- 期刊
未經冷馴化之草莓腋芽超低溫冷凍保存研究
Study on the Cryopreservation of Non-Cold Hardened Fragaria × ananassa Duch. Axillary Buds
摘要
草莓（Fragaria × ananassa Duch.）在全世界已有超過70個以上的國家進行栽培與量產，其為薔薇科（Rosaceae）多年生高經濟作物之一，因此進行種原的長期保存益顯重要，本試驗主要目的即在建立未經冷馴化草莓（「桃園1號」）腋芽最適宜的超低溫冷凍保存方法。腋芽培植體（芽體尺寸1－2 mm）在尚未浸入-196℃液態氮前之預處理結果顯示：（1）預培養於添加0.3 M蔗糖之MS基本鹽類培養基14 d後，平均存活率為96.7% ± 5.8%；（2）將填充溶液（LS）中的蔗糖濃度自0.6 M提升至1.6 M並且滲透壓保護90 min，腋芽的平均存活率仍可達到100%；以及（3）腋芽使用植物玻璃質化溶液2（PVS2）脫水處理60 min後，平均存活率達到86.7% ± 4.7%，此百分比顯著高於脫水處理180 min（46.7% ± 4.7%）和240 min（46.7% ± 17.0%）的處理組。含有腋芽的藻膠球繼代培養14 d後的存活率可達到66.7－80%。經比較後各處理之間並無顯著差異，故本研究各項結果有助於簡化草莓腋芽超低溫冷凍保存的方法。
並列摘要
Strawberry (Fragaria × ananassa Duch.) is one of the perennial crops with the high economic value. It belongs to the Rosaceae family and is distributed and cultivated in more than 70 countries. Therefore, the research on long-term storage of strawberry germplasm is important. The major objective of this study is to establish an optimum axillary bud cryopreservation method of non-cold hardened strawberry 'Taoyuan No.1'. The results obtained on pretreatment of axillary buds (bud size: 1- 2 mm) prior to plunge into liquid nitrogen (LN) at -196°C were as follows: (1) The average survival rate of buds was 96.7% ± 5.8% when the buds were precultured on MS basal medium supplemented with 0.3 M sucrose for 14 d; (2) By increasing sucrose level from 0.6 M to 1.6 M in loading solution (LS) and then osmoprotected for 90 min, the survival rate of axillary buds remained 100%; (3) The axillary buds survived was 86.7% ± 4.7% when treated with plant vitrification solution 2 (PVS2) for dehydration for 60 min. This survival percentage was significantly higher compared to PVS2 dehydration treatment for 180 min (46.7% ± 4.7%) and 240 min (46.7% ± 17.0%). After storage in LN, cryotubes were rapidly rewarmed in a water bath at 38°C for 60- 90 s. The survival rates of axillary bud beads in the range of 66.7- 80% were achieved after a subculture for 14 d. There were no significant differences in survival percentages of revived axillary buds among the storage duration in LN. These results appear promising for a simplifying cryopreservation method of strawberry axillary buds.