Skip to main content
SpringerLink
Log in

Reduction of potato mop-top virus accumulation and incidence in tubers of potato transformed with a modified triple gene block gene of PMTV

  • Published:
Molecular Breeding Aims and scope Submit manuscript

Abstract

Potato mop-top virus (PMTV) RNA 2 contains a triple gene block (TGB) that consists of a set of three overlapping genes. These genes encode three proteins necessary for viral cell-to-cell movement. The gene encoding for the second triple gene block protein, TGB2, was mutated in a region that is highly conserved among TGB2 proteins from different viruses. The mutated TGB2 gene, under transcriptional control of the 35S promoter from cauliflower mosaic virus, was transformed into potato (Solanum tuberosum cv. Hulda) by Agrobacterium-mediated transformation. Ten of the transgenic lines obtained, all of which were shown to transcribe the introduced gene, were evaluated for resistance to PMTV in a field trial. Tubers from the field were analysed for virus content by ELISA. In seven of the transgenic lines analysed there was a reduction in ELISA values relative to a non-transgenic control line. In the three lines that had the lowest ELISA readings this reduction was manifested both as a reduction in the proportion of infected tubers and as a reduction in virus levels in infected tubers. In the line showing the highest level of resistance, the total ELISA value was reduced by 79%. These results demonstrate that the introduction of a mutated TGB2 gene into potato can confer increased resistance to PMTV.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Anderson J.M., Palukaitis P. and Zaitlin M: A defective replicase gene induces resistance to cucumber mosaic virus in transgenic tobacco plants. Proc. Natl. Acad. Sci. USA 89: 8759–8763.

  2. Barker H., Reavy B. and McGeachy K.D. 1998a. High level of resistance in potato to potato mop-top virus induced by transformation with the coat protein gene. Eur. J. Plant Path. 104: 737–740.

    Google Scholar 

  3. Barker H., Reavy B., McGeachy K.D. and Dawson S. 1998b. Transformation of Nicotiana benthamiana with the potato mop-top virus coat protein gene produces a novel resistance phenotype mediated by the coat protein. Mol. Plant-Microbe Interact. 11: 626–633.

    Google Scholar 

  4. Beachy R.N. 1997. Mechanism and applications of pathogenderived resistance in transgenic plants. Curr. Opin. Biotechnol. 8: 215–220.

    Google Scholar 

  5. Beck E., Ludwig G., Auerswald E.A., Reiss B. and Schaller H. 1982. Nucleotide sequence and exact localization of the neomycin phosphotransferase gene from transposon Tn5. Gene 19: 327–336.

    Google Scholar 

  6. Beck D.L., Guilford P.J., Voot D.M., Andersen M.T. and Forster R.L.S. 1991. Triple gene block proteins of white clover mosaic potexvirus are required for transport. Virology 183: 695–702.

    Google Scholar 

  7. Beck D.L., Van Dollewerd C.J., Lough T.J., Balmori E., Voot D.M., Andersen M.T., O'Brien I.E.W. and Forster R.L.S. 1994. Disruption of virus movement confers broad-spectrum resistance against systemic infection by plant viruses with a triple gene block. Proc. Natl. Acad. Sci. USA 91: 10310–10314.

    Google Scholar 

  8. Collier G.H., Wurr D.C.E. and Huntington V.C. 1978. The effect of calcium nutrition on the incidence of internal rust spot in potato. J. Agric. Sci. 91: 249–243.

    Google Scholar 

  9. Davies H.V. and Millard P. 1985. Fractionation and distribution of calcium in sprouting and non-sprouting potato tubers. Ann Bot. 56: 745–754.

    Google Scholar 

  10. Erhardt M., Herzog E., Lauber E., Fritsch C., Guilley H., Jonard G. and Bouzoubaa S. 1999. Transgenic plants expressing the TGB1 protein of peanut clump virus complement movement of TGB1-defective peanut clump virus but not of TGB1-defective beet necrotic yellow vein virus. Plant Cell Rep. 18: 614–619.

