Rudraksha (Elaeocarpus ganitrus), is an inheritable shrub and a valuable tree species belongs to the genus Elaeocarpus with significant religious, medicinal, and ecological importance. Irrespective its high demand, its natural propagation is seriously restricted by low seed germination, persistent dormancy, and poor seed viability. The experiment intended to develop reproducible and an efficient method to optimize and develop In vitro micro propagation protocol for E. ganitrus through controlled analysis of explant selection, culture media composition, sterilization procedures, plant growth regulators (PGRs), and environmental conditions. Nodular stem sections were found to be the most responsive, and optimal sterilization was achieved using 0.1% mercuric chloride for 3-5 minutes. Maximum shoot proliferation was observed on Murashige and Skoog (MS) medium supplemented with 1.5 mg/L BAP in combination with 0.5 mg/L 2,4-D. Rooting was best achieved on half-strength MS medium containing 1.0 mg/L IBA. Acclimatization success reached up to 80% under controlled conditions. The optimized protocol provides a reliable and reproducible method for large-scale propagation and conservation of E. ganitrus.