title: Evaluation of the new vacuum infiltration vitrification (viv) cryopreservation technique for native Australian plant shoot tips.
abstract: BACKGROUND
  The application of a vacuum during the incubation in cryoprotective agents such as PVS2 allows for increased penetration, reducing total incubation times required before vitrification and post-cryopreservation regeneration is achieved.
  
  
  OBJECTIVE
  This study compared a conventional droplet-vitrification protocol to the new vacuum infiltration vitrification protocol in four Australian plant species.
  
  
  METHODS
  The new vacuum infiltration vitrification applied an 80 kPa vacuum during incubations in loading solution and PVS2. Infiltration of the cryoprotective agents into shoot tips was determined by differential scanning calorimetry measuring ice formation in the thermographs comparing a range of loading solution and PVS2 incubation times.
  
  
  CONCLUSIONS
  The application of the vacuum infiltration vitrification technique resulted in a significantly reduced PVS2 incubation time for cryogenic survival and regeneration for all four species, reducing the time needed to adequately protect shoot tips by half to a quarter when compared to a conventional droplet-vitrification technique.
authors.affiliation: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845.Australia. Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia.
authors.forename: Bryn
authors.initials: B
authors.lastname: Funnekotter
authors.affiliation: Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia. School of Plant Biology, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley WA 6009, Australia.
authors.forename: Susan E
authors.initials: SE
authors.lastname: Whiteley
authors.affiliation: Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia. School of Plant Biology, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley WA 6009, Australia.
authors.forename: Shane R
authors.initials: SR
authors.lastname: Turner
authors.affiliation: Botanic Gardens and Parks Authority, Fraser Avenue, West Perth WA 6005.Australia. School of Plant Biology, Faculty of Natural and Agricultural Sciences, University of Western Australia, Crawley WA 6009, Australia.
authors.forename: Eric
authors.initials: E
authors.lastname: Bunn
authors.affiliation: School of Biomedical Sciences, CHIRI Biosciences, Curtin University, GPO Box U1987, Perth, WA 6845, Australia.
authors.forename: Ricardo L
authors.initials: RL
authors.lastname: Mancera
mesh_terms: D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry; D018521:Plant Physiological Phenomena; D018520:Plant Shoots / Q000502:physiology*; D014618:Vacuum; D058989:Vitrification
chemical_list: D003451:Cryoprotective Agents