Eukaryotic Cell
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Eukaryotic Cell, May 2008, p. 872-880, Vol. 7, No. 5
1535-9778/08/$08.00+0     doi:10.1128/EC.00461-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

The Heterotrophic Dinoflagellate Crypthecodinium cohnii Defines a Model Genetic System To Investigate Cytoplasmic Starch Synthesis{triangledown} ,{dagger}

Philippe Deschamps,1 Delphine Guillebeault,2 Jimi Devassine,1 David Dauvillée,1 Sophie Haebel,3 Martin Steup,4 Alain Buléon,5 Jean-Luc Putaux,6 Marie-Christine Slomianny,1 Christophe Colleoni,1 Aline Devin,1 Charlotte Plancke,1 Stanislas Tomavo,1 Evelyne Derelle,2 Hervé Moreau,2 and Steven Ball1*

CNRS, UMR8576, Cité Scientifique, 59655 Villeneuve d'Ascq, France,1 Observatoire Océanologique, Laboratoire Arago, UMR7628 CNRS-Université Paris VI, BP 44, F-66651 Banyuls-sur-Mer Cedex, France,2 Center of Mass Spectrometry of Biopolymers of the University of Potsdam,3 Plant Physiology, Institute of Biochemistry and Biology, University of Potsdam, 14476 Golm, Germany,4 Institut National de la Recherche Agronomique, Centre de Recherches Agroalimentaires, Rue de la Géraudière, BP71627, 44316 Nantes Cedex 03, France,5 Centre de Recherches sur les Macromolécules Végétales, ICMG-CNRS, BP 53, F-38041 Grenoble Cedex 9, France6

Received 24 December 2007/ Accepted 20 February 2008

The nature of the cytoplasmic pathway of starch biosynthesis was investigated in the model heterotrophic dinoflagellate Crypthecodinium cohnii. The storage polysaccharide granules were shown to be composed of both amylose and amylopectin fractions with a chain length distribution and crystalline organization very similar to those of green algae and land plant starch. Preliminary characterization of the starch pathway demonstrated that C. cohnii contains multiple forms of soluble starch synthases and one major 110-kDa granule-bound starch synthase. All purified enzymes displayed a marked substrate preference for UDP-glucose. At variance with most other microorganisms, the accumulation of starch in the dinoflagellate occurs during early and mid-log phase, with little or no synthesis witnessed when approaching stationary phase. In order to establish a genetic system allowing the study of cytoplasmic starch metabolism in eukaryotes, we describe the isolation of marker mutations and the successful selection of random recombinant populations after homothallic crosses.

* Corresponding author. Mailing address: CNRS, UMR8576, USTL, Bâtiment C9, Cité Scientifique, 59655 Villeneuve d'Ascq, France. Phone: 33 3 2043 6543. Fax: 33 3 2043 6555. E-mail: steven.ball{at}univ-lille1.fr

{triangledown} Published ahead of print on 29 February 2008.

{dagger} Supplemental material for this article may be found at http://ec.asm.org/.

Eukaryotic Cell, May 2008, p. 872-880, Vol. 7, No. 5
1535-9778/08/$08.00+0     doi:10.1128/EC.00461-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.





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