Isolation and Regeneration of Protoplasts from Ectocarpus
- Susana M. Coelho1,2,4,
- Delphine Scornet1,2,
- Sylvie Rousvoal1,2,
- Nick Peters1,2,
- Laurence Dartevelle1,2,
- Akira F. Peters2,3 and
- J. Mark Cock1,2
- 1UPMC Université Paris 06, The Marine Plants and Biomolecules Laboratory, UMR 7139, Station Biologique de Roscoff, 29682 Roscoff Cedex, France
- 2CNRS, UMR 7139, Laboratoire International Associé Dispersal and Adaptation in Marine Species, Station Biologique de Roscoff, 29682 Roscoff Cedex, France
- 3Bezhin Rosko, 29250 Santec, France
Abstract
This article describes how to obtain isolated cells with no surrounding cell wall by enzymatic digestion of Ectocarpus filaments. The resultant protoplasts are totipotent and regenerate to produce individual algae under appropriate culture conditions. The yield of protoplasts and their capacity to regenerate are highly dependent on the Ectocarpus strain used, the stage of the life cycle, and the culture conditions. The highest yields are obtained with young gametophyte filaments cultivated at low density. The naked, wall-less cells produced by this protocol can be used for several applications, including studies of cell wall regeneration, investigation of the role of the cell wall in determining cell fate, and as a source of naked cells for the development of methods for introducing diverse molecules into the cell.
Footnotes
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↵4 Correspondence: coelho{at}sb-roscoff.fr
- © 2012 Cold Spring Harbor Laboratory Press