Production of Nitidine from Callus Cultures of Toddalia asiatica

Abstract

Toddalia asiatica(Rutaceae) is a woody liana used traditionally in the treatment of malaria, sprains, cough, fever, neuralgia, epilepsy, dyspepsia and other disease conditions. The extracts of the plant have been reported to have anticancer and anti-HIV activities. These activities are mainly attributed to benzophenanthridine alkaloid i.e., nitidine.  The yield of the nitidine in intact plant is too low (0.002% w/w). Plant tissue culture has been used as an alternative method for increasing the production of secondary metabolites such as nitidine. In similar attempt the present study was carried out to initiate callus cultures from T.asiatica and evaluate further strategies to improve the production of nitidine. The leaves of T. asiatica were incubated in MS medium supplemented with different combinations of growth hormones. MS medium containing NAA (2mg/L) and Kinetin (1mg/L) was found to yield good friable callus after six weeks of incubation. The pooled callus was extracted and observed for the presence of nitidine. The identification was done by TLC and HPLC by using the standard sample. The callus was found to produce 0.026% w/w of nitidine which was 10 fold more than that of intact plant.  The conditions required for growth of callus and production of nitidine were optimized and it is the first report.