Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1998 Jun;10(6):937–946. doi: 10.1105/tpc.10.6.937

Initiation and maintenance of virus-induced gene silencing

MT Ruiz 1, O Voinnet 1, DC Baulcombe 1
PMCID: PMC144041  PMID: 9634582

Abstract

The phytoene desaturase (PDS) gene of Nicotiana benthamiana was silenced in plants infected with potato virus X (PVX) vectors carrying PDS inserts, and a green fluorescent protein (GFP) transgene was silenced in plants infected with PVX-GFP. This virus-induced gene silencing (VIGS) is post-transcriptional and cytoplasmic because it is targeted against exons rather than introns of PDS RNA and against viral RNAs. Although PDS and GFP RNAs are most likely targeted through the same mechanism, the VIGS phenotypes differed in two respects. PDS mRNA was targeted by VIGS in all green tissue of the PVX-PDS-infected plant, whereas PVX-PDS was not affected. In contrast, VIGS of the GFP was targeted against PVX-GFP. Initially, VIGS of the GFP was initiated in all green tissues, as occurred with PDS VIGS. However, after 30 days of infection, the GFP VIGS was no longer initiated in newly emerging leaves, although it was maintained in tissue in which it had already been initiated. Based on these analyses, we propose a model for VIGS in which the initiation of VIGS is dependent on the virus and maintenance of it is virus independent.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

