Production of oleanolic acid and ursolic acid by cell suspension culture of Hedyotis diffusa silld.

• Main Authors: Kung-Yu Liu, 劉孔煜
• Other Authors: Chin-Wen Ho
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/41751511062409799931

碩士 === 大同大學 === 生物工程學系(所) === 97 === The stem of Hedyotis diffusa willd was used as explant to establish the in vitro culture system. For callus induction, shoot tips from regenerated plantlet were cultured on modified MS medium and the white friable callus proliferated from shoot tips. The green embryogenic callus formed when white callus were subcultured on MS medium supplemented with 0.002 mgl-1 thidiazuron. Plantlets regenerated when embryogenic callus was subcultured on MS medium without plant growth regulator. For cell suspention culture system, the white callus was cultured in modified MS medium and subcultured every month. The content of oleanolic and ursolic acid of four kinds of callus (white callus、green callus、white suspention cell and green suspention cell) were examined by high performance liquid chromatography. The green cell has the highest content of oleanolic and ursolic acid (538±77 μg/g DW) and the growth index of green cell line ( inoculum : 2 g/10 ml ) was 2.2 times within 12 days. In order to enhance the accumulation of oleanolic and ursolic acid in cell, cell was cultured in MS medium with different culture factor and precursor elicitor. The highest content of oleanolic and ursolic acid ( 1591±206 μg/g DW) in cell was shown when cell cultured in MS medium added with 12 mgl-1 methyl jasmonate after 12 days. The production of oleanolic and ursolic acid in cell was also promoted when medium added 50 gl-1 sucrose ( 1017±167 μg/g DW) or 50 mgl-1 pectin supplemented(1356 ±83 μg/g DW).