The micropropagation ofGentiana cerinaandG. corymbiferawas investigated. Cultures were initiated from axillary shoots or seed. Seeds ofG. corymbiferagerminated on a Murashige & Skoog (MS) medium containing 100 mg/litre gibberellic acid (GA3) with 54% of the seed germinating within 70 days. In the absence of GA3germination did not exceed 5%. Both species proliferated shoots on MS medium supplemented with 6‐benzylaminopurine (BAP). ForG. cerinathere was no significant difference in proliferation rates for BAP concentrations between 0.05 and 0.5 mg/litre. In contrastG. corymbiferagave highest multiplication rates on 0.2 mg/litre BAP. Addition of 1 mg/litre GA3to the medium gave improved proliferation compared to treatments in which GA3was absent. The best treatment forG. cerinaresulted in a shoot multiplication rate in excess of 7‐fold after 50 days whereas forG. corymbiferathis increase was more than 3‐fold. Root initiation occurred on MS medium supplemented with indole‐3‐butyric acid (IBA). In both species the frequency of explants with roots increased with increasing IBA concentration, but at IBA ≥ 1 mg/litre and 3 mg/ litre forG. cerinaandG. corymbiferarespectively a high proportion of the plants developed a basal callus. Survival of theG. cerinaplants during acclimatisation was related to IBA concentration but survival was unrelated to IBA concentration forG. corymbifera. G. cerinasurvival rates decreased from 80% with up to 0.3 mg/litre IBA to c. 5% at the highest rate of IBA. ForG. corymbiferaaverage survival of plants after acclimatisation was just over 20%. Thus root initiation with 0.3 mg/litre IBA can be recommended for both species.