University of Oulu

Edesi, J., Tolonen, J., Ruotsalainen, A.L. et al. Cryopreservation enables long-term conservation of critically endangered species Rubus humulifolius. Biodivers Conserv 29, 303–314 (2020) doi:10.1007/s10531-019-01883-9

Cryopreservation enables long-term conservation of critically endangered species Rubus humulifolius

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Author: Edesi, Jaanika1,2; Tolonen, Jonne1; Ruotsalainen, Anna Liisa1;
Organizations: 1Ecology and Genetics Unit, University of Oulu, P.O. Box 3000, 90014, Oulu, Finland
2Production Systems, Natural Resources Institute (Luke), 57200, Savonlinna, Finland
Format: article
Version: published version
Access: open
Online Access: PDF Full Text (PDF, 1 MB)
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Language: English
Published: Springer Nature, 2020
Publish Date: 2019-12-16


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.

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Series: Biodiversity and conservation
ISSN: 0960-3115
ISSN-E: 1572-9710
ISSN-L: 0960-3115
Volume: 29
Pages: 303 - 314
DOI: 10.1007/s10531-019-01883-9
Type of Publication: A1 Journal article – refereed
Field of Science: 1183 Plant biology, microbiology, virology
Funding: Open access funding provided by University of Oulu including Oulu University Hospital. Our sincere thanks to Aino Hämäläinen, Sirpa Lehtola, Tuomas Kauppila (from Botanical Gardens, University of Oulu, Finland) and Taina Uusitalo (Ecology and Genetics Unit, University of Oulu, Finland) for taking care of the plant material. The study was partly funded by LIFE+ 2011 (BIO/FI/917 ESCAPE).
Copyright information: © The Author(s) 2019. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (, which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.