Fluorescently Tagged Potato virus Y: A Versatile Tool for Functional Analysis of Plant-Virus Interactions

Abstract

Potato virus Y (PVY) is an economically important plant virus that infects Solanaceous crops such as tobacco and potato. To date, studies into the localization and movement of PVY in plants have been limited to detection of viral RNA or proteins ex vivo. Here, a PVY N605 isolate was tagged with green fluorescent protein (GFP), characterized and used for in vivo tracking. In Nicotiana tabacum cv. Xanthi, PVY N605-GFP was biologically comparable to nontagged PVY N605, stable through three plant-to-plant passages and persisted for four months in infected plants. GFP was detected before symptoms and fluorescence intensity correlated with PVY RNA concentrations. PVY N605-GFP provided in vivo tracking of long-distance movement, allowing estimation of the cell-to-cell movement rate of PVY in N. tabacum cv. Xanthi (7.1 ± 1.5 cells per hour). PVY N605-GFP was adequately stable in Solanum tuberosum cvs. Désirée and NahG-Désirée and able to infect S. tuberosum cvs. Bintje and Bea, Nicotiana benthamiana, and wild potato relatives. PVY N605-GFP is therefore a powerful tool for future studies of PVY-host interactions, such as functional analysis of viral and plant genes involved in viral movement.

Comparison of transmission efficiency of various isolates of Potato virus Y among three aphid vectors

“…Furthermore, although a very small percentage of plants was infected after transmission using PA, the recipient plant titer tended to be high in those infected plants, which might be a result of individual host reaction (plant cell‐virus interaction). Potato virus Y may be distributed in plants with no obvious pattern (Rupar et al., ), and virus particles may accumulate in a particular part of a plant, part of a leaf, or particular cells within a leaf (Dietrich & Maiss, ; Cervantes & Alvarez, ). Moreover, Moury et al.…”

A Recombinant Potato virus Y Infectious Clone Tagged with the Rosea1 Visual Marker (PVY-Ros1) Facilitates the Analysis of Viral Infectivity and Allows the Production of Large Amounts of Anthocyanins in Plants

“…Nonetheless, infection efficiency of agroinoculated PVY-Ros1 may be improved by the co-expression of tombusvirus p19 silencing suppressor (Saxena et al, 2011). Recently, a PVY clone tagged with GFP has been proposed as a versatile tool for the functional analysis of plant-virus interaction (Rupar et al, 2015). Based on direct visual detection, PVY-Ros1 must be able to complement this tool with new experimental possibilities.…”

Time-Sampled Population Sequencing Reveals the Interplay of Selection and Genetic Drift in Experimental Evolution of Potato Virus Y

“…The absolute number of copies for each dilution point of the standard curve was determined by the QX100 Droplet Digital PCR system (Bio-Rad) according to a protocol previously described (47), using the same reaction mixture composition and conditions as in the respective qPCRs. The samples were analyzed by RT-qPCR using the AgPath-ID one-step RT-PCR kit (Applied Biosystems) on an ABI 7900 HT Fast instrument (Applied Biosystems) according to a previously described procedure (48), with a manual fluorescence threshold of 0.1 for both assays. Two dilutions (10× and 100×) of isolated RNA, using two technical replicates, were analyzed by both assays to control the dynamics of the replication for each sample.…”