Articles
Experience in large-scale cryopreservation and links to applied research for safe storage of plant germplasm
Article number
1113_36
Pages
239 – 250
Language
English
Abstract
Cryopreservation as a method to store plant organs in liquid nitrogen is a widely applied routine procedure.
It is suitable for preserving germplasm endangered by genetic erosion, especially for vegetatively propagated material, species that cannot produce storable seeds, and pollen representing vulnerable populations.
The IPK applies cryopreservation for large collections of potato (>1400 accessions), garlic, mint shoot tips and Allium pollen.
Various methods are routinely applied, such as dimethyl sulfoxide (DMSO) droplet freezing, vitrification, and droplet vitrification.
In contrast to well-responding genotypes, however, there are many types, which do not or only poorly respond.
The reasons for such differences are unknown and more basic research is needed to elucidate underlying mechanisms.
The effects of various cryoprotectants on the state of water in the tissues are measured by differential scanning calorimetry (DSC). In some methods like in DMSO droplet freezing, the presence of freezable water is proved by DSC and tolerated as survival and regrowth rates show.
In addition, differences in the structure between meristem and surrounding cells cause differential survival of these tissues in cryopreservation.
Likewise viruses are unequally distributed between the various tissue types and may be eliminated by liquid nitrogen treatment.
Recently, at the IPK studies on components of cryo-stress were performed.
Reactive oxygen species and changes in the proteome and metabolite patterns were investigated.
Oxygen was found to play a crucial role in stress-mediated injury and regeneration after rewarming from cryopreservation.
The elucidation of components involved in this process allows improvements in large-scale cryopreservation protocols for the valuable germplasm in genebanks.
It is suitable for preserving germplasm endangered by genetic erosion, especially for vegetatively propagated material, species that cannot produce storable seeds, and pollen representing vulnerable populations.
The IPK applies cryopreservation for large collections of potato (>1400 accessions), garlic, mint shoot tips and Allium pollen.
Various methods are routinely applied, such as dimethyl sulfoxide (DMSO) droplet freezing, vitrification, and droplet vitrification.
In contrast to well-responding genotypes, however, there are many types, which do not or only poorly respond.
The reasons for such differences are unknown and more basic research is needed to elucidate underlying mechanisms.
The effects of various cryoprotectants on the state of water in the tissues are measured by differential scanning calorimetry (DSC). In some methods like in DMSO droplet freezing, the presence of freezable water is proved by DSC and tolerated as survival and regrowth rates show.
In addition, differences in the structure between meristem and surrounding cells cause differential survival of these tissues in cryopreservation.
Likewise viruses are unequally distributed between the various tissue types and may be eliminated by liquid nitrogen treatment.
Recently, at the IPK studies on components of cryo-stress were performed.
Reactive oxygen species and changes in the proteome and metabolite patterns were investigated.
Oxygen was found to play a crucial role in stress-mediated injury and regeneration after rewarming from cryopreservation.
The elucidation of components involved in this process allows improvements in large-scale cryopreservation protocols for the valuable germplasm in genebanks.
Authors
E.R.J. Keller, M. Grübe, M.-R. Hajirezaei, M. Melzer, H.-P. Mock, H. Rolletschek, A. Senula, K. Subbarayan
Keywords
potato, garlic, mint, DSC, hypoxy, proteomics, ultrastructure
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