Vinblastine and Vincristine Alkaloids Production From Callus of Catharanthus roseus (L.) G. Don under Some abiotic factors

Abstract

This experiment was conducted in Faculty of Science labs, Kufa University, carried out during 2013-2014 to study many experiments to induce callus tissues from leaves of  (Catharanthus roseus ( L. ) G. Don in vitro using MS medium supplemented with Dichlorophenoxy actic acid (2,4-D) at difference  concentrations (0.5, 1, 1.5) mg/l with the interaction of  (BA) benzyl adenine at concentrations of (0.5, 1, 1.5) mg/l. Identically callus fresh weight, then use it in the next experiments and investigate the effect of abiotic factors (drought and sucrose) on vinblastine and vincristine production  from callus tissue, and leaves of  intact  plant. This study include determination the catalase activity, proline and total soluble carbohydrates content as responsible to the variety of the stresses in callus as well as quantities and qualities determination of vinblastine and vincristine using TLC (Thin layer chromatography ) and HPLC(high performance liquid chromatography). Results  reveled  that the highest fresh weight of callus (3.276) g was for (1mg/l 2,4-D + 1mg/l BA ) combination treatment. However, the results showed that the additions of abiotic  factors to the callus cultural  medium cause difference results in fresh and dry weights of callus in accumulation phase, reducing the fresh and dry weight in all treatments expect sucrose 40 g/l treatment which gave increase in fresh and dry weight. Also, this results showed that all treatments increase total soluble carbohydrates, proline content and catalase  enzyme activity with significant difference compared with control treatment and significant increase of alkaloids production with the superiority of (vinblastine and vincristine)  content in Callus with abiotic factors stresses than the content in control treatment( without any factor).