Author for correspondence: e-mail firstname.lastname@example.org.
INTRACELLULAR LOCALIZATION OF AN ENDOGENOUS CELLULOSE SYNTHASE OF MICRASTERIAS DENTICULATA (DESMIDIALES, CHLOROPHYTA) BY MEANS OF TRANSIENT GENETIC TRANSFORMATION1
Article first published online: 20 JUL 2010
© 2010 Phycological Society of America
Journal of Phycology
Volume 46, Issue 4, pages 839–845, August 2010
How to Cite
Vannerum, K., Abe, J., Sekimoto, H., Inzé, D. and Vyverman, W. (2010), INTRACELLULAR LOCALIZATION OF AN ENDOGENOUS CELLULOSE SYNTHASE OF MICRASTERIAS DENTICULATA (DESMIDIALES, CHLOROPHYTA) BY MEANS OF TRANSIENT GENETIC TRANSFORMATION. Journal of Phycology, 46: 839–845. doi: 10.1111/j.1529-8817.2010.00867.x
Received 30 July 2009. Accepted 12 March 2010.
- Issue published online: 2 AUG 2010
- Article first published online: 20 JUL 2010
- cellulose synthase;
- intracellular localization;
- Micrasterias denticulata;
- microparticle bombardment;
- transient genetic transformation
The desmid Micrasterias denticulata Bréb. is useful for the study of streptophyte cell wall biology and morphology. However, no tools to analyze cell biological processes in vivo in this species are available. In the present study, transient gene expression under the control of the chl a/b–binding protein gene of the Closterium peracerosum–strigosum–littorale complex (CpCAB1) promotor was achieved for M. denticulata and illustrated by the intracellular localization of an endogenous cellulose synthase (MdCesA1). A transformation efficiency of 1/5,000 cells was achieved following microparticle bombardment. The free green fluorescent protein (GFP) signal was detected both in the nucleus and in the cytoplasm. The MdCesA1-GFP fusion protein, on the other hand, occurred at the plasma membrane in particles concentrated at the lobe indentations, the lobe tips, and, to a lesser extent, along the lobe sides. Hence, the multipolar growth mechanism of the cell is reflected. In addition, the margins of cytoplasmic compartments, most likely dictyosomes, were labeled, in accordance with the known secretory pathway of cellulose synthase complexes. Besides intracellular localization studies, the utility of the system for overexpression phenotyping is discussed.