Plant tissue culture plays an important role in the production and conservation of rare plant species. An efficient in vitro plant regeneration protocol through direct organogenesis is an important need for the conservation of a valuable medicinal plant. Ruellia tuberosa nodal explants were used for in vitro regeneration on Murashige and Skoog (MS) medium containing 3% sucrose and different concentrations and combinations of plant growth regulators. A maximum shoot length of eight centimeter was obtained from nodal explants when cultured on MS medium supplemented with kinetin and BAP at 2.0mg/L concentration. The highest percentage of shoot induction (90%) was obtained in 2.0 mg/L in kinetin with a mean of 8.0±0.89 shoots. The maximum root length (3.63cm) was obtained with 95% and 90% of NAA and IBA respectively at 1 mg/l concentration. Rooted plantlets were transferred to plastic cups and grown in the green house. The survival rate was 90%.
