雷公藤組織培養與相關學術應用領域之探討

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Title:	雷公藤組織培養與相關學術應用領域之探討 Study on plant tissue culture of Tripterygium wilfordii Hook f. and its application in related academic fields
Authors:	楊宗政
Contributors:	王姿文嘉南藥理科技大學：生物科技系暨研究所
Keywords:	癒傷組織抗氧化物雷公藤農桿菌 Antioxidants Triptygium wilfordi i (TWHF) Agrobacterium
Date:	2010
Issue Date:	2010-10-12 15:10:49 (UTC+8)
Abstract:	本研究分為二大部分：第一部份為探討雷公藤癒傷組織於不同培養基條件之生長效率，癒傷組織抗氧化能力以及藉由與農桿菌共培養(co-cultivation)探討基因表現並確認其轉殖效率。本研究首先評估植物除了調節荷爾蒙外，我們嘗試在誘導培養基中加入15％椰子水，結果發現可以防止雷公藤癒傷組織褐化的產生，有加入椰子水比未加入椰子水平均重量增加18.1％，我們更一步將癒傷組織分別置於不同光度下（700、900、1100 ux）培養30 天後，結果顯示在700 lux 增加64.4％最佳。在雷公藤癒傷組織抗氧化能力實驗中，我們利用DPPH（diphenyl picrylhydrayl）測其抗自由基的活性，結果顯示雷公藤癒傷組織平均抗氧化值為0.2205 相對於dl-α-tocopherol 濃度為3.8578mM。另外，利用雷公藤癒傷組織與農桿菌共培養探討基因表現並確認其轉殖效率，但由於雷公藤癒傷組織無法再生芽體，此實驗需要更進一步討論後進行。第二部份為針對全國各大專院校從事生物科技研究現況進行調查，此研究目的是以問卷調查方式，針對國內大專院校從事植物及中草藥的研究方向，以期能提供生物科技研究人員及生物科技之相關產業，評估目前市場機會及發展目標。最後，在全國各大專院校從事植物組織培養、植物基因轉殖、中草藥臨床應用及經費來部分，我們拜訪全國17 所大專院校78 位教授。結果顯示，植物組織培養方面以蘭花培植改良佔27％為最多，是台灣最具前瞻性的生技產業；在植物基因轉殖方面，以阿拉伯芥為模型植物研究佔25％，顯示出國內學者基礎研究之重視；中草藥研究佔受訪人數56％，其中以提升免疫力靈芝佔8％，複方中草藥佔10％，對傳統中藥及天然物來源之新藥設計開發及臨床藥效分析，使得中草藥在國際性形象的提升和卓越的貢獻。 In this study, we first intended to evaluate the effects of factors other than plant hormone regulators. Addition of 15% coconut water in our callus induction medium would prevent brownish callus and promote the growth of TWHF to 15% (dry weight) when compared with those of medium without coconut water. We further investigate the influence of light intensities (700, 900, 1100 lux) in callus culture of TWHF for 30 days. Among the treatments, the highest growth rate with 64.4% increase was observed when callus was cultured under 700 lux. The anti-oxidant activity of TWHF callus was estimated by measuring the inhibition of free radical diphenyl picrylhydrazyl (DPPH). The results showed that the average antioxidant value of TWHF calli was 0.2205 and this value is equivalent to dl-α-tocopherol 3.8578mM concentration. We attempted to establish Agrobacterium-mediated genetic transformation of TWHF through indirect organogenesis. However, the failure of regeneration from putative callus transformants, more modification of protocol would be needed to achieve this goal. Finally, we underwent a market survey by visiting 78 professors (among 17 universities) who conducted the researches in plant biotechnology to provide the information for biotech industry in Taiwan. The results show that 27% of studies in plant tissue culture are related to orchid biotechnology and this is with the most profit potential in Taiwan. Twenty-five percent of plant genetic transformation adopts Arabidopsis as materials implying the importance of plant basic science for researchers in Taiwan. Fifty-six percent of respondents are interested in Chinese herb. Around 8% of these studies investigate the immunomodulation role of Reishi mushroom and 10% of the cases in this category intend to find different functions of traditional Chinese prescription. The results of this survey suggest that Chinese herb would be the candidates for new drug development in Taiwan.
Relation:	校內校外均不公開，學年度：98，101頁
Appears in Collections:	[Dept. of Biotechnology (including master's program)] Dissertations and Theses

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