A simple method for cryopreservation of shoot tips of Arabidopsis genotypes

Cryo Letters. 2017 Sep/Oct;38(5):364-371.

Abstract

BACKGROUND: The use of the model plant Arabidopsis could be a valuable tool to elucidate the basic mechanisms involved in plant cryopreservation.

Objective: A simple and powerful protocol, independent of Arabidopsis genotypes, was established using a PVS2 protocol.

Materials and methods: Two PVS2 (a and b), one PVS3 droplet-vitrification and one DMSO droplet-freezing protocol were tested with alternating temperatures during the growing phase of donor plants.

Results: PVS2 protocols, including cold acclimation of donor plants, resulted in highest recovery. The PVS2a protocol was successfully applied to a collection of different Arabidopsis genotypes with an average recovery of 94%. In addition, Differential Scanning Calorimetry confirmed the occurrence of glass transitions in the PVS3 and PVS2 protocols.

Conclusion: The PVS2a protocol is suitable to screen the large collection of Arabidopsis mutants and transgenic lines with the aim to identify cellular functions associated with cryopreservation tolerance.

MeSH terms

  • Arabidopsis / drug effects
  • Arabidopsis / genetics*
  • Arabidopsis / physiology*
  • Cryopreservation / methods*
  • Cryoprotective Agents / pharmacology
  • Crystallization
  • Ecotype
  • Freezing
  • Genotype
  • Plant Shoots / drug effects
  • Plant Shoots / physiology*
  • Vitrification

Substances

  • Cryoprotective Agents