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JXB Advance Access published online on May 31, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri187
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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 November 19, 2004
Accepted April 7, 2005

RESEARCH PAPER

Lygodium japonicum fern accumulates copper in the cell wall pectin

Haruyoshi Konno 1*, Takako Nakato 1, Susumu Nakashima 1, and Kenji Katoh 2

1 Research Institute for Bioresources, Okayama University, Kurashiki, Okayama 710-0046, Japan
2 Shionogi Research Laboratories, Shionogi & Co., Ltd., Osaka 553-0002, Japan

* To whom correspondence should be addressed.
Haruyoshi Konno, E-mail: hakonno{at}rib.okayama-u.ac.jp


  Abstract

The present work reports the results of a study on the growth kinetics and characterization of matrix polysaccharides in the cell walls of Lygodium japonicum prothallium grown in the presence of copper (Cu). When the prothallium was cultured in the media containing 0.2 mM or 0.4 mM CuSO4, it showed a rapid accumulation of Cu with a maximum uptake of Cu measured in the cells up to 20 d of culture. The maximum rate of Cu uptake into the prothallium was greater for 0.4 mM Cu-treated cells (17.2 µmol g-1 DW) than for 0.2 mM Cu-treated cells (3.2 µmol g-1 DW). Cell walls were isolated from both untreated control and Cu-treated cells and then extracted sequentially with cyclohexane-trans-1,2-diaminetetra-acetate (CDTA), Na2CO3, 1 M KOH, and 4 M KOH. The amount of pectin solubilized from 0.4 mM Cu-treated cell walls decreased to 53% of its level in the control, whereas the amount of hemicellulose solubilized from the Cu-treated cell walls represented 82% of that from control cell walls. When the polysaccharides were fractionated by anion-exchange chromatography into four carbohydrate components, considerable increases in fractions PI-3 and PII-3 eluted with 0.5 M NaCl were observed in CDTA-soluble (PI) and Na2CO3-soluble (PII) pectic polymers from Cu-treated cell walls. Fractions PI-3 and PII-3 were composed predominantly of uronic acid (more than 71% of total sugars). Approximately 66% of Cu within the cell walls was released from the 0.4 mM Cu-treated cells with the endo-pectate-lyase treatment, suggesting that most of the Cu that accumulated into the Lygodium prothallium is tightly bound to the homogalacturonan of the cell wall pectin.

Keywords: Copper accumulation; endo-pectate lyase; fern (pteridophyte); homogalacturonan; Lygodium japonicum; pectin structure; phytoremediation; prothallium.
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