1887

Abstract

One possible environmental risk related to the utilization of virus-resistant transgenic plants expressing viral sequences is the emergence of new viruses generated by recombination between the viral transgene mRNA and the RNA of an infecting virus. This hypothesis has been tested recently for cucumber mosaic virus (CMV) by comparing the recombinant populations in transgenic and non-transgenic plants under conditions of minimal selection pressure in favour of the recombinants. Equivalent populations were observed in transgenic and non-transgenic plants but, in both, there was a strongly dominant hotspot recombinant which was shown recently to be nonviable alone , suggesting that its predominance could be reduced by applying an increased selection pressure in favour of viable recombinants. Partially disabled I17F-CMV mutants were created by engineering 6 nt deletions in five sites in the RNA3 3′-non-coding region (3′-NCR). One mutant was used to inoculate transgenic tobacco plants expressing the coat protein and 3′-NCR of R-CMV. A total of 22 different recombinant types were identified, of which 12 were, as expected, between the transgene mRNA and the mutated I17F-CMV RNA3, while 10 resulted from recombination between the mutated RNA3 and I17F-CMV RNA1. Twenty recombinants were of the aberrant type, while two, including the dominant one detected previously under conditions of minimal selection pressure, were homologous recombinants. All recombinants detected were very similar to ones observed in nature, suggesting that the deployment of transgenic lines similar to the one studied here would not lead to the emergence of new viruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.013771-0
2009-11-01
2019-11-20
Loading full text...

Full text loading...

/deliver/fulltext/jgv/90/11/2798.html?itemId=/content/journal/jgv/10.1099/vir.0.013771-0&mimeType=html&fmt=ahah

