Vinblastine and Vincristine Production on Madagascar Periwinkle (Catharanthus roseus (L.) G. Don) Callus Culture Treated with Polethylene Glycol

Abstract

Vinblastine and vincristine are secondary metabolites from Madagascar periwinkles that have a very high economic value as chemotherapy drugs. These compounds are naturally produced in a very low quantity in planta . One promising alternative method for vinblastine and vincristine production is to use a treatment that can trigger plant stress response in vitro . This study has been done to evaluate the effect of drought stress using polyethylene glycol (PEG) on vinblastine a nd vincristine production in the C. roseus callus culture , which were grown on medium Zenk supplemented with plant growth regulators (PGR) 1 μM NAA + 10 μM Kinetin to induce laticifer and idioblast differentiation. 13 -week-old callus culture s were then treated with 0% , 6% , 9%, a nd 12% (w/v) PEG4000 each for 0 , 24 , 48, and 72 hours . Biochemical analysis was performed using HPLC to determine the levels of vinblastine and vincristine, while the presence of differentiated cells (idioblasts and laticifers) was determined using a histochemical method. Protein profiles of the culture were determined by SDS-Page. The results showed that drought treatment with PEG4000 , until the concentration was 12% (w/v), did not significantly affect the production of vinblastine and vincristine, but might affect terpenoid production. Histochemical analysis confirmed the presence of idioblasts, non-elongated laticifers, and laticifers that were producing and accumulating terpenoids highest in t he 12% PEG treatment. PEG treatments also did not change the protein profile of callus.