Journal of Taiwan Agricultural Research

Author:Uei-Chern Chen, Hsiao-Sung Chan, Choi-Yi Lee, Chin-Yi Tsao, Jen-Chyi Liu, Yahn-Chir Lee, and Chi-Ni Hsia*

Abstract:

    Leaf explants of Salvia miltiorrhiza were inoculated with Agrobacterium rhizogenes strain R1601 to induce formation of hairy roots.  Root cultures were established and maintained in a semi-solid or a liquid B5 medium.  They were used to study effect of source of inoculum, size of inoculum, type of media and incubation period on production of root biomass and tanshinones in liquid culture.  Results showed that root tips were superior to other root segments as inoculum for production of root biomass and tanshinones in liquid cultures.  The treatments of 10, 15 and 20 root tips/20 mL B5 medium significantly increased root growth and production of tanshinones, compared to the treatment of 5 root tips/20 mL B5 medium.  Concentration of sucrose and NH₄⁺ in B5 liquid cultures rapidly reduced to 9% and 10%, respectively, in 3 weeks, resulting in a decrease in biomass accumulation.  This indicates that carbohydrate and nitrogen might be important limiting factors for hairy root growth of S. miltiorrhiza.  Production of dry weight biomass in liquid B5 medium peaked at 5.8 g L^-1 after 6 weeks, whereas production of tanshinones peaked at 64 g L^-1 after 8 weeks.

Key words:Danshen, Salvia miltiorrhiza, Time-course study, Nitrogen source, Carbohydrate source, Secondary metabolite, Tanshinones, Hairy root culture

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