Development of callus and cell suspension culture from the leaf of Adhatoda vasica Nees using economical growth media

Adhatoda vasica Nees, commonly called Vasaka or Arusha has high medicinal value owing to its rich flavonoid and alkaloid contents. The alkaloid content of A. vasica is known to vary with genotype, and hence vegetative method is recommended for its propagation. Effective medium for A. vasica propagation is an area of interest to researchers. Suspension culture techniques have demonstrated that alkaloids can be recovered from the callus and the suspension cell cultures. In this context, we tried to emphasis upon the use of economically viable organic vermicompost and its extracts along with coelomic fluid as plant tissue culture medium without involving expensive chemicals. In the present study, an organic experimental economical medium being standardized using vermicompost, its extracts along with coelomic fluid and friable soft callus was obtained from leaf explants of A. vasica Nees without any chemical supplementation. Direct shoot formation with leaves was documented on MS medium supplemented with 2 mg/L BAP and 3mg/L IBA. Cell suspension medium is standardized using vermicompost extract and the coelomic fluid (3:1 ratio). Suspension cell culture showed cell separation and multiplication of callus. Presence of total phenolics in callus and in vivo plants exhibited no significant variation, Flavonoid content was significantly higher in the callus and suspension cell extracts, Statistically Student's t-test comparative analysis have shown significance at one per cent level where P <= 0.05 for phenols and flavonoids. Alkaloids were detected on TLC plate, under UV light at 245 and 365 nm, respectively. Rf values of vasicine was found to be 0.4 which corresponds to that of standard vasicine and 0.60 for vasicinone, respectively. HPLC confirmed the presence of vasicine at 5.461 min. This study enables analysis of callus and suspension cell culture for the presence of various secondary metabolites.