title: Cryopreservation of Lomandra sonderi (Asparagaceae) shoot tips using droplet-vitrification.
abstract: A cryopreservation protocol was developed for Lomandra sonderi (Asparagaceae), an endemic plant of southwest Western Australia used for mine site restoration. Thermal analysis of L. sonderi shoot tips using differential scanning calorimetry was used to detect the formation of ice in shoot tips and consequently allowed optimisation of the time of incubation in plant vitrification solution 2 (PVS2), which attempted to minimise phytotoxicity of, and excessive dehydration by, its cryoprotective components. Sugar pretreatments did not improve survival. Use of a loading solution containing 2 M glycerol and 0.4 M sucrose prior to incubation in PVS2 improved survival of control shoot tips. Preconditioning at 20+/-1 degree C day/night alternating temperature with a 16 h photoperiod or at a constant 5 degree C temperature with a 12 h photoperiod both significantly improved both control shoot tip survival and post-cryopreservation survival. Shoot tips that recovered from liquid nitrogen immersion were successfully re-established as actively growing in vitro plantlets.
authors.affiliation: Western Australian Biomedical Research Institute, Curtin Health Innovation Research Institute, School of Biomedical Sciences, Curtin University, Perth, WA, Australia.
authors.forename: Akshay
authors.initials: A
authors.lastname: Menon
authors.forename: Bryn
authors.initials: B
authors.lastname: Funnekotter
authors.forename: Anja
authors.initials: A
authors.lastname: Kaczmarczyk
authors.forename: Eric
authors.initials: E
authors.lastname: Bunn
authors.forename: Shane
authors.initials: S
authors.lastname: Turner
authors.forename: Ricardo L
authors.initials: RL
authors.lastname: Mancera
mesh_terms: D000064:Acclimatization; D002152:Calorimetry, Differential Scanning; D003080:Cold Temperature; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry / Q000378:metabolism; D019668:Liliaceae / Q000502:physiology*; D018520:Plant Shoots / Q000502:physiology*; D058989:Vitrification; D014904:Western Australia
chemical_list: D003451:Cryoprotective Agents