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Isolation and characterization of Arabidopsis halleri and Thlaspi caerulescens phytochelatin synthases

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Abstract

The synthesis of phytochelatins (PC) represents a major metal and metalloid detoxification mechanism in various species. PC most likely play a role in the distribution and accumulation of Cd and possibly other metals. However, to date, no studies have investigated the phytochelatin synthase (PCS) genes and their expression in the Cd-hyperaccumulating species. We used functional screens in two yeast species to identify genes expressed by two Cd hyperaccumulators (Arabidopsis halleri and Thlaspi caerulescens) and involved in cellular Cd tolerance. As a result of these screens, PCS genes were identified for both species. PCS1 was in each case the dominating cDNA isolated. The deduced sequences of AhPCS1 and TcPCS1 are very similar to AtPCS1 and their identity is particularly high in the proposed catalytic N-terminal domain. We also identified in A. halleri and T. caerulescens orthologues of AtPCS2 that encode functional PCS. As compared to A. halleri and A. thaliana, T. caerulescens showed the lowest PCS expression. Furthermore, concentrations of PC in Cd-treated roots were the highest in A. thaliana, intermediate in A. halleri and the lowest in T. caerulescens. This mirrors the known capacity of these species to translocate Cd to the shoot, with T. caerulescens being the best translocator. Very low or undetectable concentrations of PC were measured in A. halleri and T. caerulescens shoots, contrary to A. thaliana. These results suggest that extremely efficient alternative Cd sequestration pathways in leaves of Cd hyperaccumulators prevent activation of PC synthase by Cd2+ ions.

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Abbreviations

Cd:

Cadmium

EMM:

Edinburgh’s minimal medium

GSH:

Glutathione

PC:

Phytochelatin

PCS:

Phytochelatin synthase

RT-PCR:

Reverse transcription polymerase chain reaction

RACE-PCR:

Rapid amplification of cDNA ends-polymerase chain reaction

WT:

Wild type

YNB:

Yeast nitrogen base

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Acknowledgments

We are grateful to P. Salis for technical help. We also thank anonymous reviewers for their helpful and constructive comments on the manuscript. This work was supported by the European Union (Research Training Network METALHOME, grant no. HPRN–CT–2002–00243, and in part through its Sixth Framework Program for RTD, contract no FOOD-CT-2006- 016253), grant from the Belgian Programme on Interuniversity Poles (Science Policy Program VI/33) and the FNRS (FRFC 2.4.583.08). A-C. C. was supported by an Alexander-von-Humboldt fellowship.

Author information

Author notes

  1. Mikael Courbot

    Present address: Jealott’s Hill International Research Centre, Syngenta Ltd, RG42 6EY, Bracknell, UK

  2. Adrian Radu Craciun

    Present address: Laborimpex, Brussels, Belgium

  3. Denis Desgain

    Present address: UNV Cuba, Havana, Cuba

  4. Anne-Claire Cazalé

    Present address: Laboratoire des Interactions Plantes Micro-organismes (LIPM), UMR CNRS-INRA 2594/441, 31320, Castanet Tolosan, France

Authors and Affiliations

  1. Faculté des Sciences, Laboratoire de Physiologie & de Génétique moléculaire des Plantes, Université Libre de Bruxelles, Campus Plaine CP242, 1050, Brussels, Belgium

    Claire-Lise Meyer, Mikael Courbot, Adrian Radu Craciun, Denis Desgain & Nathalie Verbruggen

  2. Department of Plant Physiology, University of Bayreuth, Universitätsstrasse 30, 95440, Bayreuth, Germany

    Daniel Peisker, Anne-Claire Cazalé & Stephan Clemens

  3. Institute of Molecular and Cellular Biology, Faculty of Earth and Life Sciences, Vrije Universiteit, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands

    Henk Schat

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  1. Claire-Lise Meyer
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Correspondence to Nathalie Verbruggen.

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C.-L. Meyer, D. Peisker contributed equally to this work.

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Meyer, CL., Peisker, D., Courbot, M. et al. Isolation and characterization of Arabidopsis halleri and Thlaspi caerulescens phytochelatin synthases. Planta 234, 83–95 (2011). https://doi.org/10.1007/s00425-011-1378-z

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