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Cryopreservation of embryogenic cultures of Quercus robur using desiccation and vitrification procedures

AuthorsMartínez, M. T.; Ballester, Antonio CSIC; Viéitez Martín, Ana María
KeywordsCryogenic storage
Liquid nitrogen
Somatic embryogenesis
Issue DateApr-2003
CitationCryobiology 46(2): 182-189 (2003)
AbstractOak 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 4–6 mg clumps (1.0–1.5 mm) 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 0.7 M 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 0.3 M sucrose medium followed by application of a vitrification solution (PVS2) for 60–90 min prior to rapid plunging in liquid nitrogen.
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