1887

Abstract

A full-length cDNA copy of the genome of (PLRV) was introduced into the genome of tobacco and potato plants by -mediated transformation. Transgenic lines were obtained in which the transgene was readily detected by PCR with DNA extracted from T tobacco seedlings and clonally multiplied potato plants. PLRV-specific genomic and sub- genomic RNAs, coat protein antigen and virus particles were detected in transgenic plants. Aphids fed on the transgenic tobacco plants readily transmitted PLRV to test plants. Infected transgenic tobacco plants, like non-transgenic (WT) PLRV-infected plants, displayed no symptoms of the infection but transgenic plants of potato were severely stunted. In parallel tests, the mean PLRV titres in WT tobacco plants and transgenic tobacco plants were 600 and 630 ng virus/g leaf, respectively, although differences in PLRV titres among transgenic plants were much greater than those among infected WT plants. In similar tests with potato, the mean PLRV titre of WT plants was 50 ng virus/g leaf whereas higher concentrations (up to 3400 ng virus/g leaf) accumulated in transgenic potato plants. In tissue prints of stems, PLRV was detected in similar proportions of phloem cells in transgenic and infected WT plants. In transgenic tobacco and potato plants, but not in infected WT plants, a few stem epidermal cells also contained virus. From tissue prints of transgenic tobacco leaves, it was estimated that about one in 40000 mesophyll cells contained virus, but in transgenic potato, a greater proportion of mesophyll cells was infected.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-80-11-2813
1999-11-01
2020-08-11
Loading full text...

Full text loading...

/deliver/fulltext/jgv/80/11/0802813a.html?itemId=/content/journal/jgv/10.1099/0022-1317-80-11-2813&mimeType=html&fmt=ahah

