As a medicinal plant, the importance of evening primrose (Oenothera biennis L.) is due to its unsaturated fatty acids in the seeds and roots, and also oenotherine and comfarol in the leaves. Low germination and difficulties in seed production are the main problems encountered with growing this plant in the field. As an alternative approach, an in vitro experiment was set up for the evaluation of evening primrose production via direct and indirect regeneration of the cultivars NC-1 and VNK. For callogenesis and direct regeneration, the explants from the apical bud and petiole were cultured on MS medium supplemented with 0.25, 0.75, and $1.25mg;L^{-1}$ of both BAP and Kinetin (KIN). Indirect regeneration was performed by placing apical buds, petioles, and leaf explants on MS medium supplemented with 0.5 and $1mg;L^{-1}$ 2,4-D and 0.5, 1, and $1.25mg;L^{-1}$ of both BAP and KIN. The highest shoot induction from direct regeneration was obtained with apical bud explants of VNK treated with $0.75mg;L^{-1}$ BAP. The highest callus weight (3.17 g) obtained from indirect regeneration was with petiole explants treated with $1mg;L^{-1}$ 2, 4-D and $1mg;L^{-1}$ BAP in VNK cultivars. The highest number of torpedo embryogenic clusters (23.8) was obtained from the VNK petiole explants treated with $0.5mg;L^{-1}$ 2, 4-D and $1.25mg;L^{-1}$ BAP. BAP had higher positive effects on in vitro production of evening primrose than KIN in both direct and indirect regeneration. In general, results indicated that VNK was more potent for regeneration than NC-1 and concentrations of $0.75mg;L^{-1}$ BAP for direct and $0.5mg;L^{-1}$ 2, 4-D and $1.25mg;L^{-1}$of BAP for indirect regeneration had a higher efficiency for increasing in vitro production of evening primrose.