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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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References

  • Brooks RR (1998) Plants that hyperaccumulate heavy metals. CAB International, Wallingford, UK

    Google Scholar 

  • Clemens S, Palmgren MG, Krämer U (2002) A long way ahead: understanding and engineering plant metal accumulation. Trends Plant Sci 7:309–315

    Article  CAS  PubMed  Google Scholar 

  • Delhaize E, Ryan PR, Randall PJ (1993) Aluminum torelance in wheat (Triticum aestive L.). Plant Physiol 103:659–702

    Google Scholar 

  • Ebbs S, Lau I, Ahner B, Kochian L (2002) Phytochelatin synthesis is not responsible for Cd tolerance in the Zn/Cd hyperaccumulator Thlaspi caerulescens (J. and C. Presl). Planta 214:635–640

    CAS  PubMed  Google Scholar 

  • Grassi M, Gatti G (1995) Nuclear magnetic resonance methods in environmental chemistry. Anal Chim Rome 85:487–502

    CAS  Google Scholar 

  • Grassi M, Mingazzini M (2001) 113Cd-NMR and fluorescence studies of the interactions between Cd(II) and extracellular organic matter released by Seleniastrum capricornutum. Environ Sci Technol 35:4271–4276

    Article  CAS  PubMed  Google Scholar 

  • Hall JL (2002) Cellular mechanisms for heavy metal detoxification and tolerance. J Exp Bot 53:1–11

    Article  CAS  PubMed  Google Scholar 

  • Kostelnik RJ, Bothner-By AA (1974) Cadmium-113 nuclear magnetic resonance studies of cadmium (II)-ligand binding in aqueous solutions. I. The effect of diverse ligands on the cadmium-113 chemical shift. J Magn Reson 14:141–151

    CAS  Google Scholar 

  • Krämer U, Cotter-Howells JD, Charnock JM, Baker AJM, Smith JAC (1996) Free histidine as a metal chelator in plants that accumulate nickel. Nature 378:635–638

    Article  Google Scholar 

  • Küpper H, Mijovilovich A, Klaucke WM, Kroneck PMH (2004) Tissue- and age-dependent differences in the complexation of cadmium and zinc in the cadmium/zinc hyperaccumulator Thlaspi caerulescens (Ganges ecotype) revealed by X-ray absorption spectroscopy. Plant Physiol 134:748–757

    Article  PubMed  Google Scholar 

  • Lasat MM, Baker AJM, Kochian LV (1996) Physiological Characterization of root Zn2+ absorption and translocation to shoots in Zn hyperaccumulator and nonaccumulator species of Thlaspi. Plant Physiol 112:1715–1722

    CAS  PubMed  Google Scholar 

  • Lasat MM, Pence NS, Garvin DF, Ebbs SD, Kochian LV (2000) Molecular physiology of zinc transport in the Zn hyperaccumulator Thlaspi caerulescens. J Exp Bot 51:71–79

    Article  CAS  PubMed  Google Scholar 

  • Li J, Perdue EM, Gelbaum LT (1998) Using cadmium-113 NMR spectrometry to study metal complexation by natural organic matter. Environ Sci Technol 32:483–487

    Article  CAS  Google Scholar 

  • Lombi E, Zhao FJ, Dunham SJ, McGrath SP (2000) Cadmium accumulation in populations of Thlaspi caerulescens and Thlaspi goesingense. New Phytol 145:11–20

    CAS  Google Scholar 

  • Lombi E, Zhao FJ, McGrath SP, Young SD, Sacchi GA (2001) Physiological evidence for a high-affinity cadmium transporter highly expressed in a Thlaspi caerulescens ecotype. New Phytol 149:53–60

    Article  CAS  Google Scholar 

  • Lombi E, Tearall KL, Howarth JR, Zhao FJ, Hawkesford MJ, McGrath SP (2002) Influence of iron status on cadmium and zinc uptake by different ecotypes of the hyperaccumulator Thlaspi caerulescens. Plant Physiol 128:1359–1367

    Article  CAS  PubMed  Google Scholar 

  • Ma JF, Hiradate S, Nomoto K, Iwashita T, Matsumoto H (1997a) Internal detoxification mechanism of Al in hydrangea. Identification of Al form in the leaves. Plant Physiol 113:1033–1039

    CAS  PubMed  Google Scholar 

  • Ma JF, Zheng SJ, Hiradate S, Matsumoto H (1997b) Detoxifying aluminium with buckwheat. Nature 390:569–570

    Article  PubMed  Google Scholar 

  • Ma JF, Hiradate S, Matsumoto H (1998) High aluminum resistance in buckwheat II. Oxalic acid detoxifies aluminum internally. Plant Physiol 117:753–759

    Article  CAS  PubMed  Google Scholar 

  • Ma JF, Ryan PR, Delhaize M (2001) Aluminium tolerance in plants and the complexing role of organic acids. Trends Plant Sci 6:273–278

    Article  CAS  PubMed  Google Scholar 

  • Ma JF, Ueno D, Zhao FJ, McGrath SP (2004) Subcellular localisation of Cd and Zn in the leaves of a Cd-hyperaccumulating ecotype of Thlaspi caerulescens. Planta 10.1007/s00425-004-1392-5

