Abstract
An in vitro method to improve the production of vincristine (VCR) was developed by using CaCl2 elicitor in embryogenic suspension culture of C. roseus. CaCl2 at various levels [0 mM (T0), 5 mM (T1), 25 mM (T2), 50 mM (T3), 75 mM (T4) and 100 mM (T5)] was added in MS medium as an elicitor molecule. Cultural growth in terms of absolute dry mass, packed cell volume, colony area was increased up to T2 treatment and thereafter it declined. Antioxidant enzyme activities such as Superoxide dismutase (SOD), Catalase (CAT), Ascorbate peroxidase (APX) and Glutathione reductase (GR) in response to CaCl2 treatment were assayed. We noted that as the concentration of CaCl2 was increased in medium the activity of the above said enzymes increased linearly. HPTLC analysis of embryogenic tissues and of liquid medium at periodic intervals was performed to quantify the alkaloid yield. We noted early accumulation of VCR (after 20 days of culture) in suspended cells in response to calcium elicitation. No VCR was, however, detected in liquid medium, used for tissue cultivation.
Keywords: Vincristine, colony size, calcium chloride, elicitor, HPTLC, embryogenesis, cell, lipid, enzymes, tissue.
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