Monitoring Virus Intercellular Movement from Primary Infected Cells to Neighboring Cells in Plants.
Cell-to-cell movement
Coat protein (CP)
Potyvirus
Systemic infection
Turnip mosaic virus (TuMV)
Journal
Methods in molecular biology (Clifton, N.J.)
ISSN: 1940-6029
Titre abrégé: Methods Mol Biol
Pays: United States
ID NLM: 9214969
Informations de publication
Date de publication:
2022
2022
Historique:
entrez:
14
12
2021
pubmed:
15
12
2021
medline:
20
1
2022
Statut:
ppublish
Résumé
Viral cell-to-cell movement from the primary infected cells to neighboring cells is an essential step for viruses to establish systemic infection in plants. The classic experimental design for studying this process involves the application of a reporter protein such as β-glucuronidase (GUS), green fluorescent protein (GFP), or monomeric red fluorescent protein (mRFP or mCherry). However, such experimental settings are unable to unambiguously distinguish primary and secondary infected cells. In recent years, we have developed several double-labeling potyvirus infectious clones. Upon introduction of such vectors into plant leaf tissues, primary infected cells emit dual fluorescence (green and red) whereas secondary infected cells emit only green fluorescence. In this chapter, we provide detailed protocols on (1) construction of a GFP and mCherry-tagged turnip mosaic virus infectious clone, (2) delivery of the recombinant viral clones into plant cells by agroinfiltration, (3) confocal imaging of viral cell-to-cell movement, and (4) analysis of viral systemic infection. Using this dual-color imaging system, we have revealed coat protein (CP) is essential for TuMV cell-to-cell movement. This system provides a valuable and robust tool to study plant virus cell-to-cell movement.
Identifiants
pubmed: 34905191
doi: 10.1007/978-1-0716-1835-6_7
doi:
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
63-73Informations de copyright
© 2022. Springer Science+Business Media, LLC, part of Springer Nature.
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