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Plant and Cell Physiology Advance Access published online on June 3, 2008
Plant and Cell Physiology, doi:10.1093/pcp/pcn084
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© The Author 2008. 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

Mitochondrial Dynamics in Plant Male Gametophyte Visualized by Fluorescent Live Imaging

Ryo Matsushima1, Yuuki Hamamura2, Tetsuya Higashiyama3, Shin-ichi Arimura4, Sodmergen5, Nobuhiro Tsutsumi4 and Wataru Sakamoto1

1Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan
2Department of Biological Sciences, Graduate School of Science, University of Tokyo, Hongo, Tokyo, 113-0033, Japan
3Division of Biological Science, Graduate School of Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8602, Japan
4Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo-ku, Tokyo 113-8657, Japan
5College of Life Sciences, Peking University, Beijing 100871, China

Corresponding author: Prof. Wataru Sakamoto, Research Institute for Bioresources, Okayama University, 2-20-1 Chuo, Kurashiki, 710-0046, Japan. E-mail saka{at}rib.okayama-u.ac.jp, fax 81-86-434-1206


  Abstract

Visualization of organelles in living cells is a powerful method for studying their dynamic behaviors. Here we attempted to visualize mitochondria in angiosperm male gametophyte (pollen grain from Arabidopsis thaliana) that are composed of one vegetative cell (VC) and two sperm cells (SCs). Combination of mitochondria-targeted fluorescent proteins with VC- or SC-specific expression allowed us to observe precise number and dynamic behavior of mitochondria in the respective cell types. Furthermore, live imaging of SC mitochondria during double fertilization confirmed previous observations, demonstrated by electron microscopy in other species, that sperm mitochondria enter into egg and central cells. We also attempted to visualize mutant mitochondria that were elongated due to a defect in mitochondrial division. This mutant phenotype was indeed detectable in VC mitochondria of a heterozygous F1 plant, suggesting active mitochondrial division in male gametes. Finally, we performed mutant screening and isolated a putative mitochondrial protein-transport mutant whose phenotype was detectable only in haploid cells. The transgenic materials presented in this work are useful for not only live imaging but also studying mitochondrial functions by mutant analysis.

Keywords: fertilization - fluorescent protein - mitochondria - pollen grain - sperm cell - Arabidopsis thaliana

(Received April 13, 2008; Accepted May 29, 2008)
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