Am. J. Bot. Plant Physiology
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(American Journal of Botany. 2007;94:799-808.)
© 2007 Botanical Society of America, Inc.

Phycology

Assessing genetic stability of a range of terrestrial microalgae after cryopreservation using amplified fragment length polymorphism (AFLP)1

Julia Müller, John G. Day, Keith Harding, Dominik Hepperle, Maike Lorenz and Thomas Friedl6

Albrecht-von-Haller-Institut für Pflanzenwissenschaften, Abteilung Experimentelle Phykologie und Sammlung von Algenkulturen (SAG), Universität Göttingen, Untere Karspüle 2, 37073 Göttingen, Germany; Culture Collection of Algae and Protozoa (CCAP), Scottish Association for Marine Science, Dunstaffnage Marine Laboratory, Dunbeg, Argyll, PA37 1QA, UK; DAMAR, Drum Road, Cupar Muir, Fife, KY15 5RJ, Scotland, UK; SequentiX–Digital DNA Processing, Dorfstr. 20, 18249 Klein Raden, Germany

ABSTRACT

Cryopreservation is the long-term, indefinite storage of living biological resources at ultralow temperatures. It is almost universally assumed that cryogenic storage supports genetic and phenotypic stability of organisms. However, certain components of the cryopreservation process, particularly some cryoprotective additives (CPAs) and free radical mediated cryoinjury, may potentially cause genetic alterations. Genetic integrity in cryopreserved microalgae was assessed using a very sensitive molecular fingerprinting technique, AFLP, on 28 terrestrial microalgal strains. In about half of all investigated strains the AFLP fingerprints revealed, with high levels of reproducibility, clearly detectable genomic differences after cryopreservation employing a widely used standard two-step cooling protocol. Differences ranged from a single fragment position to multiple fragment changes and were compared to differences found between wild-type and UV-light- or radioisotope-induced mutants of Parachlorella kessleri. The basis of the changes are discussed in terms of their reversibility, as may be the case if they are attributed to DNA methylation and/or whether they are true mutations that may potentially manifest in the phenotype. The possibility that cryopreservation selects for genotypically different subpopulations of microalgae is also considered.

Key Words: AFLP • algae • cryopreservation • culture collections • DNA methylation • genetic stability






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