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INTRACELLULAR LOCALIZATION OF AN ENDOGENOUS CELLULOSE SYNTHASE OF MICRASTERIAS DENTICULATA (DESMIDIALES,CHLOROPHYTA) BY MEANS OF TRANSIENT GENETIC TRANSFORMATION1 |
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| Authors: | Katrijn Vannerum Jun Abe Hiroyuki Sekimoto Dirk Inzé Wim Vyverman |
| Affiliation: | 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;2. Department of Chemical and Biological Sciences, Faculty of Science, Japan Women’s University, 2‐8‐1 Mejirodai, Bunkyo‐ku, Tokyo 112‐8681, Japan;3. Department of Plant Systems Biology, VIB, Technologiepark 927, 9052 Ghent, Belgium Department of Plant Biotechnology and Genetics, Ghent University, Technologiepark 927, 9052 Ghent, Belgium;4. Laboratory of Protistology and Aquatic Ecology, Department of Biology, Ghent University, Krijgslaan 281‐S8, 9000 Ghent, Belgium;5. Author for correspondence: e‐mail . |
| 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. |
| Keywords: | cellulose synthase intracellular localization Micrasterias denticulata microparticle bombardment overexpression transient genetic transformation |