Plant Cell, Tissue and Organ Culture

, Volume 74, Issue 2, pp 123–134

Marker-free transgenic plants

Abstract

Selectable marker genes are widely used for the efficient transformation of crop plants. In most cases, selection is based on antibiotic or herbicide resistance. Due mainly to consumer concerns, a suite of strategies (site-specific recombination, homologous recombination, transposition and co-transformation) have been developed to eliminate the marker gene from the nuclear or chloroplast genome after selection. Current efforts concentrate on systems where marker genes are eliminated efficiently soon after transformation. Alternatively, transgenic plants are produced by the use of marker genes that do not rely on antibiotic or herbicide resistance but instead promote regeneration after transformation. Here, the merits and shortcomings of different approaches and possible directions for their future development are discussed.

homologous recombination induction non-homologous end joining screenable markers site-specific recombination transformation transposition 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. Albert H, Dale EC, Lee E & Ow DW (1995) Site-specific integration of DNA into wild-type and mutant lox sites placed in the plant genome. Plant J. 7: 649-659Google Scholar
  2. Aoyama T & Chua N-H (1997) A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J. 11: 605-612Google Scholar
  3. Aziz N & Machray GC (2003) Efficient male germ line transformation for transgenic tobacco production without selection. Plant Mol. Biol. 23: 203-211Google Scholar
  4. Bibikova M, Carroll D, Segal DJ, Trautman JK, Smith J, Kim YG & Chandrasegaran S (2001) Stimulation of homologous recombination through targeted cleavage by chimeric nucleases. Mol. Cell. Biol. 21: 289-297Google Scholar
  5. Bock R (2001) Transgenic plastids in basic research and plant biotechnology. J. Mol. Biol. 312: 425-438Google Scholar
  6. Buchholz F & Stewart AF (2001) Alteration of Cre recombinase site specificity by substrate-linked protein evolution. Nat. Biotechnol. 19: 1047-1052Google Scholar
  7. Coppoolse EC, de Vroomen, Roelofs MJD, Smit J, van Gennip F, Hersmus BJM, Nijkamp HJJ & van Haaren MJJ (2003) Cre recombinase expression can result in phenotypic aberrations in plants. Plant Mol. Biol. 23: 263-279Google Scholar
  8. Corneille S, Lutz K, Svab Z & Maliga P (2001) Efficient elimination of selectable marker genes from the plastid genome by the CRE-lox site-specific recombination system. Plant J. 27: 171-178Google Scholar
  9. Dale EC & Ow DW(1991) Gene transfer with subsequent removal of the selection gene from the host genome. Proc. Natl. Acad. Sci. USA 23: 10558-10562Google Scholar
  10. Dale PJ, Clarke B & Fontes EMG (2002) Potential for the environmental impact of transgenic crops. Nat. Biotech. 20: 567-574Google Scholar
  11. Daley M, Knauf VC, Summerfelt KR & Turner JC (1998) Co-transformation with one Agrobacterium tumefaciens strain containing two binary plasmids as a method for producing marker-free transgenic plants. Plant Cell Rep. 17: 489-496Google Scholar
  12. Daniell H (2002) Molecular strategies for gene containment in transgenic crops. Nat. Biotech. 20: 581-586Google Scholar
  13. Daniell H, Khan MS & Allison L (2002) Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology. Trends Plant Sci. 7: 84-91Google Scholar
  14. Davies GJ, Kilby J, Riou-Khamlichi C & Murray JAH (1999) Somatic and germinal inheritance of an FLP-mediated deletion in transgenic tobacco. J. Exp. Bot. 50: 1447-1456Google Scholar
  15. De Block M & Debrouwer D (1991) Two T-DNAs co-transformed into Brassica napus by a double Agrobacterium infection are mainly integrated at the same locus. Theor. Appl. Genet. 82: 257-263Google Scholar
  16. De Neve M, De Buck S, Jacobs A, Van Montagu M & Depicker A (1997) T-DNA integration patterns in co-transformed plant cells suggest that T-DNA repeats originate from co-integration of separate T-DNAs. Plant J. 11: 15-29Google Scholar
  17. Depicker A, Herman L, Jacobs S, Schell J & van Montagu M (1985) Frequencies of simultaneous transformation with different T-DNAs and their relevance to the Agrobacterium plant cell interaction. Mol. Gen. Genet. 201: 477-484Google Scholar
  18. Ebinuma H & Komamine A (2001) MAT (Multi-Auto-Transformation) Vector System. The oncogenes of Agrobacterium as positive markers for regeneration and selection of marker-free transgenic plants. In Vitro Cell. Dev. Biol. Plant 37: 103-113Google Scholar
  19. Ebinuma H, Sugita K, Matsunaga E & Yamakado M (1997a) Selection of marker-free transgenic plants using the isopentenyl transferase gene. Proc. Natl. Acad. Sci. USA 94: 2117-2121Google Scholar
  20. Ebinuma H, Sugita K, Matsunaga E, Yamakado M & Komamine A (1997b) Principle of MAT vector. Plant Biotechnol. 14: 133-139Google Scholar
  21. Ebinuma H, Sugita E, Matsunaga E, Endo S, Yamada K & Komamine A (2001) Systems for the removal of a selection marker and their combination with a positive marker. Plant Cell Rep. 20: 383-392Google Scholar
  22. Endo S, Sugita K, Sakai M, Tanaka H & Ebinuma H (2002) Single-step transformation for generating marker-free transgenic rice using the ipt-type MAT vector system. Plant J. 30: 115-122Google Scholar
  23. Gidoni D, Bar M, Leshem B, Gilboa N, Mett A & Feiler J (2001) Embryonal recombination and germline inheritance of recombined FRT loci mediated by constitutively expressed FLP in tobacco. Euphytica 121: 145-156Google Scholar
  24. Gleave AP, Mitra DS, Mudge SR & Morris BA (1999) Selectable marker-free transgenic plants without sexual crossing: transient expression of cre recombinase and use of a conditional lethal dominant gene. Plant Mol. Biol. 40: 223-235Google Scholar
  25. Goldsbrough AP, Lastrella CN & Yoder JI (1993) Transposition mediated re-positioning and subsequent elimination of marker genes from transgenic tomato. Bio/technology 11: 1286-1292Google Scholar
  26. Gorbunova V & Levy AA (1999) How plants make ends meet: DNA double-strand break repair. Trends Plant Sci. 4: 263-269Google Scholar
  27. Gorbunova V & Levy AA (2000) Analysis of extrachromosomal Ac/Ds transposableelements. Genetics 155: 349-359Google Scholar
  28. Haldrup A, Petersen SG & Okkels FT (1998) The xylose isomerase gene from Thermoanaerobacterim thermosulfurogenes allows effective selection of transgenic plant cells using D-xylose as the selection agent. Plant Mol. Biol. 37: 287-296Google Scholar
  29. Hajdukiewicz PT, Gilbertson L & Staub JM (2001) Multiple pathways for Cre/ lox-mediated recombination in plastids. Plant J. 27: 161-70Google Scholar
  30. Hare P & Chua N-H (2002) Eviction of selectable marker genes from transgenic plants. Nat. Biotech. 20: 575-580Google Scholar

Copyright information

© Kluwer Academic Publishers 2003

Authors and Affiliations

  1. 1.Botany IIUniversität KarlsruheKarlsruheGermany

Personalised recommendations