Skip to main content
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1994 Oct 25;91(22):10310–10314. doi: 10.1073/pnas.91.22.10310

Disruption of virus movement confers broad-spectrum resistance against systemic infection by plant viruses with a triple gene block.

D L Beck 1, C J Van Dolleweerd 1, T J Lough 1, E Balmori 1, D M Voot 1, M T Andersen 1, I E O'Brien 1, R L Forster 1
PMCID: PMC45009  PMID: 7937946

Abstract

White clover mosaic virus strain O (WClMV-O), species of the Potexvirus genus, contains a set of three partially overlapping genes (the triple gene block) that encodes nonvirion proteins of 26 kDa, 13 kDa, and 7 kDa. These proteins are necessary for cell-to-cell movement in plants but not for replication. The WClMV-O 13-kDa gene was mutated (to 13*) in a region of the gene that is conserved in all viruses known to possess triple-gene-block proteins. All 10 13* transgenic lines of Nicotiana benthamiana designed to express the mutated movement protein were shown to be resistant to systemic infection by WClMV-O at 1 microgram of WClMV virions per ml, whereas all plants from susceptible control lines became systemically infected. Of the 13* transgenic lines, 3 selected for their abundant seed supply were shown to be resistant to systemic infection when challenged by inoculation with three different WClMV strains (O, M, and J) or with WClMV-O RNA at 10 micrograms/ml. Most plants were also resistant to systemic infection at inoculum concentrations up to 250 micrograms of WClMV virions per ml. In addition, the three 13* transgenic plant lines were found to be resistant to systemic infection with two other members of the Potexvirus group, potato virus X and narcissus mosaic virus, and the Carlavirus potato virus S but not to be resistant to tobacco mosaic virus of the Tobamovirus group. These results indicate that virus resistance can be engineered into transgenic plants by expression of dominant negative mutant forms of triple-gene-block movement proteins.

