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Efficient Double-Stranded RNA Production Methods for Utilization in Plant Virus Control

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Plant Virology Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1236))

Abstract

Double-stranded RNA (dsRNA) is an inducer molecule of the RNA silencing (RNA interference, RNAi) pathway that is present in all higher eukaryotes and controls gene expression at the posttranscriptional level. This mechanism allows the cell to recognize aberrant genetic material in a highly sequence specific manner. This ultimately leads to degradation of the homologous target sequence, rendering the plant cell resistant to subcellular pathogens. Consequently, dsRNA-mediated resistance has been exploited in transgenic plants to convey resistance against viruses. In addition, it has been shown that enzymatically synthesized specific dsRNA molecules can be applied directly onto plant tissue to induce resistance against the cognate virus. This strongly implies that dsRNA molecules are applicable as efficacious agents in crop protection, which will fuel the demand for cost-effective dsRNA production methods. In this chapter, the different methods for dsRNA production—both in vitro and in vivo—are described in detail.

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Acknowledgment

This work was supported by: (a) a grant to FT from the Spanish Ministry of Science and Technology (BIO2009-10172), (b) the Academy of Finland (grants 250113, 256069, and 272507 to M.M.P. and grants 256197, 255342, and 256518 to D.H.B.), and (c) the COST FA0806 to A.E.V., M.C.H., P.L.S., D.H.B., M.M.P.

Author information

Authors and Affiliations

  1. Laboratory of Plant Breeding and Biometry, Agricultural University of Athens, 75 Iera Odos, Athens, 11855, Greece

    Andreas E. Voloudakis

  2. Department of Phytopathology, Laboratory of Bacteriology, Benaki Phytopathological Institute, 8 Stefanou Delta street, Kifissia, 14561, Greece

    Maria C. Holeva

  3. Department of Biosciences and Institute of Biotechnology, University of Helsinki, 56, (Viikinkaari 5), Helsinki, 00014, Finland

    L. Peter Sarin & Dennis H. Bamford

  4. Max Planck Institute for Molecular Biomedicine, Von-Esmarch-Straße 54, Münster, 48149, Germany

    L. Peter Sarin

  5. Departamento de Biología Medioambiental, Centro de Investigaciones Biológicas, CSIC, Ramiro de Maeztu 9, Madrid, 28040, Spain

    Marisol Vargas

  6. Department of Biosciences, University of Helsinki, 56, (Viikinkaari 5), Helsinki, 00014, Finland

    Minna M. Poranen

  7. Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Ramiro de Maeztu 9, Madrid, 28040, Spain

    Francisco Tenllado

Authors
  1. Andreas E. Voloudakis
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  2. Maria C. Holeva
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  3. L. Peter Sarin
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  4. Dennis H. Bamford
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  5. Marisol Vargas
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  6. Minna M. Poranen
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  7. Francisco Tenllado
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Corresponding author

Correspondence to Andreas E. Voloudakis .

Editor information

Editors and Affiliations

  1. Hokkaido University, Sapporo, Japan

    Ichiro Uyeda

  2. Hokkaido University, Sapporo, Japan

    Chikara Masuta

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Voloudakis, A.E. et al. (2015). Efficient Double-Stranded RNA Production Methods for Utilization in Plant Virus Control. In: Uyeda, I., Masuta, C. (eds) Plant Virology Protocols. Methods in Molecular Biology, vol 1236. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-1743-3_19

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  • DOI: https://doi.org/10.1007/978-1-4939-1743-3_19

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-1742-6

  • Online ISBN: 978-1-4939-1743-3

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