Abstract: In this research, we aimed to establish an effective in vitro propagation protocol for Hypericum heterophyllum Vent., an endemic Turkish species. The seeds were surface sterilised and transferred to Murashige and Skoog basal medium supplemented with BA (benzyladenine, 0.1, 1 mg L-1) and 2,4-D (2,4-dichlorophenoxyacetic acid, 0.1, 1 mg L-1) under the photoperiod (8/16 h light/dark period, 1200 μmol m-2s-1). Culturing seeds on MS basal medium supplemented with 1 mg L-1 BA and 0.1 mg L-1 2,4-D resulted in greenish and compact callus induction. MS basal medium supplemented with 4.4 mg L-1 BA was used for shoot induction, while the same basal medium supplemented with 1 mg L-1 IAA (indolacetic acid) was employed for the rooting. Regenerated plants were easily acclimatized in greenhouse conditions.