Harmal (Peganum harmala) is a wild plant with a distinguished medicinal potential due to the presence of high levels of phytoalkaloids such as harmaline, harmine and harmalol. Recently, this plant was reported to posses to antimicrobial, antifungal, anti-inflammatory, anti cancerous hypothermic and hallucinogenic activities. Meanwhile, short span of seed viability, uncontrolled extensive collection and drought are the main constraints of a sustainable propagation and supply of this important herb. In this study in vitro propagation of Peganum harmala L. was successfully achieved via in vitro propagation techniques. Different types of cytokinin (BA, kinetin, 2ip, or TDZ) at different concentrations (0.0, 0.5, 1.0, 1.5 or 2.0 mg/L) were experimented in terms of their potential promoting new microshoots. The effects of different types of auxins (IBA, NAA or IAA) on rooting of Harmal microshoots were studied at different levels (0.0, 0.5, 1.0, 1.5 or 2.0 mg/L). All explants developed new microshoots, and the maximum number of shoots (10.2) and length (8.5 cm) were obtained on medium containing (1.5 mg/L) BA. Meanwhile, no rooting was observed in the explants grown either in the control or media supplemented with (0.5, 1.0 or 2.0 mg/L) IBA, while maximum root number (8.0) and root length (3.6 cm) were obtained on MS media supplemented with 1.0 mg/L NAA. Moreover, rooted plantlets were successfully acclimatized and recorded 80% survival rate after being transferred to the greenhouse conditions.