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Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs

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Viroids

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2316))

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Abstract

Viroids are considered the most minimalistic group of pathogens. Despite their presumed inability to encode for proteins, viroids induce several diseases in plants of primary economic importance. The production of viroid derived siRNAs (vd-siRNAs) of 21–24 nt, accompanies viroid infections in plants and results from the activation of the RNA silencing mechanism and specifically the function of Dicer endonucleases. A comprehensive set of experiments for the study and thorough analysis of viroid-infected plants has been developed. Here we present a detailed experimental plan including optimized protocols for plant infection by agroinfiltration, RNA extraction, and northern blot hybridization for the detection of both viroid genomic RNA and vd-siRNAs.

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Acknowledgments

The authors would like to acknowledge support of this work by project “PlantUP” [MIS MIS5002803] which is implemented under the Action “Reinforcement of the Research and Innovation Infrastructure,” funded by the Operational Programme “Competitiveness, Entrepreneurship and Innovation” (NSRF 2014-2020) and co-financed by Greece and the European Union (European Regional Development Fund). This work was also supported by the grant “Emblematic Action for Research in the Cretan Agrofood sector: Four Institutions, Four References” (2018ΣΕ01300000) held by the General Secretary for Research and Technology of Greece.

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Authors and Affiliations

  1. Biology Department, University of Crete, Heraklion, Greece

    Konstantina Katsarou, Nikoleta Kryovrysanaki & Kriton Kalantidis

  2. IMBB/FoRTH, Heraklion, Greece

    Konstantina Katsarou, Nikoleta Kryovrysanaki & Kriton Kalantidis

Authors
  1. Konstantina Katsarou
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  2. Nikoleta Kryovrysanaki
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  3. Kriton Kalantidis
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Corresponding author

Correspondence to Kriton Kalantidis .

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Editors and Affiliations

  1. Department of Microbiology and Plant Pathology, University of California, Riverside, Riverside, CA, USA

    Ayala L. N. Rao

  2. Department of Microbiology and Plant Pathology, University of California, Riverside, Riverside, CA, USA

    Irene Lavagi-Craddock

  3. Department of Microbiology and Plant Pathology, University of California, Riverside, Riverside, CA, USA

    Georgios Vidalakis

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Katsarou, K., Kryovrysanaki, N., Kalantidis, K. (2022). Detection of Viroid RNA and vd-siRNA in N. benthamiana Plants: Northern Blot Analyses for Viroid and vd-siRNAs. In: Rao, A.L.N., Lavagi-Craddock, I., Vidalakis, G. (eds) Viroids. Methods in Molecular Biology, vol 2316. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1464-8_24

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  • DOI: https://doi.org/10.1007/978-1-0716-1464-8_24

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-1463-1

  • Online ISBN: 978-1-0716-1464-8

  • eBook Packages: Springer Protocols

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