Authors: Bryn Funnekotter Anja Kaczmarczyk Shane R Turner Eric Bunn Wenxu Zhou Steven Smith Gavin Flematti Ricardo L Mancera
Publish Date: 2013/03/04
Volume: 114, Issue: 1, Pages: 83-96
Abstract
Cell membranes are the primary sites of cryopreservation injury and measuring changes to membrane composition arising from cold acclimation may assist with providing a rationale for optimising cryopreservation methods Shoot tips from two southwest Western Australian species Grevillea scapigera and Loxocarya cinerea and Arabidopsis thaliana reference species were subjected to cryopreservation using the droplet vitrification protocol Two preconditioning regimes involving a constant temperature 23 °C CT with a 12 h light/dark cycle or an alternating temperature AT regime 20/10 °C with a 12 h light/dark cycle were compared Soluble sugars sterols and phospholipids present in the shoot tips were analysed Use of AT preconditioning acclimation resulted in a modest decrease in cryotolerance in A thaliana increased cryotolerance in G scapigera and increased survival in the nonfrozen control explants of L cinerea in comparison to CT preconditioning Increased cryotolerance was accompanied by a higher total sugar sterol and phospholipid content as well as an increase in strong hydrating phospholipid classes such as phosphatidylcholine The double bond index of bound fatty acyl chains of phospholipids was greater after AT preconditioning mostly due to a higher amount of monoenes in A thaliana and trienes in G scapigera and L cinerea These findings suggest that AT preconditioning treatments for in vitro plants can have a positive influence on cryotolerance for some plant species and this may be related to observed changes in the overall composition of cell membranes However alternative factors eg oxidative stress may be equally important with other species eg L cinereaThis work was funded by Australian Research Council Linkage Project LPO884027 BF holds an Australian Postgraduate Award and a Curtin Research Scholarship Alcoa of Australia Ltd and BHP Billiton Worsley Alumina Pty Ltd are gratefully acknowledged for their generous financial support Analytical work was supported by Matthew Timmins and Ricarda Fenske Metabolomics Australia
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