Kim H H, Yoon J W, P Y E, Cho E G, Sohn J K, Kim T K, Engelmann F
National Institute of Agricultural Biotechnology, RDA, Suwon 441-707, Korea.
Cryo Letters. 2006 Jul-Aug;27(4):223-34.
The applicability of cryopreservation protocols to a broad range of genotypes is a key issue for genebanks. We tried to identify the critical factors causing differences in survival of cryopreserved shoot tips using potato varieties coming from cultivated and wild species. The droplet-vitrification method, a combination of droplet-freezing and solution-based vitrification, was selected from several protocols. High survival after freezing was observed after dehydration with PVS2 for 20 min, cooling shoot tips placed in a droplet of PVS2 solution on aluminum foil strips by immersing the foil strips in liquid nitrogen, warming them by plunging the foil strips into a 0.8 M sucrose solution (at 40 degrees C) for 30 s and unloading in 0.8 M sucrose for 30 min. This optimized protocol was successfully applied to 12 accessions with survival ranging between 64.0 and 94.4%.
冷冻保存方案对广泛基因型的适用性是基因库的一个关键问题。我们试图利用来自栽培种和野生种的马铃薯品种，确定导致冷冻保存茎尖存活率差异的关键因素。从几种方案中选择了滴液玻璃化法，即滴冻法和基于溶液的玻璃化法的结合。在用PVS2脱水20分钟后观察到冷冻后有较高的存活率，将茎尖置于铝箔条上的一滴PVS2溶液中，通过将铝箔条浸入液氮中进行冷却，将铝箔条迅速放入40℃的0.8M蔗糖溶液中30秒进行升温，并在0.8M蔗糖中卸载30分钟。这个优化方案成功应用于12个种质，存活率在64.0%至94.4%之间。
