Abstract: Almarah is the most important cultivar of Damask rose in Syria. This study aimed to determine the best protocol for its in vitro micro-propagation. The experiments were conducted at Damascus University in Syria. Effects of different concentrations of benzyladenine BA (0, 0.5, 1 and 2 mg L-1) and gibberellic acid GA3 (0, 1 and 2 mg L-1) were evaluated on proliferation of the explants. To evaluate in vitro rooting, proliferated micro-shoots were cultured on a MS/2 medium supplemented with 3 g L-1 activated charcoal and (0, 1, 2 and 3 mg L-1) concentrations of Indol-3-Butyric Acid IBA. The experimental design of the all stages was a Complete Randomized Design. Means were analyzes with Duncan’s test at p≤0.05 using SPSS. BA increased the proliferation of the explants up to 2 mg L-1. The interaction of BA with GA3 significantly increased proliferation rate of the explants. The highest number of proliferated explants was obtained in the presence of 2 and 2 mg L-1 of GA3 and BA, respectively. The highest percentage of rooted explants was obtained in the presence of 3 mg L-1 IBA in the rooting medium but the number and length of roots significantly increased in response to increasing IBA concentration in the medium up to 2 mg L-1. Only 8% of the plantlets lost during hardening. For the first time in Syria a mass in vitro production protocol of Syrian Damask rose has been achieved. The interaction of BA (2 mg L-1) with (GA3 2 mg L-1) is highly recommended for proliferation rate of the explants while adding IBA (2 mg L-1) to the MS/2 rooting medium is highly recommended for high quality in vitro rooting.