The aim of this study was to develop methods for the in vitro propagation of Zantedeschia rehmannii. Zantedeschia rehmannii tuber fragments (1 cm2) containing eyes were soaked for 30 s in a solution containing 100 mg dm−3 L-ascorbic acid (AA) before transfer to culture vessels containing an MS medium supplemented with BAP (0 to 3 mg dm−3). Cultures were maintained in darkness. Soaking explants in an L-ascorbic acid solution improved the establishment of explants. Culture initiation should be conducted on media supplemented with 3 mg dm−3 BAP. On a multiplication stage, adventitious shoots were placed on MS media supplemented with cytokinin: BAP (0.5 to 5 mg dm−3), KIN (0.5 to 5 mg dm−3), TDZ (0.1 to 1 mg dm−3) and 2iP (2.5 to 15 mg dm−3) or BAP (0.5 to 7.5 mg dm−3) with IAA (0.5 to 2 mg dm−3). The highest coefficient of multiplication for Zantedeschia was obtained on the medium with the addition of 2.5 mg dm−3 BAP, which positively affected the shoot length (3.41 cm) and the number of adventitious shoots (4.13). Rooting took place on media supplemented with IBA, IAA and NAA at a concentration of 0.1 to 2 mg dm−3. The most numerous and the longest roots were found in plants placed on a medium with the addition of 0.1 mg dm−3 IBA.