Production of lithospermic acid B and rosmarinic acid in hairy root cultures of Salvia miltiorrhiza

Hairy root cultures of Salvia miltiorrhiza were established by infecting st erile plantlets with Agrobacterium rhizogenes ATCC 15834, and the transform ation was proved by direct detection of the inserted T-DNA by the polymeras e chain reaction. As determined by HPLC, these hairy root cultures had the ability to produce lithospermic acid B (LAB), rosmarinic acid (RA) and othe r related phenolic compounds, the water-soluble active components of the pl ant. The effect of five different basal media, MS, MS-NH4 (MS without ammon ium nitrate), B5, WPM and 6,7-V on the root growth and phenolic compound pr oduction was studied. It was found that MS-NH, and 6,7-V media were superio r to MS, B5 and WPM media in terms of both root growth and phenolic compoun d production. The time course of biomass accumulation and phenolic compound formation was also examined in the culture using MS-NH4 medium. During cul tivation, the content of RA in the roots was stable being approximately 0.4 8% of dry weight while the content of LAB fluctuated between 0.73% and 1.61 % of dry weight, and decreased gradually at the stationary phase of growth. The highest production of LAB and RA was about 64 mg L-1 and 23 mg L-1, re spectively.