Optimisation of the second phase of a two phase growth system for anthocyanin accumulation in callus cultures of Rudbeckia hirta

A two-phase growth system composed of modified Schenk-Hildebrandt medium and enriched Miller's medium was used to grow callus of R. hirta L. The second phase of the system has been optimised and the callus synthesised a 12-compound set of anthocyanins, comprising 4.97% of tissue mass (tubular flowers of the plant in nature only contain one compound comprising 0.28%). The beneficial effect of high content of cysteine on anthocyanin accumulation was noted. The predominant element of the anthocyanin set obtained in vitro is cyanidin-3-O-(6-O-malonyl-beta -D-glucopyranoside), a relatively stable compound.