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標題: 台農13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. 13 ) Treated by Vitrification
作者: 陳玟君
Chen, Wen-Chun
關鍵字: Cryopreservation;超低溫冷凍保存
出版社: 生命科學系所
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因為植物種原歧異度的快速消逝及種原所必須存在的價值,植物種原的保存已經變成全球關注的焦點。本研究以台農13號鳳梨(Ananas comosus (L.) No.13)無菌組織培養苗為實驗材料,以不同濃度蔗糖進行不同天數的預培養,以期成功以玻璃化法進行種原超低溫冷凍保存。結果顯示未經預培養前處理的植株頂芽,以LS 處理60分鐘,PVS2處理時間兩小時,其存活率為63.3%,而經過各種前處理之後,其存活率可提升至73.3%~100%,其中以0.2M長期預培養30天的植株,以LS 處理60分鐘,PVS2處理2及3小時其存活率皆可達100% 。根據生理分析測定,推測經過高濃度蔗糖長期預培養之植株,可以促使台農13號鳳梨降低相對含水量,適應高滲透逆境,並且累積溶質(可溶糖及可溶蛋白)提升了耐凍性及耐旱性。在高濃度蔗糖預培養期間形成之滲透逆境,可以誘導膜上的cation-chloride cotrasporter累積,增加離子通過細胞,幫助細胞轉換成更適應冷凍脫水之生理狀態。由本研究得知,以適當濃度蔗糖預培養適當天數前處理,可讓台農13號鳳梨使用玻璃化法超低溫冷凍保存後有最佳的存活率。企望本研究能供為發展鳳梨種原超低溫冷凍保存技術之參考依據。

Conservation of plant genetic resources has become an important issue of common global concern which is due to rapid vanishment of plant genetic diversity and the enormous potential value of genetic resources.
In this study, pineapple (Ananas comosus (L.) No.13) were used as the aseptic plantlets for testing material which precultured with different sucrose concentrations for different durations, and expects that it could be a successful vitrification procedure for cryopreservation. The results demonstrated that when the shoot tip of plantlets do not have preculture with a loading solution (LS) 60 minutes and dehydration with a plant vitrification(PVS2) for two hours, the highest survival rate was 65%, while after each kind of pretreatment, the survival percentage may promote to 73.3%~100%. After 0.2M long-term preculture 30 day, plantlet treated with a loading solution (LS) 60minutes and dehydrated with a plant vitrification(PVS2) for two hours or three hours, the highest survival rate was 100%.
Base on physiological measurements, the results suggest that preculture with high concentration sucrose may accelerate pineapple plantlets to reduce relative water content, and adapt osmotic stress, which also accumulates solutes (soluble sugar and soluble protein) to improve the tolerance of dehydration and freezing stress. In highly concentrated sucrose preculyure period forms the osmotic stress, may induce on the membrane cation-chloride cotrasporter to accumulate, the increase ion through the cell, helps the cell to transform adapts physiology of condition the freezing dehydration.Known by this research, the shoot tip of pineapple plantlets preculture with high concentration of sucrose adequately can make pineapple (Tainung No.13) has the best survival percentage after cryopreservation by vitrification. This study can be as an important reference for developing pineapple cryopreservation technology.
其他識別: U0005-2708200619480400
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