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Overexpression of Arabidopsis phytochelatin synthase in tobacco plants enhances Cd2+ tolerance and accumulation but not translocation to the shoot

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Abstract

Phytochelatins (PCs) are metal binding peptides involved in heavy metal detoxification. To assess whether enhanced phytochelatin synthesis would increase heavy metal tolerance and accumulation in plants, we overexpressed the Arabidopsis phytochelatin synthase gene (AtPCS1) in the non-accumulator plant Nicotiana tabacum. Wild-type plants and plants harbouring the Agrobacterium rhizogenes rolB oncogene were transformed with a 35S AtPCS1 construct. Root cultures from rolB plants could be easily established and we demonstrated here that they represent a reliable system to study heavy metal tolerance. Cd2+ tolerance in cultured rolB roots was increased as a result of overexpression of AtPCS1, and further enhanced when reduced glutathione (GSH, the substrate of PCS1) was added to the culture medium. Accordingly, HPLC analysis showed that total PC production in PCS1-overexpressing rolB roots was higher than in rolB roots in the presence of GSH. Overexpression of AtPCS1 in whole seedlings led to a twofold increase in Cd2+ accumulation in the roots and shoots of both rolB and wild-type seedlings. Similarly, a significant increase in Cd2+ accumulation linked to a higher production of PCs in both roots and shoots was observed in adult plants. However, the percentage of Cd2+ translocated to the shoots of seedlings and adult overexpressing plants was unaffected. We conclude that the increase in Cd2+ tolerance and accumulation of PCS1 overexpressing plants is directly related to the availability of GSH, while overexpression of phytochelatin synthase does not enhance long distance root-to-shoot Cd2+ transport.

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Abbreviations

DAG:

Days after germination

GSH:

Glutathione

PC:

Phytochelatin

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Acknowledgements

Special thanks are given to Dr. Annette Pickford for helpful comments during manuscript revision. We thank Dr. Adele Figliolia (INP Rome, Italy) for helpful discussions, Prof. Rita Biasi and Dr. Patricia Gutierrez (University of Viterbo, Italy) for their help in ANOVA analysis. This work was partially supported by grants from Istituto Pasteur Fondazione Cenci-Bolognetti, and MIUR (FIRB, PRIN, Centro di Eccellenza in Biologia e Medicina Molecolare).

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  1. Vincenzo Censi

    Present address: Istituto di Ricerche di Biologia Molecolare P. Angeletti, IRBM, Via Pontina Km 30,6, 00040, Pomezia (Rome), Italy

Authors and Affiliations

  1. Dipartimento di Genetica e Biologia Molecolare, Universitá La Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy

    Mirella Pomponi & Paolo Costantino

  2. Istituto Biologia e Patologia Molecolari, CNR, Universitá La Sapienza, Piazzale Aldo Moro 5, 00185, Rome, Italy

    Vincenzo Censi, Valentina Di Girolamo & Maura Cardarelli

  3. Istituto di Scienze delle Produzioni Alimentari CNR (ISPA), Sez. di Lecce via Monteroni, 73100, Lecce, Italy

    Angelo De Paolis

  4. Dipartimento di Biologia Evolutiva e Funzionale, Universitá di Parma, Viale delle Scienze 11/A, 43100, Parma, Italy

    Luigi Sanità di Toppi

  5. Istituto Sperimentale per la Nutrizione delle Piante, Via della Navicella 2/4, 00184, Rome, Italy

    Rita Aromolo

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  1. Mirella Pomponi
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Correspondence to Maura Cardarelli.

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Pomponi, M., Censi, V., Di Girolamo, V. et al. Overexpression of Arabidopsis phytochelatin synthase in tobacco plants enhances Cd2+ tolerance and accumulation but not translocation to the shoot. Planta 223, 180–190 (2006). https://doi.org/10.1007/s00425-005-0073-3

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