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JXB Advance Access published online on May 9, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri167
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© The Author [2005]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved. For Permissions, please e-mail: journals.permissions@oupjournals.org
Received January 19, 2005
Accepted March 16, 2005

RESEARCH PAPER

Suppressed expression of the apoplastic ascorbate oxidase gene increases salt tolerance in tobacco and Arabidopsis plants

Atsuko Yamamoto 1, Md. Nazmul H. Bhuiyan 2, Rungaroon Waditee 2, Yoshito Tanaka 1, Muneharu Esaka 3, Kazuko Oba 4, André T. Jagendorf 5, and Teruhiro Takabe 6*

1 Graduate School of Environmental and Human Sciences, Meijo University, Nagoya, 468-8502, Japan
2 Research Institute, Meijo University, Tempaku-ku, Nagoya, 468-8502, Japan
3 Applied Biological Science, Hiroshima University, Higashi-Hiroshima 739-8528, Japan
4 Faculty of Domestic Science, Nagoya Women's University, Nagoya 467-8610, Japan
5 Department of Plant Biology, Cornell University, Ithaca, NY 14853, USA
6 Graduate School of Environmental and Human Sciences, Meijo University, Nagoya, 468-8502, Japan; Research Institute, Meijo University, Tempaku-ku, Nagoya, 468-8502, Japan

* To whom correspondence should be addressed.
Teruhiro Takabe, E-mail: takabe{at}ccmfs.meijo-u.ac.jp


  Abstract

Transgenic tobacco plants expressing the ascorbate oxidase (AAO) gene in sense and antisense orientations, and an Arabidopsis mutant in which the T-DNA was inserted into a putative AAO gene, were used to examine the potential roles of AAO for salt-stress tolerance in plants. AAO activities in the transgenic tobacco plants expressing the gene in sense and antisense orientations were, respectively, about 16-fold and 0.2-fold of those in the wild type. Under normal growth conditions, no significant differences in phenotypes were observed, except for a delay in flowering time in the antisense plants. However, at high salinity, the percentage germination, photosynthetic activity, and seed yields were higher in antisense plants, with progressively lower levels in the wild type and the sense plants. The redox state of apoplastic ascorbate in sense plants was very low even under normal growth conditions. Upon salt stress, the redox state of symplastic and apoplastic ascorbate decreased among the three types of plants, but was lowest in the sense plants. The hydrogen peroxide contents in the symplastic and apoplastic spaces were higher in sense plants, progressively lower in the wild type, followed by the antisense plants. The Arabidopsis T-DNA inserted mutant exhibited very low ascorbate oxidase activity, and its phenotype was similar to that of antisense tobacco plants. These results suggest that the suppressed expression of apoplastic AAO under salt-stress conditions leads to a relatively low level of hydrogen peroxide accumulation and a high redox state of symplastic and apoplastic ascorbate which, in turn, permits a higher seed yield.

Keywords: Abiotic stress; antisense expression; apoplast; ascorbate oxidase; hydrogen peroxide; salt stress.
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