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Plant and Cell Physiology Advance Access published online on October 29, 2009
Plant and Cell Physiology, doi:10.1093/pcp/pcp143
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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

Dynamic Aspects of Ion Accumulation by Vesicle Traffic under Salt Stress in Arabidopsis

Kohei Hamaji1,*, Megumi Nagira1,3,*, Katsuhisa Yoshida1, Miwa Ohnishi1,2, Yoshihisa Oda4, Tomohiro Uemura4, Tatsuaki Goh1, Masa H. Sato5, Miyo Terao-Morita6, Masao Tasaka6, Sei-ichiro Hasezawa7, Akihiko Nakano4,8, Ikuko Hara-Nishimura9, Masayoshi Maeshima10, Hidehiro Fukaki1,2 and Tetsuro Mimura1,2

1 Graduate School of Science, Kobe University, Nada-ku, Kobe, 657-8501, Japan.
2 Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Agency (JST), Chuou-ku, Tokyo, 113-0027 Japan.
3 Faculty of Science, Nara-Women's University, Nara, 630-8506 Japan
4 Graduate School of Science, the University of Tokyo, Hongo, Bunkyo-ku, Tokyo, 113-0033 Japan.
5 Graduate School of Life and Environmental Sciences, Kyoto Prefectural University, Sakyo-ku, Kyoto, 606-8522, Japan
6 Graduate School of Biological Sciences, Nara institute of Science and Technology, Ikoma, Nara, 630-0192 Japan.
7 Graduate School of Frontier Sciences, the University of Tokyo, Kashiwa, Chiba, 277-8562 Japan.
8 RIKEN Advanced Science Institute, Wako, Saitama, 351-0198 Japan
9 Graduate School of Science, Kyoto University, Sakyo-ku, Kyoto, 606-8502, Japan.
10 Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya, 464-8601 Japan.

Corresponding author: Tetsuro Mimura Graduate School of Science, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan Telephone / Fax: +81-78-803-5708 E-mail: mimura{at}kobe-u.ac.jp


  Abstract

The intracellular membrane dynamics of Arabidopsis cells under high salt treatment were investigated. When Arabidopsis was treated with high levels of NaCl in hydroponic culture, root tip cells showed rapid changes in the vacuolar volume, decrease in the number of small acid compartments, active movement of vesicles and accumulation of Na+ both in the central vacuole and in the vesicles around the main vacuole observed with the Na+-dependent fluorescence of Sodium Green. Detailed observation of Arabidopsis suspension-cultured cells under high salt treatment showed a similar pattern of response to that observed in root tip cells. Immunostaining of suspension-cultured cells with antibodies against AtNHX1 clearly showed the occurrence of dotted fluorescence in the cytoplasm only under salt treatment. We also confirmed the existence of AtNHX1 in the vacuolar membrane isolated from suspension-cultured cells with immunofluorescence.

Knockout of vacuolar Qa-SNARE protein; VAM3/SYP22 caused an increase in salt tolerance. In mutant plants, the distribution of Na+ between roots and shoots differed from that of wild type plants, with Na+ accumulating more in roots and less in the shoots of the mutant plants. The role of vesicle traffic under salt stress is discussed.

Keywords: Arabidopsis suspension-cultured cells - Na+/H+ antiporter - NHX - Salt tolerance - SNARE - Vacuole

* K.H. and M.N. contributed equally to this work.

(Received August 7, 2009; Accepted October 14, 2009)
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