- 學位論文
石斛蘭之體胚發生與植株再生
Somatic Embryogenesis and Plant Regeneration of Dendrobium
摘要
源於石斛蘭（Dendrobium ‘Chiengmai Pink’）重覆再生系統之胚性團塊，其外層細胞形成大量體胚。各個體胚皆可以獨立發育成一個完整的植株。比較固態與液態培養，體胚之發育與增殖較適合在固態下培養。培養基含TDZ會明顯減少抽芽比率，但鮮重之增殖倍率在統計上並沒有差異。 石斛蘭（Dendrobium ‘Chiengmai Pink’）之幼葉片段培養於含cytokinin（2iP, BA, kinetin, zeatin and TDZ）的1/2 MS基礎培養基，約30-45天，不經由癒傷組織，直接在切口附近形成多量體胚（somatic embryos）。最好的處理是含1 mg/l TDZ的培養基，有66.8 %的培植體產生平均13.3個體胚。將這些體胚培養在不含生長調節劑的培養基，經2-3個月，發育成株高約3-4公分的小植株。 以石斛蘭（Dendrobium ‘Chiengmai Pink’）無菌瓶苗之假球莖片段培養於1 mg/l TDZ之1/2 MS水晶洋菜培養基（GelriteTM-gelled 1/2 MS basal medium），一個月後，形成芽體、類原球體和綠色塊狀物。芽體從假球莖片段中抽出，有66.8 %的培植體產生平均3.36個芽。類原球體和綠色塊狀物在假球莖片段切口處形成，有35 %的培植體產生類原球體。根培植體並沒有任何形態發生，且在一個月內就褐化了。
並列摘要
Embryogenic masses from the repetitive regeneration system of Dendrobium ‘Chiengmai Pink’ formed large amount of somatic embryos from outer cell layers. Each somatic embryo could independently develop into an intact plantlet. Compare with solid and liquid culture, embryo development and proliferation is more suitable on the solid medium. Medium containing TDZ significantly reduced shooting rate, but it was no statistical different on the proliferation rate of fresh weight. Young leaf segments of Dendrobium ‘Chiengmai Pink’ directly formed somatic embryos from leaf regions near cut ends without an intervening callus within 30-45 days when cultured on a 1/2 MS basal medium supplemented with cytokinins (2iP, BA, kinetin, zeatin and TDZ). The best response was found at 1 mg/l TDZ. In this treatment, 66.8 % of leaf explants formed 13.3 somatic embryos. Somatic embryos developed into plantlets (3-4 cm in height) with several roots and shoots when cultured on a medium devoid of plant growth regulator after 2-3 months of culture. Pseudobulb segments taken from in vitro young plantlets of Dendrobium ‘Chiengmai Pink’ formed shoots, protocrom-like-bodies(PLB) and green masses within one month when cultured on a GelriteTM-gelled 1/2 MS basal medium containing 1 mg/l TDZ. In the best treatment, shoots formed within pseudobulb segments and 55 % of explants produced 3.36 shoots. PLBs and green masses formed from the cut side, 35 % of explants produced PLBs. Root explants turned brown within one month and no morphogenesis was observed.