Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/21527
標題: 台農16號鳳梨玻璃化法超低溫冷凍保存前處理流程之探討
Study of Pretreatment Protocols on the Cryopreservation of Pineapple ( Tainung No. 16 ) Treated by Vitrification
作者: 劉淑芬
Liu, Shu Fen
關鍵字: cryopreservation;超低溫冷凍保存;vitrification;pineapple;preculture;玻璃化法;鳳梨;預培養
出版社: 生命科學系
摘要: 
本研究以台農16號鳳梨(Ananas comosus (L.) Merr. cv. Tainung No. 16 )組織培養苗為材料,探討高濃度蔗糖培養基預培養與冷凍保護劑(PVS2)處理對鳳梨頂芽超低溫冷凍保存之貢獻。並且分析鳳梨對高濃度蔗糖培養基預培養處理的生理反應,希望經由生理的變化趨勢,了解預培養與超低溫冷凍保存後存活率的關係。
台農16號鳳梨組織培養苗以不同濃度蔗糖預培養基進行短期、中期和長期預培養,配合LS處理60分鐘,PVS2處理0~8小時,進行超低溫冷凍保存。台農16號鳳梨經0.2M蔗糖長期預培養處理,超低溫冷凍保存之存活率可達100%。根據生理分析測定,推測長時期以0.2M蔗糖培養基馴化鳳梨,可促使台農16號鳳梨適應高滲透逆境,累積足夠的溶質,如蔗糖、蘋果酸和澱粉,增加鳳梨的耐旱性與耐凍性。PVS2的最適處理時間為2~4小時,有最佳的超低溫冷凍保存之存活率。因此,玻璃化法冷凍保存流程之前處理中,以適當高濃度蔗糖長期預培養與適當時間之冷凍保護劑處理,為鳳梨超低溫冷凍保存成功關鍵之兩步驟。

This study investigates effects of preculturing high concentration of sucrose and effects of cryoprotectant (PVS2) on pineapple (Ananas comosus (L.) Merr. cv. Tainung No. 16 ) apices cryopreservation. The physiological responses of pineapple to understand the relations between preculture and cryopreserved survival rate were analyzed.
The apices from in vitro-grown plantlets of pineapple TN16 were precultured for short-,mid-and long-term on hormone-free MS medium containing different sucrose concentrations. The precultured apices were treated with a loading solution (LS) for 60 minutes and then dehydrated with a plant vitrification solution (PVS2) for 0 to 8 hours, and following cryopreservation. The highest survival rate of 100% was obtained when pineapple plantlets were long-term precultured on 0.2M sucrose medium. Based on the physiological measurements, the results suggest acclimation of pineapple plantlets occurred during long-term precultured on 0.2M sucrose medium, by accumulation of compatible solutes such as sucrose, malic acid and starch to improve the tolerance to dehydration and freezing stress. The best period of PVS2 treatment was 2~4 hours and had great effects on survival rates of cryopreservation. In conclusion, preculture with high concentrations of sucrose and time of pretreatment with cryoprotectant were two key steps to achieve the successfully cryopreservation of pineapple.
URI: http://hdl.handle.net/11455/21527
Appears in Collections:生命科學系所

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