Stachys inflata Benth., an important medicinal plant in the Lamiaceae family, is highly valued for its secondary metabolites and therapeutic properties. However, its overexploitation in the wild and limited distribution poses significant conservation concerns. This study aimed to develop an efficient propagation protocol for S. inflata through callus induction and shoot regeneration. Leaf explants were cultured on Murashige and Skoog (MS) medium supplemented with various plant growth regulators (PGRs). Callus induction was achieved using combinations of 2,4-D and BAP, with the highest efficiency observed at 0.5 mg/l 2,4-D + 0.1 mg/l BAP. For indirect shoot regeneration, the combination of 2 mg/l BAP + 0.5 mg/l IBA was optimal, resulting in 100% regeneration frequency and the highest number of shoots per callus explant. Node explants showed favorable responses to several PGR combinations, with 2 mg/l Kinetin inducing the highest number and length of shoots. Rooting of regenerated shoots was successfully induced by 0.5 mg/l IBA, resulting in excellent root formation. Acclimatized plantlets were successfully transferred to the greenhouse, demonstrating healthy growth and survival. This study provides a valuable in vitro propagation protocol for S. inflata, with potential applications for its mass propagation, conservation, and sustainable use in medicinal and aromatic plant industries.
