- 期刊
大葉桉以玻璃質化法進行超低溫保存之研究
Cryopreservation of Eucalyptus robusta by Vitrification Technique
摘要
本試驗嘗試利用大葉桉(Eucalyptus robusta)的芽體與懸浮培養細胞為材料，利用玻璃質化法進行超低溫保存之研究，檢測冰凍保護劑PVS2(plant vitrification solution 2)的毒性，並進行抗凍劑LS(loading solution)試驗及不同濃度PVS2處理的效果，嘗試找出較佳的配方與處理方式，以提高芽體與懸浮培養細胞冰凍後的活力。試驗結果顯示，浸泡PVS2溶液超過15分鐘將使大葉桉懸浮細胞活性降低至35%以下；LS處理可以降低PVS2對大葉桉懸浮細胞的毒性，使懸浮細胞在PVS2處理15分鐘後，仍可維持將近60%的活性。以50% PVS2處理大葉桉懸浮細胞5分鐘，再以100% PVS2處理10分鐘，細胞活力可達68%，解凍後能誘導出癒合組織。芽體經50% PVS2處理60分鐘，再以100% PVS2處理60分鐘，細胞活力可達82%。
並列摘要
This study was to investigate the cryopreservation by vitrification technique; Eucalyptus robusta buds and suspension cultured cell were used in this study. The toxicity of plant vitrification solution 2 (PVS2) and treatment by different concentrations of PVS2 were also studied in order to find out the optimal formula for the improvement of cell viability. After soaking in PVS2 more then 15 minutes, the viability of the suspension cultured cells of Eucalyptus robusta decreased to less than 35%. The loading solution (LS) treatment reduce the toxicity of PVS2 to the suspension cultured cells of Eucalyptus robusta, after soaking in PVS2 more then 15 minutes, with 60% cell viability maintained. Placing the suspension cultured cells of Eucalyptus robusta into 50% PVS2 for 5 minutes, then into 100% PVS2 for 10 minutes, then cell viability retain at 68%, with callus being induced. Placing the buds of Eucalyptus robusta into 50% PVS2 for 60 minutes, then into 100% PVS2 for 60 minutes, the cell viability could be maintained up to 82%.