High efficiency adventitious indirect shoot organogenesis and plant Regeneration from callus of Centella asiatica (L.) -An Important Antijaundice Medicinal Plant

An efficient protocol was designed for successful regeneration of Centella asiatica (L.) from in vitro derived callus through somatic embryogenesis. Nodal explants were isolated and cultured on MS medium fortified with 0.5 - 1.0 mg/l 2,4- D alone and in combination with BAP. The above combination induced profuse, compact, light green to greenish coloured calli. It was observed that the increased concentration of 2,4- D induced the direct shoot organogenesis occasionally. Some differences in the morphology of callus such as changes in the colour and texture was also observed with increasing the concentration of 2,4- D. Maximum frequency of callus induction was noticed on 2.0 mg/l 2,4- D and 0.5 mg/l BAP which was greenish colour, friable and granular in appearance. The calli were separated and further cultured on fresh media containing BAP alone and in combination with auxins such as NAA and IAA. The maximum regeneration frequencies of shoots were recorded on BAP 2.5 mg/l + 0.5 NAA mg/l. The well regenerated healthy micro shoots were separated and transferred to rooting medium for profuse rooting. MS medium augmented with IBA 1.0 mg/l showed maximum rooting frequency with high mean root number and root length. The well rooted plants were transferred to field conditions. Survival frequency was found to be 80%. This reproducible protocol can be used for further regeneration and genetic transformation studies.