Genebank Department, Leibniz Institute of Plant Genetics and Crop Plant Research (IPK), Seeland, Germany.
Methods Mol Biol. 2021;2180:647-661. doi: 10.1007/978-1-0716-0783-1_35.
Cryopreservation of shoot tips facilitates long-term storage of plant genetic resources which can otherwise only be propagated vegetatively. The vitrification approach using the cryoprotectant plant vitrification solution 3 (PVS3, 50% sucrose and 50% glycerol) is easy to handle, has shown to produce high regrowth percentages in a number of potato, mint, garlic, and shallot accessions, and is, thus, highly suitable for routine cryopreservation of plant genetic resources. In the current chapter, the vitrification procedure is described for potato, mint, garlic, and shallot and includes details about modifications for the different plant species. Special emphasis is given on the preparation of the different culture media, solutions, the culture conditions prior and post-cryopreservation, and the preparation of the shoot tips from different sources. Furthermore, protocols to introduce plants into in vitro culture and methods to estimate cryopreservation success are provided.
茎尖的超低温保存有助于长期保存植物遗传资源,否则这些资源只能通过营养繁殖来进行繁殖。使用植物玻璃化溶液 3(PVS3,50%蔗糖和 50%甘油)的玻璃化方法易于操作,已在许多马铃薯、薄荷、大蒜和青葱品种中显示出较高的再生率,因此非常适合植物遗传资源的常规超低温保存。在本章中,描述了马铃薯、薄荷、大蒜和青葱的玻璃化程序,包括针对不同植物物种的修改细节。特别强调了不同培养基、溶液的制备、超低温保存前后的培养条件,以及从不同来源制备茎尖的方法。此外,还提供了将植物引入体外培养的方案和评估超低温保存成功的方法。