1887

Abstract

A full-length cDNA clone of the RNA genome of the cucurbit potyvirus zucchini yellow mosaic virus (ZYMV) was constructed downstream from a bacteriophage T7 RNA polymerase promoter. A single extra guanosine residue not present in ZYMV RNA was added to the 5′ and 3′ ends. Capped (mGpppG) ZYMV RNA transcripts were infectious in 10 of 91 test plants; uncapped RNA transcripts were not infectious. The appearance of symptoms in plants inoculated with the infectious transcript was delayed for more than a week compared to plants inoculated with native viral RNA. The progeny virions recovered from infected plants had the same biological properties (aphid non-transmissibility and typical symptoms) as the parental virus. The progeny virions also reacted positively with ZYMV antiserum and ZYMV-specific probes by dot blot hybridization. The authenticity of the progeny virus was verified by identifying a specific molecular marker (C substituted for T in the 3′ non-coding region) using nucleotide sequence analysis.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-72-11-2639
1991-11-01
2024-04-24
Loading full text...

Full text loading...

/deliver/fulltext/jgv/72/11/JV0720112639.html?itemId=/content/journal/jgv/10.1099/0022-1317-72-11-2639&mimeType=html&fmt=ahah

References

  1. Ahlquist P., French R., Janda M., Loesch-Fries L. S. 1984; Multicomponent RNA plant virus infection derived from cloned viral cDNA. Proceedings of the National Academy of Sciences, U.S.A. 81:7066–7070
    [Google Scholar]
  2. Antignus Y., Raccah B., Gal-On A., Cohen S. 1989; Biological and serological characterization of zucchini yellow mosaic and watermelon mosaic virus-2 isolates in Israel. Phytoparasitica 17:289–297
    [Google Scholar]
  3. Atreya C. D., Raccah B., Pirone T. P. 1990; A point mutation in the coat protein abolishes aphid transmissibility of a potyvirus. Virology 178:161–165
    [Google Scholar]
  4. Carrington J. C., Freed D. D. 1990; Cap-independent enhancement of translation by a plant potyvirus 5′ nontranslation region. Journal of Virology 64:1590–1597
    [Google Scholar]
  5. Dawson W. O., Beck D. L., Knorr D. A., Grantham G. L. 1986; cDNA cloning of the complete genome of tobacco mosaic virus and production of infectious transcripts. Proceedings of the National Academy of Sciences, U.S.A. 83:1832–1836
    [Google Scholar]
  6. Domier L. L., Franklin K. M., Hunt A. G., Rhoads R. E., Shaw J. G. 1989; Infectious in vitro transcripts from cloned cDNA of a potyvirus, tobacco vein mottling virus. Proceedings of the National Academy of Sciences, U.S.A. 86:3509–3513
    [Google Scholar]
  7. Dougherty W. G., Carrington J. C. 1988; Expression and function of potyviral gene products. Annual Review of Phytopathology 26:123–143
    [Google Scholar]
  8. Eggen R., Verver J., Wellink J., Jong A. D., Goldbach R., van Kammen A. 1989; Improvements of the infectivity of in vitro transcripts from cloned cowpea mosaic virus cDNA: impact of terminal nucleotide sequences. Virology 173:447–455
    [Google Scholar]
  9. French R., Ahlquist P. 1988; Characterization and engineering of sequences controlling in vivo synthesis of brome mosaic virus subgenomic RNA. Journal of Virology 62:2411–2420
    [Google Scholar]
  10. Gal-On A., Antignus Y., Rosner A., Raccah B. 1990a; Nucleotide sequence of zucchini yellow mosaic virus capsidencoding gene and its expression in Escherichia coli. Gene 87:273–277
    [Google Scholar]
  11. Gal-On A., Antignus Y., Rosner A., Raccah B. 1990b; Nucleotide sequence comparison between zucchini yellow mosaic virus (ZYNV) strains differing in their multiplication rate and transmissibility by aphids. In Abstracts of the 8th International Congress of Virology, Berlin, Germany p 468
    [Google Scholar]
  12. Heaton L. A., Carrington J. C., Morris T. J. 1989; Turnip crinkle virus infection from RNA synthesized in vitro. Virology 170:214–218
    [Google Scholar]
  13. Korneluk R. G., Quan F., Gravel R. A. 1985; Rapid and reliable dideoxy sequencing of double-stranded DNA. Gene 40:317–323
    [Google Scholar]
  14. Kunkel T. A., Roberts J. D., Zakour R. A. 1987; Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods in Enzymology 154:367–382
    [Google Scholar]
  15. Laemmli U. K. 1970; Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature, London 227:680–685
    [Google Scholar]
  16. Lisa V., Boccardo G., D’Agostino G., Dellavalle G., D’Aquillo M. 1981; Characterization of a potyvirus that causes zucchini yellow mosaic virus. Phytopathology 571:667–672
    [Google Scholar]
  17. Maniatis T., Fritsch E. F., Sambrook J. 1982 Molecular Cloning: A Laboratory Manual New York: Cold Spring Harbor Laboratory;
    [Google Scholar]
  18. Meshi T., Ishikawa M., Motoyoshi F., Semba K., Okada Y. 1986; In vitro transcription of infectious RNAs from full-length cDNAs of tobacco mosaic virus. Proceedings of the National Academy of Sciences, U.S.A. 83:5043–5047
    [Google Scholar]
  19. Milne R. G., Luisoni E. 1977; Rapid immuno-electron microscopy of virus preparations. Methods in Virology 6:265–281
    [Google Scholar]
  20. Petty I. T. D., Hunter B. G., Wei N., Jackson A. O. 1989; Infectious barley mosaic virus RNA transcribed in vitro from full-length genomic cDNA clones. Virology 171:342–349
    [Google Scholar]
  21. Riechmann J. L., Lain S., García J. A. 1989; The genome-linked protein and 5′ end RNA sequence of plum pox potyvirus. Journal of General Virology 70:2785–2789
    [Google Scholar]
  22. Riechmann J. L., Lain S., Garcia J. A. 1990; Infectious in vitro transcripts from plum pox potyvirus cDNA clone. Virology 177:710–716
    [Google Scholar]
  23. Rosner A., Bar-Joseph M., Moskovitz M., Mevarech M. 1983; Diagnosis of specific viral RNA sequences in plant extracts by hybridization with polynucleotide kinase-mediated, 32P-labeled, double stranded RNA probe. Phytopathology 73:609–702
    [Google Scholar]
  24. Sanger F., Nicklen S., Coulson A. R. 1977; DNA sequencing with chain-terminating inhibitors. Proceedings of the National Academy of Sciences, U.S.A. 74:5463–5467
    [Google Scholar]
  25. Thomas P. S. 1980; Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proceedings of the National Academy of Sciences, U.S.A. 77:5201–5205
    [Google Scholar]
  26. van der werf S., Bradley J., Wimmer E., Studier F. W., Dunn J. J. 1986; Synthesis of infectious poliovirus RNA by purified T7 RNA polymerase. Proceedings of the National Academy of Sciences, U.S.A. 83:2330–2334
    [Google Scholar]
  27. Vos P., Jaegle M., Wellink J., Verver J., Eggen R., van Kammen A., Goldbach R. 1988; Infectious RNA transcripts derived from full-length DNA copies of the genomic RNAs of cowpea mosaic virus. Virology 165:33–41
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-72-11-2639
Loading
/content/journal/jgv/10.1099/0022-1317-72-11-2639
Loading

Data & Media loading...

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