References

  1. An G., Ebert P. R., Mitra A., Ha S. B.. 1988; Binary vectors. Plant Molecular Biology Manual A3 1–19 Dordrecht: Kluwer Academic;
    [Google Scholar]
  2. Angell S. M., Baulcombe D. C.. 1997; Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. EMBO Journal16:3675–3684
    [Google Scholar]
  3. Atabekov J. G., Taliansky M. E., Drampyan A. H., Kaplan I. B., Turka I. E.. 1984; Systemic infection by a phloem- restricted virus in parenchyma cells in mixed infection. Biologicheskie Nauki10:28–31 (in Russian
    [Google Scholar]
  4. Barker H.. 1987; Invasion of non-phloem tissue in Nicotiana clevelandii by potato leafroll luteovirus is enhanced in plants also infected with potato Y potyvirus. Journal of General Virology68:1223–1227
    [Google Scholar]
  5. Barker H., Harrison B. D.. 1985; Restricted multiplication of potato leafroll virus in resistant potato genotypes. Annals of Applied Biology107:205–212
    [Google Scholar]
  6. Barker H., Solomon R. M.. 1990; Evidence of simple genetic control in potato of ability to restrict potato leaf roll virus concentration in leaves. Theoretical and Applied Genetics80:188–192
    [Google Scholar]
  7. Barker H., Woodford J. A. T.. 1992; Spread of potato leafroll virus is decreased from plants of potato clones in which virus accumulation is restricted. Annals of Applied Biology121:345–354
    [Google Scholar]
  8. Barker H., Reavy B., Kumar A., Webster K. D., Mayo M. A.. 1992; Restricted virus multiplication in potatoes transformed with the coat protein gene of potato leafroll luteovirus: similarities with a type of host gene- mediated resistance. Annals of Applied Biology120:55–64
    [Google Scholar]
  9. Barker H., Reavy B., Webster K. D., Jolly C. A., Kumar A., Mayo M. A.. 1993; Relationship between transcript production and virus resistance in transgenic tobacco expressing the potato leafroll virus coat protein gene. Plant Cell Reports13:54–58
    [Google Scholar]
  10. Bevan M.. 1984; Binary Agrobacterium vectors for plant transformation. Nucleic Acids Research12:8711–8721
    [Google Scholar]
  11. Derrick P. M., Barker H.. 1997; Short and long distance spread of potato leafroll luteovirus: effects of host genes and transgenes conferring resistance to virus accumulation in potato. Journal of General Virology78:243–251
    [Google Scholar]
  12. Kaido M., Mori M., Mise K., Okuno T., Furusawa I.. 1995; Inhibition of brome mosaic virus (BMV) amplification in protoplasts from transgenic tobacco plants expressing replicable BMV RNAs. Journal of General Virology76:2827–2833
    [Google Scholar]
  13. Kawchuk L. M., Martin R. R., Rochon D. M., McPherson J.. 1989; Identification and characterization of the potato leafroll virus putative coat protein gene. Journal of General Virology70:783–788
    [Google Scholar]
  14. Keese P., Martin R. R., Kawchuk L. M., Waterhouse P. M., Gerlach W. L.. 1990; Nucleotide sequences of an Australian and a Canadian isolate of potato leafroll luteovirus and their relationships with two European isolates. Journal of General Virology71:719–724
    [Google Scholar]
  15. Kubo S., Harrison B. D., Robinson D. J., Mayo M. A.. 1975; Tobacco rattle virus in tobacco mesophyll protoplasts: infection and virus multiplication. Journal of General Virology27:293–304
    [Google Scholar]
  16. Kunkel T. A., Roberts J. D., Zakour R. A.. 1987; Rapid and efficient site-specific mutagenesis without phenotypic selection. Methods in Enzymology154:367–382
    [Google Scholar]
  17. Leiser R.-M., Ziegler-Graff V., Reutenauer A., Herrbach E., Lemaire O., Guilley H., Richards K., Jonard G.. 1992; Agro-infection as an alternative to insects for infecting plants with beet western yellows luteovirus. Proceedings of the National Academy of Sciences USA89:9136–9140
    [Google Scholar]
  18. Mayo M. A., Ziegler-Graff V.. 1996; Molecular biology of luteoviruses. Advances in Virus Research46:416–468
    [Google Scholar]
  19. Mayo M. A., Robinson D. J., Jolly C. A., Hyman L.. 1989; Nucleotide sequence of potato leafroll luteovirus RNA. Journal of General Virology70:1037–1051
    [Google Scholar]
  20. Murphy G., Kavanagh T.. 1988; Speeding-up the sequencing of double-stranded DNA. Nucleic Acids Research16:5198
    [Google Scholar]
  21. Mutterer J. D., Ziegler-Graff V., Richards K. E., Smith H. G., Barker H.. 1999; Agro-infection as a means of transmitting luteoviruses to host plants for study of genome function. In The Luteoviridae , chapter 4 Edited by Wallingford: CAB International; (in press)
    [Google Scholar]
  22. Naidu R. A., Mayo M. A., Reddy S. V., Jolly C. A., Torrance L.. 1997; Diversity among the coat proteins of luteoviruses associated with chickpea stunt disease in India. Annals of Applied Biology130:37–47
    [Google Scholar]
  23. Prüfer D., Schmitz J., Tacke E., Kull B., Rohde W.. 1997; In vivo expression of a full- length cDNA copy of potato leafroll virus (PLRV) in protoplasts and transgenic plants. Molecular and General Genetics253:609–614
    [Google Scholar]
  24. Reavy B., Sandgren M., Barker H., Heino P., Oxelfelt P.. 1997; A coat protein transgene from a Scottish isolate of potato mop-top virus mediates strong resistance against Scandinavian isolates which have similar coat protein genes. European Journal of Plant Pathology103:829–834
    [Google Scholar]
  25. Roberts I. M.. 1986; Practical aspects of handling, preparing and staining samples containing plant virus particles for electron microscopy. In Developments and Applications in Virus Testing pp.213–243 Edited by Jones R. A. C., Torrance L.. Wellesbourne: Association of Applied Biologists;
    [Google Scholar]
  26. Sambrook J., Fritsch E. F., Maniatis T.. 1989; Molecular Cloning: A Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  27. Schmitz J., Stussi-Garaud C., Tacke E., Prüfer D., Rohde W., Rohfritsch O.. 1997; In situ localization of the putative movement protein (pr17) from potato leafroll luteovirus (PLRV) in infected and transgenic potato plants. Virology235:311–322
    [Google Scholar]
  28. Töpfer R., Matzeit V., Gronenborn B., Schell J., Steinbiss H. H.. 1987; A set of plant expression vectors for transcriptional and translational fusions. Nucleic Acids Research15:5890
    [Google Scholar]
  29. Waterhouse P. M., Gildow F. E., Johnstone G. R.. 1988; Luteovirus group. AAB Descriptions of Plant Viruses no. 339 pp. 9 Wellesbourne: Association of Applied Biologists;
    [Google Scholar]
  30. Webster K. D., Barker H.. 1998; Detection and quantification of transcript RNA in transgenic plants using digoxigenin-labelled cDNA probes.. In Plant Virology Protocols: From Virus Isolation to Transgenic Resistance , Methods in Molecular Biology Series pp437–445 Edited by Foster G. D., Taylor S. Totowa, NJ: Humana Press;
    [Google Scholar]
  31. Wisniewski L. A., Powell P. A., Nelson R. S., Beachy R. N.. 1990; Local and systemic spread of tobacco mosaic virus in transgenic tobacco. Plant Cell2:559–567
    [Google Scholar]
  32. Yamaya J., Yoshioka M., Meshi T., Okada Y., Ohno T.. 1988; Expression of tobacco mosaic virus RNA in transgenic plants. Molecular and General Genetics211:520–525
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-80-11-2813
Loading
/content/journal/jgv/10.1099/0022-1317-80-11-2813
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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