Abstract

Ex situ storage plays an important role in the conservation of plant biodiversity. Cryopreservation at ultra-low temperatures (− 196 °C) is the only long-term ex situ preservation method for plant species that cannot be stored in seed banks. In the present study, we developed a cryopreservation protocol for micropropagated Rubus humulifolius (Rosaceae) plants representing currently critically endangered population of the species in Finland. Abscisic acid (ABA) has been found to increase the freezing tolerance of several plant species. Thus, we studied the effect of a 10-day pretreatment with 0, 2 or 4 mg/l ABA in comparison to freshly dissected buds. We also studied how the duration of in vitro subculture affects cryopreservation result. The ABA pretreatment had divergent effect on control and cryopreserved buds: the regeneration of non-cryopreserved control buds increased from 51% to 70%, 90% or 87% while the regeneration of cryopreserved buds decreased from 52% to 35%, 6% or 9% after 0, 2 or 4 mg/l ABA pre-treatments, respectively. Buds from plants subcultured for 1 month had 63% survival, which, however, decreased to 29% or nil% after 2 or 4 months subculture. The regenerated plants were successfully transferred from in vitro to in vivo conditions in common garden. Growing in garden is needed for future restoration of the species in wild. Cryostorage and other ex situ conservation actions carried out in botanical gardens may be of increasing importance as a tool to maintain plant biodiversity in the future.

中文翻译：

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低温保存可以对极度濒危物种悬钩子（Rubus humulifolius）进行长期保存

摘要

异地存储在植物生物多样性保护中起着重要作用。对于无法存储在种子库中的植物物种，超低温（-196°C）的低温保存是唯一的非原生境长期保存方法。在本研究中，我们开发了微繁殖的悬钩子悬钩子的冷冻保存方案（蔷薇科）植物代表了目前该物种的极度濒危种群。已发现脱落酸（ABA）可提高几种植物的抗冻性。因此，我们研究了与新鲜解剖的芽相比，用0、2或4 mg / l ABA进行10天预处理的效果。我们还研究了体外亚培养的持续时间如何影响冷冻保存结果。ABA预处理对对照和低温保存的芽有不同的作用：非低温保存的对照芽的再生从51％增加到70％，90％或87％，而低温保存的芽的再生从52％减少到35％，6％或0、2或4 mg / l ABA预处理后分别达到9％。传代培养1个月的植物的芽具有63％的存活率，但是，在传代2或4个月后，其存活率降低至29％或为零。将再生的植物成功地从体外条件转移到普通花园中的体内条件下。将来在野外恢复该物种需要在花园中种植。冷冻箱及其他作为未来维持植物生物多样性的工具，在植物园中进行的非原生境保护行动可能越来越重要。