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The tyrosyl-DNA phosphodiesterase gene family in Medicago truncatula Gaertn.: bioinformatic investigation and expression profiles in response to copper- and PEG-mediated stress

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Abstract

The Tdp1 gene encoding tyrosyl-DNA phosphodiesterase has been extensively investigated in animal cells, due to the role of this enzyme in the repair of topoisomerase I-DNA covalent lesions. In contrast, information in this regard is totally missing in plants. We report for the first time in plants on the Tdp1 gene family from barrel medic (Medicago truncatula Gaertn.), composed of two members, hereby named MtTdp1α and MtTdp1β. The expression profiles of MtTdp1α and MtTdp1β genes were evaluated in plantlets grown in vitro using copper and polyethylene glycol (PEG 6000) as stress agents. In situ detection of reactive oxygen species (ROS) was carried out by histochemical staining, while the level of oxidative DNA damage, quantified in terms of 7,8-dihydro-8-oxoguanine (8-oxo-dG), increased up to 7.4- and 6.7-fold in response to copper and PEG 6000 treatments, respectively. Quantitative real-time polymerase chain reaction revealed that both Tdp1 genes were significantly up-regulated in response to copper and PEG. The Tdp1 genes were also significantly up-regulated during seed rehydration, an aspect of seed physiology in which DNA repair is a key component. Thus, the Tdp1 genes might be used as novel tools for improving stress tolerance in crops. The expression patterns of the barrel medic top1α and top1β genes, encoding distinct isoforms of DNA topoisomerase I, were also analyzed and discussed to acquire additional information on their specific functions, closely related to that of the Tdp1 gene in animal cells.

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Abbreviations

Tdp:

Tyrosyl-DNA phosphodiesterase

ROS:

Reactive oxygen species

8-oxo-dG:

7, 8-Dihydro-8-oxoguanine

PEG:

Polyethylene glycol

QRT-PCR:

Quantitative real-time polymerase chain reaction

top1:

Gene encoding DNA topoisomerase I

topo I:

DNA topoisomerase I

Mt:

Medicago truncatula

DAB:

3, 3′-Diaminobenzidine

NBT:

Nitroblue tetrazolium

APX:

Ascorbate peroxidase

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Acknowledgments

This research was supported by Fondo di Ateneo per la Ricerca-University of Pavia. The authors would like to thank Drs. Sergio Arcioni and Andrea Porceddu (IGV-CNR, Perugia, Italy) for providing R108-1 Medicago truncatula genotype.

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Authors and Affiliations

  1. Dipartimento di Genetica e Microbiologia, Via Ferrata 1, 27100, Pavia, Italy

    Anca Macovei, Alma Balestrazzi & Daniela Carbonera

  2. C.R.A. Centro di Ricerca per le Produzioni Foraggere e Lattiero-Casearie, Viale Piacenza 29, 26900, Lodi, Italy

    Massimo Confalonieri

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  1. Anca Macovei
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  2. Alma Balestrazzi
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  4. Daniela Carbonera
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Correspondence to Daniela Carbonera.

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Macovei, A., Balestrazzi, A., Confalonieri, M. et al. The tyrosyl-DNA phosphodiesterase gene family in Medicago truncatula Gaertn.: bioinformatic investigation and expression profiles in response to copper- and PEG-mediated stress. Planta 232, 393–407 (2010). https://doi.org/10.1007/s00425-010-1179-9

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