| 摘要註: |
在本論文中，實驗設計將不同品系蝴蝶蘭Doritaenopsis I-Hsin Ice Coke “K68502”、Doritaenopsis Malibu Peewee “K63357” 和Phalaenopsis Tropican Lady “K40311” 的不定芽培植體 (adventitious bud explants) 培養在玻璃瓶和可程控生物反應器內。測試不同的淹灌頻率、通氣量、更換培養液頻率和搭配額外添加polyvinylpyrrolidone (PVP) 處理培養六週後，比較不定芽數目和鮮重的增生倍率，並分析反應器內部二氧化碳濃度變化、培養液養份消耗和培植體的抗氧化相關表現。根據實驗結果，試圖尋找培養的最佳條件，以改善一些不易培養但具商業價值品系的增生問題。實驗結果顯示，過高的通氣量 (每分鐘40 ml) 或淹灌頻率 (每小時五分鐘)會增加葉片的H2O2 和superoxide dismutase (EC1.15.1.1)活性，因而限制芽體的增生。以每兩小時淹灌四分鐘，每分鐘通氣4 ml，每三週更換培養液，前三週添加0.02% (w/v) PVP 是蝴蝶蘭不定芽增生的最佳條件，而且芽數和鮮重的增生效率皆遠優於傳統玻璃瓶。此外，由於三週培養後，培養液內仍有未消耗的醣類及其他養份，包括NO3-、phosphate、malate、NH4+和K+等。因此，未來可嘗試降低更換培養液頻率，以降低蝴蝶蘭的生產成本。 In this thesis, the experiments were designed to culture the adventitious bud explants of different Phalaenopsis cultivars, such as Doritaenopsis I-Hsin Ice Coke ”K68502”, Doritaenopsis Malibu Peewee “K63357” and Phalaenopsis Tropican Lady “K40311”, into glass vessels and a programmable bioreactor with various treatments, including immersion frequency, airflow rate, replacing medium frequency, and adding polyvinylpyrrolidone (PVP), for six weeks. Comparing the propagation number and biomass of adventitious buds, analyzing the changes of CO2 concentration and the consumptions of medium nutrients inside the bioreactors, and the anti-oxidation related expressions of explants, then, according to these results, try to look for the optimum culture condition in order to improve the propagation issue for some Phalaenopsis cultivars with difficult culture characteristics but possess commercial value.The results revealed that a higher airflow rate (40 ml/min) or immersion frequency (5min/1hr) increased the levels of H2O2 and superoxide dismutase (EC1.15.1.1) activity of the leaf, so, limited the propagation of adventitious buds. The optimum culture condition of adventitious buds was 4 ml per minute airflow rate, 4 min immersion every two hours,replacing medium every three weeks, and simultaneously adding 0.02% (w/v) PVP for beginning three weeks, thus, the proliferation efficiency of biomass and bud number both were prominently superior to those of traditional glass vessels. Moreover, due to the residual sucrose and other nutrients, including nitrate, phosphate, malate, ammonium, potassium, etc, all were not completely consumed after three weeks cultured, in the future, the frequency of replacing medium could try to be decreased, and lower the production cost of Phalaenopsis. |