RT Journal Article SR Electronic(1) A1 Nurkiyanova, Kulpash M. A1 Ryabov, Eugene V. A1 Commandeur, Ulrich A1 Duncan, George H. A1 Canto, Tomas A1 Gray, Stewart M. A1 Mayo, Mike A. A1 Taliansky, Michael E. YR 2000 T1 Tagging Potato leafroll virus with the jellyfish green fluorescent protein gene JF Journal of General Virology, VO 81 IS 3 SP 617 OP 626 DO https://doi.org/10.1099/0022-1317-81-3-617 PB Microbiology Society, SN 1465-2099, AB A full-length cDNA corresponding to the RNA genome of Potato leafroll virus (PLRV) was modified by inserting cDNA that encoded the jellyfish green fluorescent protein (GFP) into the P5 gene near its 3′ end. Nicotiana benthamiana protoplasts electroporated with plasmid DNA containing this cDNA behind the 35S RNA promoter of Cauliflower mosaic virus became infected with the recombinant virus (PLRV-GFP). Up to 5% of transfected protoplasts showed GFP-specific fluorescence. Progeny virus particles were morphologically indistinguishable from those of wild-type PLRV but, unlike PLRV particles, they bound to grids coated with antibodies to GFP. Aphids fed on extracts of these protoplasts transmitted PLRV-GFP to test plants, as shown by specific fluorescence in some vascular tissue and epidermal cells and subsequent systemic infection. In plants agroinfected with PLRV-GFP cDNA in pBIN19, some cells became fluorescent and systemic infections developed. However, after either type of inoculation, fluorescence was mostly restricted to single cells and the only PLRV genome detected in systemically infected tissues lacked some or all of the inserted GFP cDNA, apparently because of naturally occurring deletions. Thus, intact PLRV-GFP was unable to move from cell to cell. Nevertheless, PLRV-GFP has novel potential for exploring the initial stages of PLRV infection., UL https://www.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-81-3-617