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Methods Mol Biol ; 2180: 647-661, 2021.
Article in English | MEDLINE | ID: mdl-32797441

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.


Subject(s)
Cell Culture Techniques/methods , Cryopreservation/methods , Cryoprotective Agents/pharmacology , Garlic/cytology , Mentha/cytology , Plant Shoots/cytology , Shallots/cytology , Solanum tuberosum/cytology , Cell Proliferation , Cells, Cultured , Garlic/drug effects , Glycerol/chemistry , Mentha/drug effects , Plant Shoots/drug effects , Shallots/drug effects , Solanum tuberosum/drug effects , Sucrose/chemistry
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