Title

臺灣白花蝴蝶蘭莖頂玻璃化法超低溫冷凍保存流程之探討

Translated Titles

Investigation on the Protocol for Cryopreservation by Vitrification of Shoot Tips of Phalaenopsis aphrodite subsp. formosana

Authors

江徽之

Key Words

臺灣白花蝴蝶蘭 ; 莖頂 ; 玻璃化法 ; 超低溫冷凍保存 ; Phalaenopsis aphrodite subsp. formosana ; shoot tip ; vitrification ; cryopreservation

PublicationName

中興大學生命科學系所學位論文

Volume or Term/Year and Month of Publication

2012年

Academic Degree Category

碩士

Advisor

廖松淵

Content Language

繁體中文

Chinese Abstract

本試驗以原生種蘭花-臺灣白花蝴蝶蘭(Phalaenopsis aphrodite subsp. formosana)實生瓶苗,進行莖頂的玻璃化法超低溫冷凍保存,藉由各種生理分析,探討預培養處理及冷凍保護劑處理對冷凍保存後存活率的影響,以及預培養過程對莖頂生理狀態的影響。 生長至適當大小的蝴蝶蘭植株進行預培養後,切取1.5-2 mm莖頂,進行玻璃化法超低溫冷凍保存,依序測試不同PVS2處理時間、不同LS處理時間、不同預培養蔗糖濃度及不同預培養天數。試驗結果顯示,經0.4 M蔗糖濃度預培養7天後,以LS處理120 min,再以PVS2處理120 min,經液態氮保存至少一週後回溫,以回復培養基回復生長一個月後,存活率可達72%。 臺灣白花蝴蝶蘭植株經預培養後,分析莖頂水分生理及內含物變化,觀察到隨著預培養蔗糖濃度及預培養天數增加,莖頂組織內相對含水量、水分潛勢及滲透潛勢皆有逐漸下降的趨勢,累積於莖頂內的可溶醣及可溶性蛋白濃度顯著增加,顯示蝴蝶蘭經預培養後,可透過滲透調節改變莖頂生理狀態,提高對冷凍保護劑的耐受力及耐凍性,進而提升冷凍保存後的存活率。 總結上述,臺灣白花蝴蝶蘭玻璃化法超低溫冷凍保存的最佳流程為以0.4 M蔗糖濃度進行預培養7天,切取莖頂以LS處理120 min搭配PVS2處理120 min,經液態氮保存後,可獲得最高存活率。

Topic Category 生命科學院 > 生命科學系所
生物農學 > 生物科學
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