痲瘋樹之組織培養研究

Study on tissue culture of Jatropha curcas

王立釩(Li-Fan Wang)

指導教授 ： 蔡新聲

朝陽科技大學/理工學院/應用化學系生化科技碩士班/碩士(2014年)

摘要

痲瘋樹 (Jatropha curcas. L.) 屬於大戟科 (Euphorbiaceae) 落葉灌木或小喬木，原產熱帶美洲，葉子及樹汁被廣泛應用於傳統療法，其有效成分麻瘋樹酮 (jatrophone)、痲瘋樹三酮 (jatrophatrione) 及痲瘋樹鹼 (jatrophain)等皆具有抗癌作用。近年來，全球面臨能源枯竭和環境氣候變遷等問題，因此替代能源的開發成為重要議題。由於痲瘋樹生長快速，果實生產期可長達 50 年，植株不僅耐旱還可種植於貧瘠土地並具有水土保持等功能。痲瘋樹全株都有毒，其種子含有豐富的脂肪及各種碳水化合物，果實部分可萃取出生物柴油，萃取物可製造肥皂及含氮有機肥料，剩餘枝葉及果實經過脫毒後也可做為動物飼料。痲瘋樹果實含油率在 30% 到 61% 之間，迄今為止已發現的野生品種含油率最高為 61.8%，因此痲瘋樹是一種具有潛力並可成為未來替代石化燃料的能源作物，本研究乃利用組織培養技術建立痲瘋樹之大量繁殖系統，以無菌苗作為培植體，針對培養基成分組成進行試驗，探討培植體部位、基礎鹽類、植物生長調節劑種類及其濃度等條件對痲瘋樹大量繁殖效率之影響。結果顯示芽體誘導之最佳條件為利用頂芽培養於 MS 鹽類、2 mg/L BA、3% 蔗糖、0.9% Agar、pH = 5.7 ± 0.1，配合四層藥包紙之透氣處理 6 週後誘導之芽體數為4.1個，其結果約為未經透氣處理的 2 倍；添加 TDZ 誘導芽體可獲得最高之誘導率 90% 和芽體數為 3.3 個，誘導過程會大量產生癒合組織，而影響芽體生長；將增殖之組培苗培養於未添加植物生長調節劑之培養基，並添加活性碳可降低癒合組織產生，有助於增生芽生長；使用植物生長調節劑 IBA 誘導芽體發根可獲得細長的根系，添加 NAA 處理的根系則粗壯且較短，以添加 0.5 mg/L IBA 的誘導率 93% 為最高；馴化已完成發根且健康的無菌苗，移植存活率可達 100%。

關鍵字

痲瘋樹 ； 大戟科 ； 組織培養 ； 芽體誘導 ； 能源作物

並列摘要

Jatropha curcas L. Commonly known as Barbados Nut, Purging Nut or Physic Nut belongs to the family Euphorbiaceae. The plant is a perennial tree or large shrub, native to the tropical America. The plant is grown in low or high rainfall areas, used to reclaim land, improve the environment, reduce soil erosion, and is an alternative to the use of grains as biofuel feedstock. In past, Metsiyen in the Haitian culture dates back this as a medicinal crop and used it to ‘calm the stomach’. However, presently, it is widely used as an “Energy crop”. In recent years, the world is facing energy depletion and environmental issues such as climate change, alternative energy sources therefore have become an important issues. The plant produces many useful products; e.g. its seeds contain 30-61% of oil, which is an important raw material necessary to produce biofuel. Therefore, Jatropha is a potential alternative to fossil fuels and can be the future of energy crops. In the present study, tissue culture techniques have been used to establish in vitro propagation protocol of Jatropha curcas L.
In order to optimize an in vitro propagation protocol, experiments were carried out with different plant parts (leaf, terminal bud, stem and root), different basal media (MS, WPM, B5 and N6), plant growth regulators (BA, Kinetin and TDZ), and ventilating container closures (disposable papers and aluminum foil). Also, cotyledon, leaf, stem and root explants of in vitro derived seedlings were used as explants to induce callus in Jatropha curcas L. The results showed that the maximum response (83%) of adventitious bud formation was obtained in terminal bud used as explant cultured on MS basal medium supplemented with 1 mg/L BA, 3% sucrose and 0.9% agar. Among the four basal medium, MS supported the maximum adventitious bud (83%) in the terminal bud explant. Out of three cytokinins tested for adventitious bud formation, though TDZ resulted in the maximum response (90%) and a higher average number of buds (3.3 buds/explant), however buds did not grow, and remained stunted and induced callus. Among the five BA concentrations, a supplement of BA 2.0 mg/L in the medium resulted in the maximum adventitious bud formation (88%) followed by BA 1.0 mg/L (83%). Among the two ventilating container closures tested, the maximum average adventitious bud formation (4.1/explant) was recorded with AF3 + DP3 (2 layers of aluminum foil as container closure for 3 weeks of culture, then exchanging container closure with 4 layers of dispense paper for the next 3 weeks of culture). A supplement of 0.2% active charcoal in the medium prevented callus formation and promoted growth of buds in the culture. Induction of roots in 93% of in vitro shoots was achieved on MS basal medium supplemented with 0.5 mg/L IBA. Rooted shoots survived (100%) on a mixture of Peat: Vermiculite: Perlite (1:1:1 v/v).Thus, in the present study, we have developed an in vitro propagation protocol for Jatropha curcas L.

並列關鍵字

Euphorbiaceae ； tissue culture ； bud induction ； Energy crops ； Jatropha

參考文獻

胡文若、王瑞章、孫文章、陳俊仁，「包舌蘭無菌播種技術」，台南

陳國峰、何政坤、郭幸榮，「農桿根群菌促進台灣紅豆杉成熟樹插穗

文，大同大學生物工程學系，台北(2007)。

莊錦華、李哖，「菸鹼酸、椰子汁與香蕉泥對台灣一葉蘭種子發芽與

趙大衛，選修生物，翰林，台南 (2012)。