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Study of Pretreatment Protocols on the Cryopreservation of Pineapple ( Tainung No. 16 ) Treated by Vitrification
Liu, Shu Fen
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。
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.
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