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

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Plant Virology Protocols

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.

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Correspondence to Andreas E. Voloudakis .

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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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