1887

Abstract

Infectious full-length clones of Beet necrotic yellow vein virus (BNYVV) and Beet soil-borne mosaic virus (BSBMV), both genus Benyvirus, were used for fluorescent labelling with the objective to study their interaction in coinfection and superinfection experiments. Fluorescent labelling was achieved by replacing a part of the RNA2 encoded coat protein read-through domain with either GFP or mRFP fluorescent marker proteins. This resulted in a translational fusion comprising the coat and the fluorescent protein. The labelled viruses were infectious and moved systemically in Nicotiana benthamiana, producing wild-type-like symptoms. Virus particles could be observed by electron microscopy, demonstrating that the viral read-through domain is dispensable for particle formation. Coinfection experiments revealed a spatial separation of differentially labelled populations of both identical and different Benyvirus species after N. benthamiana agro-inoculation. Identical observations were obtained when Tobacco rattle virus (TRV) was differentially labelled and used for coinfection. In contrast, coinfections of BSBMV with Potato virus X (PVX) or TRV resulted in many co-infected cells lacking spatial separation. Micro-projectile co-bombardment of N. benthamiana leaves revealed that two differently labelled populations of the same virus co-infected only a few cells before starting to separate. In superinfection experiments with N. benthamiana, BSBMV and BNYVV were unable to establish a secondary infection in plants that were previously infected with BNYVV or BSBMV. Taken together, this is the first work to describe the interaction between two economically important Benyviruses using fluorescence-labelled full-length clones.

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2018-07-30
2020-01-29
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References

  1. Gilmer D, Ratti C. Benyvirus. In King AMQ, Adams MJ, Carstens EB, Lefkowitz EJ. (editors) Virus Taxonomy: Classification and Nomenclature of Viruses Ninth Report of the International Committee on Taxonomy of Viruses Elsevier Inc; 2012; pp.1133–1138
    [Google Scholar]
  2. Peltier C, Hleibieh K, Thiel H, Klein E, Bragard C et al. Molecular biology of the Beet necrotic yellow vein virus. Plant viruses 2008;2:14–24
    [Google Scholar]
  3. Heidel GB, Rush CM, Kendall TL, Lommel SA, French RC. Characteristics of Beet soilborne mosaic virus, a furo-like virus infecting sugar beet. Plant Disease 1997;81:1070–1076 [CrossRef]
    [Google Scholar]
  4. Lee L, Telford EB, Batten JS, Scholthof KB, Rush CM. Complete nucleotide sequence and genome organization of Beet soilborne mosaic virus, a proposed member of the genus Benyvirus. Arch Virol 2001;146:2443–2453 [CrossRef][PubMed]
    [Google Scholar]
  5. Keskin B. Polymyxa betae n. sp., ein Parasit in den Wurzeln von Beta vulgaris Tournefort, besonders während der Jugendentwicklung der Zuckerrübe. Arch Microbiol 1964;49:348–374
    [Google Scholar]
  6. Tamada T, Kondo H. Biological and genetic diversity of plasmodiophorid-transmitted viruses and their vectors. J Gen Plant Pathol 2013;79:307–320 [CrossRef]
    [Google Scholar]
  7. Workneh F, Villanueva E, Steddom K, Rush CM. Spatial association and distribution of Beet necrotic yellow vein virus and Beet soil-borne mosaic virus in sugar beet fields. Plant Dis 2003;87:707–711 [CrossRef]
    [Google Scholar]
  8. Ratti C, Hleibieh K, Bianchi L, Schirmer A, Autonell CR et al. Beet soil-borne mosaic virus RNA-3 is replicated and encapsidated in the presence of BNYVV RNA-1 and -2 and allows long distance movement in Beta macrocarpa. Virology 2009;385:392–399 [CrossRef][PubMed]
    [Google Scholar]
  9. D'Alonzo M, Delbianco A, Lanzoni C, Autonell CR, Gilmer D et al. Beet soil-borne mosaic virus RNA-4 encodes a 32 kDa protein involved in symptom expression and in virus transmission through Polymyxa betae. Virology 2012;423:187–194 [CrossRef][PubMed]
    [Google Scholar]
  10. Syller J. Facilitative and antagonistic interactions between plant viruses in mixed infections. Mol Plant Pathol 2012;13:204–216 [CrossRef][PubMed]
    [Google Scholar]
  11. Syller J, Grupa A. Antagonistic within-host interactions between plant viruses: molecular basis and impact on viral and host fitness. Mol Plant Pathol 2016;17:769–782 [CrossRef][PubMed]
    [Google Scholar]
  12. Dietrich C, Maiss E. Fluorescent labelling reveals spatial separation of potyvirus populations in mixed infected Nicotiana benthamiana plants. J Gen Virol 2003;84:2871–2876 [CrossRef][PubMed]
