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Plant and Cell Physiology Advance Access published online on February 2, 2005
Plant and Cell Physiology, doi:10.1093/pcp/pci047
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Plant and Cell Physiology 2005 © The Japanese Society of Plant Physiologists (JSPP); all rights reserved.
Received August 19, 2004
Revised December 27, 2004
Accepted January 7, 2005

Regular Paper

High Intracellular Phosphorus Contents Exhibit a Correlationship with Arsenate Resistance in Chlamydomonas Mutants

Isao Kobayashi 1, Shoko Fujiwara 2, Kosuke Shimogawara 3, Chiseko Sakuma 1, Yasuo Shida 1, Toshikazu Kaise 1, Hideaki Usuda 3, and Mikio Tsuzuki 2*

1 School of Life Science, Tokyo University of Pharmacy and Life Science, Horinouchi, Hachioji, Tokyo 192-0392, Japan
2 School of Life Science, Tokyo University of Pharmacy and Life Science, Horinouchi, Hachioji, Tokyo 192-0392, Japan; CREST, Japan Science and Technology Agency, Kawaguchi, Saitama, 332-0392 Japan
3 Laboratory of Chemistry, Teikyo University School of Medicine, Ohtsuka, Hachioji, Tokyo 192-0395, Japan

* To whom correspondence should be addressed.
Mikio Tsuzuki, E-mail: mtsu{at}ls.toyaku.ac.jp


  Abstract

Pi in the medium relieved toxicity of arsenate against cellular growth of Chlamydomonas reinhardtii. To investigate the relationship between intracellular P contents and arsenate resistance, we determined the intracellular P contents of arsenate-sensitive and arsenate-resistant mutants, which had been generated by random insertional mutagenesis. All of thirteen arsenate-resistant mutants showed higher P contents than the parent strain, while arsenate-sensitive mutants with high P contents were not found. In one of the arsenate-resistant mutants, AR3, the intracellular P content was about twice that in the wild type during growth in the absence of arsenate. Arsenate incorporation in AR3 was suppressed within 10 min after the addition of 1 mM arsenate, while Pi incorporation continued even after arsenate uptake ceased. Whereas the P content of the wild type decreased to half in the presence of 0.5 mM arsenate, almost the same degree (about 50%) of decrease was observed in AR3 cells grown in the presence of as much as 3 mM arsenate. AR3, in which PTB1, a homologue of a Pi transporter gene, had been disrupted, exhibited a higher activity of a high-affinity Pi transporter, suggesting that it may be due to a compensatory transport activity. These data suggest that the intracellular level of P is one of the important factors of arsenate resistance.

Keywords: Arsenate resistance; Arsenate uptake; Chlamydomonas reinhardtii; NMR; Pi transporter; Pi uptake.
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