First published online August 14, 2003; 10.1104/pp.103.022673
Plant Physiology 133:361-367 (2003)
© 2003 American Society of Plant Biologists
CELL BIOLOGY AND SIGNAL TRANSDUCTION
Dehydroascorbate Uptake Activity Correlates with Cell Growth and Cell Division of Tobacco Bright Yellow-2 Cell Cultures
Nele Horemans*,
Geert Potters,
Leen De Wilde and
Roland J. Caubergs
University of Antwerp, Department of Biology, Plant Physiology Groenenborgerlaan 171, B-2020, Antwerp, Belgium
Recently, ascorbate (ASC) concentration and the activity of a number of enzymes from the ASC metabolism have been proven to correlate with differences in growth or cell cycle progression. Here, a possible correlation between growth and the activity of a plasma membrane dehydroascorbate (DHA) transporter was investigated. Protoplasts were isolated from a tobacco (Nicotiana tabacum) Bright Yellow-2 cell culture at different intervals after inoculation and the activity of DHA transport was tested with 14C-labeled ASC. Ferricyanide (1 mM) or dithiothreitol (1 mM) was included in the test to keep the external 14C-ASC in its oxidized respectively reduced form. Differential uptake activity was observed, correlating with growth phases of the cell culture. Uptake of DHA in cells showed a peak in exponential growth phase, whereas uptake in the presence of dithiothreitol did not. The enhanced DHA uptake was not due to higher endogenous ASC levels that are normally present in exponential phase because preloading of protoplasts of different ages did not affect DHA uptake. Preloading was achieved by incubating cells before protoplastation for 4 h in a medium supplemented with 1 mM DHA. In addition to testing cells at different growth phases, uptake of DHA into the cells was also followed during the cell cycle. An increase in uptake activity was observed during M phase and the M/G1 transition. These experiments are the first to show that DHA transport activity into plant cells differs with cell growth. The relevance of the data to the action of DHA and ASC in cell growth will be discussed.
* Corresponding author; e-mail neho{at}ruca.ua.ac.be; fax: 32-3-218-04-17.
Received February 26, 2003;
returned for revision April 21, 2003;
accepted June 14, 2003.
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