Chapter 35
Cryopreservation of Plant Shoot Tips of Potato, Mint, Garlic,
and Shallot Using Plant Vitrification Solution 3
Angelika Senula and Manuela Nagel
Abstract
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.
Key words Cryoprotection, Long-term storage, PVS3, Plant genetic resources, Tissue culture, Ultralow temperature
1 Introduction
Cryopreservation is an important technology to maintain plant
genetic resources which otherwise would be lost in nature. In
principle, any totipotent tissue might be suitable for germplasm
maintenance [1]. However, herbaceous plant species are commonly
maintained by preserving shoot tips. These highly organized structures include the terminal portion of a shoot (1–2 mm) comprising
the meristem, the primordial and developing leaves, and adjacent
stem and tend to be genetic stable [2]. The largest collections of
cryopreserved shoot tips are located in Belgium, Colombia, Korea,
Peru, and Germany and maintain banana [3], cassava, Allium [4],
and potato [5, 6] genetic resources.
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_35, © Springer Science+Business Media, LLC, part of Springer Nature 2021
647
Cryopreservation of Plant Shoot Tips of Potato, Mint, Garlic,
and Shallot Using Plant Vitrification Solution 3
Angelika Senula and Manuela Nagel
Abstract
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.
Key words Cryoprotection, Long-term storage, PVS3, Plant genetic resources, Tissue culture, Ultralow temperature
1 Introduction
Cryopreservation is an important technology to maintain plant
genetic resources which otherwise would be lost in nature. In
principle, any totipotent tissue might be suitable for germplasm
maintenance [1]. However, herbaceous plant species are commonly
maintained by preserving shoot tips. These highly organized structures include the terminal portion of a shoot (1–2 mm) comprising
the meristem, the primordial and developing leaves, and adjacent
stem and tend to be genetic stable [2]. The largest collections of
cryopreserved shoot tips are located in Belgium, Colombia, Korea,
Peru, and Germany and maintain banana [3], cassava, Allium [4],
and potato [5, 6] genetic resources.
Willem F. Wolkers and Harrie ¨ tte Oldenhof (eds.), Cryopreservation and Freeze-Drying Protocols, Methods in Molecular Biology,
vol. 2180, https://doi.org/10.1007/978-1-0716-0783-1_35, © Springer Science+Business Media, LLC, part of Springer Nature 2021
647