PLANT CELL BIOTECHNOLOGY AND MOLECULAR BIOLOGY

Gloriosa superba is an important medicinal plant with tuberous roots belongs to the family Liliaceae. The V- shaped root tuber of the plant possessed several secondary metabolites like colchicine, gloriosine etc. and colchicine is one of the major alkaloid that used for curing various ailments like gout, cancer, asthma etc. The plant is currently threatened due to the unscientific collection of the plant for medicinal purposes. This threatened plant can be conserved through the in vitro techniques like in vitro tuberization and the callus induction. The present study mainly focused in vitro induction of roots and quantification of colchicine from both the in vitro regenerated root tubers and callus of Gloriosa superba along with its comparison with the normal root tubers. For the establishment of in vitro root tubers and callus, the explants such as node and leaf were inoculated in MS medium at different hormonal concentrations. The colchicine was estimated and quantified through various chromatographic technique like HPTLC and HPLC. The results showed that the MS medium with NAA (2.5 mgl^-1) + BAP (0.5 mgl^-1) induced maximum root tubers which are white in colour and nearly less than 1 cm long. All the in vitro tubers were sub cultured in the MS medium with different hormone concentration for the growth and elongation of the roots. Maximum callus induction were noticed in the MS medium with the hormone combination NAA (0.5 mgl^-1) + BAP (2.5 mgl^-1) within 21 days of inoculation. HPLC results showed that the naturally inhabited root tuber possessed 5.853 mg of colchicine, the callus possessed only 0.0664 mg and the in vitro regenerated root tubers possessed 0.3 mg of colchicine. Suspension culture with elicitors will be helpful for the enhancement of bioactive compounds especially colchicine from both the in vitro root tubers as well as the callus and could be used for the development of new pharmaceutical drugs in future.