馬鈴薯莖頂組織超低溫冷凍保存之研究

Study on Cryopreservation using Potato Shoot Tip

詹庭筑(Ting-Zhu Zhan) ； 陸柏憲(Po-Hsien Lu)

《作物、環境與生物資訊》 16卷3期 (2019/09) Pp. 162-172

https://doi.org/10.30061/CEB.201909_16(3).0005

摘要

本試驗以馬鈴薯（Solanum tuberosum）台農一號（TNG1）及大西洋（Atlantic）之試管苗莖頂組織為研究材料，利用藻膠包埋玻璃質化法（encapsulation-vitrification） 進行超低溫冷凍保存（cryopreservation）之研究。本試驗之目的在於研究馬鈴薯適宜進行超低溫冷凍保存之處理條件，有助於建立馬鈴薯種原長期保存之整體流程。經試驗結果顯示，凍存前之預處理，以填充溶液（LS）滲透保護處理90 min，再以植物玻璃質化溶液2（PVS2）脫水處理180 min，馬鈴薯莖頂組織有最佳之存活率。經凍存1 h後以KG回復培養基進行回復生長，台農一號及大西洋存活率分別可達70.48%與76.92%，皆顯著高於IBK回復培養基（23.28%、14.09%），顯示KG回復培養基對馬鈴薯莖頂組織有較佳之回復生長條件。此外，本試驗進一步利用冷凍鋁片法（aluminum cryo-plate）進行超低溫冷凍保存，結果顯示台農一號及大西洋之存活率可達76.29%與88.39%，而再生率則達59.29%與60.92%。綜合整體試驗結果顯示，馬鈴薯莖頂組織可利用藻膠包埋玻璃質化法與冷凍鋁片法進行超低溫冷凍保存，兩者於凍存後皆有高存活率，且經回溫及回復生長後培植體皆可成功再生成完整植株。若以冷凍鋁片法進行馬鈴薯種原長期保存，可確保培植體有較高之冷凍效率，為理想且頗具應用潛力之超低溫冷凍保存技術。

關鍵字

冷凍鋁片法 ； 超低溫冷凍保存 ； 藻膠包埋玻璃質化法 ； 馬鈴薯

並列摘要

In this study, the shoot tips of potato (Solanum tuberosum) cultivars, Tainung No.1 (TNG1) and Atlantic, were used for the study of cryopreservation with the method of encapsulation-vitrification. The purpose of the experiment was to study the optimal processing conditions of cryopreservation and to establish the whole process of long-term preservation of potato shoot tips. With the application of encapsulation-vitrification, the results showed the optimal survival rate by treated with LS permeation protection was 90 min, dehydrated with plant vitrification solution 2 (PVS2) was 180 min before cryopreservation. The survival rates of TNG1 and Atlantic were 70.48% and 76.92% respectively after being frozen by liquid nitrogen for 1 h, with KG recovery medium. Both survival rates were significantly higher than IBK recovery medium (23.28%, 14.09%). The regeneration rate of the two varieties could reach 31.90% and 53.85% respectively. It was shown that the KG recovery medium was much better for the recovery growth of the potato shoot tips. In addition, the result of further utilizing the aluminum cryo-plate showed that the survival rates of TNG1 and Atlantic could reach 76.29% and 88.39%, while the regeneration rates were 59.29% and 60.92%, respectively. Overall, the shoot tips of potato would be cryopreserved by encapsulation-vitrification and aluminum cryo-plate. Both of them showed high survival rates after cryopreservation, and explants could successfully regenerate to complete plants after warming and recovery growth. It demonstrates that the germplasm resources preservation of potato is an ideal and potential cryopreservation technology by being applied the method of aluminum cryo-plate.

並列關鍵字

Aluminum cryo-plate ； Cryopreservation ； Encapsulation-vitrification ； Potato