    Google Scholar 

  11. Gilmer D., Gafny R., Ding B., Wolf S., LucasW.J. and Beachy R.N. 1992. Efficient cell-to-cell movement of beet necrotic yellow vein virus requires 3′ proximal genes located on RNA2. Virology 189: 40–47.

    Google Scholar 

  12. Herzog E., Hemmer O., Hauser S., Meyer G., Bouzoubaa S. and Fritsch C. 1998. Identification of genes involved in replication and movement of peanut clump virus. Virology 248: 312–322.

    Google Scholar 

  13. Hofvander P., Persson P.T., Tallberg A. and Wikström O. 1991. Genetically engineered modification of potato to form amylopectin-type starch. PCT application, WO 92/11376.

  14. Jones R.A.C. and Harrison B.D. 1969. The behaviour of potato mop-top virus in soil, and evidence for its transmission by Spongospora subterranea (Wallr.) Lagerh. Ann. Appl. Biol. 63: 1–17.

    Google Scholar 

  15. Lapidot M., Gafny R., Ding B., Wolf S., Lucas W.J. and Beachy R.N. 1993. A dysfunctional movement protein of tobacco mosaic virus that partially modifies the plasmodesmata and limits virus spread in transgenic plants. Plant J. 4: 959–970.

    Google Scholar 

  16. Lauber E., Bleykasten-Grosshans C., Erhardt M., Bouzoubaa S., Jonard G., Richards K.E. and Guilley H. 1998. Cell-to-cell movement of beet necrotic yellow vein virus. I. Heterologous complementation experiments provide evidence for specific interactions among the triple gene block proteins. Mol. Plant-Microbe Interact. 11: 618–625.

    Google Scholar 

  17. Lough T.J., Shash K., Xoconostle-Cazares B., Hofstra K.R., Beck D.L., Balmori E., Forster R.L.S. and Lucas W.J. 1998. Molecular dissection of the mechanism by which potexvirus triple gene block proteins mediate cell-to-cell transport of infectious RNA. Mol. Plant-Microbe Interact. 11: 801–814.

    Google Scholar 

  18. McGeachy K.D. and Barker H. 2000. Potato mop-top virus RNA can move long distance in the absence of coat protein: evidence from resistant, transgenic plants. Mol. Plant-Microbe Interact. 13: 125–128.

    Google Scholar 

  19. Powell-Abel P., Nelson R.S., De B., Hoffman N., Rogers S.G., Fraley R.T. and Beachy R.N. 1986. Delay of disease development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738–743.

    Google Scholar 

  20. Pringle C.R. 1999. Virus taxonomy - 1999. Arch. Virol. 144: 421–429.

    Google Scholar 

  21. Reavy B., Arif M., Kashiwazaki S., Webster K.D. and Barker H. 1995. Immunity to potato mop-top virus in Nicotiana benthamiana plants expressing the coat protein gene is effective against fungal inoculation of the virus. Mol. Plant-Microbe Interact. 8: 286–291.

    Google Scholar 

  22. Reavy B., Sandgren M., Barker H., Heino P. and Oxelfelt P. 1997. A coat protein transgene from a Scottish isolate of potato mop-top virus mediates strong resistance against Scandinavian isolates which have similar coat protein genes. Eur. J. Plant Path. 103: 829–834.

    Google Scholar 

  23. Rowhani A., Biardi L., Routh G. and Daubert S.D. 1998. Development of a sensitive colorimetric PCR assay for detection of virus in woody plants. Plant Dis. 8: 880–884.

    Google Scholar 

  24. Sambrook J., Fritsch E.F. and Maniatis T. 1989. Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Plainview, NY.

    Google Scholar 

  25. Sandgren M. 1995. Potato mop-top virus (PMTV): distribution in Sweden, development of symptoms during storage and cultivar trials in field and glasshouse. Potato Res. 38: 387–397.

