R. damascena, renowned for its aromatic essence, holds immense significance in various industries, including perfumery and cosmetics. However, its propagation presents challenges due to its recalcitrant nature. This study aimed to investigate micropropagation in vitro from single nodes as an alternative to traditional cutting methods, focusing on enhancing plant material preservation. Nodal explants were subjected to different auxin treatments (Indole-3-butyric-acid at T1:0.1, T2:0.5, and T3:1 mg/L; Indole-3-acetic-acid at T4:0.1, T5:0.5, and T6:1 mg/L; and 1-Naphthaleneacetic-acid at T7:0.5, T8:1, and T9:1.5 mg/L) to assess rooting efficiency and subsequent plant development. Results revealed a significant increase in rooting rate, with the highest rooting rate observed in the T3 treatment (97.22%) with 1 mg/L of IBA after 12 weeks of incubation. Moreover, the mortality rate varied significantly among treatments, with the highest rate observed in the control group (55.56%). The bud break rate was significantly higher in the T3 treatment compared to other treatments (100%). Correlations between morphological traits unveiled intricate relationships, highlighting the influence of auxin type and concentration on various parameters such as mortality and bud break rate. During the acclimatization process, a substrate composed of perlite, peat, and sand in a ratio of 3:1:1 (v/v/v) was utilized. The IBA-treated plants demonstrated superior growth, with an average apical growth of (2.84 cm) and a leaf area of (20.95 cm²) after 6 weeks. Our findings provide valuable insights into optimizing micropropagation techniques for R. damascena, thereby contributing to sustainable cultivation practices and meeting the increasing demand for this prized botanical resource.