Newly developed encapsulation-dehydration protocol for plantcryopreservation.

A simplified and efficient encapsulation-dehydration protocol, which is a compromise between vitrification and encapsulation-dehydration, was presented for plant cryopreservation. In this protocol, during the encapsulation process, the apices precultured with 0.3 M sucrose for 16 h were simultaneously osmoprotected with a mixture of 02 M glycerol plus 0.4 M sucrose for 1 h. These encapsulated apices were directly dehydrated with dry silica gel prior to a plunge into LN without the pretreatment of 0.8M sucrose for 16 h. This protocol produced much higher rates of recovery growth in the three plant species tested (wasabi, chrysanthemum, and mint) than those cryopreserved by the conventional encapsulation-dehydration. This protocol also considerably reduced the time needed for the cryogenic procedure. Thus, this new protocol appears promising for cryopreservation of shoot apices and other explants.