INTRACELLULAR LOCALIZATION OF AN ENDOGENOUS CELLULOSE SYNTHASE OF MICRASTERIAS DENTICULATA (DESMIDIALES, CHLOROPHYTA) BY MEANS OF TRANSIENT GENETIC TRANSFORMATION

Authors

  • Katrijn Vannerum,

    1. Laboratory of Protistology and Aquatic Ecology, Department of Biology, Ghent University, Krijgslaan 281-S8, 9000 Ghent, Belgium
      Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium
      Department of Plant Biotechnology and Genetics, Ghent University, Technologiepark 927, 9052 Ghent, Belgium
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  • Jun Abe,

    1. Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan
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  • Hiroyuki Sekimoto,

    1. Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, 2-8-1 Mejirodai, Bunkyo-ku, Tokyo 112-8681, Japan
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  • Dirk Inzé,

    1. Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium
      Department of Plant Biotechnology and Genetics, Ghent University, Technologiepark 927, 9052 Ghent, Belgium
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  • Wim Vyverman

    1. Laboratory of Protistology and Aquatic Ecology, Department of Biology, Ghent University, Krijgslaan 281-S8, 9000 Ghent, Belgium
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  • Received 30 July 2009. Accepted 12 March 2010.

Abstract

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.

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