Production Of Artemisia Annua L. Cell Biomass And Artemisinin Via Manipulation Of Culture Medium, Abiotic Culture Condition And Elicitation
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Date
2014-02
Authors
Lo, Kiah Yann
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Publisher
Universiti Sains Malaysia
Abstract
Due to the expensive, low yield and complexity of direct synthesis of artemisinin, in vitro plant cell culture techniques could be considerably the most applicable alternatives for artemisinin production. This study was carried out to establish and optimize in vitro cell suspension culture from elite clone of Artemisia annua L. of Vietnam origin for cell biomass and secondary metabolites production. Friable green calli were successfully induced from the aseptic leaf pieces on solidified MS medium supplemented with 0.5 mg/L BA, 0.5 mg/L NAA, 0.5 g/L casein hydrolysate and 30 g/L sucrose after 35 days of culture. More calli were induced when the leaf pieces were placed with abaxial surface in contact with the medium. Long term maintenance of the cell line was achieved by transferring callus of 0.50 g/ 40 ml gelled medium onto fresh medium at four weeks interval and cultured under continuous illumination. By using at least four months old calli, A. annua L. cell suspension culture was established with the same callus induction medium but without the agar. Strategies to formulate cell growth and secondary metabolite production medium were carried out by manipulating medium nutrients, abiotic culture conditions, and elicitation. Results indicated that initial inoculum density of 0.50 g/ 25 ml liquid medium required 16 days to reach the maximum cell biomass and artemisinin production. Clumping of cells was reduced by filtering the cells through 850 μm mesh sieve during subculture process when necessary. The optimal abiotic growth conditions which stimulated highest cell biomass and secondary metabolites at 16 days culture period were by culturing the cells under continuous illumination on shaker with speed of 100 rpm and temperature of 24 –28 °C.
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Keywords
Cell biomass and artemisinin , via manipulation of culture medium