Three types of culture systems, solid medium system, liquid shake flask system, and temporary immersion system (TIS) were used for testing their efficiency in propagating Curcuma zedoaria plantlets. The proliferation medium used in shoot multiplication was the Murashige and Skoog medium supplemented with 0.5 mg/L 6-benzylaminopurine (BA) and 0.5 mg/L Indole-3-butyric acid (IBA). Among the three systems used, the liquid shake flask system significantly induced more shoot formation and larger shoots from the shoot explants of C. zedoaria. Meanwhile, divided shoot explants produced significantly higher number of shoots than the undivided shoot explants. The in vitro plantlets derived from the three different culture systems produced healthy and morphologically similar to the mother plants after acclimatization and being transferred to the field.