Skip to main content
The Plant Cell logoLink to The Plant Cell
. 1995 Aug;7(8):1157–1172. doi: 10.1105/tpc.7.8.1157

Identification of tomato bushy stunt virus host-specific symptom determinants by expression of individual genes from a potato virus X vector.

H B Scholthof 1, K B Scholthof 1, A O Jackson 1
PMCID: PMC160941  PMID: 7549478

Abstract

In this study, we analyzed the influence of two nested genes (p19 and p22) of tomato bushy stunt virus (TBSV) on disease symptoms in systemically infected plants and in local lesion hosts. The contribution of individual genes was determined by bioassays with an infectious clone of wild-type TBSV, with p19/p22 mutant derivatives, and by expression of individual TBSV genes from a heterologous potato virus X (PVX) vector. Our results showed that TBSV genes could be expressed at high levels from the PVX vector. The subcellular localization of these proteins as well as the ability of PVX-expressed p22 to trans complement TBSV cell-to-cell movement defective mutants indicate that the exogenously expressed proteins are functionally active. Inoculation studies with TBSV mutants and the PVX derivatives demonstrated that p19 induced a generalized necrosis upon systemic infection of Nicotiana benthamiana and N. clevelandii. In addition, p19 elicited the formation of local necrotic lesions in N. tabacum; however, in N. glutinosa and N. edwardsonii, the local lesion response was activated by p22. These results show that the p19 and p22 proteins of TBSV are important symptom determinants and that closely related plant species may contain different resistance genes that selectively respond to individual TBSV proteins.

Full Text

The Full Text of this article is available as a PDF (3.6 MB).

