Environment damage has brought about the elimination of many plant species, and numerous more are endangered. Similarly, ornamental and endemic Centaurea fenzlii Reichardt that grows in Turkey is threatened that necessities measures for its immediate conservation. In vitro culture techniques are widely used to conserve uncommon, threatened plants. This study reports a proficient technique for multiple micro-shoot regeneration of C. fenzlii. A low recurrence (35%) germination was acquired on MS medium supplemented with 2.60 mu M GA(3). Juvenile leaves were utilized as an explants source. The leaves were regenerated on MS medium that contained 1.11, 2.22, 4.44, 8.88 mu M BAP + 2.685, 5.37, 10.64 mu M NAA. The best callus regeneration percentage was seen on MS medium that used 4.44 mu M BAP + 10.64 mu M NAA. Callus regeneration percentage extended between 21.33 to 66.87. The most noteworthy micro-shoot regeneration percentage was gotten on MS medium containing 8.88 mu M BAP + 2.685 mu M NAA. Micro-shoot regeneration percentage run between 5.66 to 36.87%. The highest number of micro-shoots per explants was noted on MS medium containing 4.44 mu M BAP + 10.64 mu M NAA. Also number of micro-shoots per explants extended between 1.66 to 5.87. The rooting percentage of C. fenzlii was low with a few roots. The best rooting was noted following seven weeks of culture on 4.90 M IBA. IBA appears to act as stimulant in rooting of C. fenzlii.