Bio/Technology
10, 422 - 429 (1992)
doi:10.1038/nbt0492-422
Increased Resistance to Potato Virus X and Preservation of Cultivar Properties in Transgenic Potato Under Field ConditionsErik Jongedijk1, Antoon A. J. M. de Schutter2, Ton Stolte3, Peter J. M. van den Elzen1
& Ben J. C. Cornelissen3
1MOGEN International nv, Einsteinweg 97, 2333 CB Leiden, The Netherlands.
2De Schutter Farming Partnership, Olsterweg 30, 8256 RV Biddinghuizen, The Netherlands
3Foundation Inspection Service Noordzeepolders of the Dutch Inspection Service (NAK), P.O. Box 1115, 7801 BC Emmeloord, The Netherlands. During the last three years we performed field trials to assess levels of resistance against potato virus X (PVX) and changes in intrinsic properties of the potato cultivars Bintje and Escort upon the introduction of the PVX coat protein (CP) gene. Analysis of leaf and tuber samples collected in the field at two week intervals revealed a stable expression of the PVX CP gene throughout the growing season. This resulted in a large decrease in PVX incidence among clonal progeny obtained from previously infected Bintje and Escort clones. Based on evaluation of 50 defined morphological characteristics, tuber yield and grading, 81.8% of the Escort and 17.9% of the Bintje derived transgenic clones proved to be true to type. Overall lightsprout morphology was a useful critenon for the early detection of deviant transgenic clones. Using the polymerase chain reaction (PCR) with convergent primers spanning transgenic sequences, true to type clones could be distinguished unambiguously from the corresponding untransformed cultivars. Clear distinctions between independent transgenic clones could be made by inverted PCR (IPCR) diagnosis revealing integration− specific border fragments. These results demonstrate the commercial feasibility of improving potato cultivars by selectively adding new traits while preserving intrinsic properties, and the possibility of unambiguously identifying independent transgenic cultivars.
REFERENCES
- Harris, P.M. (Ed.) 1978. The potato Crop: The Scientific Basis for Improvement. Chapman & Hall, London, UK.
- Beemster, A.B.R. and de Bokx, J.A. 1987. Survey of properties and symptoms p. 84−140. In: Viruses of Potatoes and Seed-Potato Production, 2nd ed. J. A. de Bokx and J. P. H. van der Want, (Eds.). PUDOC, Wageningen, The Netherlands.
- Rochow, W.F. and Ross, F.A. 1995. Virus multiplication in plants double infected by potato viruses X and Y. Viroloy 1: 10−27.
- Kaniewski, W., Lawson, C., Sammons, B., Haley, L., Hart, J., Delannay, X. and Tumer, N.E. 1990. Field resistance of transgenic Russet potato to effects of infection potato virus X and potato virus Y. Bio/Technology 8: 750−754.
- Powell Abel, P., Nelson, R.S., De, B., Hoffman, N., Rogers, S.G., Fraley, R.T. and Beachy, R.N. 1986. Delay of symptom development in transgenic plants that express the tobacco mosaic virus coat protein gene. Science 232: 738−743.
- Tumer, N.E., O'Connell, K.M., Nelson, R.S., Sanders, P.R., Beachy, R.N., Fraley, R.T. and Shah, D.M. 1987. Expression of alfalfa mosaic virus coat protein gene confers cross-protection in transgenic tobacco and tomato plants. EMBO J. 6: 1181−1188.
- Loesch-Fries, L.S., Merlo, D., Zinnen, T., Burhop, L., Hill, K., Krahn, K., Jarvis, N., Nelson, S. and Halk, E. 1987. Expression of alfalfa mosaic virus RNA 4 in transgenic plants confers virus resistance. EMBO J. 6: 1845−1851.
- Van Dun, M.P., Bol, J.F. and Van Vloten-Doting, L. 1987. Expression of alfalfa mosaic virus and tobacco rattle virus coat protein genes in transgenic tobacco plants. Virology 159: 299−305.
- Hemenway, C., Fang, R.X., Kaniewski, W., Chua, N.-H. and Tumer, N.E. 1988. Analysis of the mechanism of protection in transgenic plants expressing potato virus X protein or its antisense RNA. EMBO J. 7: 1273−1280.
- Cuozzo, M., O'Connell, K.M., Kaniewski, W., Fang, R.X., chua, N-H. and Turner, N.E. 1988. Viral protection in transgenic tobacco plants expressing the cucumber mosaic virus coat protein or its antisense RNA. Bio/Technology 6: 549−557.
