Molecular Breeding

, Volume 11, Issue 4, pp 315–323 | Cite as

A selection system for transgenic plants based on galactose as selective agent and a UDP-glucose:galactose-1-phosphate uridyltransferase gene as selective gene

  • Morten Joersbo
  • Kirsten Jørgensen
  • Janne Brunstedt


A new selection system based on galactose as selective agent and a UDP-glucose:galactose-1-phosphate uridyltransferase gene as selective gene is presented. A broad range of plant species, including agronomically important crops such as maize and rice, is sensitive to low dosages of galactose. The toxicity of galactose is believed to be due to accumulation of galactose-1-phosphate, generated by endogenous galactokinase after uptake. Here, it is demonstrated that this toxicity can be sufficiently alleviated by the Agrobacterium tumefaciens-mediated introduction of the E. coli UDP-glucose:galactose-1-phosphate uridyltransferase (galT) gene, driven by a 35S-promoter, to allow transgenic shoots of potato and oil seed rape to regenerate on galactose containing selection media, resulting in high transformation frequencies (up to 35% for potato). Analysis of genomic DNA and UDP-glucose:galactose-1-phosphate uridyltransferase activity in randomly selected potato transformants confirmed the presence and active expression of the galT gene. The agricultural performance of transgenic potatoes was evaluated by monitoring the phenotype and tuber yield for two generations and these characters were found to be indistinguishable from non-transgenic controls. Thus, the galactose selection system provides a new alternative being distinct from conventional antibiotic and herbicide selection systems as well as so-called positive selection systems where the selective agent has a beneficial effect.

Antibiotic-independent selection Carbohydrate selection Oil seed rape Plant transformation Potato 


