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Monitoring the Intracellular Trafficking of Virus-Induced Structures and Intercellular Spread of Viral Infection in Plants Using Endomembrane Trafficking Pathway-Specific Chemical Inhibitor and Organelle-Selective Fluorescence Dye

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Plant-Virus Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2724))

Abstract

Infection by positive-strand RNA viruses induces extensive remodeling of the host endomembrane system in favor of viral replication and movement. The integral membrane protein 6K2 of potyviruses induces the formation of membranous virus replication vesicles at the endoplasmic reticulum exit site (ERES). The intracellular trafficking of 6K2-induced vesicles along with microfilaments requires the vesicular transport pathway, actomyosin motility system, and possibly post-Golgi compartments such as endosomes as well. Recent studies have shown that endocytosis is essential for the intracellular movement of potyviruses from the site of viral genome replication/assembly site to plasmodesmata (PD) to enter neighboring cells. In this chapter, we describe a detailed protocol of how to use endomembrane trafficking pathway-specific chemical inhibitors and organelle-selective fluorescence dye to study the trafficking of potyviral proteins and potyvirus-induced vesicles and to unravel the role of endocytosis and the endocytic pathway in potyvirus infection in Nicotiana benthamiana plants.

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Acknowledgments

The protocol described here is based on the work supported by grants from Agriculture and Agri-Food Canada and the Natural Sciences and Engineering Research Council of Canada to A.W.

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Authors and Affiliations

  1. London Research and Development Centre, Agriculture and Agri-Food Canada, London, ON, Canada

    Yinzi Li & Aiming Wang

Authors
  1. Yinzi Li
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  2. Aiming Wang
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Corresponding author

Correspondence to Aiming Wang .

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Editors and Affiliations

  1. Departamento de Bioquimica e Biologia Molecular, Universidade Federal de Viçosa, Viçosa, Minas Gerais, Brazil

    Elizabeth P.B. Fontes

  2. Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland

    Kristiina Mäkinen

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© 2024 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Li, Y., Wang, A. (2024). Monitoring the Intracellular Trafficking of Virus-Induced Structures and Intercellular Spread of Viral Infection in Plants Using Endomembrane Trafficking Pathway-Specific Chemical Inhibitor and Organelle-Selective Fluorescence Dye. In: Fontes, E.P., Mäkinen, K. (eds) Plant-Virus Interactions. Methods in Molecular Biology, vol 2724. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3485-1_10

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  • DOI: https://doi.org/10.1007/978-1-0716-3485-1_10

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-3484-4

  • Online ISBN: 978-1-0716-3485-1

  • eBook Packages: Springer Protocols

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