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Cryopreservation-recalcitrance in microalgae: novel approaches to identify and avoid cryo-injury

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Abstract

Standard two-step freezing protocols areunsatisfactory for Euglena gracilis and manyother microalgae, particularly those with larger cellsizes, complex morphologies and/or those susceptibleto environmental stress. Using techniques that allowmechanisms of injury and sites of damage to beidentified (e.g. monitoring oxygen evolving capacity,detection of •OH, microscopic visualisation ofintracellular ice and structural/ultrastructuraldamage), it is possible to improve conventionalcryopreservation methodologies. In E. gracilisthis has resulted in the development of protocolswhich increased post-thaw viability levels from 0 to20%. Alternative cryoprotection strategies testedincluded vitrification and encapsulation/dehydration.Vitrification was unsuccessful due to the hightoxicity of the solutions. Encapsulation/dehydration,with or without two-step cooling were suitable forcryopreservation of E. gracilis, the latterresulted in the highest levels of post-thaw viability(40%) and viability was maintained after 12 monthsstorage.

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Authors and Affiliations

  1. Culture Collection of Algae and Protozoa, Centre for Ecology & Hydrology Windermere, Far Sawrey, NERC, Ambleside, Cumbria, LA22, 0LP, UK

    John G. Day

  2. Department of Soil, Crop and Atmospheric Sciences, Cornell University, Ithaca, NY, 14853, USA

    Roland A. Fleck

  3. Plant Conservation Biotechnology Group, Division of Molecular and Life Sciences, School of Science and Engineering, University of Abertay Dundee, Bell Street, Dundee, DD1 1HG, Scotland, UK

    Erica E. Benson

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  1. John G. Day
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  2. Roland A. Fleck
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  3. Erica E. Benson
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Day, J.G., Fleck, R.A. & Benson, E.E. Cryopreservation-recalcitrance in microalgae: novel approaches to identify and avoid cryo-injury. Journal of Applied Phycology 12, 369–377 (2000). https://doi.org/10.1023/A:1008107229005

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