References

  1. Aaziz, R. & Tepfer, M. ( 1999; ). Recombination between genomic RNAs of two cucumoviruses under conditions of minimal selection pressure. Virology 263, 282–289.[CrossRef]
    [Google Scholar]
  2. Ahlquist, P., Dasgupta, R. & Kaesberg, P. ( 1981; ). Near identity of 3′ RNA secondary structure in bromoviruses and cucumber mosaic virus. Cell 23, 183–189.[CrossRef]
    [Google Scholar]
  3. Blanchard, C. L., Boyce, P. M. & Anderson, B. J. ( 1996; ). Cucumber mosaic virus RNA 5 is a mixed population derived from the conserved 3′-terminal regions of genomic RNAs 2 and 3. Virology 217, 598–601.[CrossRef]
    [Google Scholar]
  4. Boccard, F. & Baulcombe, D. ( 1993; ). Mutational analysis of cis-acting sequences and gene function in RNA3 of cucumber mosaic virus. Virology 193, 563–578.[CrossRef]
    [Google Scholar]
  5. Canto, T., Choi, S. K. & Palukaitis, P. ( 2001; ). A subpopulation of RNA 1 of Cucumber mosaic virus contains 3′ termini originating from RNAs 2 or 3. J Gen Virol 82, 941–945.
    [Google Scholar]
  6. Chen, Y. K., Goldbach, R. & Prins, M. ( 2002; ). Inter- and intramolecular recombinations in the cucumber mosaic virus genome related to adaptation to alstroemeria. J Virol 76, 4119–4124.[CrossRef]
    [Google Scholar]
  7. de Wispelaere, M. & Rao, A. L. ( 2009; ). Production of cucumber mosaic virus RNA5 and its role in recombination. Virology 384, 179–191.[CrossRef]
    [Google Scholar]
  8. de Wispelaere, M., Gaubert, S., Trouilloud, S., Belin, C. & Tepfer, M. ( 2005; ). A map of the diversity of RNA3 recombinants appearing in plants infected with Cucumber mosaic virus and Tomato aspermy virus. Virology 331, 117–127.[CrossRef]
    [Google Scholar]
  9. Felden, B., Florentz, C., Giege, R. & Westhof, E. ( 1994; ). Solution structure of the 3′-end of brome mosaic virus genomic RNAs. Conformational mimicry with canonical tRNAs. J Mol Biol 235, 508–531.[CrossRef]
    [Google Scholar]
  10. Fernandez-Cuartero, B., Burgyan, J., Aranda, M. A., Salanki, K., Moriones, E. & Garcia-Arenal, F. ( 1994; ). Increase in the relative fitness of a plant virus RNA associated with its recombinant nature. Virology 203, 373–377.[CrossRef]
    [Google Scholar]
  11. Jacquemond, M. & Lot, H. ( 1981; ). L'ARN satellite du virus de la mosaïque du concombre. I. Comparaison de l'aptitude à induire la nécrose de la tomate d'ARN satellites isolés de plusieur souches du virus. Agronomie 1, 927–932 (in French).[CrossRef]
    [Google Scholar]
  12. Joshi, R. L., Joshi, S., Chapeville, F. & Haenni, A. L. ( 1983; ). tRNA-like structures of plant viral RNAs: conformational requirements for adenylation and aminoacylation. EMBO J 2, 1123–1127.
    [Google Scholar]
  13. Masuta, C., Ueda, S., Suzuki, M. & Uyeda, I. ( 1998; ). Evolution of a quadripartite hybrid virus by interspecific exchange and recombination between replicase components of two related tripartite RNA viruses. Proc Natl Acad Sci U S A 95, 10487–10492.[CrossRef]
    [Google Scholar]
  14. Moreno, I. M., Bernal, J. J., García de Blas, B., Rodriguez-Cerezo, E. & García-Arenal, F. ( 1997; ). The expression level of the 3a movement protein determines differences in severity of symptoms between two strains of tomato aspermy cucumovirus. Mol Plant Microbe Interact 10, 171–179.[CrossRef]
    [Google Scholar]
  15. Palukaitis, P., Roossinck, M. J., Dietzgen, R. G. & Francki, R. I. ( 1992; ). Cucumber mosaic virus. Adv Virus Res 41, 281–348.
    [Google Scholar]
  16. Pierrugues, O., Guilbaud, L., Fernandez-Delmond, I., Fabre, F., Tepfer, M. & Jacquemond, M. ( 2007; ). Biological properties and relative fitness of inter-subgroup cucumber mosaic virus RNA 3 recombinants produced in vitro. J Gen Virol 88, 2852–2861.[CrossRef]
    [Google Scholar]
  17. Raj, S. K., Kumar, S. & Choudhari, S. ( 2007; ). Identification of Tomato aspermy virus as the cause of yellow mosaic and flower deformation of chrysanthemums in India. Australas Plant Dis Notes 2, 1–2.[CrossRef]
    [Google Scholar]
  18. Raj, S. K., Kumar, S., Choudhari, S. & Verma, D. K. ( 2009; ). Biological and molecular characterization of three isolates of Tomato aspermy virus infecting chrysanthemums in India. J Phytopathol 157, 117–125.[CrossRef]
    [Google Scholar]
  19. Rietveld, K., Plejj, C. W. & Bosch, L. ( 1983; ). Three-dimensional models of the tRNA-like 3′ termini of some plant viral RNAs. EMBO J 2, 1079–1085.
    [Google Scholar]
  20. Shi, B., Ding, S. & Symons, R. H. ( 1997; ). Two novel subgenomic RNAs derived from RNA 3 of tomato aspermy cucumovirus. J Gen Virol 78, 505–510.
    [Google Scholar]
  21. Shi, B. J., Symons, R. H. & Palukaitis, P. ( 2007; ). The cucumovirus 2b gene drives selection of inter-viral recombinants affecting the crossover site, the acceptor RNA and the rate of selection. Nucleic Acids Res 36, 1057–1071.[CrossRef]
    [Google Scholar]
  22. Sivakumaran, K., Bao, Y., Roossinck, M. J. & Kao, C. C. ( 2000; ). Recognition of the core RNA promoter for minus-strand RNA synthesis by the replicases of Brome mosaic virus and Cucumber mosaic virus. J Virol 74, 10323–10331.[CrossRef]
    [Google Scholar]
  23. Suzuki, M., Hibi, T. & Masuta, C. ( 2003; ). RNA recombination between cucumoviruses: possible role of predicted stem-loop structures and an internal subgenomic promoter-like motif. Virology 306, 77–86.[CrossRef]
    [Google Scholar]
  24. Tepfer, M. ( 2002; ). Risk assessment of virus-resistant transgenic plants. Annu Rev Phytopathol 40, 467–491.[CrossRef]
    [Google Scholar]
  25. Teycheney, P. Y., Aaziz, R., Dinant, S., Salánki, K., Tourneur, C., Balázs, E., Jacquemond, M. & Tepfer, M. ( 2000; ). Synthesis of (−)-strand RNA from the 3′ untranslated region of plant viral genomes expressed in transgenic plants upon infection with related viruses. J Gen Virol 81, 1121–1126.
    [Google Scholar]
  26. Thompson, J. R., Buratti, E., de Wispelaere, M. & Tepfer, M. ( 2008; ). Structural and functional characterization of the 5′ region of subgenomic RNA5 of cucumber mosaic virus. J Gen Virol 89, 1729–1738.[CrossRef]
    [Google Scholar]
  27. Turturo, C., Friscina, A., Gaubert, S., Jacquemond, M., Thompson, J. R. & Tepfer, M. ( 2008; ). Evaluation of potential risks associated with recombination in transgenic plants expressing viral sequences. J Gen Virol 89, 327–335.[CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.013771-0
Loading
/content/journal/jgv/10.1099/vir.0.013771-0
Loading

Data & Media loading...

Most Cited This Month

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error