An efficient plant regeneration protocol through two-stage culture of nodal segment has been reported here for a
valuable medicinal plant, Bacopa monnieri. Multiple shoots with large number of shoot buds were induced from
nodal explant on Murashige and Skoog’s (1962) (MS) medium fortified with (1.0-5.0 mg L-1) N6- benzyladenine
(BA). Of the different concentrations of BA tested, 3.0 mg L-1 BA offered the best result, where the nodes
swelled and an average of 6.5 shoots with numerous buds/node were recorded. The swollen nodes, cultured on
MS + 3.0 mg L-1 BA, were sub-cultured either on MS or MS supplemented with 1.0 mg L-1 GA3 medium. The
best result (114.2 shoots/ node) with an average shoot length of 6.4 cm was observed on MS media supplemented
with 1.0 mg L-1 GA3. Cent per cent acclimatization of healthy in vitro regenerated shoots was obtained by ex
vitro rooting in plastic pots containing garden soil, which saves time and tissue culture cost by abolishing the in
vitro rooting step.