How does metabolic rate in plant shoot tips change after cryopreservation?

Cryopreservation allows the long-term storage of plant germplasm, but can cause damage to plant tissues, which must be repaired for survival to occur. This repair process is fuelled by the metabolic function of mitochondria; however, little is known about how metabolic function is affected by the cryopreservation process in plants. We compared metabolic rates of shoot tips of two Australian native species, Androcalva perlaria and Anigozanthos viridis. Overall, cryopreservation resulted in a significant reduction in the metabolic rates of shoot tips from both species, even in tissues that regenerated after cryopreservation. Metabolic rate did not increase within 48 h after of thawing, even in shoot tips which later regenerated. When examined in isolation, both pre-treatment on desiccation medium and exposure to cryoprotective agents significantly decreased metabolic rates in regenerating shoot tips of A. viridis, however both caused a significant increase in shoot tips of A. perlaria, suggesting diversity of response to cryopreservation stresses across species. Measurements of shoot tip metabolic rate during cryopreservation will inform investigations into cellular energy production and provide critical information on the state of shoot health after exposure to different cryoprotective treatments, which could play a useful role in guiding protocol optimisation for threatened species to maximise post-cryopreservation regeneration.

Review: The case for studying mitochondrial function during plant cryopreservation.

Cryopreservation has several advantages over other ex situ conservation methods, and indeed is the only viable storage method for the long term conservation of most plant species. However, despite many advances in this field, it is increasingly clear that some species are ill-equipped to overcome the intense stress imposed by the cryopreservation process, making protocol development incredibly difficult using traditional trial and error methods. Cryobiotechnology approaches have been recently recognised as a strategic way forward, utilising intimate understanding of biological systems to inform development of more effective cryopreservation protocols. Mitochondrial function is a model candidate for a cryobiotechnological approach, as it underpins not only energy provision, but also several other key determinants of germplasm outcome, including stress response, reduction-oxidation status, and programmed cell death. Extensive research in animal cell and tissue cryopreservation has established a clear link between mitochondrial health and cryopreservation survival, but also indicates that mitochondria are routinely subject to damage from multiple aspects of the cryopreservation process. Evidence is already emerging that mitochondrial dysfunction may also occur in plant cryopreservation, and this research can be greatly expanded by using considered applications of innovative technologies. A range of mitochondria-targeted prophylactic and therapeutic interventions already exist with potential to improve cryopreservation outcomes through mitochondrial function.