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An efficient in vitro micropropagation system was developed for direct shoot growth of Ocimum basilicum, an important medicinal plant, using nodal explants. The excised nodes were cultured on Murashige and Skoog (MS) medium containing two plant growth regulators (6-benzyladenine and 2-isopentanyl adenine) with various combinations and concentrations for the study of shoot induction. Addition of L-glutamine was essential to induce sprouting of axillary buds. The best condition for shoot growth was with 6-benzyladenine (BA) 10.0 μM+ L-glutamine 30 mg/L in MS medium. The optimum shoot formation frequency was 100% and about 13.4±1.80 shoots were obtained from each explant after 8 weeks of culture. Shoots (more than 4 cm) were rooted most effectively in 5.0 μM indole-3-butyric acid (IBA) supplemented with half-strength MS medium. The plantlets thus obtained hardened off and were transferred to natural soil, where they grew well and attained sexual maturity.