Selected References

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

  1. Allison R. F., Janda M., Ahlquist P. Infectious in vitro transcripts from cowpea chlorotic mottle virus cDNA clones and exchange of individual RNA components with brome mosaic virus. J Virol. 1988 Oct;62(10):3581–3588. doi: 10.1128/jvi.62.10.3581-3588.1988. [DOI] [PMC free article] [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. Boyer J. C., Haenni A. L. Infectious transcripts and cDNA clones of RNA viruses. Virology. 1994 Feb;198(2):415–426. doi: 10.1006/viro.1994.1053. [DOI] [PubMed] [Google Scholar]
  4. 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]
  5. Cruz S. S., Baulcombe D. C. Molecular analysis of potato virus X isolates in relation to the potato hypersensitivity gene Nx. Mol Plant Microbe Interact. 1993 Nov-Dec;6(6):707–714. doi: 10.1094/mpmi-6-707. [DOI] [PubMed] [Google Scholar]
  6. Dalmay T., Rubino L., Burgyán J., Russo M. Replication and movement of a coat protein mutant of cymbidium ringspot tombusvirus. Mol Plant Microbe Interact. 1992 Sep-Oct;5(5):379–383. doi: 10.1094/mpmi-5-379. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Dolja V. V., McBride H. J., Carrington J. C. Tagging of plant potyvirus replication and movement by insertion of beta-glucuronidase into the viral polyprotein. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10208–10212. doi: 10.1073/pnas.89.21.10208. [DOI] [PMC free article] [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. Hamamoto H., Sugiyama Y., Nakagawa N., Hashida E., Matsunaga Y., Takemoto S., Watanabe Y., Okada Y. A new tobacco mosaic virus vector and its use for the systemic production of angiotensin-I-converting enzyme inhibitor in transgenic tobacco and tomato. Biotechnology (N Y) 1993 Aug;11(8):930–932. doi: 10.1038/nbt0893-930. [DOI] [PubMed] [Google Scholar]
  11. Hearne P. Q., Knorr D. A., Hillman B. I., Morris T. J. The complete genome structure and synthesis of infectious RNA from clones of tomato bushy stunt virus. Virology. 1990 Jul;177(1):141–151. doi: 10.1016/0042-6822(90)90468-7. [DOI] [PubMed] [Google Scholar]
  12. Hillman B. I., Hearne P., Rochon D., Morris T. J. Organization of tomato bushy stunt virus genome: characterization of the coat protein gene and the 3' terminus. Virology. 1989 Mar;169(1):42–50. doi: 10.1016/0042-6822(89)90039-1. [DOI] [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. Jupin I., Guilley H., Richards K. E., Jonard G. Two proteins encoded by beet necrotic yellow vein virus RNA 3 influence symptom phenotype on leaves. EMBO J. 1992 Feb;11(2):479–488. doi: 10.1002/j.1460-2075.1992.tb05078.x. [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. Keen N. T. Gene-for-gene complementarity in plant-pathogen interactions. Annu Rev Genet. 1990;24:447–463. doi: 10.1146/annurev.ge.24.120190.002311. [DOI] [PubMed] [Google Scholar]
  17. Knorr D. A., Mullin R. H., Hearne P. Q., Morris T. J. De novo generation of defective interfering RNAs of tomato bushy stunt virus by high multiplicity passage. Virology. 1991 Mar;181(1):193–202. doi: 10.1016/0042-6822(91)90484-S. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McLean M. A., Hamilton R. I., Rochon D. M. Symptomatology and movement of a cucumber necrosis virus mutant lacking the coat protein protruding domain. Virology. 1993 Apr;193(2):932–939. doi: 10.1006/viro.1993.1202. [DOI] [PubMed] [Google Scholar]
  19. Otsuki Y., Shimomura T., Takebe I. Tobacco mosaic virus multiplication and expression of the N gene in necrotic responding tobacco varieties. Virology. 1972 Oct;50(1):45–50. doi: 10.1016/0042-6822(72)90344-3. [DOI] [PubMed] [Google Scholar]
  20. Padgett H. S., Beachy R. N. Analysis of a tobacco mosaic virus strain capable of overcoming N gene-mediated resistance. Plant Cell. 1993 May;5(5):577–586. doi: 10.1105/tpc.5.5.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Petty I. T., Donald R. G., Jackson A. O. Multiple genetic determinants of barley stripe mosaic virus influence lesion phenotype on Chenopodium amaranticolor. Virology. 1994 Jan;198(1):218–226. doi: 10.1006/viro.1994.1024. [DOI] [PubMed] [Google Scholar]
  22. 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]
  23. Pfitzner U. M., Pfitzner A. J. Expression of a viral avirulence gene in transgenic plants is sufficient to induce the hypersensitive defense reaction. Mol Plant Microbe Interact. 1992 Jul-Aug;5(4):318–321. doi: 10.1094/mpmi-5-318. [DOI] [PubMed] [Google Scholar]
  24. Qiu S. G., Schoelz J. E. Three regions of cauliflower mosaic virus strain W260 are involved in systemic infection of solanaceous hosts. Virology. 1992 Oct;190(2):773–782. doi: 10.1016/0042-6822(92)90915-c. [DOI] [PubMed] [Google Scholar]
  25. Rommens C. M., Salmeron J. M., Baulcombe D. C., Staskawicz B. J. Use of a gene expression system based on potato virus X to rapidly identify and characterize a tomato Pto homolog that controls fenthion sensitivity. Plant Cell. 1995 Mar;7(3):249–257. doi: 10.1105/tpc.7.3.249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Schoelz J. E., Wintermantel W. M. Expansion of Viral Host Range through Complementation and Recombination in Transgenic Plants. Plant Cell. 1993 Nov;5(11):1669–1679. doi: 10.1105/tpc.5.11.1669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Scholelz J. E., Shepherd R. J. Host range control of cauliflower mosaic virus. Virology. 1988 Jan;162(1):30–37. doi: 10.1016/0042-6822(88)90391-1. [DOI] [PubMed] [Google Scholar]
  28. Scholthof K. B., Hillman B. I., Modrell B., Heaton L. A., Jackson A. O. Characterization and detection of sc4: a sixth gene encoded by sonchus yellow net virus. Virology. 1994 Oct;204(1):279–288. doi: 10.1006/viro.1994.1532. [DOI] [PubMed] [Google Scholar]
  29. Spratt B. G., Hedge P. J., te Heesen S., Edelman A., Broome-Smith J. K. Kanamycin-resistant vectors that are analogues of plasmids pUC8, pUC9, pEMBL8 and pEMBL9. Gene. 1986;41(2-3):337–342. doi: 10.1016/0378-1119(86)90117-4. [DOI] [PubMed] [Google Scholar]
  30. Stanley J. Geminiviruses: plant viral vectors. Curr Opin Genet Dev. 1993 Feb;3(1):91–96. doi: 10.1016/s0959-437x(05)80347-8. [DOI] [PubMed] [Google Scholar]
  31. Weber H., Schultze S., Pfitzner A. J. Two amino acid substitutions in the tomato mosaic virus 30-kilodalton movement protein confer the ability to overcome the Tm-2(2) resistance gene in the tomato. J Virol. 1993 Nov;67(11):6432–6438. doi: 10.1128/jvi.67.11.6432-6438.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES