Cryopreservation of Plant Genetic Resources

  • Daisuke TanakaEmail author
  • Takao Niino
  • Matsuo Uemura
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 1081)


Cryopreservation encompasses several interconnect disciplines including physiology and cryophysics. This chapter reviews the current techniques for cryopreservation of plant genetic resources (PGRs). Vitrification is an effective ice crystal avoidance mechanism for hydrated cells and tissues. With any cryopreservation method, whole or partial parts of specimens which are sufficiently dehydrated can be vitrified by rapid cooling in liquid nitrogen (LN). Techniques discussed are the vitrification protocol, encapsulation-vitrification protocol, droplet vitrification protocol (DV), vitrification protocol using cryo-plates (V cryo-plate), and air dehydration protocol using cryo-plates (D cryo-plate). In these DV, V, and D cryo-plate protocols, specimens to be cryopreserved are immersed directly into LN on aluminum foil strips or cryo-plates; removal from LN to rewarming solution results in a high level of plant regrowth with ultrarapid cooling and warming. The protocols were applied to a wide array of plant species including wild and multi-ploid species, although fine tuning of the protocols was required for successful application to specific plant species and lines. These three protocols efficiently complement each other and appear highly promising to facilitate large-scale cryobanking of PGRs in genebanks. Cryo-scanning electron microscopy makes it possible to examine the cellular and water behavior in plant tissues when immersed in LN. It has been verified that tissues cryopreserved by the process of vitrification and the cryo-plate protocols are cryopreservation methods for reliable long-term preservation of PGRs.


Cryopreservation Vitrification D cryo-plate V cryo-plate Intracellular ice formation Plant genetic resources 



Aluminum cryo-plate


Cryo-scanning electron micro-scopy

D cryo-plate

Air dehydration protocol using cryo-plates


Droplet vitrification protocol


Encapsulation-vitrification pro-tocol


Liquid nitrogen


Loading solution

MS medium

Murashige and Skoog medium


Plant genetic resources


Plant vitrification solution 2

V cryo-plate

Vitrification protocol using cryo-plates


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Copyright information

© Springer Nature Singapore Pte Ltd. 2018

Authors and Affiliations

  1. 1.Genetic Resources CenterNational Agriculture and Food Research OrganizationTsukubaJapan
  2. 2.Gene Research CenterUniversity of TsukubaTsukubaJapan
  3. 3.United Graduate School of Agricultural Sciences and Department of Plant-biosciences, Faculty of AgricultureIwate UniversityMoriokaJapan

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