkish province of Ankara. Biotechnological approaches are desired as an alternative to break deep seed dormancy and micro propagation of C. tchihatcheffii. The seed dormancy was broken by horizontally shaking them in 2 mg/L GA(3) suspension under dark conditions for 72 hours. Stem nodes from one week old in vitro germinated seedlings were cultured on MS medium containing 0.3, 0.6, 0.9, 1.2, 1.5 mg/L Kinetin + 0.1 mg/L NAA (5 combinations), 0.3, 0.6,0.9, 1.2, 1.5 mg L-1 BAP + 0.1 mg/L NAA (5 combinations) and MS medium used as control for 45 days. Changing concentrations of Kinetin + NAA were the most effective in regeneration. Maximum number of (4.50) shoots per explant with induction of floral buds, and roots was noted on MS medium containing 0.9 mg/L Kin + 0.1 mg/L NAA under 16 h light photo period after 14-16 days of culture. These plants were transferred to pots, where most of the buds converted to flowers and set seeds. A comparison showed no abnormality noted in plants regenerated under in vitro conditions and those growing in situ.