Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures
Somatic embryogenesis (SE) is considered the most effective method for vegetative propagation of Norway spruce (<i>Picea abies</i> L. Karst). For mass propagation, a storage method that is able to handle large quantities of embryogenic tissues (ETs) reliably and at a low cost is required...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-12-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/24/15516 |
| _version_ | 1797457313853865984 |
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| author | Saila A. Varis Susanna Virta Itziar A. Montalbán Tuija Aronen |
| author_facet | Saila A. Varis Susanna Virta Itziar A. Montalbán Tuija Aronen |
| author_sort | Saila A. Varis |
| collection | DOAJ |
| description | Somatic embryogenesis (SE) is considered the most effective method for vegetative propagation of Norway spruce (<i>Picea abies</i> L. Karst). For mass propagation, a storage method that is able to handle large quantities of embryogenic tissues (ETs) reliably and at a low cost is required. The aim of the present study was to compare freezing at −80 °C in a freezer to cryopreservation using liquid nitrogen (LN) as a method for storing Norway spruce ETs. The possibility of simplifying both the pre-treatment and thawing processes in cryopreservation was also studied. The addition of abscisic acid (ABA) to the pre-treatment media and using polyethylene glycol PEG4000 instead of PEG6000 in a cryoprotectant solution were tested. Both the pre-and post-treatments on semi-solid media could be simplified by reducing the number of media, without any loss of genotype or embryo production capacity of ETs. On the contrary, the storage of ETs in a freezer at −80 °C instead of using LN was not possible, and the addition of ABA to the pre-treatment media did not provide benefits but increased costs. The lower regeneration rate after using PEG4000 instead of PEG6000 in a cryoprotectant solution in cryovials was unexpected and unwanted. The simplified pre-and post-treatment protocol will remarkably reduce the workload and costs in the mass-cryopreservation of future forest regeneration materials and in thawing the samples for mass propagations, respectively. |
| first_indexed | 2024-03-09T16:20:28Z |
| format | Article |
| id | doaj.art-311a9ab3f0f443a1a1abc47e89747cf4 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T16:20:28Z |
| publishDate | 2022-12-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-311a9ab3f0f443a1a1abc47e89747cf42023-11-24T15:22:51ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-12-0123241551610.3390/ijms232415516Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic CulturesSaila A. Varis0Susanna Virta1Itziar A. Montalbán2Tuija Aronen3Natural Resources Institute Finland (Luke), Vipusenkuja 5, FI-57200 Savonlinna, FinlandNatural Resources Institute Finland (Luke), Vipusenkuja 5, FI-57200 Savonlinna, FinlandDepartment of Forestry Science, NEIKER, 01080 Arkaute, SpainNatural Resources Institute Finland (Luke), Vipusenkuja 5, FI-57200 Savonlinna, FinlandSomatic embryogenesis (SE) is considered the most effective method for vegetative propagation of Norway spruce (<i>Picea abies</i> L. Karst). For mass propagation, a storage method that is able to handle large quantities of embryogenic tissues (ETs) reliably and at a low cost is required. The aim of the present study was to compare freezing at −80 °C in a freezer to cryopreservation using liquid nitrogen (LN) as a method for storing Norway spruce ETs. The possibility of simplifying both the pre-treatment and thawing processes in cryopreservation was also studied. The addition of abscisic acid (ABA) to the pre-treatment media and using polyethylene glycol PEG4000 instead of PEG6000 in a cryoprotectant solution were tested. Both the pre-and post-treatments on semi-solid media could be simplified by reducing the number of media, without any loss of genotype or embryo production capacity of ETs. On the contrary, the storage of ETs in a freezer at −80 °C instead of using LN was not possible, and the addition of ABA to the pre-treatment media did not provide benefits but increased costs. The lower regeneration rate after using PEG4000 instead of PEG6000 in a cryoprotectant solution in cryovials was unexpected and unwanted. The simplified pre-and post-treatment protocol will remarkably reduce the workload and costs in the mass-cryopreservation of future forest regeneration materials and in thawing the samples for mass propagations, respectively.https://www.mdpi.com/1422-0067/23/24/15516abscisic acidlong-term storagepolyethylene glycol<i>Picea abies</i>pre-treatmentsomatic embryogenesis |
| spellingShingle | Saila A. Varis Susanna Virta Itziar A. Montalbán Tuija Aronen Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures International Journal of Molecular Sciences abscisic acid long-term storage polyethylene glycol <i>Picea abies</i> pre-treatment somatic embryogenesis |
| title | Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures |
| title_full | Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures |
| title_fullStr | Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures |
| title_full_unstemmed | Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures |
| title_short | Reducing Pre- and Post-Treatments in Cryopreservation Protocol and Testing Storage at −80 °C for Norway Spruce Embryogenic Cultures |
| title_sort | reducing pre and post treatments in cryopreservation protocol and testing storage at 80 °c for norway spruce embryogenic cultures |
| topic | abscisic acid long-term storage polyethylene glycol <i>Picea abies</i> pre-treatment somatic embryogenesis |
| url | https://www.mdpi.com/1422-0067/23/24/15516 |
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