In-vitro Micropropagation of Adhatoda vasica Nees. (Basok) for Commercial Exploitation in Bangladesh

Abstract

To enhance the production of the major phytochemical active compounds of Adhatoda vasica at large scale, wide cultivation and their conservation as a local valuable medicinal plant attempts were made to establish a protocol in vitro micropropagation for large scale production of Adhatoda vasica. Nodal segments were used as explants for regeneration of multiple shoot and juvenile leaves were used as explants for callus induction. Among the different growth regulator formulations MS medium supplemented with BAP (10.0 mg/l) was found best for multiple shoot formation, in which 93.33% explants produced multiple shoots, 7 days required for initiation of shooting, the nodal explants remained green and viable for further subculture. After 2 months, maximum number of multiple shoots 10.6 ± 1.82, highest length of plantlet 5.2 ± 2.20 cm and maximum number of leaves 15.6 ± 1.80 were recorded. Maximum 100% callus formation was observed on MS medium supplemented with IAA (0.05 mg/l) + NAA (0.05 mg/l) + BAP (1.0 mg/l). 30 explants were inoculated for callus induction; callus initiation started from 14 days, 30 explants showed callus formation and gave soft, green coloured, fast growing callus. Media containing IBA (1.0 mg/l) with GA3 (1.0 mg/l) also showed better response, 30 explants were inoculated; callus initiation started from 14 days, 28 explants showed callus formation, 93.33% of responsive explants and callus gives light green colour. Best callus mediated shooting found from MS + 10.0 mg/l BAP medium, 10 explants were transferred, 80% explants showed regeneration, 10 days required for initiation of shooting, mean number of multiple shoot 4.0 ± 1.70, mean length of plantlet 3.0 ± 1.56 cm and mean number of leaves were 11.0 ± 1.45. Rooting response of in vitro raised shoots from nodal explants on MS + IBA (2.5 mg/l) + IAA (0.5 mg/l) + NAA (0.5 mg/l) performed best; root formation of explants was 80%, 12 days required for initiation of rooting from shoot regenerated from nodal explants and root formation of explants was 60%, 13 days required for initiation of rooting from callus mediated shoot. Well rooted plantlets were transferred to plastic pots containing soil for hardening. All acclimatized plantlets were then transferred to garden soil. The ultimate survival rate of the transferred plantlets to soil was about 78%.