- Nelson, R.S., McCormick, S.H., Delannay, X., Dubé, P., Leyton, J., Anderson, E.J., Koniewska, M., Proksch, R.K., Horsch, R.B., Rogers, S.G., Fraley, R.T. and Beachy, R.N. 1988. Virus tolerance, plant growth and field performance of transgenic tomato plants expressing coat protein from tobacco mosaic virus. Bio/Technology 6: 403−409.
- Beachy, R.N., Loesch-Fries, S. and Tumer, N. 1990. Coat protein-mediated resistance against virus infection. Ann. Rev. Phytopathol. 28: 451−474.
- Hill, K.K., Jarvis-Eagan, N., Halk, E.L., Krahn, K.J., Liao, L.W., Matheson, R.S., Merlo, D.J., Nelson, S.E., Rashka, K.E. and Loesch-Fries, L. 1991. The development of virus-resistant alfalfa, Medicago sativa L. Bio/Technology 9: 373−377.
- Ling, K., Namba, S., Gonsalves, C., Slightom, J.L. and Gonsalves, D. 1991. Protection against detrimental effects of potyvirus infection in transgenic tobacco plants expressing the papaya ringspot virus coat protein gene. Bio/Technology 9: 752−758.
- Hoekema, A., Huisman, M.J., Molendijk, L.,, van den Elzen, P.J.M., and Cornelissen, B.J.C. 1989. The genetic engineering of two commercial potato cultivars for resistance potato virus X. Bio/Technology 7: 273−278.
- Lawson, C., Kaniewski, W., Haley, L., Rozman, R., Newell, C., Sanders, P. and Tumer, N.E. 1990. Engineering resistance to mixed virus infection in a commercial potato cultivar: Resistance to potato virus X and potato virus Y in transgenic Russet Burbank. Bio/Technology8: 127−134.
- Kawchuk, L.M., Martin, R.R. and McPherson, J. 1991. Sense and antisense RNA-mediated resistance to potato leafroll virus in Russet Burbank potato plants. Molecular Plant-Microbe Interactions 4: 247−253.
- Van der Wilk, F., Postumus-Lutke Wilink, D., Huisman, M.J., Huttinga, H. and Goldbach, R. 1991. Expression of the potato leafroll luteovirus coat protein gene in transgenic potato plants inhibits viral infection. Plant Mol. Biol. 17: 431−439.
- Huisman, M.J., Linthorst, H.J.M., Bol, J.F. and Cornelissen, B.J.C. 1988. The complete nucleotide sequence of potato virus X and its homologies at the amino acid level with various plus-stranded RNA viruses. J. Gen. Virol. 69: 1789−1798.
- Parlevliet, J.E., Van der Zaag, D.E. and Bakker, J.J. 1991. 66th descriptive Variety List of agronomic crops 1991. Leiter-Nypels, Maastricht, The Netherlands.
- International Union for the Protection of New Varieties of Plants (UPOV). 1986. Guidelines for the conduct of tests for distinctness, homogeneity and stability of potato (Solanum tuberosum).
- Karp, A. and Bright, S.W.J. 1985. On the causes and origins of somaclonal variation. Oxford Surveys of Plant Molecular and Cell Biology 2: 199−234.
- Lee, M. and Phillips, R.L. 1988. The chromosomal basis of somaclonal variation. Ann. Rev. Plant Physiol. Plant Mol. Biol. 39: 413−437.
- Houwing, A., Suk, R. and Ros, B. 1986. Generation of lightsprouts suitable for potato variety identification by means of artificial light. Acta Horticulturae 182: 359−363.
- Winer, B.J., 1971. Statistical Principles in Experimental Design, 2nd ed. McGraw-Hill, New York.
- Clark, M.F. and Adams, A.N. 1977. Characteristics of the microplate method of enzyme-linked immunosorbent assay for the detection of plant viruses. J. Gen. Virol. 34: 475−483.
- Van Slogteren, G.M.S., Hoge, H.C., Hooykaas, P.J.J. and Schilperoort, R.A. 1983. Clonal analyses of heterogeneous crown gall tumor tissues induced by wild type and shooter mutant strains of Agrobacterium tumefaciens expression of T-DNA genes. Plant Mol. Biol. 2: 321−333.
- Does, M.P., Dekker, B.M.M., De Groot, M.J.A. and Offringa, R.1991. A quick method to estimate the T-DNA copy number in transgenic plants at an early stage after transformation, using inverted PCR. Plant Molec. Biol. 17: 151−153.
|