    [Google Scholar]
  13. Tatineni S, French R. The coat protein and NIa protease of two Potyviridae family members independently confer super-infection exclusion. J Virol 2016;90:10886–10905 [CrossRef][PubMed]
    [Google Scholar]
  14. Gutiérrez S, Pirolles E, Yvon M, Baecker V, Michalakis Y et al. The multiplicity of cellular infection changes depending on the route of cell infection in a plant virus. J Virol 2015;89:9665–9675 [CrossRef][PubMed]
    [Google Scholar]
  15. Julve JM, Gandía A, Fernández-del-Carmen A, Sarrion-Perdigones A, Castelijns B et al. A coat-independent superinfection exclusion rapidly imposed in Nicotiana benthamiana cells by tobacco mosaic virus is not prevented by depletion of the movement protein. Plant Mol Biol 2013;81:553–564 [CrossRef][PubMed]
    [Google Scholar]
  16. Takahashi T, Sugawara T, Yamatsuta T, Isogai M, Natsuaki T et al. Analysis of the spatial distribution of identical and two distinct virus populations differently labeled with cyan and yellow fluorescent proteins in coinfected plants. Phytopathology 2007;97:1200–1206 [CrossRef][PubMed]
    [Google Scholar]
  17. González-Jara P, Tenllado F, Martínez-García B, Atencio FA, Barajas D et al. Host-dependent differences during synergistic infection by Potyviruses with potato virus X. Mol Plant Pathol 2004;5:29–35 [CrossRef][PubMed]
    [Google Scholar]
  18. Vance VB. Replication of potato virus X RNA is altered in coinfections with potato virus Y. Virology 1991;182:486–494[PubMed]
    [Google Scholar]
  19. Wisler GC, Lewellen RT, Sears JL, Wasson JW, Liu H-Y et al. Interactions between Beet necrotic yellow vein virus and Beet soil-borne mosaic virus in sugar beet. Plant Dis 2003;87:1170–1175 [CrossRef]
    [Google Scholar]
  20. Folimonova SY. Superinfection exclusion is an active virus-controlled function that requires a specific viral protein. J Virol 2012;86:5554–5561 [CrossRef][PubMed]
    [Google Scholar]
  21. González-Jara P, Fraile A, Canto T, García-Arenal F. The multiplicity of infection of a plant virus varies during colonization of its eukaryotic host. J Virol 2009;83:7487–7494 [CrossRef][PubMed]
    [Google Scholar]
  22. Tamada T, Kusume T. Evidence that the 75K readthrough protein of beet necrotic yellow vein virus RNA-2 is essential for transmission by the fungus Polymyxa betae. J Gen Virol 1991;72:1497–1504 [CrossRef][PubMed]
    [Google Scholar]
  23. Gilmer D, Bouzoubaa S, Hehn A, Guilley H, Richards K et al. Efficient cell-to-cell movement of beet necrotic yellow vein virus requires 3' proximal genes located on RNA 2. Virology 1992;189:40–47 [CrossRef][PubMed]
    [Google Scholar]
  24. Chiba S, Hleibieh K, Delbianco A, Klein E, Ratti C et al. The benyvirus RNA silencing suppressor is essential for long-distance movement, requires both zinc-finger and NoLS basic residues but not a nucleolar localization for its silencing-suppression activity. Mol Plant Microbe Interact 2013;26:168–181 [CrossRef][PubMed]
    [Google Scholar]
  25. Lauber E, Guilley H, Tamada T, Richards KE, Jonard G. Vascular movement of beet necrotic yellow vein virus in Beta macrocarpa is probably dependent on an RNA 3 sequence domain rather than a gene product. J Gen Virol 1998;79:385–393 [CrossRef][PubMed]
    [Google Scholar]
  26. Peltier C, Klein E, Hleibieh K, D'Alonzo M, Hammann P et al. Beet necrotic yellow vein virus subgenomic RNA3 is a cleavage product leading to stable non-coding RNA required for long-distance movement. J Gen Virol 2012;93:1093–1102 [CrossRef][PubMed]
    [Google Scholar]
  27. Chiba S, Miyanishi M, Andika IB, Kondo H, Tamada T. Identification of amino acids of the beet necrotic yellow vein virus p25 protein required for induction of the resistance response in leaves of Beta vulgaris plants. J Gen Virol 2008;89:1314–1323 [CrossRef][PubMed]
    [Google Scholar]
  28. Koenig R, Jarausch W, Li Y, Commandeur U, Burgermeister W et al. Effect of recombinant beet necrotic yellow vein virus with different RNA compositions on mechanically inoculated sugarbeets leaves of Beta vulgaris plants. J Gen Virol 1991;89:1314–1323
    [Google Scholar]
  29. Tamada T, Abe H. Evidence that beet necrotic yellow vein virus RNA-4 is essential for efficient transmission by the fungus Polymyxa betae. J Gen Virol 1989;70:3391–3398 [CrossRef]
    [Google Scholar]
  30. Schmidlin L, Link D, Mutterer J, Guilley H, Gilmer D. Use of a Beet necrotic yellow vein virus RNA-5-derived replicon as a new tool for gene expression. J Gen Virol 2005;86:463–467 [CrossRef][PubMed]
