title: Cryopreservation of zygotic embryonic axes and somatic embryos of European chestnut.
abstract: For Castanea sativa (European chestnut), a species with recalcitrant seeds that is not easily propagated vegetatively, cryopreservation is one of the most promising techniques for maintaining genetic resource diversity and for conservation of selected germplasms. Long-term conservation of selected seeds and valuable embryogenic lines can be achieved through the cryopreservation of zygotic embryonic axes and somatic embryos, respectively. This chapter describes methods for the desiccation-based cryostorage of zygotic embryonic axes, and the vitrification-based cryopreservation of somatic embryos. For zygotic embryonic axes, the highest post-thaw survival and plantlet recovery rates are obtained by desiccation in a laminar flow hood to 20-25% moisture content, followed by direct immersion in liquid nitrogen. For somatic embryos, embryogenesis resumption rates of over 60% are achieved by preculture of embryo clumps for 3 days on solid medium containing 0.3 M sucrose, incubation in PVS2 vitrification solution for 60 min at 0°C, and direct immersion in liquid nitrogen. Plantlet recovery from cryostored embryogenic lines requires proliferation of the thawed embryos and subsequent maturation before germination and conversion into plantlets.
authors.affiliation: Instituto de Investigaciones Agrobiológicas de Galicia, CSIC, Santiago de Compostela, Spain. amvieitez@iiag.csic.es
authors.forename: Ana M
authors.initials: AM
authors.lastname: Vieitez
authors.forename: M Carmen
authors.initials: MC
authors.lastname: San-José
authors.forename: Elena
authors.initials: E
authors.lastname: Corredoira
mesh_terms: D015925:Cryopreservation; D003470:Culture Media; D003890:Desiccation; D029962:Fagaceae / Q000196:embryology*; D018525:Germination; D012639:Seeds / Q000254:growth & development*; D058989:Vitrification
chemical_list: D003470:Culture Media