Cryopreservation of axillary buds of a Eucalyptus grandis x eucalyptus camaldulensis hybrid.

This paper presents preliminary results from a study to develop methodology for the cryopreservation of axillary buds from an in vitro hybrid Eucalyptus grandis (W. Hill ex Maiden.) x Eucalyptus camaldulensis (Dehnnh.), maintained for use in a genetic modification programme. Axillary buds were encapsulated in an alginate gel, precultured on media containing elevated levels of sucrose, or a combination of sucrose and glycerol. Encapsulated buds were then dehydrated by evaporation prior to a two-step freezing process in liquid nitrogen. Eighteen percent of shoot explants survived freezing when sucrose alone was used as a protectant against dehydration and cryopreservation. Significantly higher survival (49%) was obtained with the incorporation of glycerol into the protocol. Following cryopreservation, shoots appeared to develop normally, with no evidence of adventitious meristems.