Biochemical Studies During Sequential stages of Root and Shoot Differentiation in Callus cultures of Cardiospermum halicacabum L.

Present investigation was carried out on Cardiospermum halicacabum L. (Ballon Vine) an important medicinal plant. Leaf explants were found to be the best for induction and growth of callus. Depending on fresh and dry weight of callus, high growth value callus was obtained on MS medium supplemented with 2, 4-D (1.0 mg/l) in combination with Kn (1.0 mg/l). This callus was selected for further studies of root and shoot organogenesis as well as biochemical studies. Regeneration of roots (15 days) from callus was observed on MS medium + BAP (0.5 mg/l) + NAA (0.3 mg/l) while shoots (16 days) on MS medium + BAP (0.5 mg/l) + NAA (0.1 mg/l). Starch content and reducing sugars were high in control or undifferentiated callus, which further increases significantly in root and shoot differentiating cultures. Contents of total soluble sugars, total soluble proteins and total phenols were lower in the control callus which increases in the root and shoot differentiating cultures. The activities of enzymes i.e. a-amylase, acid protease and peroxidase increased up to appearance of green patches (8^th day) and were reached at peak on 12^th day, the day that coincide with the visual appearance of roots and shoot. Conversely, the acid invertase activity decreases till the appearance of roots and shoots.