Differential responses of Mimusops elengi and Manilkara zapota seeds and embryos to cryopreservation

Several dehydration protocols were evaluated for their ability to cryopreserve intact seeds and excised embryonic axes of Mimusops elengi and Manilkara zapota (Sapotaceae). Both interspecific and intraspecific variations in cryotolerance were found. M. zapota embryonic axes were more tolerant of cryopreservation than those of M. elengi, and showed higher desiccation tolerance, higher post-thawing survival and development, and a much wider range of moisture contents for cryopreservation. Maximum development rates were 94% and 27% for M. zapota and M. elengi, respectively. Intact seeds of both species tolerated desiccation to low moisture levels, but were sensitive to liquid nitrogen exposure, and cryopreserved seeds failed to germinate. Assessment of developing embryos excised from cryopreserved seeds associated nonviability of cotyledons and plumules with germination failure. Other structures survived at variable rates; most hypocotyls and radicles (up to 76% and 98% for M. elengi and M. zapota, respectively) were viable. The different cryotolerance between hypocotyls and cotyledons is a critical cause for failure in cryopreservation, contributing to the difficulty in developing protocols for such intermediate oily seeds

Advances in understanding the fundamental aspects required for successful cryopreservation of Australian flora

Australia is host to an amazing diversity of species, many of which require conservation efforts. In vitro culture provides a tool for not only conserving these threatened species but allows for their propagation from limited starting material. Cryopreservation provides the greatest long-term storage option for in vitro cultures and as a conservation tool for other germplasm. However, while cryopreservation has proven capable of delivering viable long-term storage with some plant taxa, the process of deriving protocols is still largely an incremental process. The key to faster and more intuitive optimising of cryopreservation protocols lies with continuing to develop a better understanding of key factors, including issues with plant physiology (such as genetic stability, the composition of the proteome and metabolome, cell membrane characteristics, and antioxidant defences) and how the stresses imposed by cryopreservation (such as the excision damage, desiccation, cryoprotective agent toxicity, ice crystal damage, and cooling to cryogenic temperatures) interact and contribute to the cryocapability of a species. This review focuses on the advances that have been made towards understanding cryogenic stress and how this has led to improved cryopreservation protocols, in the context of cryopreserving Australian flora