1887

Abstract

Function of the melon necrotic spot virus (MNSV) genome-encoded proteins (p29, p89, p7A, p7B and p42) has been studied. Protein-expression mutants of an infectious, full-length cDNA clone of a Spanish MNSV-Al isolate and a recombinant green fluorescent protein (GFP)-expressing virus were used in infection bioassays on melon plants. Results revealed that p29 and p89 are both essential for virus replication, whereas small proteins p7A and p7B are sufficient to support viral movement between adjacent cells operating . It is also demonstrated that, in addition to its structural role as coat protein, p42 is an important factor controlling symptoms and is required for systemic transport. Moreover, both p42 and p7B, among all of the MNSV-encoded proteins, were able to delay RNA silencing in transient-expression assays on GFP-transgenic plants. Finally, the presence of p42 also produced an enhancing effect on local spread similar to that of potyviral helper component proteinase (HC-Pro), probably due to its RNA silencing-suppression ability.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.81793-0
2006-08-01
2019-11-17
Loading full text...

Full text loading...

/deliver/fulltext/jgv/87/8/2371.html?itemId=/content/journal/jgv/10.1099/vir.0.81793-0&mimeType=html&fmt=ahah

References

  1. Anandalakshmi, R., Marathe, R., Ge, X., Herr, J. M., Jr, Mau, C., Mallory, A., Pruss, G., Bowman, L. & Vance, V. B. ( 2000; ). A calmodulin-related protein that suppresses posttranscriptional gene silencing in plants. Science 290, 142–144.[CrossRef]
    [Google Scholar]
  2. Astruc, N., Marcos, J. F., Macquaire, G., Candresse, T. & Pallás, V. ( 1996; ). Studies on the diagnosis of hop stunt viroid in fruit trees: identification of new hosts and application of a nucleic acid extraction procedure based on non-organic solvents. Eur J Plant Pathol 102, 837–846.[CrossRef]
    [Google Scholar]
  3. Baulcombe, D. C., Chapman, S. & Santa Cruz, S. ( 1995; ). Jellyfish green fluorescent protein as a reporter for virus infections. Plant J 7, 1045–1053.[CrossRef]
    [Google Scholar]
  4. Brigneti, G., Voinnet, O., Li, W.-X., Ji, L.-H., Ding, S.-W. & Baulcombe, D. C. ( 1998; ). Viral pathogenicity determinants are suppressors of transgene silencing in Nicotiana benthamiana. EMBO J 22, 6739–6746.
    [Google Scholar]
  5. Campbell, R. N. & Sim, S. T. ( 1994; ). Host specificity and nomenclature of Olpidium bornovanus (=Olpidium radicale) and comparisons to Olpidium brassicae. Can J Bot 72, 1136–1143.[CrossRef]
    [Google Scholar]
  6. Campbell, R. N., Wipf-Scheibel, C. & Lecoq, H. ( 1996; ). Vector-assisted seed transmission of melon necrotic spot virus in melon. Phytopathology 86, 1294–1298.[CrossRef]
    [Google Scholar]
  7. Cañizares, M. C., Marcos, J. F. & Pallás, V. ( 2001; ). Molecular variability of twenty-one geographically distinct isolates of carnation mottle virus (CarMV) and phylogenetic relationships within the Tombusviridae family. Arch Virol 146, 2039–2051.[CrossRef]
    [Google Scholar]
  8. Carrington, J. C., Heaton, L. A., Zuidema, D., Hillman, B. I. & Morris, T. J. ( 1989; ). The genome structure of turnip crinkle virus. Virology 170, 219–226.[CrossRef]
    [Google Scholar]
  9. Cohen, Y., Gisel, A. & Zambryski, P. C. ( 2000; ). Cell-to-cell and systemic movement of recombinant green fluorescent protein-tagged turnip crinkle viruses. Virology 273, 258–266.[CrossRef]
    [Google Scholar]
  10. Covelli, L., Coutts, R. H. A., Di Serio, F., Citir, A., Açıkgöz, S., Hernández, C., Ragozzino, A. & Flores, R. ( 2004; ). Cherry chlorotic rusty spot and Amasya cherry diseases are associated with a complex pattern of mycoviral-like double-stranded RNAs. I. Characterization of a new species in the genus Chrysovirus. J Gen Virol 85, 3389–3397.[CrossRef]
