Journal of Taiwan Agricultural Research

Author:Uei-Chern Chen, Chi-Ni Hsia, Mau-Shing Yeh, Chin-Yi Tsao, and Hsin-Sheng Tsay*

Abstract:

    Bupleurum kaoi, a native medicinal plant in Taiwan which contains saikosaponins higher than other commercial Bupleurum species, was chosen for producing saikosaponins in vitro.  Cell sus-pension cultures of B. kaoi were cultured in a half-strength MS liquid medium containing 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) and used for studying effect of speed of shaker, size of fasks and inoculum density on cell proliferation as well as studying effect of elicitors, yeast extract (YE), abscisic acid (ABA), and jasmonic acid (JA), on saikosaponin production.  Results showed that cell proliferation of B. kaoi in suspension cultures was higher using shaker speed at 80 rpm than that of 100 rpm with peak period of cell growth at 3-4 weeks.  Cell proliferation of B. kaoi was also af-fected by inoculum density and fask size, with the optimum inoculum density of 12% by adding 6 mL  packed cell volume (PCV) cultures in 250-mL fasks.  Three elicitors (YE, ABA, and JA) tested in 3-week-old suspension cultures of B. kaoi were found reducing cell fresh weight and dry weight sig-nifcantly.  However, one week after the treatments of elicitors, saikosaponin contents and its produc-tivity increased 1.9-2.0 folds and 1.6-1.8 folds, respectively.  Further study using 500 mg/L YE as elicitor in cultures of B. kaoi for two weeks, similar increasing tendency on saikosaponin contents and its productivity was found with 2.1-fold and 2.8-fold, respectively.  This study concludes that adding 500 mg/L YE into cell suspension cultures of B. kaoi after the lag phase of cell cycle for one to two weeks would not reduce cell growth but increase saikosaponin production.

Key words:Bupleurum kaoi, Medicinal plant, Cell suspension culture, Saikosaponin, Elicitor

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