  • McGrath SP, Zhao FJ (2003) Phytoextraction of metals and metalloids from contaminated soils. Curr Opin Biotechnol 14:277–282

    Article  CAS  PubMed  Google Scholar 

  • Persans MW, Nieman K, Salt DE (2001) Functional activity and role of cation-efflux family members in Ni hyperaccumulation in Thlaspi goesingense. Proc Natl Acad Sci USA 98:9995–10000

    Article  CAS  PubMed  Google Scholar 

  • Robinson BH, Leblanc M, Petit D, Brooks RR, Kirkman JH, Gregg PEH (1998) The potential of Thlaspi caerulescens for phytoremediation of contaminated soils. Plant Soil 203:47–56

    Article  CAS  Google Scholar 

  • Roosens N, Verbruggen N, Meerts P, Ximénez-Embún P, Smith JAC (2003) Natural variation of cadmium tolerance and its relationship to metal hyperaccumulation for seven populations of Thlaspi caerulescens from Western Europe. Plant Cell Environ 26:1657–1672

    Article  CAS  Google Scholar 

  • Sadler PJ, Viles JH (1996) 1H and 113Cd NMR investigations of Cd2+ and Zn2+ binding sites on serum albumin: competition with Ca2+, Ni2+, Cu2+, and Zn2+. Inorg Chem 35:4490–4496

    Article  CAS  PubMed  Google Scholar 

  • Salt DE, Prince RC, Baker AJM, Raskin I, Pickering IJ (1999) Zinc ligands in the metal hyperaccumulator Thlaspi caerulescens as determined using X-ray absorption spectroscopy. Environ Sci Technol 33:713–717

    Article  CAS  Google Scholar 

  • Sanità di Toppi L, Gabbrielli R (1999) Response to cadmium in higher plants. Environ Exp Bot 41:105–130

    Article  Google Scholar 

  • Sarret G, Saumitou-Laprade P, Bert V, Proux O, Hazemann JL, Traverse A, Marcus MA, Manceau A (2002) Forms of zinc accumulated in the hyperaccumulator Arabidopsis halleri. Plant Physiol 130:1815–1862

    Article  CAS  PubMed  Google Scholar 

  • Schat H, Llugany M, Vooijs R, Hartley-Whitaker J, Bleeker PM (2002) The role of phytochelatins in constitutive and adaptive heavy metal tolerances in hyperaccumulator and non-hyperaccumulator metallophytes. J Exp Bot 53:2381–2392

    Article  CAS  PubMed  Google Scholar 

  • Schutzendubel A, Schanz P, Teichmann T, Gross K, Langenfeld-Heyser R, Godbold D, Polle A (2001) Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in Scots pine roots. Plant Physiol 127:887–898

    Article  CAS  PubMed  Google Scholar 

  • Shen RF, Iwashita T, Ma JF (2004) Form of Al changes with Al concentration in leaves of buckwheat. J Exp Bot 55:131–136

    Article  CAS  PubMed  Google Scholar 

  • Shen ZG, Zhao FJ, McGrath SP (1997) Uptake and transport of zinc in the hyperaccumulator Thlaspi caerulescens and the non-hyperaccumulator Thlaspi ochroleucum. Plant Cell Environ 20:898–906

    Article  CAS  Google Scholar 

  • Somashekaraiah BV, Padmaja K, Prasad ARK (1992) Phytotoxicity of cadmium ions on germinating seedlings of mung bean (Phaseolus vulgaris): involvement of lipid peroxides in chlorophyll degradation. Physiol Plant 85:85–89

    Article  CAS  Google Scholar 

  • Ueno D, Zhao FJ, Ma JF (2004) Interactions between Cd and Zn in relation to their hyperaccumulation in Thlaspi caerulescens. Soil Sci Plant Nutr 50:591–597

    CAS  Google Scholar 

  • Van Assche F, Clijsters H (1990) Effects of metals on enzyme activity in plants. Plant Cell Environ 13:195–206

    CAS  Google Scholar 

  • Zha HG, Jiang RF, Zhao FJ, Vooijs R, Schat H, Barker JHA, McGrath SP (2004) Co-segregation analysis of cadmium and zinc accumulation in Thlaspi caerulescens interecotypic crosses. New Phytol 163:299–312

    Article  CAS  Google Scholar 

  • Zhao FJ, Lombi E, Breedon T, McGrath SP (2000) Zinc hyperaccumulation and cellular distribution in Arabidopsis halleri. Plant Cell Environ 23:507–514

    Article  CAS  Google Scholar 

  • Zhao FJ, Hamon RE, Lombi E, McLaughlin MJ, McGrath SP (2002) Characteristics of cadmium uptake in two contrasting ecotypes of the hyperaccumulator Thlaspi caerulescens. J Exp Bot 53:535–543

    Article  CAS  PubMed  Google Scholar 

Download references

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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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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  • DOI: https://doi.org/10.1007/s00425-005-1491-y

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