    [Google Scholar]
  11. Díaz, J. A., Bernal, J. J., Moriones, E. & Aranda, M. A. ( 2003; ). Nucleotide sequence and infectious transcripts from a full-length cDNA clone of the carmovirus Melon necrotic spot virus. Arch Virol 148, 599–607.[CrossRef]
    [Google Scholar]
  12. Díaz, J. A., Nieto, C., Moriones, E., Truniger, V. & Aranda, M. A. ( 2004; ). Molecular characterization of a Melon necrotic spot virus strain that overcomes the resistance in melon and nonhost plants. Mol Plant Microbe Interact 17, 668–675.[CrossRef]
    [Google Scholar]
  13. Furuki, I. ( 1981; ). Epidemiological Studies on Melon Necrotic Spot (Technical Bulletin 14). Shizuoka Agricultural Experiment Station, Shizuokaken, Japan.
  14. García-Castillo, S., Sánchez-Pina, M. A. & Pallás, V. ( 2003; ). Spatio-temporal analysis of the RNAs, coat and movement (p7) proteins of Carnation mottle virus in Chenopodium quinoa plants. J Gen Virol 84, 745–749.[CrossRef]
    [Google Scholar]
  15. Gosalvez, B., Navarro, J. A., Lorca, A., Botella, F., Sánchez-Pina, M. A. & Pallás, V. ( 2003; ). Detection of melon necrotic spot virus in water samples and melon plants by molecular methods. J Virol Methods 113, 87–93.[CrossRef]
    [Google Scholar]
  16. Hacker, D. L., Petty, I. T. D., Wei, N. & Morris, T. J. ( 1992; ). Turnip crinkle virus genes required for RNA replication and virus movement. Virology 186, 1–8.[CrossRef]
    [Google Scholar]
  17. Hearne, P. Q., Knorr, D. A., Hillman, B. I. & Morris, T. J. ( 1990; ). The complete genome structure and synthesis of infectious RNA from clones of tomato bushy stunt virus. Virology 177, 141–151.[CrossRef]
    [Google Scholar]
  18. Herranz, M. C., Sanchez-Navarro, J.-A., Sauri, A., Mingarro, I. & Pallás, V. ( 2005; ). Mutational analysis of the RNA-binding domain of the Prunus necrotic ringspot virus (PNRSV) movement protein reveals its requirement for cell-to-cell movement. Virology 339, 31–41.[CrossRef]
    [Google Scholar]
  19. Hibi, T. & Furuki, I. ( 1985; ). Melon necrotic spot virus. In CMI/AAB Descriptions of Plants Viruses (no. 302). Wellesbourne, UK: Association of Applied Biologists.
  20. Huang, M., Koh, D. C.-Y., Weng, L.-J., Chang, M.-L., Yap, Y. K., Zhang, L. & Wong, S.-M. ( 2000; ). Complete nucleotide sequence and genome organization of Hibiscus chlorotic ringspot virus, a new member of the genus Carmovirus: evidence for the presence and expression of two novel open reading frames. J Virol 74, 3149–3155.[CrossRef]
    [Google Scholar]
  21. Hull, R. ( 2002; ). Virus movement through the plant and effects on plant metabolism. In Matthews' Plant Virology, 4th edn, pp. 373–436. San Diego: Academic Press.
  22. Kakani, K., Sgro, J.-Y. & Rochon, D. ( 2001; ). Identification of specific cucumber necrosis virus coat protein amino acids affecting fungus transmission and zoospore attachment. J Virol 75, 5576–5583.[CrossRef]
    [Google Scholar]
  23. Kasschau, K. D. & Carrington, J. C. ( 1998; ). A counterdefensive strategy of plant viruses: suppression of posttranscriptional gene silencing. Cell 95, 461–470.[CrossRef]
    [Google Scholar]
  24. Knoester, M., van Loon, L. C., van den Heuvel, J., Hennig, J., Bol, J. F. & Linthorst, H. J. M. ( 1998; ). Ethylene-insensitive tobacco lacks nonhost resistance against soil-borne fungi. Proc Natl Acad Sci U S A 95, 1933–1937.[CrossRef]
    [Google Scholar]
  25. Kubo, C., Nakazono-Nagaoka, E., Hagiwara, K., Kajihara, H., Takeuchi, S., Matsuo, K., Ichiki, T. U. & Omura, T. ( 2005; ). New severe strains of Melon necrotic spot virus: symptomatology and sequencing. Plant Pathol 54, 615–620.[CrossRef]
    [Google Scholar]
  26. Lange, L. & Insunza, V. ( 1977; ). Root-inhabiting Olpidium species: the O. radicale complex. Trans Br Mycol Soc 69, 377–384.[CrossRef]
    [Google Scholar]
