Cryopreservation of in vitro-grown meristems of strawberry (Fragaria × ananassa Duch.) by encapsulation-vitrification

Abstract

Alginate-coated meristems from in vitro-grown strawberry (Fragaria × ananassa Duch.) were successfully cryopreserved following dehydration by a vitrification solution. Excised meristems from cold-hardened plantlets at 4 °C for two weeks in the dark were encapsulated into alginate-gel beads containing a mixture of 2 M glycerol plus 0.4 M sucrose. These encapsulated and cryoprotected meristems were dehydrated with a highly concentrated vitrification solution (designated PVS2) for 2 hr at 0 °C prior to a plunge into liquid nitrogen. Successfully encapsulated vitrified meristems remained green and then developed shoots within one week after plating without intermediary callus formation. The average rate of shoot formation of encapsulated vitrified meristems amounted to nearly 90%. The cryogenic protocol was successfully applied to four cultivars of strawberry. It was also confirmed that encapsulated vitrified meristems cooled to - 196 °C produced higher shoot formation than encapsulated dried meristems. Besides, the recovery growth was much earlier than the latter. The encapsulation-vitrification method is easy to handle and produces high levels of shoot formation. Thus, the protocol promises for cryopreservation of strawberry germplasm.