朵丽蝶兰拟原球体增殖与再生之研究.doc

朵麗蝶蘭擬原球體增殖與再生之研究 Studies on Protocrom-like-body Multiplication and Plantlet Regeneration of Doritaenopsis Orchid 研究生：黃雅文 Huang, Ya-Wen 指導教授：陳福旗 Chen, Fure-Chyi 【摘要】 本研究之目的在針對朵麗蝶蘭Doritaenopsis Tinny Ruby × Dtps. Minho Princess擬原球體（PLBs）經物理性處理後，置於誘導培養基中培養，以探討最適合誘導PLBs形成條件，並探討不同培養基成分對PLBs增殖之影響。擬原球體經物理處理後，具頂端分生組織的培植體通常會發育為小植株，基部的培植體則會形成二次擬原球體，進行四次之繼代培養，擬原球體之平均增殖率為17.4倍。在培養基中添加硝酸銀，或培植體浸泡硝酸銀能提高擬原球體發生與增殖。白化擬原球體經切割處理後，培養在缺乏tryptone的培養基，會導致擬原球體的發生率下降，濃度提高可以改善擬原球體的發生率。橫切的基部培植體，培養於添加洋菜之擬原球體發生率為97.5％；而不經切割的培植體，在培養基添加水晶洋菜則可促進25.6％擬原球體發生率。培養基添加ethephon處理之擬原球體發生率較ACC高；添加硝酸銀可促使擬原球體發生與增殖。培養基添加乙烯促進劑與抑制劑，瓶內乙烯的濃度會隨培養時間而有上升的趨勢，ethephon處理的乙烯生成量最高達3.391ppm。組織切片的結果顯示，擬原球體起源於表皮或次表皮細胞，將具頂端分生組織的擬原球體移至再生培養基，於光照環境下，可再生為小植株。 關鍵字：朵麗蝶蘭、擬原球體（protocorm-like bodies, PLBs）、ACC（1-aminocyclopropane-1-carboxylic acid）、AVG（aminoethoxyvinyl glycine）、硝酸銀（silver nitrate）、ethephonDoritaenopsis Tinny Ruby × Dtps. Minho Princess. Sectioned PLBs explants possessing apical meristem were regenerated into single plantlets, while sectioned PLBs without apical meristem could produce more secondary PLBs. The average proliferation rate of PLBs formation was 17.4 for four subcultures. Silver nitrate was effective in stimulating the formation of PLBs at the wound edges of explants. Without tryptone PLBs formation was decreased. Higher level of tryptone was promotive for PLB proliferation. In transversely bisected PLBs the formation rate of secondary PLBs reached 97.5％with ethephon effect higher than the use of ACC. Ethylene inhibitor AVG decreased PLB formation. Higher level of silver nitrate promoted PLBs formation. The ethylene content in flasks containing explants increased with culture time, no matter how the medium contained ethylene precursors or inhibitors. The highest ethylene reached 3.391ppm in medium supplementing ethephon. Histological sections indicated that secondary PLBs were induced from epidermal or sub-epidermal cells. Plantlets were obtained after transferring PLBs onto regeneration medium