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Cold acclimation and cryoprotectants in a freeze-tolerant Antarctic nematode, Panagrolaimus davidi
Authors:D A Wharton  K F Judge  M R Worland
Institution:(1) Department of Zoology, University of Otago, PO Box 56, Dunedin, New Zealand e-mail: david.wharton@stonebow.otago.ac.nz Fax: +64-3-4797584, NZ;(2) British Antarctic Survey, Natural Environmental Research Council, High Cross, Madingley Road, Cambridge, CB3 OET, UK, GB
Abstract:Panagrolaimus davidi is a freeze-tolerant Antarctic nematode which survives extensive intracellular freezing. This paper describes the development of culture techniques which provide clean samples, with a high degree of freeze tolerance and in sufficient quantities for the analysis of potential cryoprotectants. Cultures grown at 20 °C survived a short-term freezing stress but survival declined with the time spent frozen. Acclimation of cultures at 5 °C enhanced the long-term survival of freezing. Starvation, however, reduced the nematode's ability to survive short-term freezing. The principal cryoprotectants detected by gas chromatography were trehalose and glycerol. The levels of trehalose, but not those of glycerol, increased significantly after acclimation. Trehalose may stabilise membranes and protect them against the dehydrating effects of the osmotic stresses resulting from freeze concentration effects but other factors, such as recrystallisation inhibition, may be involved in long-term survival. Accepted: 7 March 2000
Keywords:Liquid culture  Freeze tolerance  Intracellular freezing  Trehalose  Glycerol
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