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

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. Desiree and NahG-Desiree and able to infect S. tuberosum cvs. Bintje and Bea, Nicotiana benthamiana, and wild pot...

[1]  S. Elena,et al.  Within-Host Spatiotemporal Dynamics of Plant Virus Infection at the Cellular Level , 2014, PLoS genetics.

[2]  J. Laliberté,et al.  6K2-induced vesicles can move cell to cell during turnip mosaic virus infection , 2013, Front. Microbiol..

[3]  S. Chapman,et al.  Replication and trafficking of a plant virus are coupled at the entrances of plasmodesmata , 2013, The Journal of cell biology.

[4]  Johannes E. Schindelin,et al.  Fiji: an open-source platform for biological-image analysis , 2012, Nature Methods.

[5]  G. Grondin,et al.  Impact on the Endoplasmic Reticulum and Golgi Apparatus of Turnip Mosaic Virus Infection , 2012, Journal of Virology.

[6]  H. Scholthof,et al.  Host impact on the stability of a plant virus gene vector as measured by a new fluorescent local lesion passaging assay. , 2012, Journal of virological methods.

[7]  J. Daròs,et al.  Visual Tracking of Plant Virus Infection and Movement Using a Reporter MYB Transcription Factor That Activates Anthocyanin Biosynthesis1[W] , 2012, Plant Physiology.

[8]  Thierry Candresse,et al.  Top 10 plant viruses in molecular plant pathology. , 2011, Molecular plant pathology.

[9]  A. Kladnik,et al.  Distribution of Potato virus Y in potato plant organs, tissues, and cells. , 2011, Phytopathology.

[10]  T. Natsuaki,et al.  An infectious full-length cDNA clone of potato virus YNTN-NW, a recently reported strain of PVY that causes potato tuber necrotic ringspot disease , 2011, Archives of Virology.

[11]  J. Lorenzen,et al.  Potato virus Y: An Evolving Concern for Potato Crops in the United States and Canada. , 2010, Plant disease.

[12]  P. Forterre Defining Life: The Virus Viewpoint , 2010, Origins of Life and Evolution of Biospheres.

[13]  M. Ghanim,et al.  A simple, rapid and inexpensive method for localization of Tomato yellow leaf curl virus and Potato leafroll virus in plant and insect vectors. , 2009, Journal of virological methods.

[14]  V. Beneš,et al.  The MIQE guidelines: minimum information for publication of quantitative real-time PCR experiments. , 2009, Clinical chemistry.

[15]  E. Kok,et al.  PVY(NTN) elicits a diverse gene expression response in different potato genotypes in the first 12 h after inoculation. , 2009, Molecular plant pathology.

[16]  K. Oparka,et al.  Live-cell imaging of viral RNA genomes using a Pumilio-based reporter. , 2009, The Plant journal : for cell and molecular biology.

[17]  K. Verbanac,et al.  Real-time PCR to determine transgene copy number and to quantitate the biolocalization of adoptively transferred cells from EGFP-transgenic mice. , 2008, BioTechniques.

[18]  J. Valkonen,et al.  Three heterologous proteins simultaneously expressed from a chimeric potyvirus: infectivity, stability and the correlation of genome and virion lengths. , 2008, Virus research.

[19]  K. Gruden,et al.  Single-step RT real-time PCR for sensitive detection and discrimination of Potato virus Y isolates. , 2008, Journal of virological methods.

[20]  Shih-Shun Lin,et al.  Effectiveness and stability of heterologous proteins expressed in plants by Turnip mosaic virus vector at five different insertion sites. , 2007, Virus research.

[21]  E. Balázs,et al.  The role of the coat protein region in symptom formation on Physalis floridana varies between PVY strains. , 2007, Virus research.

[22]  M. Pezzotti,et al.  Stability of Potato virus X expression vectors is related to insert size: implications for replication models and risk assessment , 2007, Transgenic Research.

[23]  J. Randles,et al.  Single-stranded DNA of Tomato leaf curl virus accumulates in the cytoplasm of phloem cells. , 2006, Virology.

[24]  W. J. Lucas,et al.  Plant viral movement proteins: agents for cell-to-cell trafficking of viral genomes. , 2006, Virology.

[25]  H. Förster,et al.  Studies on the etiology of tuber necrotic ringspot disease in potato , 1984, Potato Research.

[26]  E. Holmes,et al.  A phylogenetic survey of recombination frequency in plant RNA viruses , 2006, Archives of Virology.

[27]  J. Valkonen,et al.  A novel insertion site inside the potyvirus P1 cistron allows expression of heterologous proteins and suggests some P1 functions. , 2005, Virology.

[28]  C. Kerlan,et al.  Characterization of Potato virus Y (PVY) molecular determinants involved in the vein necrosis symptom induced by PVYN isolates in infected Nicotiana tabacum cv. Xanthi. , 2005, The Journal of general virology.

[29]  D. Scheel,et al.  The oligopeptide elicitor Pep-13 induces salicylic acid-dependent and -independent defense reactions in potato , 2004 .

[30]  Š. Baebler,et al.  Spread of potato virus YNTN in potato cultivars (Solanum tuberosum L.) with different levels of sensitivity , 2004 .

[31]  A. Palloix,et al.  Mutations in potato virus Y genome-linked protein determine virulence toward recessive resistances in Capsicum annuum and Lycopersicon hirsutum. , 2004, Molecular plant-microbe interactions : MPMI.

[32]  J. Valkonen,et al.  Localization of a potyvirus and the viral genome-linked protein in wild potato leaves at an early stage of systemic infection. , 2003, Molecular plant-microbe interactions : MPMI.

[33]  N. V. Rusetsky,et al.  Distribution and accumulation of PVM and PVY, PVX in infected potato plants , 2001 .

[34]  P. Zambryski,et al.  Cell-to-cell and systemic movement of recombinant green fluorescent protein-tagged turnip crinkle viruses. , 2000, Virology.

[35]  N. Boonham,et al.  Detection of Ralstonia solanacearumStrains with a Quantitative, Multiplex, Real-Time, Fluorogenic PCR (TaqMan) Assay , 2000, Applied and Environmental Microbiology.

[36]  L. Simón-Buela,et al.  Virus particles of cucumber green mottle mosaic tobamovirus move systemically in the phloem of infected cucumber plants. , 1999, Molecular plant-microbe interactions : MPMI.

[37]  C. D'arcy,et al.  In Situ Localization of Barley Yellow Dwarf Virus-PAV 17-kDa Protein and Nucleic Acids in Oats. , 1998, Phytopathology.

[38]  K. Oparka,et al.  Studying the movement of plant viruses using green fluorescent protein , 1996 .

[39]  I. E. Johansen Intron insertion facilitates amplification of cloned virus cDNA in Escherichia coli while biological activity is reestablished after transcription in vivo. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[40]  A. Roberts,et al.  Assembly and movement of a plant virus carrying a green fluorescent protein overcoat. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[41]  C. Robaglia,et al.  Cell-free cloning and biolistic inoculation of an infectious cDNA of potato virus Y. , 1996, The Journal of general virology.

[42]  V. Pallás,et al.  Long-distance movement of cherry leaf roll virus in infected tobacco plants. , 1996, The Journal of general virology.

[43]  D. Baulcombe,et al.  Jellyfish green fluorescent protein as a reporter for virus infections. , 1995, The Plant journal : for cell and molecular biology.

[44]  S. Howell,et al.  Long distance movement of cauliflower mosaic virus in infected turnip plants. , 1992 .