  27. Li, W.-Z., Qu, F. & Morris, T. J. ( 1998; ). Cell-to-cell movement of turnip crinkle virus is controlled by two small open reading frames that function in trans. Virology 244, 405–416.[CrossRef]
    [Google Scholar]
  28. Lough, T. J., Emerson, S. J., Lucas, W. J. & Forster, R. L. S. ( 2001; ). Trans-complementation of long-distance movement of White clover mosaic virus triple gene block (TGB) mutants: phloem-associated movement of TGBp1. Virology 288, 18–28.[CrossRef]
    [Google Scholar]
  29. Lu, R., Folimonov, A., Shintaku, M., Li, W.-X., Falk, B. W., Dawson, W. O. & Ding, S.-W. ( 2004; ). Three distinct suppressors of RNA silencing encoded by a 20-kb viral RNA genome. Proc Natl Acad Sci U S A 101, 15742–15747.[CrossRef]
    [Google Scholar]
  30. Marcos, J. F., Vilar, M., Pérez-Payá, E. & Pallás, V. ( 1999; ). In vivo detection, RNA-binding properties and characterization of the RNA-binding domain of the p7 putative movement protein from carnation mottle carmovirus (CarMV). Virology 255, 354–365.[CrossRef]
    [Google Scholar]
  31. Más, P. & Pallás, V. ( 1995; ). Non-isotopic tissue-printing hybridization: a new technique to study long-distance plant virus movement. J Virol Methods 52, 317–326.[CrossRef]
    [Google Scholar]
  32. McLean, M. A., Campbell, R. N., Hamilton, R. I. & Rochon, D. M. ( 1994; ). Involvement of the cucumber necrosis virus coat protein in the specificity of fungus transmission by Olpidium bornovanus. Virology 204, 840–842.[CrossRef]
    [Google Scholar]
  33. Meng, C., Chen, J., Peng, J. & Wong, S.-M. ( 2006; ). Host-induced avirulence of hibiscus chlorotic ringspot virus mutants correlates with reduced gene-silencing suppression activity. J Gen Virol 87, 451–459.[CrossRef]
    [Google Scholar]
  34. Morales, M., Orjeda, G., Nieto, C. & 10 other authors ( 2005; ). A physical map covering the nsv locus that confers resistance to Melon necrotic spot virus in melon (Cucumis melo L.). Theor Appl Genet 111, 914–922.[CrossRef]
    [Google Scholar]
  35. Navarro, J. A., Botella, F., Maruhenda, A., Sastre, P., Sánchez-Pina, M. A. & Pallás, V. ( 2004; ). Comparative infection progress analysis of Lettuce big-vein virus and Mirafiori lettuce virus in lettuce crops by developed molecular diagnosis techniques. Phytopathology 94, 470–477.[CrossRef]
    [Google Scholar]
  36. Ohshima, K., Ando, T., Motomura, N., Matsuo, K. & Sako, N. ( 2000; ). Comparative study on genomes of two Japanese melon necrotic spot virus isolates. Acta Virol 44, 309–314.
    [Google Scholar]
  37. Pallás, V., Sánchez-Navarro, J. A., Más, P., Cañizares, M. C., Aparicio, F. & Marcos, J. F. ( 1998; ). Molecular diagnostic techniques and their potential role in stone fruit certification schemes. Options Méditerr 19, 191–208.
    [Google Scholar]
  38. Panavas, T., Hawkins, C. M., Panavienė, Ž. & Nagy, P. D. ( 2005; ). The role of the p33 : p33/p92 interaction domain in RNA replication and intracellular localization of p33 and p92 proteins of Cucumber necrosis tombusvirus. Virology 338, 81–95.[CrossRef]
    [Google Scholar]
  39. Panavienė, Ž., Baker, J. M. & Nagy, P. D. ( 2003; ). The overlapping RNA-binding domains of p33 and p92 replicase proteins are essential for tombusvirus replication. Virology 308, 191–205.[CrossRef]
    [Google Scholar]
  40. Qu, F. & Morris, T. J. ( 2005; ). Suppressors of RNA silencing encoded by plant viruses and their role in viral infections. FEBS Lett 579, 5958–5964.[CrossRef]
    [Google Scholar]
  41. Qu, F., Ren, T. & Morris, T. J. ( 2003; ). The coat protein of turnip crinkle virus suppresses posttranscriptional gene silencing at an early initiation step. J Virol 77, 511–522.[CrossRef]
    [Google Scholar]
  42. Rajendran, K. S. & Nagy, P. D. ( 2003; ). Characterization of the RNA-binding domains in the replicase proteins of tomato bushy stunt virus. J Virol 77, 9244–9258.[CrossRef]