    [Google Scholar]
  31. Erhardt M, Dunoyer P, Guilley H, Richards K, Jonard G et al. Beet necrotic yellow vein virus particles localize to mitochondria during infection. Virology 2001;286:256–262 [CrossRef][PubMed]
    [Google Scholar]
  32. Schmitt C, Balmori E, Jonard G, Richards KE, Guilley H. In vitro mutagenesis of biologically active transcripts of beet necrotic yellow vein virus RNA 2: evidence that a domain of the 75-kDa readthrough protein is important for efficient virus assembly. Proc Natl Acad Sci USA 1992;89:5715–5719 [CrossRef][PubMed]
    [Google Scholar]
  33. von Stetten D, Noirclerc-Savoye M, Goedhart J, Gadella TW, Royant A. Structure of a fluorescent protein from Aequorea victoria bearing the obligate-monomer mutation A206K. Acta Crystallogr Sect F Struct Biol Cryst Commun 2012;68:878–882 [CrossRef][PubMed]
    [Google Scholar]
  34. Valentin C, Dunoyer P, Vetter G, Schalk C, Dietrich A et al. Molecular basis for mitochondrial localization of viral particles during beet necrotic yellow vein virus infection. J Virol 2005;79:9991–10002 [CrossRef][PubMed]
    [Google Scholar]
  35. Ratcliff F, Harrison BD, Baulcombe DC. A similarity between viral defense and gene silencing in plants. Science 1997;276:1558–1560 [CrossRef][PubMed]
    [Google Scholar]
  36. Bergua M, Zwart MP, El-Mohtar C, Shilts T, Elena SF et al. A viral protein mediates superinfection exclusion at the whole-organism level but is not required for exclusion at the cellular level. J Virol 2014;88:11327–11338 [CrossRef][PubMed]
    [Google Scholar]
  37. Zhang XF, Sun R, Guo Q, Zhang S, Meulia T et al. A self-perpetuating repressive state of a viral replication protein blocks superinfection by the same virus. PLoS Pathog 2017;13:e1006253 [CrossRef][PubMed]
    [Google Scholar]
  38. Zhang XF, Zhang S, Guo Q, Sun R, Wei T et al. A new mechanistic model for viral cross protection and superinfection exclusion. Front Plant Sci 2018;9:40 [CrossRef][PubMed]
    [Google Scholar]
  39. Laufer M, Mohammad H, Maiss E, Richert-Pöggeler K, dall'ara M et al. Biological properties of Beet soil-borne mosaic virus and Beet necrotic yellow vein virus cDNA clones produced by isothermal in vitro recombination: Insights for reassortant appearance. Virology 2018;518:25–33 [CrossRef][PubMed]
    [Google Scholar]
  40. Campbell RE, Tour O, Palmer AE, Steinbach PA, Baird GS et al. A monomeric red fluorescent protein. Proc Natl Acad Sci USA 2002;99:7877–7882 [CrossRef][PubMed]
    [Google Scholar]
  41. Davis SJ, Vierstra RD. Soluble, highly fluorescent variants of green fluorescent protein (GFP) for use in higher plants. Plant Mol Biol 1998;36:521–528 [CrossRef][PubMed]
    [Google Scholar]
  42. Gibson DG, Young L, Chuang RY, Venter JC, Hutchison CA et al. Enzymatic assembly of DNA molecules up to several hundred kilobases. Nat Methods 2009;6:343–345 [CrossRef][PubMed]
    [Google Scholar]
  43. Inoue H, Nojima H, Okayama H. High efficiency transformation of Escherichia coli with plasmids. Gene 1990;96:23–28 [CrossRef][PubMed]
    [Google Scholar]
  44. Voinnet O, Rivas S, Mestre P, Baulcombe D. An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 2003;33:949–956 [CrossRef][PubMed]
    [Google Scholar]
  45. Draghici HK, Varrelmann M. Evidence that the linker between the methyltransferase and helicase domains of potato virus X replicase is involved in homologous RNA recombination. J Virol 2009;83:7761–7769 [CrossRef][PubMed]
    [Google Scholar]
  46. Dietrich C, Maiss E. Red fluorescent protein DsRed from Discosoma sp. as a reporter protein in higher plants. Biotechniques 2002;32:286–291[PubMed]
    [Google Scholar]
  47. Liu Y, Schiff M, Marathe R, Dinesh-Kumar SP. Tobacco Rar1, EDS1 and NPR1/NIM1 like genes are required for N-mediated resistance to tobacco mosaic virus. Plant J 2002;30:415–429 [CrossRef][PubMed]
    [Google Scholar]
  48. Ghazala W, Varrelmann M. Tobacco rattle virus 29K movement protein is the elicitor of extreme and hypersensitive-like resistance in two cultivars of Solanum tuberosum. Mol Plant Microbe Interact 2007;20:1396–1405 [CrossRef][PubMed]
    [Google Scholar]
  49. Gray DJ, Hiebert E, Lin CM, Compton ME, McColley DW et al. Simplified construction and performance of a device for particle bombardment. Plant Cell Tissue Organ Cult 1994;37:179–184 [CrossRef]
    [Google Scholar]
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