Profiling cryopreservation protocols for Ribes ciliatum using differential scanning calorimetry

DSC analysis was performed at three points in the cryopreservation process on encapsulated-dehydrated meristems of Ribes ciliatum. Meristems were excised from shoots pre-treated with either sucrose or glucose, encapsulated in alginate beads, dehydrated in sucrose solutions, air dried, and plunged in liquid nitrogen. Thermal analysis revealed glass transitions during cooling of air-desiccated meristems, however, on rewarming a small endothermic event was detected suggesting glass destabilization can occur. Interestingly, this did not occur in alginate beads or meristems when these components were cooled and rewarmed separately. The possibility exists that thermal and moisture gradients may arise within the alginate bead/tissue complex and we propose that the heterogeneous composition of the meristems and the surrounding alginate may promote ice nucleation on rewarming. The significance of this regarding the stabilization of glasses formed in alginate beads and their encapsulated meristems is discussed. This study also reports an approach to Ribes cryopreservation in which the pregrowth of shoots in 0.75M sucrose for 1 week can be used as a substitute for cold acclimation.

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