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.
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© 2011 Humana Press
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Vieitez, A.M., San-José, M.C., Corredoira, E. (2011). Cryopreservation of Zygotic Embryonic Axes and Somatic Embryos of European Chestnut. In: Thorpe, T., Yeung, E. (eds) Plant Embryo Culture. Methods in Molecular Biology, vol 710. Humana Press. https://doi.org/10.1007/978-1-61737-988-8_15
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DOI: https://doi.org/10.1007/978-1-61737-988-8_15
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