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Plant and Cell Physiology Advance Access published online on February 27, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp031
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© The Author 2009. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Microtubules regulate dynamic organization of vacuoles in Physcomitrella patens

Yoshihisa Oda1, Aiko Hirata1, Toshio Sano1,2, Tomomichi Fujita3, Yuji Hiwatashi4, Yoshikatsu Sato5, Akeo Kadota6, Mitsuyasu Hasebe4,5,7 and Seiichiro Hasezawa1,2

1. Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba, 277-8562, Japan
2. Institute for Bioinformatics Research and Development (BIRD), Japan Science and Technology Agency (JST), Chiyoda-ku, Tokyo 102-8666, Japan
3. Division of Biological Sciences, Graduate School of Science, Hokkaido University, Kita-ku Kita-10-jo-Nishi-8, Sapporo 060-0810, Hokkaido, Japan
4. Division of Evolutionary Biology, National Institute for Basic Biology, Okazaki 444-8585, Japan
5. ERATO, JST, Okazaki 444-8585, Japan
6. Department of Biological Sciences, Graduate School of Science, Tokyo Metropolitan University, Minami-Ohsawa 1-1, Hachioji, Tokyo, 1920397 Japan
7. Department of Basic Biology, School of Life Science, The Graduate University for Advanced Studies, Okazaki 444-8585, Japan

Corresponding Author: Seiichiro Hasezawa Address: Department of Integrated Biosciences, Graduate School of Frontier Sciences, The University of Tokyo, Kashiwanoha 5-1-5, Kashiwa, Chiba, 2778562, Japan. Phone: 81-4-7136-3706, E-mail: hasezawa{at}k.u-tokyo.ac.jp


  Abstract

Eukaryotic cells have developed several essential membrane components. In flowering plants, appropriate structures and distributions of the major membrane components are predominantly regulated by actin microfilaments. In this study, we have focused on the regulatory mechanism of vacuolar structures in the moss, Physcomitrella patens. High ability of P. patens for homologous recombination enabled us to stably express GFP protein- or RFP-fusion proteins and its simple body structure of P. patens enabled us to perform detailed visualization of the intra-cellular vacuolar and cytoskeletal structures. Three-dimensional analysis and high-speed time laps observations revealed surprisingly complex structures and dynamics of the vacuole, with inner sheets and tubular protrusions, and frequent rearrangements by separation and fusion of the membranes. Depolymerization of microtubules dramatically affected these structures and movements. Dual observation of microtubules and vacuolar membranes revealed that microtubules induced tubular protrusions and cytoplasmic strands of the vacuoles, indicative of interactions between microtubules and vacuolar membranes. These results demonstrate a novel function of microtubules in maintaining the distribution of vacuole and suggest a functional divergence of cytoskeletal functions in land plant evolution.

Keywords: Microtubule - Vacuole - Vacuolar membrane - Physcomitrella patens - Actin microfilament - cytoskeleton

Present addressYoshihisa Oda; Department of Biological Science, Graduate School of Science, The University of Tokyo, Hongo 7-3-1, Bunkyo-ku, Tokyo, 1130033, Japan

(Received October 30, 2008; Accepted February 19, 2009)
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