Genotoxic effects due to in vitro culture and H2O2 treatments in Petunia x hybrida cells monitored through DNA diffusion assay, FPG-SCGE and gene expression profile analyses

• Autorzy: Ventura L. , Macovei A. , Dona M. , Paparella S. , Buttafava A. , Giovannini A. , Carbonera D. , Balestrazzi A.
• Języki publikacji: EN

Abstrakty

EN

In the present work, Petunia × hybrida leaf discs maintained on regeneration medium for 8 days were used to assess the effects of genotoxic stress induced by in vitro culture. The investigation was carried out by comparing the response of intact leaves excised from Petunia × hybrida plantlets grown in vitro and the regenerating leaf discs. In situ detection by histochemical staining and alkaline-Single Cell Gel Electrophoresis (SCGE) analysis demonstrated that both reactive oxygen species accumulation and DNA damage were enhanced in explants cultured in vitro. Significant up-regulation of the PhOGG1 (8-oxoguanine DNA glycosylase/lyase), PhAPX (ascorbate peroxidase) and PhMT2 (metallothionein) genes involved in DNA repair and antioxidant defence was observed in the explants cultured in vitro, compared to intact leaves. The Petunia × hybrida leaf discs were exposed to increasing (0, 100, 150, 200 and 400 mM) doses of the model genotoxic agent hydrogen peroxide (H₂O₂) and then analysed. The DNA diffusion assay highlighted the dose- and time-dependent fluctuations of programmed cell death/necrosis events in response to H₂O₂. Leaf discs treated with increasing H₂O₂ concentration and untreated controls were analysed by FPG-SCGE to assess the level of oxidative DNA damage at different time points following treatments. The PhOGG1, PhAPX and PhMT2 genes were significantly up-regulated in response to H₂O₂, reaching the highest transcript levels with the 150 mM dose. Based on the reported data, these genes might be used as molecular indicators of the genotoxic stress response in Petunia × hybrida cells.

• Wydawca: -
• Czasopismo: Acta Physiologiae Plantarum
• Rocznik: 2014
• Tom: 36
• Numer: 02
• Opis fizyczny: p.331-341,fig.,ref.

Twórcy

autor

Ventura L.

• Dipartimento di Chimica, Universita di Pavia, Via Taramelli 12, 27100 Pavia, Italy

autor

Macovei A.

• Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Laboratori di Genetica e Microbiologia 'A. Buzzati-Traverso', Universita di Pavia, Via Ferrata1, 27100 Pavia, Italy

autor

Dona M.

• Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Laboratori di Genetica e Microbiologia 'A. Buzzati-Traverso', Universita di Pavia, Via Ferrata1, 27100 Pavia, Italy

autor

Paparella S.

• Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Laboratori di Genetica e Microbiologia 'A. Buzzati-Traverso', Universita di Pavia, Via Ferrata1, 27100 Pavia, Italy

autor

Buttafava A.

• Dipartimento di Chimica, Universita di Pavia, Via Taramelli 12, 27100 Pavia, Italy

autor

Giovannini A.

• Consiglio per la Ricerca e la Sperimentazione in Agricoltura, Unita di Ricerca per la Floricoltura e le Specie Ornamentali (CRA-FSO), Corso Inglesi 508, 18038 Sanremo, IM, Italy

autor

Carbonera D.

• Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Laboratori di Genetica e Microbiologia 'A. Buzzati-Traverso', Universita di Pavia, Via Ferrata1, 27100 Pavia, Italy

autor

Balestrazzi A.

• Dipartimento di Biologia e Biotecnologie 'Lazzaro Spallanzani', Laboratori di Genetica e Microbiologia 'A. Buzzati-Traverso', Universita di Pavia, Via Ferrata1, 27100 Pavia, Italy

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Identyfikatory

DOI

10.1007/s11738-013-1415-6