Agrobacterium rhizogenes mediated hairy root induction for increased Colchicine content in Gloriosa superba L

Gloriosa superba L. a medicinally important monocot, member of the family Liliaceae, was investigated for callusing, direct regeneration and induction of hairy root using soil bacterium Agrobacterium rhizogenes. Friable callus was initiated from corms on basal MS medium amended with 2,4-D (1 mg L) and kinetin (1 mg L). Callus generated from such effective combinations were co-cultivated with the bacterium in vitro. Infection was facilitated by acetosyringone (AS) (20 mg L) supplemented during co-cultivation. Extensive hairy roots were induced from the callus within 15-18 d. These roots were then established on MS basal broth medium. Alkaloids like colchicine and colchicoside were extracted from hairy roots and compared with authentic standard colchicines in HPLC. Statistical media optimization was employed to select effective media composition. The findings indicated that nitrogen, inoculum and ferric ions have the highest f value that influenced colchicine production (22.7 mM) and hairy root growth.