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Elimination of Grapevine rupestris stem pitting-associated virus (GRSPaV) from two Vitis vinifera cultivars by in vitro chemotherapy

Abstract

The grapevine (Vitis vinifera L.) cultivars ‘Agiorgitiko’ and ‘Malagouzia’, naturally infected with Grapevine rupestris stem pitting-associated virus (GRSPaV), were subjected to in vitro chemotherapy using the antiviral inosine 5′-monophosphate dehydrogenase inhibitors tiazofurin (TR), ribavirin (RBV) and mycophenolic acid (MPA). The chemotherapy lasted 80 days and was carried out as two consecutive treatments. Severe phytotoxicity, estimated after 40 days of culture, was observed in drug-treated explants, especially when high doses of TR were used. Phytotoxicity exhibited a cultivar- and chemical compound-dependent profile. The virus eradication status of the survived plantlets was determined by nested RT-PCR using total RNA templates, after 80 days of drug treatment and one year later, after the passage of one dormancy period, in potted plants grown in a greenhouse. Data indicated that the highest GRSPaV elimination in ‘Agiorgitiko’ was obtained with 10 μg  ml−1 TR, 30 μg  ml−1 RBV and 20 μg  ml−1 MPA. The eradication rates were lower in the case of ‘Malagouzia’, where the highest ones were achieved after treatments with 15 μg ml−1 TR and 80 μg ml−1 MPA. This is the first report on GRSPaV elimination in grapevine following treatment with antiviral compounds, which could provide an alternative to the traditional methods of virus eradication through meristem culture and thermotherapy.

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Fig. 1

Abbreviations

BA:

benzyladenine

GLRaV:

grapevine leafroll-associated virus

GRSPaV:

grapevine rupestris stem pitting-associated virus

GVA:

grapevine virus A

IMPDH:

inosin 5′-monophosphate dehydrogenase

MPA:

mycophenolic acid

NAA:

naphthaleneacetic acid

RBV:

ribavirin

RSP:

rupestris stem pitting

TR:

tiazofurin

WPM:

woody plant medium

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Acknowledgments

This study is part of a PhD thesis (F. G. Skiada) and was financed by a research project for the reinforcement of new scientists [PENED 2003, funded by 75 % of the public cost from the European Union, and 25 % from the Greek Government (Ministry of Development - General Secretariat of Research and Technology, GSRT), (Measure 8.3, E.P.A.N. - Third European Community Framework)], while the infrastructure and basic chemicals were provided by Vitro Hellas S.A. We also thank Prof. H.N. Jayaram for kindly providing the tiazofurin, and the PhD student A. Katsiani for her help in some experiments.

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Correspondence to E. P. Eleftheriou.

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Skiada, F.G., Maliogka, V.I., Katis, N.I. et al. Elimination of Grapevine rupestris stem pitting-associated virus (GRSPaV) from two Vitis vinifera cultivars by in vitro chemotherapy. Eur J Plant Pathol 135, 407–414 (2013). https://doi.org/10.1007/s10658-012-0097-z

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Keywords

  • Mycophenolic acid
  • Nested RT-PCR
  • Phytotoxicity
  • Ribavirin
  • Tiazofurin