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  1. ACNFP 1994. Report on the use of antibiotic resistance markers in genetically modified food organisms. Advisory Committee on Novel Foods and Processes. Dept. of Health and Ministry of Agriculture, Fisheries and Food, London.Google Scholar
  2. Bowen B.A. 1993. Markers for plant gene transfer. In: Kung S.D. and Wu R. (eds), Transgenic Plants. Vol. 1. Academic Press, San Diego, CA, USA, pp. 89–146.Google Scholar
  3. Cornwell T.L., Adhya S.L., Reznikoff W.S. and Frey P.A. 1987. The nucleotide sequence of the galT gene of Escherichia coli. Nucl. Acids Res. 15: 8116.Google Scholar
  4. Dellaporta S.L., Wood J. and Hicks J.B. 1983. A plant DNA mini-preparation: version II. Plant Mol. Biol. Rep. 1: 19–21.Google Scholar
  5. Dey P.M. 1980. Biochemistry of βD-galactosidic linkages in the plant kingdom. In: Tipson R.S. and Horton D. (eds), Advances in Carbohydrate Chemistry and Biochemistry. Vol. 37. Academic Press, New York, USA, pp. 342–345.Google Scholar
  6. Dressler K., Biedlingmaier S., Grossberger H., Kemmer J., Nölle U., Rodmanis-Blumer A. et al. 1982. Galactose metabolism in Petunia. Z. Pflanzenphysiol. 107: 409–418.Google Scholar
  7. Farkas G.L. 1954. Die toxische Wirkung einiger Zucker auf die Würzeln der Pflanzen. Zuckerantagonismen. Biol. Zentralblatt. 73: 506–521.Google Scholar
  8. Gamborg O.L., Miller R.A. and Ojima K. 1968. Nutrient requirements of suspension cultures of soybean root cells. Exp. Cell Res. 50: 151–158.Google Scholar
  9. Gross K.C., Pharr D.M. and Locy R.D. 1981. Growth of callus initiated from cucumber hypocotyls on galactose and galactose-containing oligosaccharides. Plant Science Lett. 20: 333–341.Google Scholar
  10. Haldrup A., Petersen S.G. and Okkels F.T. 1998. The xylose isomerase gene from Thermoanaerobacterium thermosulfogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent. Plant Mol. Biol. 37: 287–296.Google Scholar
  11. Heldt H.W. 1997. Plant Biochemistry and Molecular Biology. Oxford Univ. Press, Oxford, pp. 306–308.Google Scholar
  12. Hughes R. and Street H.E. 1974. Galactose as an inhibitor of the expansion of roots cells. Ann. Bot. 38: 555–564.Google Scholar
  13. Jefferson R.A., Burgess S.M. and Hirsh D. 1986. β-Glucuronidase from Escherichia coli as a gene-fusion marker. Proc. Natl. Acad. Sci. 83: 8447–8451.Google Scholar
  14. Jefferson R.A., Kavanagh T.A. and Bevan M.W. 1987. GUS-fusions: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J. 6: 3901–3907.Google Scholar
  15. Joersbo M. 2001. Advances in the selection of transgenic plants using non-antibiotic marker genes. Physiol. Plant. 111: 269–272.Google Scholar
  16. Joersbo M., Donaldson I., Kreiberg J., Petersen S.G., Brunstedt J. and Okkels F.T. 1998. Analysis of mannose selection used for transformation of sugar beet. Mol. Breeding 4: 111–117.Google Scholar
  17. Joersbo M., Petersen S.G. and Okkels F.T. 1999. Parameters interacting with mannose selection employed for the production of transgenic sugar beet. Physiol. Plant. 105: 109–115.Google Scholar
  18. Kay R., Chan A., Daly M. and McPherson J. 1987. Duplication of CaMV 35S promoter sequences creates a strong enhancer for plant genes. Science 236: 1299–1302.Google Scholar
  19. Knittel N., Gruber V., Hahne G. and Lénée P. 1994. Transformation of sunflower (Helianthus annuus L.): a reliable protocol. Plant Cell Rep. 14: 81–86.Google Scholar
  20. Leonard R.T. and Hodges T.K. 1980. The Plasma Membrane. In: Tolbert N.E. (ed.), The Biochemistry of Plants. Vol. 1. Academic Press, New York, pp. 163–182.Google Scholar
  21. Minocha S.C. and Halperin W. 1974. Hormones and metabolites which control tracheid differentiation, with or without concomitant effects on growth, in cultured tuber tissue of Helianthus tuberosus L. Planta 116: 319–331.Google Scholar
  22. Misra S. and Gedamu L. 1989. Heavy metal tolerant transgenic Brassica napus L. and Nicotiana tabacum L. plants. Theor. Appl. Genet. 78: 161–168.Google Scholar
  23. Murashige T. and Skoog F. 1962. A revised medium for rapid growth and bioassay with tobacco tissue cultures. Physiol. Plant. 15: 473–497.Google Scholar
  24. Neufeld E.F., Feingold D.S. and Hassid W.Z. 1994. Phosphorylation of D-galactose and L-arabinose by extracts of Phaseolus areus seedlings. J. Biol. Chem. 235: 450–459.Google Scholar
  25. Nielsen S.V.S., Poulsen G.B. and Larsen M.E. 1991. Regeneration of shoots from pea (Pisum sativum) hypocotyl explants. Physiol. Plant. 82: 99–102.Google Scholar
  26. Odell J.T., Nagy F. and Chua N.H. 1985. Identification of DNA sequences required for activity of the cauliflower mosaic virus 35S promoter. Nature 313: 810–812.Google Scholar
  27. Ordin L. and Bonner J. 1956. Effect of galactose on growth and metabolism of Avena coleoptile sections. Plant Physiol. 32: 212–215.Google Scholar
  28. Roberts R.M. and Butt V.S. 1969. Patterns of incorporation of D-galactose into cell wall polysaccharides of growing maize roots. Planta 84: 250–262.Google Scholar
  29. Roberts R.M., Heishman A. and Wicklin C. 1971. Growth inhibition and metabolite pool levels in plant tissues fed D-glu-cosamine and D-galactose. Plant Physiol. 48: 36–42.Google Scholar
  30. Smart E. and Pharr D.M. 1981. Separation and characteristics of galactose-1-phosphate and glucose-1-phosphate uridyltransferase from fruit peduncles of cucumber. Planta 153: 370–375.Google Scholar
  31. Thorpe T.A. and Meier D.D. 1972. Starch metabolism, respiration, and shoot formation in tobacco callus cultures. Physiol. Plant. 27: 365–369.Google Scholar
  32. Vancanneyt G., Schmidt R., O'Connor-Sanchez A., Willmitzer L. and Rocha-Sosa M. 1990. Construction of an intron-containing marker gene: splicing of the intron in transgenic plants and its use in monitoring early events in Agrobacterium-mediated plant transformation. Mol. Gen. Genet. 220: 245–250.Google Scholar
  33. Zöllner N. and Heuckenkamp P.U. 1970. β-D-Galactose-1-phos-phate: Bestimmung mit Uridyl-transferase. In: Bergmeyer H.U. (ed.), Methoden der enzymatischen Analyse. Vol. 2. Verlag Chemie, Weinheim, Germany, pp. 1250–1253.Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  • Morten Joersbo
    • 1
  • Kirsten Jørgensen
    • 2
  • Janne Brunstedt
    • 3
  1. 1.Danisco SeedHolebyDenmark
  2. 2.Department of Plant Biology, Plant Biochemistry LaboratoryRoyal Veterinary and Agricultural UniversityFrederiksberg CDenmark
  3. 3.Danisco InnovationCopenhagen KDenmark

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