    [Google Scholar]
  43. Rajendran, K. S. & Nagy, P. D. ( 2004; ). Interaction between the replicase proteins of Tomato bushy stunt virus in vitro and in vivo. Virology 326, 250–261.[CrossRef]
    [Google Scholar]
  44. Rajendran, K. S., Pogany, J. & Nagy, P. D. ( 2002; ). Comparison of Turnip crinkle virus RNA-dependent RNA polymerase preparations expressed in Escherichia coli or derived from infected plants. J Virol 76, 1707–1717.[CrossRef]
    [Google Scholar]
  45. Riviere, C. J. & Rochon, D. M. ( 1990; ). Nucleotide sequence and genomic organization of melon necrotic spot virus. J Gen Virol 71, 1887–1896.[CrossRef]
    [Google Scholar]
  46. Riviere, C. J., Pot, J., Tremaine, J. H. & Rochon, D. M. ( 1989; ). Coat protein of melon necrotic spot carmovirus is more similar to those of tombusviruses than those of carmoviruses. J Gen Virol 70, 3033–3042.[CrossRef]
    [Google Scholar]
  47. Robbins, M. A., Reade, R. D. & Rochon, D. M. ( 1997; ). A cucumber necrosis virus variant deficient in fungal transmissibility contains an altered coat protein shell domain. Virology 234, 138–146.[CrossRef]
    [Google Scholar]
  48. Rochon, D. M. & Tremaine, J. H. ( 1989; ). Complete nucleotide sequence of the cucumber necrosis virus genome. Virology 169, 251–259.[CrossRef]
    [Google Scholar]
  49. Roth, B. M., Pruss, G. J. & Vance, V. B. ( 2004; ). Plant viral suppressors of RNA silencing. Virus Res 102, 97–108.[CrossRef]
    [Google Scholar]
  50. Ruiz, M. T., Voinnet, O. & Baulcombe, D. C. ( 1998; ). Initiation and maintenance of virus-induced gene silencing. Plant Cell 10, 937–946.[CrossRef]
    [Google Scholar]
  51. Ryabov, E. V., van Wezel, R., Walsh, J. & Hong, Y. ( 2004; ). Cell-to-cell, but not long-distance, spread of RNA silencing that is induced in individual epidermal cells. J Virol 78, 3149–3154.[CrossRef]
    [Google Scholar]
  52. Scholthof, H. B. ( 2005; ). Plant virus transport: motions of functional equivalence. Trends Plant Sci 10, 376–382.[CrossRef]
    [Google Scholar]
  53. Thomas, C. L., Leh, V., Lederer, C. & Maule, A. J. ( 2003; ). Turnip crinkle virus coat protein mediates suppression of RNA silencing in Nicotiana benthamiana. Virology 306, 33–41.[CrossRef]
    [Google Scholar]
  54. Verchot-Lubicz, J. ( 2005; ). A new cell-to-cell transport model for potexviruses. Mol Plant Microbe Interact 18, 283–290.[CrossRef]
    [Google Scholar]
  55. Vilar, M., Esteve, V., Pallás, V., Marcos, J. F. & Pérez-Payá, E. ( 2001; ). Structural properties of carnation mottle virus p7 movement protein and its RNA-binding domain. J Biol Chem 276, 18122–18129.[CrossRef]
    [Google Scholar]
  56. Vilar, M., Saurí, A., Monné, M., Marcos, J. F., von Heijne, G., Pérez-Payá, E. & Mingarro, I. ( 2002; ). Insertion and topology of a plant viral movement protein in the endoplasmic reticulum membrane. J Biol Chem 277, 23447–23452.[CrossRef]
    [Google Scholar]
  57. Voinnet, O., Lederer, C. & Baulcombe, D. C. ( 2000; ). A viral movement protein prevents spread of the gene silencing signal in Nicotiana benthamiana. Cell 103, 157–167.[CrossRef]
    [Google Scholar]
  58. White, K. A., Skuzeski, J. M., Li, W., Wei, N. & Morris, T. J. ( 1995; ). Immunodetection, expression strategy and complementation of turnip crinkle virus p28 and p88 replication components. Virology 211, 525–534.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.81793-0
Loading
/content/journal/jgv/10.1099/vir.0.81793-0
Loading

Data & Media loading...

Supplements

vol. , part 8, pp. 2371 – 2380

Images of the abaxial side of melon cotyledons after agroinfiltration of pMOG(GFP)

List of primers used for site-directed mutagenesis of pMNSV(Al) and pMNSV(Al)-Dcp-GFP



PDF

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error