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標題: 巨峰與蜜紅葡萄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 grapevine(Vitis vinifera L.) treated by vitrification
作者: 陳瓊均
Chen, Chiung-Chun
關鍵字: cryopreservation;冷凍保存
出版社: 生命科學系所
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巨峰葡萄和蜜紅葡萄經不同高濃度蔗糖預培養3天後,以LS處理60分鐘,PVS2冷凍保護劑脫水115分鐘,進行超低溫冷凍保存。巨峰葡萄經0.6M高濃度蔗糖可達60.8%之存活率;蜜紅葡萄經0.3M高濃度蔗糖可達58.9%之存活率。二供試品種在最適預培養濃度下, LS與PVS2處理不同時間其存活率會有異,處理時間太短,無法誘導組織適當脫水,太長則易造成細胞產生過度滲透衝擊及毒性。

Effects of preculture in high concentration sucrose and cryoprotectant pretreatment on two cultivars of grapevine (Vitis vinifera L.) named cv.‘Kyoho'and cv.‘Honey Red'during cryopreservation were investigated. To understand the effects of high concentration of sucrose in preculture medium to the survival of cryopreservation from grape shoot tips, we analyzed the hydric and biochemical modification.
Precultured shoot tips with different sucrose concentrations for 3 days were loaded with LS for 60 min and dehydrated with PVS2 for 115 min at 00C before immersion into LN. The best survival rate of 60.8% and 58.9% were obtained when shoot tips were precultured on 0.6M and 0.3M sucrose for grape of cv.‘Kyoho'and cv.‘Honey Red', respectively. In the adaptable preculture median, cryoprotectant pretreatment is important, when pretreatment duration too short could led to tissue without adaptable dehydration, pretreatment duration too long could have a severely osmotic stress and toxicity.
After preculturing two cultivars of grape shoot tips on different sucrose concentrations for 3 days, the water potential and osmotic potential decreased. In cv. ‘Honey Red', the turgor pressure was significantly decreased when were precultured on 0.4M sucrose. However, cv. ‘Kyoho 'that turgor pressure was significantly decreased when were precultured on 0.7M sucrose. Preculturing with sugar-rich medium induced changes in the level of total soluble proteins. Sucrose concentration affected the accumulation of soluble sugar, and indirectly affected osmotic potential change. What kinds of soluble sugars accumulate after osmotic stress is important. Accumulation of disaccharide can enhance survival of cryopreserved shoot tips. The TTC reduced in stained shoot tips, indicated that preculturing with high concentration sucrose led to tissue vitality loss.
Thus, preculture with high concentration sucrose and the duration for cryoprotectant pretreatment play two key roles for investigating grape vitrification.
其他識別: U0005-0408200623245600
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