Cryopreservation of embryogenic cell suspensions of the Spanish grapevine cultivars ‘Albariño’ and ‘Tempranillo’

Embryogenic cell suspensions of two elite Spanish grapevine cultivars (‘Albariño’ and ‘Tempranillo’) were successfully cryopreserved by encapsulation-dehydration. The method implied the encapsulation of cells in alginate beads, subsequent culture in liquid medium with 1 M sucrose for 4 days, and desiccation for 2-4 h in the flow of a laminar-flow bench, before immersing in liquid nitrogen. With this simple method, up to 50 % of cell viability in the cryopreserved beads was measured (using the triphenyl tetrazolium method), which corresponded to vigorous growth of 100 % of beads after culture on semi-solid medium. The cryopreserved encapsulated cells were successfully used for initiation of new cell suspensions and their embryogenic capacity was studied. This cryopreservation method is an advance to store ready-to-use competent embryogenic tissue for grapevine genetic transformation projects.