title: Cryopreservation of embryogenic cultures of Quercus robur using desiccation and vitrification procedures.
abstract: Oak embryogenic cultures are generally maintained by repetitive embryogenesis. To facilitate management of embryogenic lines and limit the risks of somaclonal variation and contamination a cryopreservation protocol should be developed. In this work we investigated the ability of several pre-treatments to enable 4-6mg clumps (1.0-1.5mm) of globular-heart stage somatic embryos of Quercus robur to withstand freezing in liquid nitrogen. In the best of the two embryogenic culture lines used, 56% of clumps resumed embryogenesis after cooling when they had been pre-treated by successive pre-culture on 0.3 and 0.7M sucrose supplemented media followed by desiccation in the air flow of a laminar flow cabinet to water contents of 24-34%. In both lines, embryogenesis resumption rates of about 70% were achieved by pre-culture on 0.3M sucrose medium followed by application of a vitrification solution (PVS2) for 60-90min prior to rapid plunging in liquid nitrogen.
authors.affiliation: Instituto de Investigaciones Agrobiológicas de Galicia, CSIC, Apartado 122, 15080, Santiago de Compostela, Spain.
authors.forename: M T
authors.initials: MT
authors.lastname: Martínez
authors.forename: A
authors.initials: A
authors.lastname: Ballester
authors.forename: A M
authors.initials: AM
authors.lastname: Vieitez
mesh_terms: D002460:Cell Line; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology; D003890:Desiccation / Q000379:methods; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005615:Freezing; D005990:Glycerol / Q000494:pharmacology; D009584:Nitrogen; D029963:Quercus / Q000737:chemistry / Q000187:drug effects / Q000196:embryology*; D012639:Seeds / Q000187:drug effects* / Q000254:growth & development; D013395:Sucrose / Q000494:pharmacology; D013997:Time Factors
chemical_list: D003451:Cryoprotective Agents; D013395:Sucrose; D009584:Nitrogen; D005990:Glycerol; D004121:Dimethyl Sulfoxide