Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/22324
標題: 北蕉玻璃化法超低溫冷凍保存前處理流程之探討
Investigation of pretreatments protocol on the cryopreservation of Giant Cavendish (Musa acuminata Colla) treated by vitrification
作者: 胡馨分
Hu, Shin-Fen
關鍵字: vitrification
玻璃化法
出版社: 生命科學系所
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摘要: 本研究以北蕉(Giant Cavendish. Musa acuminata Colla)組織培養苗為材料,探討玻璃化法超低溫冷凍保存前處理流程對存活率之影響。北蕉以高濃度蔗糖培養基健化與處理冷凍保護劑LS和PVS2,及進行生理分析和切片觀察,並探討流程中各種處理對冷凍保存存活率之影響和貢獻。 北蕉組織培養苗以不同濃度蔗糖培養基進行短期預培養和長期預培養,配合LS處理0~60分鐘,PVS2處理0~3小時,進行超低溫冷凍保存。結果顯示經0.3M蔗糖培養8週後,處理LS 20分鐘、PVS2 2小時,超低溫冷凍保存之存活率可達37.5%,未經液態氮處理可達77.08%。根據生理分析測定,發現北蕉經0.3M蔗糖長期培養後之相對含水量減少、滲透潛勢降低、可溶性醣類含量和可溶性蛋白質含量之也都增加。顯示高糖培養導致北蕉莖頂脫水及累積溶質,有助於適應高滲透逆境及增加冷凍保存時的耐凍性,推論高糖預培養有助於提高超低溫冷凍保存後的存活率。 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,為北蕉超低溫冷凍保存成功關鍵之兩步驟。
This study investigates effects of cryopreservation pretreatment by vitrification on banana (Giant Cavendish. Musa acuminata Colla). The physiological responses and observed section were analyzed by preculturing plantlets with high concentration of sucrose and cryoprotectant (LS and PVS2) treatment to investigate the effects and contribution of cryopreservation. The apices from in vitro-grown banana plantlets were precultured for short- and long-time on hormone-free MS medium containing different sucrose concentrations. The precultured apices were then treated with a loading solution (LS) for 0~60 minutes and then dehydrated with a plant vitrification solution (PVS2) for 0~3 hours, and following cryopreservation. Banana plantlet cultured on 0.3M sucrose medium long-time results in best survival rates of 37.5%, and treated without LN survival rates of 77.08%. Based on the physiological measurement,banana apices relative water content reduce, osmotic potential decrease, and both soluble sugar content and soluble protein content increase. These results indicated that banana plantlet culturing with high concentration of sucrose result in dehydration of banana shoot tips and accumulation of soluble sugar which renders high osmotic potential and increased freeze-tolerance for cryopreservation. It suggsets that precultrue in high concentration of sucrose is helpful for increasing survival rates after cryopreservation. Thus preculture with high concentration sucrose and the duration for pretreatment with cryoprotectant were achieved through investigating two key steps of the vitrification protocol.
URI: http://hdl.handle.net/11455/22324
其他識別: U0005-1008200616135200
文章連結: http://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-1008200616135200
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