篇名 | High Efficient Somatic Embryo Multiplication and Embling Conversion of Dieffenbachia ‘Tiki’ |
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卷期 | 61:2 |
並列篇名 | 黛粉葉高效率體胚增殖與胚苗轉化 |
作者 | 沈榮壽 、 李哖 |
頁次 | 091-104 |
關鍵字 | primary somatic embryogenesis 、 secondary somatic embryogenesis 、 dicamba 、 thidiazuron 、 初級體胚發生 、 次級體胚發生 、 3,6-二氯甲氧基苯甲酸 、 1-苯基-3- |
出刊日期 | 201506 |
本研究探討黛粉葉體胚發生量化增殖效率與胚苗轉化再生體系。雄花序培植體於半量MS基本鹽類,添加20 g·L-1蔗糖、10 g·L-1葡萄糖及1.8 g·L-1水晶洋菜等為基礎培養基,結果顯示在2 mg·L-1 dicamba及0.5 mg·L-1 TDZ組合,具有最高之初級體胚發生頻率可達100%以及最多成熟體胚形成數達75.2個,對黛粉葉雄花序培植體高效率體胚發生的促進,具顯著的效果。以得自前一試驗均質之初級體胚,作為次級培植體來源,繼代培養於相同基本培養基添加2 mg·L-1 dicamba及TDZ (0.5、1.0 mg·L-1),培養13週後顯現最高效率的次級體胚發生頻率(100%),次級體胚形成數則高達34.3及33.0個,顯示此培養基組成可有效促進次級體胚發生和體胚量化增殖。此外,成熟次級體胚在1/2 MS添加150 mL·L-1椰子水之培養基組成之胚苗轉化顯示,大小約為4-5 mm次級體胚,培養在含有0.5%葡萄糖及1.0 mg·L-1 TDZ濃度培養基中其胚苗轉化率最高達73.3%;平均每一培植體之胚苗轉化再生植株最多達7.8個,且可發育成為具形態雙極性之獨立個體。胚苗轉化之再生植株經出瓶種植在溫室中具有良好的生長活力。
In this study, a regeneration system was developed for improving the efficiency of somatic embryogenesis and embling recovery by using male inflorescences and mature somatic embryos of Dieffenbachia Schott ‘Tiki’. The highest percentage (100%) of primary somatic embryogenesis (PSE) and the highest yield (75.2) of mature primary embryoids were obtained using a half strength modified Murashige and Skoog (MS) medium containing 2 mg·L-1 dicamba and 0.5 mg·L-1 thidiazuron (TDZ) from male inflorescence explants. The mature embryoids obtained from PSE were also used to induce secondary or repetitive somatic embryogenesis. PSEs derived from male inflorescence were cross-cut into four sections before subculturing them on the same basal medium supplemented with 2 mg·L-1 dicamba and 0.5-1.0 mg·L-1 TDZ. The highest secondary somatic embryogenesis frequency (100%) was obtained after 8 weeks of culture, and the highest yield of mature secondary embryos, 33.0-34.3 per explant, was obtained after 13 weeks of culture. In addition, embling conversion of mature secondary embryos was investigated. Green secondary embryos, 4-5 mm in size and 8-10 weeks in age, were cultured on a half strength modified MS medium supplemented with 150 mL·L-1 coconut water as a basal medium for embling conversion. The results indicated that the highest converted frequency was 73.3%, and the number of converted emblings per explant reached 7.8 when the medium contained 0.5% glucose and 1.0 mg·L-1 TDZ. Plantlets regenerated from the germinated embryoids acclimatized in the greenhouse and exhibited vigorous growth.