Skip to main content
PMC home page
The Plant Cell logoLink to The Plant Cell
. 1993 Jul;5(7):795–807. doi: 10.1105/tpc.5.7.795

Transgenic tobacco plants expressing the geminivirus BL1 protein exhibit symptoms of viral disease.

E Pascal 1, P E Goodlove 1, L C Wu 1, S G Lazarowitz 1
PMCID: PMC160317  PMID: 8364356

Abstract

Bipartite geminiviruses, such as squash leaf curl virus (SqLCV), encode two movement proteins (MPs), BR1 and BL1, that are essential for viral movement in and subsequent infection of the host plant. To elucidate the biochemical functions of these MPs and define their respective contributions to viral infection, we have generated transgenic Nicotiana benthamiana plants expressing SqLCV BR1 and BL1. Transgenic plants expressing BR1 or a truncated BL1 were phenotypically indistinguishable from wild-type N. benthamiana. In contrast, transgenic plants expressing full-length BL1, alone or in combination with BR1, were strikingly abnormal both in their growth properties and phenotypic appearance, with leaves that were mosaic and curled under, thus mimicking typical SqLCV disease symptoms in this host. BL1 was localized to the cell wall and plasma membrane fractions, whereas BR1 was predominantly in the microsomal membrane fraction. These findings demonstrate that expression of BL1 in transgenic plants is sufficient to produce viral disease symptoms, and they further suggest that BL1 and BR1 carry out distinct and independent functions in viral movement.

Full Text

The Full Text of this article is available as a PDF (3.5 MB).

Selected References

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

  1. Abel P. P., Nelson R. S., De B., Hoffmann N., Rogers S. G., Fraley R. T., Beachy R. N. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science. 1986 May 9;232(4751):738–743. doi: 10.1126/science.3457472. [DOI] [PubMed] [Google Scholar]
  2. Baughman G. A., Jacobs J. D., Howell S. H. Cauliflower mosaic virus gene VI produces a symptomatic phenotype in transgenic tobacco plants. Proc Natl Acad Sci U S A. 1988 Feb;85(3):733–737. doi: 10.1073/pnas.85.3.733. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Berna A., Gafny R., Wolf S., Lucas W. J., Holt C. A., Beachy R. N. The TMV movement protein: role of the C-terminal 73 amino acids in subcellular localization and function. Virology. 1991 Jun;182(2):682–689. doi: 10.1016/0042-6822(91)90609-f. [DOI] [PubMed] [Google Scholar]
  4. Bush D. R. Proton-Coupled Sucrose Transport in Plasmalemma Vesicles Isolated from Sugar Beet (Beta vulgaris L. cv Great Western) Leaves. Plant Physiol. 1989 Apr;89(4):1318–1323. doi: 10.1104/pp.89.4.1318. [DOI] [PMC free article] [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., Wong M. L., Shaw A. L., Prasad B. V., Zambryski P. Visualization and characterization of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell. 1992 Apr;4(4):397–411. doi: 10.1105/tpc.4.4.397. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. 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]
  9. Ding B., Haudenshield J. S., Hull R. J., Wolf S., Beachy R. N., Lucas W. J. Secondary plasmodesmata are specific sites of localization of the tobacco mosaic virus movement protein in transgenic tobacco plants. Plant Cell. 1992 Aug;4(8):915–928. doi: 10.1105/tpc.4.8.915. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Etessami P., Callis R., Ellwood S., Stanley J. Delimitation of essential genes of cassava latent virus DNA 2. Nucleic Acids Res. 1988 Jun 10;16(11):4811–4829. doi: 10.1093/nar/16.11.4811. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Feinberg A. P., Vogelstein B. "A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity". Addendum. Anal Biochem. 1984 Feb;137(1):266–267. doi: 10.1016/0003-2697(84)90381-6. [DOI] [PubMed] [Google Scholar]
  12. Gardiner W. E., Sunter G., Brand L., Elmer J. S., Rogers S. G., Bisaro D. M. Genetic analysis of tomato golden mosaic virus: the coat protein is not required for systemic spread or symptom development. EMBO J. 1988 Apr;7(4):899–904. doi: 10.1002/j.1460-2075.1988.tb02894.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holt C. A., Beachy R. N. In vivo complementation of infectious transcripts from mutant tobacco mosaic virus cDNAs in transgenic plants. Virology. 1991 Mar;181(1):109–117. doi: 10.1016/0042-6822(91)90475-q. [DOI] [PubMed] [Google Scholar]
  14. Kay R., Chan A., Daly M., McPherson J. Duplication of CaMV 35S Promoter Sequences Creates a Strong Enhancer for Plant Genes. Science. 1987 Jun 5;236(4806):1299–1302. doi: 10.1126/science.236.4806.1299. [DOI] [PubMed] [Google Scholar]
  15. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  16. Lazarowitz S. G., Lazdins I. B. Infectivity and complete nucleotide sequence of the cloned genomic components of a bipartite squash leaf curl geminivirus with a broad host range phenotype. Virology. 1991 Jan;180(1):58–69. doi: 10.1016/0042-6822(91)90009-z. [DOI] [PubMed] [Google Scholar]
  17. Lazarowitz S. G., Wu L. C., Rogers S. G., Elmer J. S. Sequence-specific interaction with the viral AL1 protein identifies a geminivirus DNA replication origin. Plant Cell. 1992 Jul;4(7):799–809. doi: 10.1105/tpc.4.7.799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. 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]
  19. Meshi T., Watanabe Y., Saito T., Sugimoto A., Maeda T., Okada Y. Function of the 30 kd protein of tobacco mosaic virus: involvement in cell-to-cell movement and dispensability for replication. EMBO J. 1987 Sep;6(9):2557–2563. doi: 10.1002/j.1460-2075.1987.tb02544.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Odell J. T., Nagy F., Chua N. H. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. 1985 Feb 28-Mar 6Nature. 313(6005):810–812. doi: 10.1038/313810a0. [DOI] [PubMed] [Google Scholar]
  21. Reed K. C., Mann D. A. Rapid transfer of DNA from agarose gels to nylon membranes. Nucleic Acids Res. 1985 Oct 25;13(20):7207–7221. doi: 10.1093/nar/13.20.7207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rogers S. G., Bisaro D. M., Horsch R. B., Fraley R. T., Hoffmann N. L., Brand L., Elmer J. S., Lloyd A. M. Tomato golden mosaic virus A component DNA replicates autonomously in transgenic plants. Cell. 1986 May 23;45(4):593–600. doi: 10.1016/0092-8674(86)90291-6. [DOI] [PubMed] [Google Scholar]
  23. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  24. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  25. Sunter G., Bisaro D. M. Transactivation of geminivirus AR1 and BR1 gene expression by the viral AL2 gene product occurs at the level of transcription. Plant Cell. 1992 Oct;4(10):1321–1331. doi: 10.1105/tpc.4.10.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Verwoerd T. C., Dekker B. M., Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 1989 Mar 25;17(6):2362–2362. doi: 10.1093/nar/17.6.2362. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Von Arnim A., Frischmuth T., Stanley J. Detection and possible functions of African cassava mosaic virus DNA B gene products. Virology. 1993 Jan;192(1):264–272. doi: 10.1006/viro.1993.1029. [DOI] [PubMed] [Google Scholar]
  28. 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]
  29. Zijlstra C., Hohn T. Cauliflower Mosaic Virus Gene VI Controls Translation from Dicistronic Expression Units in Transgenic Arabidopsis Plants. Plant Cell. 1992 Dec;4(12):1471–1484. doi: 10.1105/tpc.4.12.1471. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES