夜來香體胚形成及再生之研究

Studies on Somatic Embryogenesis and Regeneration of Polianthes tuberosa L

本試驗希藉由植物生長調節劑組合及培養環境等參試條件，探討夜來香癒合組織誘導、體胚形成及植株再生過程，以建立一個完善的植株再生系統。以‘重瓣’夜來香（Polianthes tuberosa‘Double’）之幼花瓣為培植體，培養於含有auxins與cytokinins組合之MS培養基。結果顯示，auxins之添加對夜來香癒合組織的誘導具顯著影響，施以2，4-D（1，2，與4mg／L）即顯著（P≦0.001）影響癒合組織形成率，可達93%以上；但kinetin濃度增加（0，1，與2 mg／L）反而不利癒合組織的形成率（P≦0.05）；又2，4-D組合kinetin者，可誘導細緻且鵝黃色之胚性癒合組織。將胚性癒合組織培養於TDZ 0.01與0.1 mg／L之MS培養基，置於4μmol m-2s -1的弱光環境下，可獲得體胚及次生體胚。將體胚再繼代於含TDZ 0.01 mg／L之MS培養基四週後，可獲14 %的植株再生率。’嘉農秀玉’夜來香（P. tuberosa‘Chia-Nong Bright Jewel’）之未熟種子培植體培養於2，4-D 4 mg／L組合kinetin1 mg／L之MS培養基，誘導獲得之胚性癒合組織，培養於含有NAA 0.1 mg／L與BA 4.5 mg／L之Modified MS培養基，置於4μmol m-2s -1的弱光環境，可促進體胚形成及成熟，並獲得再生植株。 To establish an efficient plant regeneration of Polianthes tuberosa L., this study is to conduct the effects of MS basal medium supplemented with plant growth regulators and culture conditions on callus induction, somatic embryogenesis and plant regeneration. The results showed that using the young petal explants of P. tuberosa ‘Double’ cultured on MS medium containing auxin significantly increased callus formation. More than 93% of the explants forming callus was achieved by culturing young petal explants on basal medium contaning 1, 2 and 4mg/L 2,4-D. The higher concentration of kinetin (0, 1, and 2 mg/L), the less callus formation. However, the basal medium containing 2, 4-D and kinetin combination seemed to be more advantage for induction of yellowish, fine and smooth type of embryogenic calli. Somatic embryo and secondary somatic embryo could be obtained by culturing embryogenic calli on basal medium containing 0.01 and 0.1 mg/L TDZ under low light intensity at 4μmol m-2s -1. Plant regeneration with the rate of 14% was derived from subculturing somatic embryo on basal medium supplemented with 0.01 mg/L TDZ for 4 weeks. Embryogenic calli could also be derived from immature seed explants of P. tuberosa ‘Chia-Nong Bright Jewel’ cultured on basal medium containing 4 mg/L 2,4-D and 1 mg/L kinetin. Somatic embryogenesis, mature and plant regeneration were developed by subculturing embryogenic calli on modified MS medium containing 0.1 mg/L NAA and 4.5 mg/L BA under low light intensity at 4μmol m-2s -1.