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S. Alagumanian*, G. Jahirhussain and M. V. Rao


An efficient protocol was achieved for rapid clonal propagation of the medicinally important plant species, Centella asiatica (L.) Urban through shoot tip proliferation and ex vitro rooting. High frequency bud break (88%) and multiple shoot formation were induced from shoot tip segments are cultured on MS medium supplemented with BAP (17.76 μM) plus GA3 (1.44 μM). Although callus free multiple shoot formation was a function of cytokinin activity alone it fastest bud break coupled with enhanced frequency of shoot development (88%) and internode elongation were dependent on the synergistic effect of GA3 (1.44 μM). By repeated subculturing of nodal segments harvested from the newly formed axenic shoots, prolific shoot cultures, free of proximal callusing and showing a high frequency multiplication rate were established within three months. The percentage of shoot multiplication as well as the number of shoots per shoot tip attained the highest values (88%, 16.8 shoots/shoot tip) during the first two culture passages; beyond this there was a gradual decline in shoot bud differentiation. Half-strength MS medium fortified with NAA (10.74 μM) induced the highest number of roots. All in vitro rooted shoots survived in field. Dipping of the basal end of shoots collected from multiplication medium in IBA (10.74 μM) solution for 10 days induced roots and its transfer to small pots facilitated the survival of all rooted shoots (100%). Rooting of ex vitro by direct transfer of shoots from multiplication medium exhibited 90% survival. Use of commercial sugar and tap water and also the omission of in vitro rooting reduce the propagation cost 50 – 70 %. The plantlets were hardened off and successfully established in the natural soil, where they grew and matured normally. The protocol enables to harvest more than 25,000 plantlets within 160 days starting from a single shoot tip explant.

Keywords: Centella asiatica, Medicinal plant, Micropropagation, Explant source, Multiple shoot-bud induction, Ex vitro rooting.

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