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Marker-free transgenic plants

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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.

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References

  • 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: 649-659

    Google Scholar 

  • Aoyama T & Chua N-H (1997) A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J. 11: 605-612

    Google Scholar 

  • Aziz N & Machray GC (2003) Efficient male germ line transformation for transgenic tobacco production without selection. Plant Mol. Biol. 23: 203-211

    Google Scholar 

  • 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: 289-297

    Google Scholar 

  • Bock R (2001) Transgenic plastids in basic research and plant biotechnology. J. Mol. Biol. 312: 425-438

    Google Scholar 

  • Buchholz F & Stewart AF (2001) Alteration of Cre recombinase site specificity by substrate-linked protein evolution. Nat. Biotechnol. 19: 1047-1052

    Google Scholar 

  • 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: 263-279

    Google Scholar 

  • 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: 171-178

    Google Scholar 

  • Dale EC & Ow DW(1991) Gene transfer with subsequent removal of the selection gene from the host genome. Proc. Natl. Acad. Sci. USA 23: 10558-10562

    Google Scholar 

  • Dale PJ, Clarke B & Fontes EMG (2002) Potential for the environmental impact of transgenic crops. Nat. Biotech. 20: 567-574

    Google Scholar 

  • 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: 489-496

    Google Scholar 

  • Daniell H (2002) Molecular strategies for gene containment in transgenic crops. Nat. Biotech. 20: 581-586

    Google Scholar 

  • Daniell H, Khan MS & Allison L (2002) Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology. Trends Plant Sci. 7: 84-91

    Google Scholar 

  • 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: 1447-1456

    Google Scholar 

  • 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: 257-263

    Google Scholar 

  • 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: 15-29

    Google Scholar 

  • 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: 477-484

    Google Scholar 

  • 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: 103-113

    Google Scholar 

  • 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: 2117-2121

    Google Scholar 

  • Ebinuma H, Sugita K, Matsunaga E, Yamakado M & Komamine A (1997b) Principle of MAT vector. Plant Biotechnol. 14: 133-139

    Google Scholar 

  • 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: 383-392

    Google Scholar 

  • 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: 115-122

    Google Scholar 

  • 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: 145-156

    Google Scholar 

  • 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: 223-235

    Google Scholar 

  • Goldsbrough AP, Lastrella CN & Yoder JI (1993) Transposition mediated re-positioning and subsequent elimination of marker genes from transgenic tomato. Bio/technology 11: 1286-1292

    Google Scholar 

  • Gorbunova V & Levy AA (1999) How plants make ends meet: DNA double-strand break repair. Trends Plant Sci. 4: 263-269

    Google Scholar 

  • Gorbunova V & Levy AA (2000) Analysis of extrachromosomal Ac/Ds transposableelements. Genetics 155: 349-359

    Google Scholar 

  • 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: 287-296

    Google Scholar 

  • Hajdukiewicz PT, Gilbertson L & Staub JM (2001) Multiple pathways for Cre/ lox-mediated recombination in plastids. Plant J. 27: 161-70

    Google Scholar 

  • Hare P & Chua N-H (2002) Eviction of selectable marker genes from transgenic plants. Nat. Biotech. 20: 575-580

    Google Scholar 

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Correspondence to Holger Puchta.

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Puchta, H. Marker-free transgenic plants. Plant Cell, Tissue and Organ Culture 74, 123–134 (2003). https://doi.org/10.1023/A:1023934807184

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  • DOI: https://doi.org/10.1023/A:1023934807184

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