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Engineering resistance to PVY in different potato cultivars in a marker-free transformation system using a ‘shooter mutant’ A. tumefaciens

  • Genetic Transformation and Hybridization
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Abstract

In this work, Potato virus Y (PVY) resistant potatoes were generated using an environmentally safe construct. For this purpose, a ‘shooter’ mutant Agrobacterium-based transformation system was used. The isopentenyl transferase gene (ipt) present on the Ti plasmid of ‘shooter’ strains enhances shoot regeneration and can be used as a phenotypic selection marker. The introduced marker-free binary vector carried a hairpin construct derived from the coat protein gene of PVY-NTN strain in order to induce gene silencing. Transformation resulted in high regeneration rates (1.4–5.7 shoots per explant). With pre-selection for the ipt + phenotype the transformation frequency was 24–53%, while without selection 12–28% of the shoots were PCR positive. The presence of the transgene was verified by Southern hybridization. In 16 of 31 challenged transformant lines PVY could be detected neither by RT-PCR nor by back inoculation. A 62.5% of these resistant lines proved to be also ipt-free. This transformation system was reproducible in four potato cultivars, suggesting that it could easily be adapted for other species.

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Abbreviations

CP:

Coat protein

gus :

Beta-glucuronidase gene

hpt :

Hygromycin phosphotransferase gene

ipt :

Isopentenyl transferase gene

nptII :

Neomycin phosphotransferase gene

PTGS:

Post-transcriptional gene silencing

PVX:

Potato virus X

PVY:

Potato virus Y

YCP:

The coat protein region of the Potato virus Y genome used in this paper

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Acknowledgements

We are grateful to Dr. Zsófia Bánfalvi and Dr. Brigitta Dudás for critical reading and editing the manuscript, to Mihály Kondrák and Apor Jeney for their help with the Southern analysis. This work was supported by the Hungarian Széchenyi NKFP-4/0023 grant (“Potato competitiveness”) and the Hungarian OTKA Science School (TS 447778). L. Palkovics was the recipient of a Bolyai scholarship (Hungarian Academy of Sciences).

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Authors and Affiliations

  1. Agricultural Biotechnology Center, Szent-Györgyi A. u. 4., 2100, Gödöllő, Hungary

    Ágnes Bukovinszki & Márta Csányi

  2. Agricultural Research Institute of the Hungarian Academy of Sciences, Brunszvik u. 2, 2462, Martonvásár, Hungary

    Zoltán Divéki & Ervin Balázs

  3. Department of Plant Pathology, Corvinus University of Budapest, Villányi út 29-43, 1118, Budapest, Hungary

    László Palkovics

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  1. Ágnes Bukovinszki
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  2. Zoltán Divéki
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  3. Márta Csányi
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  4. László Palkovics
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  5. Ervin Balázs
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Correspondence to Ágnes Bukovinszki.

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Communicated by D. Dudits

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Bukovinszki, Á., Divéki, Z., Csányi, M. et al. Engineering resistance to PVY in different potato cultivars in a marker-free transformation system using a ‘shooter mutant’ A. tumefaciens . Plant Cell Rep 26, 459–465 (2007). https://doi.org/10.1007/s00299-006-0257-8

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  • DOI: https://doi.org/10.1007/s00299-006-0257-8

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