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Identification of the form of Cd in the leaves of a superior Cd-accumulating ecotype of Thlaspi caerulescens using 113Cd-NMR

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Abstract

Thlaspi caerulescens (Ganges ecotype) is a known Cd hyperaccumulator, however, the ligands which coordinate to Cd ions in the leaves have not been identified. In the present study, the chemical form of Cd was investigated by using 113Cd-nuclear magnetic resonance (NMR) spectroscopy. Plants were grown hydroponically with a highly enriched 113Cd stable isotope. Measurements of 113Cd-NMR with intact leaves showed a signal at the chemical shift of around −16 ppm. Crude leaf sap also gave a similar chemical shift. Purification by gel filtration (Sephadex G-10), followed by cationic and anionic exchange chromatography, showed that Cd occurred only in the anionic fraction, which gave the same chemical shift as intact leaves. Further purification of the anionic fraction, combined with 113Cd- and 1H-NMR studies, revealed that only the fraction containing malate showed a chemical shift similar to the intact leaves. These results indicate that Cd was coordinated mainly with malate in the leaves of T. caerulescens. The malate concentration in the leaves was not affected by increasing Cd concentration in the solution, suggesting that malate synthesis is not induced by Cd. Because the Cd-malate complex is relatively weak, we suggest that the complex forms inside the vacuoles as a result of an efficient tonoplast transport of Cd and a constitutively high concentration of malate in the vacuoles, and that the formation of the Cd-malate complex may lead to a decrease of subsequent Cd efflux to the cytoplasm.

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Acknowledgments

The study was partly supported by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology of Japan (No. 15658021 to J. F. Ma). Rothamsted Research receives grant-aided support from the U.K. Biotechnology and Biological Sciences Research Council.

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Authors and Affiliations

  1. Faculty of Agriculture, Kagawa University, Ikenobe 2393, Miki-cho, Kita-gun,, Kagawa 761-0795, Japan

    Daisei Ueno

  2. Suntory Institute for Bioorganic Research, Wakayamadai 1-1-1, Shimamoto-cho, Mishima-gun, Osaka, 618-8503, Japan

    Takashi Iwashita

  3. Agriculture and Environment Division, Rothamsted Research, Harpenden, Hertfordshire, AL5 2JQ, UK

    Fang-Jie Zhao & Steve P McGrath

  4. Research Institute for Bioresources, Okayama University, Chuo 2-20-1,, Kurashiki 710-0046, Japan

    Jian Feng Ma

Authors
  1. Daisei Ueno
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  2. Jian Feng Ma
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  3. Takashi Iwashita
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  4. Fang-Jie Zhao
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  5. Steve P McGrath
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Correspondence to Jian Feng Ma.

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Ueno, D., Ma, J.F., Iwashita, T. et al. Identification of the form of Cd in the leaves of a superior Cd-accumulating ecotype of Thlaspi caerulescens using 113Cd-NMR. Planta 221, 928–936 (2005). https://doi.org/10.1007/s00425-005-1491-y

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