Gentiana spp. are ornamental as well as valuable medicinal plants. This research was conducted to investigate the effects of different cytokinin and auxin treatments on shoot propagation and rooting of both G. lutea and G. boissieri, and to establish the most appropriate micropropagation protocols for these species. For this purpose, MS media containing benzyl adenine (BA) (1, 2, 3, and 4 mg/l), zeatin (0.5, 1, 2, and 3 mg/l), and kinetin (0.5, 1, 2, and 3 mg/l) were used for shoot propagation, and the optimal cytokine applications were determined by examining shoot number per explant (SN/E) and shoot length (SL). To determine rooting ability, shoots were transferred to MS media containing indole acetic acid (IAA), indole butyric acid (IBA), and naphthalene acetic acid (NAA) at different concentrations (1, 2, 3, and 4 mg/l). The optimum auxin applications for root and plant development were determined by examining the rooting percentage (%, RP), root number per shoot (RN /S), root length (RL), shoot length (SL), and plant fresh weight (FW). As a result of the study, it was found that the media containing 3 mg/l and 2 mg/l BA were the most suitable shoot propagation media for G. lutea and G. boissieri, respectively. The media containing 4 mg/l IBA and 1 mg/l AgNO3 gave the highest rooting and growth parameters for G. lutea, while G. boissieri had the best rooting and growth values in media supplemented with 2 mg/l NAA. In addition, G. boissieri was found to be superior to G. lutea in both shoot propagation and rooting.