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Somatic embryogenesis efficiently eliminates viroid infections from grapevines

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Abstract

Indirect somatic embryogenesis is effective at eliminating the most important viruses affecting grapevines. Accordingly, this technique was tested as a method for eradicating two widespread viroids, Grapevine yellow speckle viroid 1 (GYSVd-1) and Hop stunt viroid (HSVd), from four grapevine cultivars. Both viroids were detected by RT-PCR in grapevine floral explants used for initiating embryogenic cultures, as well as in undifferentiated cells of embryogenic and non-embryogenic calli from anthers and ovaries. In contrast, somatic embryos differentiated from these infected calli were viroid-free, and viroids were not detected in embryo-derived plantlets even 3 years after their transfer to greenhouse conditions. A wider spatial distribution of HSVd than GYSVd-1 within proliferating calli was revealed by in situ hybridization, whereas no hybridization signal was detected in the somatic embryos. In addition, GYSVd-1 and HSVd were localised in the nucleus of infected cells, conclusively showing the nuclear accumulation of representative members of Apscaviroid and Hostuviroid genera, which has been only an assumption so far. Somatic embryogenesis was compared to in vitro thermotherapy, a technique routinely used for virus eradication. After thermotherapy, HSVd and GYSVd-1 were detected in all in vitro plantlets of the cultivar Roussan, and in all lines analysed after 3 years of culture in greenhouse. The high efficiency with which somatic embryogenesis may eliminate viroids and viruses from several infected grapevine cultivars, should allow the availability of virus- and viroid-free material, which would be useful not only for sanitary selection but also for basic research on plant-virus and plant-viroid interactions in grapevine.

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Acknowledgements

Our thanks go to A. Schneider and F. Mannini for providing the mother plants. This research was partly funded by Regione Piemonte (Tech4Wine Project) and by the Dipartimento Agroalimentare of the CNR of Italy (A. Leone and D. Mariotti 2008 Award for Advanced Research in Agriculture to the last author).

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Authors and Affiliations

  1. Istituto di Virologia Vegetale, UOS Grugliasco, Consiglio Nazionale delle Ricerche, Via L. da Vinci 44, 10095, Grugliasco, TO, Italy

    Giorgio Gambino, Rosalina Vallania & Ivana Gribaudo

  2. Istituto di Virologia Vegetale, UOS Bari, Consiglio Nazionale delle Ricerche, Via Amendola 165/A, 70126, Bari, Italy

    Beatriz Navarro & Francesco Di Serio

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  1. Giorgio Gambino
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  2. Beatriz Navarro
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  3. Rosalina Vallania
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  4. Ivana Gribaudo
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  5. Francesco Di Serio
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Correspondence to Ivana Gribaudo or Francesco Di Serio.

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Gambino, G., Navarro, B., Vallania, R. et al. Somatic embryogenesis efficiently eliminates viroid infections from grapevines. Eur J Plant Pathol 130, 511–519 (2011). https://doi.org/10.1007/s10658-011-9770-x

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