Abstract
Within the Bunyaviridae virus family, members of the genus Tospovirus are unique in their ability to infect plants. A characteristic genetic difference between tospoviruses and the animal-infecting members of this virus family is the occurrence of an additional gene, denoted NSM, located on the genomic M RNA segment. This gene has previously been implicated in the cell-to-cell movement of this virus during systemic infection. Transgenic tobacco plants have been obtained expressing the NSM protein of tomato spotted wilt virus (TSWV), the type member of the tospoviruses, from a constitutive promoter. Detectable amounts of the NSM protein could be observed in plants from nine different lines. The protein was only detectable in fractions enriched for cell wall material. More detailed immunogold labelling studies revealed specific association of NSM protein with plasmodesmata. Plants accumulating the NSM protein to detectable levels developed aberrations in growth, resulting in a significant reduction of size and accelerated senescence. In addition, these plants are restricted in their capacity to produce flowers. The results presented provide additional evidence that the NSM protein, by modifying plasmodesmata, represents the cell-to-cell movement function of tospoviruses. Furthermore, the phenotype of the NSM transgenic plants suggests involvement of the NSM gene product in TSWV symptom expression
Similar content being viewed by others
References
Avila, A.C. de, Huguenot, C., Resende, R. de O., Kitajima, E.W., Goldbach, R.W. and Peters, D. (1990) Serological differentiation of 20 isolates of tomato spotted wilt virus. J. Gen. Virol. 71, 2801-7.
Avila, A.C. de De Haan, P., Smeets, M.L.L., Resende, R. de O., Kormelink, R., Kitajima, E.W., Goldbach, R.W. and Peters, D. (1992) Distinct levels of relationships between tospovirus isolates. Arch. Virol. 128, 211-27.
Avila, A.C. de De Haan, P., Kormelink, R., Resende, R. de O., Goldbach, R.W. and Peters, D. (1993) Classification of tospoviruses based on phylogeny of nucleoprotein gene sequences. J. Gen. Virol. 74, 153-59.
Bevan, M. (1984) Binary Agrobacteriumvectors for plant transformation. Nucl. Acids Res. 12, 8711-22.
De Haan P., Wagemakers, L., Peters, D. and Goldbach, R. (1990) The S RNA segment of tomato spotted wilt virus has an ambisense character. J. Gen. Virol. 71, 1001-7.
De Haan, P., Kormelink, R., Resende, R. de O., Van Poelwijk, F., Peters, D. and Goldbach, R. (1991) Tomato spotted wilt virus L RNA encodes a putative RNA polymerase. J. Gen. Virol. 72, 2207-16.
Ding, B., Haudensfield, J.S., Hull, R.J., Wolf, S., Beachy, R.N. and Lucas, W.J. (1992) Secondary plasmodesmata are specific sites of localization of the tobacco mosaic virus movement protein in transgenic plants. Plant Cell 4, 915-28.
Ditta, G., Stanfield, S., Corbin, D. and Helsinki, D.R. (1980) Broad host range cloning system for Gram-negative bacteria: construction of a gene bank for Rhizobium meliloti. Proc. Natl Acad. Sci. USA 77, 7347-51.
Gallie, D.R., Sleat, D.E., Watts, J.W. and Turner, P.C. (1987) A comparison of eukaryotic viral 5′-leader sequences as enhancers of mRNA expression in vivo. Nucl. Acids Res. 15, 8693-711.
Gielen, J.J.L., De Haan, P., Kool, A.J., Peters, D., Van Grinsven, M.Q.J.M. and Goldbach, R.W. (1991) Engineered resistance to tomato spotted wilt virus, a negative-strand RNA virus. Bio/ Technology 9, 1363-7.
Goldbach, R. and Peters, D. (1994) Possible causes of the emergence of tospovirus diseases. Sem. Virol. 5, 113-20.
Horsch, R.B., Fry, J.E., Hoffmann, N.L., Eichholtz, D., Rogers, S.G. and Fraley, R.T. (1985) A simple method for transferring genes into plants. Science 227, 1229-31.
Kim, K.S. and Lee, K.W. (1992) Geminivirus-induced macrotubules and their suggested role in cell-to-cell movement. Phytopathology 82, 664-9.
Kormelink, R., De Haan, P., Meurs, C., Peters, D. and Goldbach, R. (1992) The nucleotide sequence of the M RNA segment of tomato spotted wilt virus, a bunyavirus with two ambisense RNA segments. J. Gen. Virol. 73, 2795-804.
Kormelink, R., Storms, M., Van Lent, J., Peters, D. and Goldbach, R. (1994) Expression and subcellular localization of the NSM protein of tomato spotted wilt virus (TSWV), a putative viral movement protein. Virology 200, 56-65.
Lucas, W.J., Ding, B. and Van der Schoot, C. (1993) Plasmodesmata and the supracellular nature of plants. New Phytol. 125, 435-76.
Murphy, F.A., Fauquet, C.M., Bishop, D.H.L., Ghabrial, S.A., Jarvis, A.W., Martilli, G.P., Mayo, M.A. and Summers, M.D. (eds.) (1995) Sixth Report of the International Committee on Taxonomy of Viruses. Wien, Austria: Springer-Verlag.
Pascal, E., Goodlove, P.E., Wu, L.C. and Lazarowitz, S.G. (1993) Transgenic tobacco plants expressing the geminivirus BL1 protein exhibit symptoms of viral disease. Plant Cell 5, 795-807.
Sambrook, J., Fritsch, E.F. and Maniatis, T. (1989) Molecular cloning: a Laboratory Manual, 2nd ed. Cold Spring Harbor, New York: Cold Spring Habor Laboratory Press.
Storms, M.M.H., Kormelink, R., Peters, D., Van Lent, J.W.M. and Goldbach, R.W. (1995) The nonstructural NSm protein of tomato spotted wilt virus induces tubular structures in plant and insect cells. Virol. 214, 485-93.
Van Lent, J., Wellink, J. and Goldbach, R. (1990) Evidence for the involvement of the 58K and 48K proteins in the intercellular movement of cowpea movement virus. J. Gen. Virol. 71, 219-23.
Vaquero, C., Turner, A.P., Demangeat, G., Sanz, A., Serra, M.T., Roberts, K. and Garcia-Luque, I. (1994) The 3a protein from cucumber mosaic virus increases the gating capacity of plasmodesmata in transgenic tobacco plants. J. Gen. Virol. 75, 3193-7.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Prins, M., Storms, M.M., Kormelink, R. et al. Transgenic tobacco plants expressing the putative movement protein of tomato spotted wilt tospovirus exhibit aberrations in growth and appearance. Transgenic Res 6, 245–251 (1997). https://doi.org/10.1023/A:1018450426401
Issue Date:
DOI: https://doi.org/10.1023/A:1018450426401