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Influence of Medium Components on Callus Proliferation and Saikosaponin Content of Bupleurum kaoi Liu, Chao et Chuang
Callus of Bupleurum kaoi grown on various strength of MS basal medium containing 0.2 mg/L 2, 4-D, 0.5 g/L peptone, 0.5 g/L casein hydrolysate and 100 mL/L coconut water for (A) 2 weeks, (B) 4weeks, (C) 6weeks, and (D) 8 weeks of culture. (E) Browning in callus cultured in 1/4 × MS and 1/2 × MS treatments for 6 weeks. (F) Callus were classified from 0 to 5 according to their increasing browning.
Callus of Bupleurum kaoi grown on various strength of MS basal medium containing 0.2 mg/L 2, 4-D, 0.5 g/L peptone, 0.5 g/L casein hydrolysate and 100 mL/L coconut water for (A) 2 weeks, (B) 4weeks, (C) 6weeks, and (D) 8 weeks of culture. (E) Browning in callus cultured in 1/4 × MS and 1/2 × MS treatments for 6 weeks. (F) Callus were classified from 0 to 5 according to their increasing browning.

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

Abstract:

    Medium components and their best concentrations of cytokinin, sucrose, Difco Bacto-agar, peptone, casein hydrolysate, coconut water were separately tested for callus growth of Bupleurum kaoi Liu, Chao et Chuang. Saikosaponins production of callus tissues was monitored.  A half-strength MS salt medium supplemented with 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D) was used as a basal medium (D2 medium) for callus growth.  The results showed that additional N6-benzyladenine (BA) and N-furfuryladenine (kinetin) inhibited callus growth in all concentration tested in this study.  Other than cytokinin, all factors could increase callus growth at a proper range of concentrations.  It was concluded that 3% sucrose, 0.9% Difco Bacto-agar, 0.5 g/L peptone, 0.5 g/L casein hydrolysate, and 100 mL/L coconut water had the best effects on callus growth of B. kaoi.  Various strengths of MS basal salts in combination with the aforementioned components at the best concentrations were further tested.  Among four strengths of MS tested, half-strength MS treatment had the highest fresh and dry weight of callus growth after 4 weeks of culture.  However, the highest saikosaponin a, c, d contents examined by high performance liquid chromatography (HPLC) was obtained from callus grown on the full-strength MS basal medium.

Key words:Bupleuri Radix, Medicinal plant, Cell culture, Secondary metabolite, High performance liquid chromatography

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Journal of
Taiwan Agricultural Research
Vol. 73, No.1 (2024) ISSN: 2790-086X Vol. 73, No.1 (2024)
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