    Google Scholar 

  26. Sandgren M. 1996. On spraing in potato. Ph.D. thesis, Swedish University of Agricultural Sciences, Uppsala.

    Google Scholar 

  27. Savenkov E.I., Sandgren M. and Valkonen J.P.T. 1999. Complete sequence of RNA 1 and the presence of tRNA-like structures in all RNAs of potato mop-top virus, genus Pomovirus. J. Gen. Virol. 80: 2779–2784.

    Google Scholar 

  28. Scott K.P., Kashiwazaki S., Reavy B. and Harrison B.D. 1994. The nucleotide sequence of potato mop-top virus RNA 2: a novel type of genome organization for a furovirus. J. Gen. Virol. 75: 3561–3568.

    Google Scholar 

  29. Seppänen P., Puska R., Honkanen J., Tyulkina L.G., Fedorkin O., Morozov S.Yu. and Atabekov J.G. 1997. Movement protein derived resistance to triple gene block-containing plant viruses. J. Gen. Virol. 78: 1241–1246.

    Google Scholar 

  30. Silva G.H., Chase R., Hammerschmidt M.L., Vitosh M.L. and Kitchen R.B. 1991. Irrigation, nitrogen and gypsum effects on specific gravity and internal defects on Atlantic potatoes. Am. Potato J. 68: 751–765.

    Google Scholar 

  31. Solovyev A.G., Savenkov E.I., Grdzelishvili V.Z., Kalinina N.O., Morozov S.Yu., Schiemann J. and Atabekov J.G. 1999. Movement of hordeivirus hybrids with exchanges in the triple gene block. Virology 253: 278–287.

    Google Scholar 

  32. Solovyev A.G., Stroganova T.A., Zamyatnin A.A., Fedorkin O.N., Schiemann J. and Morozov S.Yu. 2000. Subcellular sort-ing of small membrane-associated triple gene block proteins: TGBp3-assisted targeting of TGBp2. Virology 269: 113–127.

    Google Scholar 

  33. Visser R.G.F. 1991. Regeneration and transformation of potato by Agrobacterium tumefaciens. In: Lindsey K. (ed.), Plant Tissue Culture Manual, Kluwer Academic Publishers, Dordrecht, Netherlands. pp. B5/1–B5/9.

    Google Scholar 

  34. Visser R.G.F., Stolte A. and Jacobsen E. 1991. Expression of a chimaeric granule-bound starch synthase-GUS gene in transgenic potato plants. Plant Mol. Biol. 17: 691–699.

    Google Scholar 

  35. Walkerpeach C.R. and Velten J. 1994. Agrobacteriummediated gene transfer to plant cells: cointegrate and binary vector systems. In: Gelvin S.B. and Schilperoort R.A. (eds.), Plant Molecular Biology Manual, 2nd ed. Kluwer Academic Publishers, Dordrecht, Netherlands, pp. B1/1–B1/19.

    Google Scholar 

  36. Yang Y., Ding B., Baulcombe D.C. and Verchot J. 2000. Cell-to-cell movement of the 25K protein of potato virus X is regulated by three other viral proteins. Mol. Plant-Microbe Interact. 13: 599–605.

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Svalöf Weibull AB, 268 81 Svalöv, Sweden Plant Science Sweden AB, Herman Ehles väg 2-4, 268 31, Svalöv, Sweden

    Margareta Melander

  2. Sweden Scandinavian Biotechnology Research AB, Svalöf Weibull AB, 268 81 Svalöv, P.O. Box 166, 230 53, Alnarp, Sweden

    Michael Lee

  3. Department of Plant Biology, Swedish University of Agricultural Sciences, P.O. Box 7080, 750 07, Uppsala, Sweden

    Maria Sandgren

Authors
  1. Margareta Melander
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Michael Lee
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Sandgren
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Melander, M., Lee, M. & Sandgren, M. Reduction of potato mop-top virus accumulation and incidence in tubers of potato transformed with a modified triple gene block gene of PMTV. Molecular Breeding 8, 197–206 (2001). https://doi.org/10.1023/A:1013719726848

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1013719726848

Search

Navigation