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. Angell S. M., Baulcombe D. C. Consistent gene silencing in transgenic plants expressing a replicating potato virus X RNA. EMBO J. 1997 Jun 16;16(12):3675–3684. doi: 10.1093/emboj/16.12.3675. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baulcombe D. C., Chapman S., Santa Cruz S. Jellyfish green fluorescent protein as a reporter for virus infections. Plant J. 1995 Jun;7(6):1045–1053. doi: 10.1046/j.1365-313x.1995.07061045.x. [DOI] [PubMed] [Google Scholar]
  4. Baulcombe D. C. Mechanisms of Pathogen-Derived Resistance to Viruses in Transgenic Plants. Plant Cell. 1996 Oct;8(10):1833–1844. doi: 10.1105/tpc.8.10.1833. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Baulcombe D. C. RNA as a target and an initiator of post-transcriptional gene silencing in transgenic plants. Plant Mol Biol. 1996 Oct;32(1-2):79–88. doi: 10.1007/BF00039378. [DOI] [PubMed] [Google Scholar]
  6. Chalfie M., Tu Y., Euskirchen G., Ward W. W., Prasher D. C. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. doi: 10.1126/science.8303295. [DOI] [PubMed] [Google Scholar]
  7. Chapman S., Kavanagh T., Baulcombe D. Potato virus X as a vector for gene expression in plants. Plant J. 1992 Jul;2(4):549–557. doi: 10.1046/j.1365-313x.1992.t01-24-00999.x. [DOI] [PubMed] [Google Scholar]
  8. Depicker A., Montagu M. V. Post-transcriptional gene silencing in plants. Curr Opin Cell Biol. 1997 Jun;9(3):373–382. doi: 10.1016/s0955-0674(97)80010-5. [DOI] [PubMed] [Google Scholar]
  9. Donson J., Kearney C. M., Hilf M. E., Dawson W. O. Systemic expression of a bacterial gene by a tobacco mosaic virus-based vector. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7204–7208. doi: 10.1073/pnas.88.16.7204. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. English J. J., Mueller E., Baulcombe D. C. Suppression of Virus Accumulation in Transgenic Plants Exhibiting Silencing of Nuclear Genes. Plant Cell. 1996 Feb;8(2):179–188. doi: 10.1105/tpc.8.2.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Genschik P., Parmentier Y., Durr A., Marbach J., Criqui M. C., Jamet E., Fleck J. Ubiquitin genes are differentially regulated in protoplast-derived cultures of Nicotiana sylvestris and in response to various stresses. Plant Mol Biol. 1992 Dec;20(5):897–910. doi: 10.1007/BF00027161. [DOI] [PubMed] [Google Scholar]
  12. Giuliano G., Bartley G. E., Scolnik P. A. Regulation of carotenoid biosynthesis during tomato development. Plant Cell. 1993 Apr;5(4):379–387. doi: 10.1105/tpc.5.4.379. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Goodwin J., Chapman K., Swaney S., Parks T. D., Wernsman E. A., Dougherty W. G. Genetic and biochemical dissection of transgenic RNA-mediated virus resistance. Plant Cell. 1996 Jan;8(1):95–105. doi: 10.1105/tpc.8.1.95. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Haseloff J., Siemering K. R., Prasher D. C., Hodge S. Removal of a cryptic intron and subcellular localization of green fluorescent protein are required to mark transgenic Arabidopsis plants brightly. Proc Natl Acad Sci U S A. 1997 Mar 18;94(6):2122–2127. doi: 10.1073/pnas.94.6.2122. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kavanagh T., Goulden M., Santa Cruz S., Chapman S., Barker I., Baulcombe D. Molecular analysis of a resistance-breaking strain of potato virus X. Virology. 1992 Aug;189(2):609–617. doi: 10.1016/0042-6822(92)90584-c. [DOI] [PubMed] [Google Scholar]
  16. Kumagai M. H., Donson J., della-Cioppa G., Harvey D., Hanley K., Grill L. K. Cytoplasmic inhibition of carotenoid biosynthesis with virus-derived RNA. Proc Natl Acad Sci U S A. 1995 Feb 28;92(5):1679–1683. doi: 10.1073/pnas.92.5.1679. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lindbo J. A., Silva-Rosales L., Proebsting W. M., Dougherty W. G. Induction of a Highly Specific Antiviral State in Transgenic Plants: Implications for Regulation of Gene Expression and Virus Resistance. Plant Cell. 1993 Dec;5(12):1749–1759. doi: 10.1105/tpc.5.12.1749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mann V., Pecker I., Hirschberg J. Cloning and characterization of the gene for phytoene desaturase (Pds) from tomato (Lycopersicon esculentum). Plant Mol Biol. 1994 Feb;24(3):429–434. doi: 10.1007/BF00024111. [DOI] [PubMed] [Google Scholar]
  19. Metzlaff M., O'Dell M., Cluster P. D., Flavell R. B. RNA-mediated RNA degradation and chalcone synthase A silencing in petunia. Cell. 1997 Mar 21;88(6):845–854. doi: 10.1016/s0092-8674(00)81930-3. [DOI] [PubMed] [Google Scholar]
  20. Pecker I., Chamovitz D., Linden H., Sandmann G., Hirschberg J. A single polypeptide catalyzing the conversion of phytoene to zeta-carotene is transcriptionally regulated during tomato fruit ripening. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4962–4966. doi: 10.1073/pnas.89.11.4962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pruss G., Ge X., Shi X. M., Carrington J. C., Bowman Vance V. Plant viral synergism: the potyviral genome encodes a broad-range pathogenicity enhancer that transactivates replication of heterologous viruses. Plant Cell. 1997 Jun;9(6):859–868. doi: 10.1105/tpc.9.6.859. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rommens C. M., Salmeron J. M., Oldroyd G. E., Staskawicz B. J. Intergeneric transfer and functional expression of the tomato disease resistance gene Pto. Plant Cell. 1995 Oct;7(10):1537–1544. doi: 10.1105/tpc.7.10.1537. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Sijen T., Wellink J., Hiriart J. B., Van Kammen A. RNA-Mediated Virus Resistance: Role of Repeated Transgenes and Delineation of Targeted Regions. Plant Cell. 1996 Dec;8(12):2277–2294. doi: 10.1105/tpc.8.12.2277. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Smith H. A., Swaney S. L., Parks T. D., Wernsman E. A., Dougherty W. G. Transgenic plant virus resistance mediated by untranslatable sense RNAs: expression, regulation, and fate of nonessential RNAs. Plant Cell. 1994 Oct;6(10):1441–1453. doi: 10.1105/tpc.6.10.1441. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tanzer M. M., Thompson W. F., Law M. D., Wernsman E. A., Uknes S. Characterization of Post-Transcriptionally Suppressed Transgene Expression That Confers Resistance to Tobacco Etch Virus Infection in Tobacco. Plant Cell. 1997 Aug;9(8):1411–1423. doi: 10.1105/tpc.9.8.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Tenllado F., García-Luque I., Serra M. T., Díaz-Ruíz J. R. Nicotiana benthamiana plants transformed with the 54-kDa region of the pepper mild mottle tobamovirus replicase gene exhibit two types of resistance responses against viral infection. Virology. 1995 Aug 1;211(1):170–183. doi: 10.1006/viro.1995.1389. [DOI] [PubMed] [Google Scholar]

Articles from The Plant Cell are provided here courtesy of Oxford University Press

RESOURCES