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Plant and Cell Physiology Advance Access published online on April 28, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp063
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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

Live imaging of chloroplast FtsZ1 filaments, rings, spirals, and motile dot structures in the AtMinE1 mutant and overexpressor of Arabidopsis thaliana

Makoto T. Fujiwara1,2, Kohsuke Sekine2, Yoshiharu Y. Yamamoto1,4, Tomoko Abe1, Naoki Sato2 and Ryuuichi D. Itoh3

1 RIKEN, Hirosawa 2-1, Wako, Saitama 351-0198, Japan
2 Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Meguro, Tokyo 153-8902, Japan
3 Department of Chemistry, Biology and Marine Science, Faculty of Science, University of the Ryukyus, Senbaru 1, Nishihara, Okinawa 903-0213, Japan

Corresponding authors:Dr. Ryuuichi D. Itoh, Department of Chemistry, Biology and Marine Science, Faculty of Science, University of the Ryukyus, Senbaru 1, Nishihara, Okinawa 903-0213, Japan, Tel: 81-98-895-8898 / Fax: 81-98-895-8576 / E-mail: ryuitoh{at}sci.u-ryukyu.ac.jp Makoto T. Fujiwara, Department of Life Sciences, Graduate School of Arts and Sciences, University of Tokyo, Komaba 3-8-1, Tokyo 153-8902, Japan, Tel: 81-3-5454-4947 / Fax: 81-3-5454-6998 / E-mail: mtf1{at}mac.com


  Abstract

Chloroplast division involves the tubulin-related GTPase FtsZ that assembles into a ring structure (Z-ring) at the mid-chloroplast division site, which is where invagination and constriction of the envelope membranes occur. Z-ring assembly is usually confined to the mid-chloroplast site by a well balanced counteraction of the stromal proteins MinD and MinE. The in vivo mechanisms by which FtsZ nucleates at specific sites, polymerises into a protofilament and organises a closed ring of filament bundles remain largely unknown. To clarify the dynamic aspects of FtsZ, we developed a living cell system for simultaneous visualisation of various FtsZ configurations, utilising the Arabidopsis thaliana overexpressor and mutant of the MinE (AtMinE1) gene, which were modified to weakly express green fluorescent protein (GFP) fused to AtFtsZ1-1. Time-lapse observation in the chloroplasts of both plants revealed disorderly movement of the dots and short filaments of FtsZ. The short filaments often appeared to emanate from the dots and to converge with a long filament, producing a thick cable. In the AtMinE1 overexpressor, we also observed spirals along the longitudinal axis of the organelle that often rolled the closed rings together. In the atminE1 mutant, we visualised the ‘isolated’ rings with a maximum diameter of ~2 mm that did not encircle the organelle periphery, but appeared to be suspended in the stroma. Our observations further demonstrated heterogeneity in chloroplast shapes and concurrently altered configurations of FtsZ in the mutant.

Keywords: chloroplast division - FtsZ filament - FtsZ ring - GFP - Min system - plastid division

4 Present address: Faculty of Applied Biological Sciences, Gifu University, Yanagido 1-1, Gifu 501-1193, Japan

(Received March 7, 2009; Accepted April 24, 2009)
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