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. 1987 Jul;6(7):1845–1851. doi: 10.1002/j.1460-2075.1987.tb02442.x

Expression of alfalfa mosaic virus RNA 4 in transgenic plants confers virus resistance

L S Loesch-Fries 1, D Merlo 1, T Zinnen 1, L Burhop 1, K Hill 1, K Krahn 1, N Jarvis 1, S Nelson 1, E Halk 1
PMCID: PMC553567  PMID: 16453779

Abstract

Agrobacterium-mediated transfer from a binary vector was used to produce transgenic Nicotiana tabacum plants that expressed coat protein of the plant virus, alfalfa mosaic virus (AMV). Expression levels of the chimeric gene, which was under the control of the cauliflower mosaic virus 19S promoter, were determined in primary transformed plants, in the progeny from self-fertilization and in the progeny from crosses to normal tobacco. RNA transcripts that were of the expected size as well as a protein of the Mr and antigenicity of AMV coat protein accumulated in the transgenic plants. Plants that expressed the highest levels of coat protein developed fewer primary infections following inoculation with two strains of AMV and developed systemic infection slower than did plants that did not express coat protein. Resistance was specifically against virions of the AMV strains. AMV RNA and the unrelated virus, tobacco mosaic virus, were as infectious on progeny that expressed coat protein as they were on progeny that did not. The relationship between the virus resistance expressed by these transgenic plants and that observed in virus cross-protection is discussed.

Keywords: alfalfa mosaic virus, cross-protection, transgenic tobacco, virus coat protein, virus resistance

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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. 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]
  3. Avrameas S. Coupling of enzymes to proteins with glutaraldehyde. Use of the conjugates for the detection of antigens and antibodies. Immunochemistry. 1969 Jan;6(1):43–52. doi: 10.1016/0019-2791(69)90177-3. [DOI] [PubMed] [Google Scholar]
  4. Balázs E., Bouzoubaa S., Guilley H., Jonard G., Paszkowski J., Richards K. Chimeric vector construction for higher-plant transformation. Gene. 1985;40(2-3):343–348. doi: 10.1016/0378-1119(85)90059-9. [DOI] [PubMed] [Google Scholar]
  5. Beck E., Ludwig G., Auerswald E. A., Reiss B., Schaller H. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene. 1982 Oct;19(3):327–336. doi: 10.1016/0378-1119(82)90023-3. [DOI] [PubMed] [Google Scholar]
  6. Bol J. F., Kraal B., Brederode F. T. Limited proteolysis of alfalfa mosaic virus: influence on the structural and biological function of the coat protein. Virology. 1974 Mar;58(1):101–110. doi: 10.1016/0042-6822(74)90144-5. [DOI] [PubMed] [Google Scholar]
  7. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  8. Christou P., Platt S. G., Ackerman M. C. Opine synthesis in wild-type plant tissue. Plant Physiol. 1986 Sep;82(1):218–221. doi: 10.1104/pp.82.1.218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dhaese P., De Greve H., Gielen J., Seurinck L., Van Montagu M., Schell J. Identification of sequences involved in the polyadenylation of higher plant nuclear transcripts using Agrobacterium T-DNA genes as models. EMBO J. 1983;2(3):419–426. doi: 10.1002/j.1460-2075.1983.tb01439.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ditta G., Stanfield S., Corbin D., Helinski D. R. Broad host range DNA cloning system for gram-negative bacteria: construction of a gene bank of Rhizobium meliloti. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7347–7351. doi: 10.1073/pnas.77.12.7347. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Franck A., Guilley H., Jonard G., Richards K., Hirth L. Nucleotide sequence of cauliflower mosaic virus DNA. Cell. 1980 Aug;21(1):285–294. doi: 10.1016/0092-8674(80)90136-1. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Mazodier P., Cossart P., Giraud E., Gasser F. Completion of the nucleotide sequence of the central region of Tn5 confirms the presence of three resistance genes. Nucleic Acids Res. 1985 Jan 11;13(1):195–205. doi: 10.1093/nar/13.1.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Murray M. G., Thompson W. F. Rapid isolation of high molecular weight plant DNA. Nucleic Acids Res. 1980 Oct 10;8(19):4321–4325. doi: 10.1093/nar/8.19.4321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Pinck L., Hirth L. The replicative RNA and the viral RNA synthesis rate in tobacco infected with alfalfa mosaic virus. Virology. 1972 Aug;49(2):413–425. doi: 10.1016/0042-6822(72)90494-1. [DOI] [PubMed] [Google Scholar]
  16. Priefer U. B., Simon R., Pühler A. Extension of the host range of Escherichia coli vectors by incorporation of RSF1010 replication and mobilization functions. J Bacteriol. 1985 Jul;163(1):324–330. doi: 10.1128/jb.163.1.324-330.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  18. Van Vloten-Doting L., Jaspars E. M. The uncoating of alfalfa mosaic virus by its own RNA. Virology. 1972 Jun;48(3):699–708. doi: 10.1016/0042-6822(72)90154-7. [DOI] [PubMed] [Google Scholar]

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