DITERPENOID PRODUCTION IN HAIRY ROOT CULTURES OF SALVIA-MILTIORRHIZA

Transformed root cultures of Salvia miltiorrhiza have been established by infecting pieces of sterile grown plants with Agrobacterium rhizog enes strains LBA 9402, ATCC 15834, TR 105, R 1601 and A 4 1027. These strains showed different abilities to induce hairy roots on this plant . A promoting effect of acetosyringone on the Agrobacterium-mediated h airy root initiation in Salvia miltiorrhiza was also observed. All tra nsformed root cultures were assayed for opines by TLC, and were positi ve for agropine and mannopine. A sensitive quantitative HPLC procedure was developed for the simultaneous quantification of tanshinone I, ta nshinone II(A), tanshinone II(B), tanshinone V, dihydrotanshinone I, c ryptotanshinone, tanshinone VI, and the colourless diterpene ferrugino l. These seven major tanshinones and ferruginol were found not only in the roots, but also in the liquid medium. Apparently, the hairy roots released secondary metabolites into the culture medium. The main prod ucts in the hairy root cultures were the tanshinones which are the act ive principles of this plant species. The effects of culture condition s on hairy root growth and the diterpene production have also been stu died. The most apparent effect was seen when ammonium nitrate levels o f the medium were modified.