Hirai D
Central Agricultural Experiment Station, Hokkaido Research Organization, Hokkaido, Japan.
Cryo Letters. 2011 Jul-Aug;32(4):287-96.
The droplet vitrification method was improved for maneuverability by embedding shoot tips in gelled droplets before osmoprotection. This newly modified cryopreserving method -gelled droplet vitrification - was compared with other PVS2-based cryopreservation methods using potato shoot tips. Survival rates of each cryogenic procedure held at 25 degree C were about 40 percent by cryotube-vitrification procedures (vitrification and encapsulation vitrification methods) and about 70 percent by PVS2-droplet procedures (droplet vitrification and gelled droplet vitrification methods). Much higher cooling rates of PVS2-droplet procedures than cryotube- vitrification procedures increased their survival rates. The gelled droplet vitrification method was applied to shoot tips of 26 potato cultivars and six wild potatoes. After a little modifications of the conditions for preculture, osmoprotection and dehydration, all cultivars and wild potatoes produced high enough survival rates to be of value to genebanks and all surviving shoot tips developed normal shoots within 3 weeks.
通过在渗透保护前将茎尖嵌入凝胶化液滴中,改进了液滴玻璃化法的可操作性。这种新改良的冷冻保存方法——凝胶化液滴玻璃化法——与其他基于PVS2的使用马铃薯茎尖的冷冻保存方法进行了比较。在25摄氏度下,通过冷冻管玻璃化程序(玻璃化法和包埋玻璃化法),每个低温处理程序的存活率约为40%,而通过PVS2液滴程序(液滴玻璃化法和凝胶化液滴玻璃化法)的存活率约为70%。PVS2液滴程序比冷冻管玻璃化程序更高的冷却速率提高了它们的存活率。凝胶化液滴玻璃化法应用于26个马铃薯品种和6个野生马铃薯的茎尖。在对预培养、渗透保护和脱水条件进行一些修改后,所有品种和野生马铃薯都产生了足够高的存活率,对基因库具有价值,并且所有存活的茎尖在3周内都发育出了正常的芽。