- 學位論文
白花蛇舌草（Hedyotis diffusa willd.）細胞懸浮培養生產齊墩果酸、熊果酸之研究
Production of oleanolic acid and ursolic acid by cell suspension culture of Hedyotis diffusa silld.
摘要
以田間白花蛇舌草（Hedyotis diffusa willd.）莖段為培植體建立無菌培養系統，利用再生植株之莖頂為培植體，可誘導出白色鬆軟癒合組織，白色癒合組織繼代至含0.002 mgl-1 thidiazuron培養基中，可產生綠色緊實之胚性癒合組織，將胚性癒合組織繼代至不含生長調節劑之MS培養基可再生大量小植株。以白色癒合組織建立細胞懸浮培養，每個月繼代一次進行細胞增殖，白色的懸浮細胞逐漸轉成綠色懸浮細胞。分析不同來源之癒合組織及細胞，以懸浮培養之綠色細胞中齊墩果酸與熊果酸含量較高。細胞不同接種量進行生長曲線測試，結果以2 g/ 10 ml medium之接種量培養12天，細胞鮮重可增加2.2倍為最佳。為促進細胞中齊墩果酸與熊果酸之產量，以此接種量搭配不同培養因子、誘引劑及前驅物處理，培養週期為12天，培養基中提高蔗糖濃度至50 gl-1時，細胞中齊墩果酸與熊果酸含量增加為1017±167 μg/g DW，或者添加低濃度50 mgl-1果膠累積也可達1356±83 μg/g DW，所有處理以添加12 mgl-1 methyl jasmonate累積量最高，齊墩果酸及熊果酸含量1591±206 μg/g DW，為不加誘引劑累積量的3倍。
並列摘要
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).
參考文獻
延伸閱讀
- Liang, Z. T., Jiang, Z. H., Fong, D. W. F., & Zhao, Z. Z. (2009). 高效液相色譜法測定白花蛇舌草及其代用品中齊墩果酸和熊果酸的含量. 藥物食品分析, 17(2), 69-77. https://www.airitilibrary.com/Article/Detail?DocID=10219498-200904-17-2-69-77-a
- 張志宇（2014）。數株Saccharomyces cerevisiae 細胞壁粗區分物之抗發炎及抑制腫瘤細胞增生功效研究〔碩士論文，國立臺灣大學〕。華藝線上圖書館。https://doi.org/10.6342/NTU.2014.02619
- 黃柏欣（2013）。壹、越南產黃毛假鷹爪果實之化學成分研究貳、中草藥萃取物抑制白菜炭疽菌之活性研究參、沉香葉部甲醇萃取物之二氯甲烷層抗癌活性成分研究〔碩士論文，國立虎尾科技大學〕。華藝線上圖書館。https://doi.org/10.6827/NFU.2013.00088
- Hsu, H. F., Wu, Y. C., Chen, L. C., & Houng, J. Y. (2009). Induction of Apoptosis of A549 Lung Cancer Cell Line by Dehydrocostus Lactone Isolated from Glossogyne Tenuifolia. 藥物食品分析, 17(2), 107-115. https://www.airitilibrary.com/Article/Detail?DocID=10219498-200904-17-2-107-115-a
- 王懷俐（2009）。Study of lipid production from selected oleaginous microorganism applied as biodiesel feedstock〔碩士論文，元智大學〕。華藝線上圖書館。https://doi.org/10.6838/YZU.2009.00323