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Expression of the PsMT A1 gene in white poplar engineered with the MAT system is associated with heavy metal tolerance and protection against 8-hydroxy-2′-deoxyguanosine mediated-DNA damage

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Abstract

Marker-free transgenic white poplar (Populus alba L., cv ‘Villafranca’) plants, expressing the PsMT A1 gene from Pisum sativum for a metallothionein-like protein, were produced by Agrobacterium tumefaciens-mediated transformation. The 35SCaMV-PsMT A1 -NosT cassette was inserted into the ipt-type vector pMAT22. The occurrence of the abnormal ipt-shooty phenotype allowed the visual selection of transformants, while the yeast site-specific recombination R/RS system was responsible for the excision of the undesired vector sequences with the consequent recovery of normal marker-free transgenic plants. Molecular analyses confirmed the presence of the 35SCaMV-PsMT A1 -NosT cassette and transgene expression. Five selected lines were further characterized, revealing the ability to withstand heavy metal toxicity. They survived 0.1 mM CuCl2, a concentration which strongly affected the nontransgenic plants. Moreover, root development was only slightly affected by the ectopic expression of the transgene. Reactive oxygen species were accumulated to a lower extent in leaf tissues of multi-auto-transformation (MAT)-PsMTA1 plants exposed to copper and zinc, compared to control plants. Tolerance to photo-oxidative stress induced by paraquat was another distinctive feature of the MAT-PsMTA1 lines. Finally, low levels of DNA damage were detected by quantifying the amounts of 8-hydroxy-2′-deoxyguanosine in leaf tissues of the transgenic plants exposed to copper.

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Acknowledgments

This research was supported by funds from the Italian Ministry for Education, University and Scientific Research, to D. Carbonera (PRIN 2005055337), to S.Botti (PRIN 2005055337_005), to M. Racchi (PRIN 2005055337_004) and to G. Lingua (PRIN 2005055337_003). Authors gratefully acknowledge Dr Keiko Yamada (Nippon Paper Industries, Tokyo, Japan) for supplying the pMAT22 vector.

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Author notes

  1. Samantha Zelasco

    Present address: C.R.A.-Centro di Ricerca per l’Olivicoltura e l’Industria Olearia (CRA-OLI), C/da Li Rocchi Vermicelli, Rende, 87036, Cosenza, Italy

Authors and Affiliations

  1. Dipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, Italy

    Alma Balestrazzi, Silvia Botti & Daniela Carbonera

  2. C.R.A.-Unità di Ricerca per le Produzioni Legnose fuori Foresta, Casale Monferrato, Italy

    Samantha Zelasco & Paolo Calligari

  3. Dipartimento di Biologia Evoluzionistica Sperimentale, Università di Bologna, Bologna, Italy

    Stefania Biondi & Cinzia Franchin

  4. Dipartimento di Biotecnologie Agrarie, Università di Firenze, Firenze, Italy

    Milvia Racchi & Adelaide Turchi

  5. Dipartimento di Scienze dell’Ambiente e della Vita, Università del Piemonte Orientale, Alessandria, Italy

    Guido Lingua & Graziella Berta

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  1. Alma Balestrazzi
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  2. Silvia Botti
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Correspondence to Daniela Carbonera.

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Communicated by A. Feher.

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Balestrazzi, A., Botti, S., Zelasco, S. et al. Expression of the PsMT A1 gene in white poplar engineered with the MAT system is associated with heavy metal tolerance and protection against 8-hydroxy-2′-deoxyguanosine mediated-DNA damage. Plant Cell Rep 28, 1179–1192 (2009). https://doi.org/10.1007/s00299-009-0719-x

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  • DOI: https://doi.org/10.1007/s00299-009-0719-x

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