Published: Vol 4, Iss 20, Oct 20, 2014 DOI: 10.21769/BioProtoc.1269 Views: 7912
Reviewed by: Hiromasa SaitohTie LiuArsalan Daudi
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Abstract
Here, we present a simple and rapid protocol to measure the extent of cell-to-cell movement of RNA viruses in planta. To do that, the green fluorescent protein (GFP) gene was incorporated into the genome of Melon necrotic spot virus (MNSV) as a coat protein (CP) fusion protein using the Thosea asigna virus 2A catalytic peptide (TaV 2a) (Serra-Soriano et al., 2014). TaV 2a allows the co-translational cleavage of the fusion protein resulting in the independent expression of both proteins (Kim et al., 2011). Viral infection was initiated by agro-infiltration of Cucumis melo leaves. At 6-7 days post-infiltration, fluorescent infection foci images were taken with a fluorescent stereo microscope and infection areas were measured using FIJI software.
Keywords: Plant virusMaterials and Reagents
Equipment
Software
Procedure
Recipes
Acknowledgments
This work was funded by grant BIO2011-25018 from the Spanish Ministerio de Economia y Competitividad and by Prometeo Program GV2011/003 from the Generalitat Valenciana. J.A.N. and M.S. are the recipients of a postdoctoral contract and a PhD fellowship from the Ministerio de Educacion y Ciencia of Spain.
References
Article Information
Copyright
© 2014 The Authors; exclusive licensee Bio-protocol LLC.
How to cite
Navarro, J. A., Serra-Soriano, M. and Pallás, V. (2014). A Protocol to Measure the Extent of Cell-to-cell Movement of RNA Viruses in Planta. Bio-protocol 4(20): e1269. DOI: 10.21769/BioProtoc.1269.
Category
Plant Science > Plant immunity > Perception and signaling
Plant Science > Plant immunity > Disease bioassay
Microbiology > Microbe-host interactions > In vivo model
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