Full text

PDF
10310

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. A simple and general method for transferring genes into plants. Science. 1985 Mar 8;227(4691):1229–1231. doi: 10.1126/science.227.4691.1229. [DOI] [PubMed] [Google Scholar]
  2. Anderson J. M., Palukaitis P., Zaitlin M. A defective replicase gene induces resistance to cucumber mosaic virus in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1992 Sep 15;89(18):8759–8763. doi: 10.1073/pnas.89.18.8759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beck D. L., Forster R. L., Bevan M. W., Boxen K. A., Lowe S. C. Infectious transcripts and nucleotide sequence of cloned cDNA of the potexvirus white clover mosaic virus. Virology. 1990 Jul;177(1):152–158. doi: 10.1016/0042-6822(90)90469-8. [DOI] [PubMed] [Google Scholar]
  4. Beck D. L., Guilford P. J., Voot D. M., Andersen M. T., Forster R. L. Triple gene block proteins of white clover mosaic potexvirus are required for transport. Virology. 1991 Aug;183(2):695–702. doi: 10.1016/0042-6822(91)90998-q. [DOI] [PubMed] [Google Scholar]
  5. Citovsky V., Knorr D., Schuster G., Zambryski P. The P30 movement protein of tobacco mosaic virus is a single-strand nucleic acid binding protein. Cell. 1990 Feb 23;60(4):637–647. doi: 10.1016/0092-8674(90)90667-4. [DOI] [PubMed] [Google Scholar]
  6. Citovsky V., Zambryski P. How do plant virus nucleic acids move through intercellular connections? Bioessays. 1991 Aug;13(8):373–379. doi: 10.1002/bies.950130802. [DOI] [PubMed] [Google Scholar]
  7. Clark M. F., Adams A. N. Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J Gen Virol. 1977 Mar;34(3):475–483. doi: 10.1099/0022-1317-34-3-475. [DOI] [PubMed] [Google Scholar]
  8. De Jong W., Ahlquist P. A hybrid plant RNA virus made by transferring the noncapsid movement protein from a rod-shaped to an icosahedral virus is competent for systemic infection. Proc Natl Acad Sci U S A. 1992 Aug 1;89(15):6808–6812. doi: 10.1073/pnas.89.15.6808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Deom C. M., Lapidot M., Beachy R. N. Plant virus movement proteins. Cell. 1992 Apr 17;69(2):221–224. doi: 10.1016/0092-8674(92)90403-y. [DOI] [PubMed] [Google Scholar]
  10. Deom C. M., Oliver M. J., Beachy R. N. The 30-kilodalton gene product of tobacco mosaic virus potentiates virus movement. Science. 1987 Jul 24;237(4813):389–394. doi: 10.1126/science.237.4813.389. [DOI] [PubMed] [Google Scholar]
  11. Deom C. M., Schubert K. R., Wolf S., Holt C. A., Lucas W. J., Beachy R. N. Molecular characterization and biological function of the movement protein of tobacco mosaic virus in transgenic plants. Proc Natl Acad Sci U S A. 1990 May;87(9):3284–3288. doi: 10.1073/pnas.87.9.3284. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Deom C. M., Wolf S., Holt C. A., Lucas W. J., Beachy R. N. Altered function of the tobacco mosaic virus movement protein in a hypersensitive host. Virology. 1991 Jan;180(1):251–256. doi: 10.1016/0042-6822(91)90029-b. [DOI] [PubMed] [Google Scholar]
  13. Devereux J., Haeberli P., Smithies O. A comprehensive set of sequence analysis programs for the VAX. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 1):387–395. doi: 10.1093/nar/12.1part1.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Fitchen J. H., Beachy R. N. Genetically engineered protection against viruses in transgenic plants. Annu Rev Microbiol. 1993;47:739–763. doi: 10.1146/annurev.mi.47.100193.003515. [DOI] [PubMed] [Google Scholar]
  15. Forster R. L., Bevan M. W., Harbison S. A., Gardner R. C. The complete nucleotide sequence of the potexvirus white clover mosaic virus. Nucleic Acids Res. 1988 Jan 11;16(1):291–303. doi: 10.1093/nar/16.1.291. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Giesman-Cookmeyer D., Lommel S. A. Alanine scanning mutagenesis of a plant virus movement protein identifies three functional domains. Plant Cell. 1993 Aug;5(8):973–982. doi: 10.1105/tpc.5.8.973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gilmer D., Bouzoubaa S., Hehn A., Guilley H., Richards K., Jonard G. Efficient cell-to-cell movement of beet necrotic yellow vein virus requires 3' proximal genes located on RNA 2. Virology. 1992 Jul;189(1):40–47. doi: 10.1016/0042-6822(92)90679-j. [DOI] [PubMed] [Google Scholar]
  18. Gleave A. P. A versatile binary vector system with a T-DNA organisational structure conducive to efficient integration of cloned DNA into the plant genome. Plant Mol Biol. 1992 Dec;20(6):1203–1207. doi: 10.1007/BF00028910. [DOI] [PubMed] [Google Scholar]
  19. Habili N., Symons R. H. Evolutionary relationship between luteoviruses and other RNA plant viruses based on sequence motifs in their putative RNA polymerases and nucleic acid helicases. Nucleic Acids Res. 1989 Dec 11;17(23):9543–9555. doi: 10.1093/nar/17.23.9543. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Herskowitz I. Functional inactivation of genes by dominant negative mutations. Nature. 1987 Sep 17;329(6136):219–222. doi: 10.1038/329219a0. [DOI] [PubMed] [Google Scholar]
  21. Inokuchi Y., Hirashima A. Interference with viral infection by defective RNA replicase. J Virol. 1987 Dec;61(12):3946–3949. doi: 10.1128/jvi.61.12.3946-3949.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kanyuka K. V., Vishnichenko V. K., Levay K. E., Kondrikov DYu, Ryabov E. V., Zavriev S. K. Nucleotide sequence of shallot virus X RNA reveals a 5'-proximal cistron closely related to those of potexviruses and a unique arrangement of the 3'-proximal cistrons. J Gen Virol. 1992 Oct;73(Pt 10):2553–2560. doi: 10.1099/0022-1317-73-10-2553. [DOI] [PubMed] [Google Scholar]
  23. Longstaff M., Brigneti G., Boccard F., Chapman S., Baulcombe D. Extreme resistance to potato virus X infection in plants expressing a modified component of the putative viral replicase. EMBO J. 1993 Feb;12(2):379–386. doi: 10.1002/j.1460-2075.1993.tb05669.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Malyshenko S. I., Kondakova O. A., Nazarova JuV, Kaplan I. B., Taliansky M. E., Atabekov J. G. Reduction of tobacco mosaic virus accumulation in transgenic plants producing non-functional viral transport proteins. J Gen Virol. 1993 Jun;74(Pt 6):1149–1156. doi: 10.1099/0022-1317-74-6-1149. [DOI] [PubMed] [Google Scholar]
  25. Meshi T., Motoyoshi F., Maeda T., Yoshiwoka S., Watanabe H., Okada Y. Mutations in the tobacco mosaic virus 30-kD protein gene overcome Tm-2 resistance in tomato. Plant Cell. 1989 May;1(5):515–522. doi: 10.1105/tpc.1.5.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Morozov SYu, Dolja V. V., Atabekov J. G. Probable reassortment of genomic elements among elongated RNA-containing plant viruses. J Mol Evol. 1989 Jul;29(1):52–62. doi: 10.1007/BF02106181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Niesbach-Klösgen U., Guilley H., Jonard G., Richards K. Immunodetection in vivo of beet necrotic yellow vein virus-encoded proteins. Virology. 1990 Sep;178(1):52–61. doi: 10.1016/0042-6822(90)90378-5. [DOI] [PubMed] [Google Scholar]
  28. Petty I. T., French R., Jones R. W., Jackson A. O. Identification of barley stripe mosaic virus genes involved in viral RNA replication and systemic movement. EMBO J. 1990 Nov;9(11):3453–3457. doi: 10.1002/j.1460-2075.1990.tb07553.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Sumi S., Tsuneyoshi T., Furutani H. Novel rod-shaped viruses isolated from garlic, Allium sativum, possessing a unique genome organization. J Gen Virol. 1993 Sep;74(Pt 9):1879–1885. doi: 10.1099/0022-1317-74-9-1879. [DOI] [PubMed] [Google Scholar]
  30. Truve E., Aaspôllu A., Honkanen J., Puska R., Mehto M., Hassi A., Teeri T. H., Kelve M., Seppänen P., Saarma M. Transgenic potato plants expressing mammalian 2'-5' oligoadenylate synthetase are protected from potato virus X infection under field conditions. Biotechnology (N Y) 1993 Sep;11(9):1048–1052. doi: 10.1038/nbt0993-1048. [DOI] [PubMed] [Google Scholar]
  31. Valkonen J. P., Pehu E., Jones M. G., Gibson R. W. Resistance in Solanum brevidens to both potato virus Y and potato virus X may be associated with slow cell-to-cell spread. J Gen Virol. 1991 Feb;72(Pt 2):231–236. doi: 10.1099/0022-1317-72-2-231. [DOI] [PubMed] [Google Scholar]
  32. Wolf S., Deom C. M., Beachy R. N., Lucas W. J. Movement protein of tobacco mosaic virus modifies plasmodesmatal size exclusion limit. Science. 1989 Oct 20;246(4928):377–379. doi: 10.1126/science.246.4928.377. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES