Optimizing conditions for succesful plant regeneration from cryopreserved castanea sativa shoot tips

Abstract

European chestnut is currently being ravaged by ink and blight diseases. Cryopreservation can constitute a complementary approach to field collections for long-term conservation of germplasm. A two-step vitrification methodology (LS-PVS2) was successful in cryopreserving micropropagated Castanea sativa shoot apices so that they could be recovered for plant regeneration. In order to optimize shoot performance after cryopreservation different parameters, such as explant type, duration of PVS2 treatment, sucrose content in preculture medium, loading pre-treatment, and sequence of media used after thawing, were studied.