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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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
Aoyama T & Chua N-H (1997) A glucocorticoid-mediated transcriptional induction system in transgenic plants. Plant J. 11: 605-612
Aziz N & Machray GC (2003) Efficient male germ line transformation for transgenic tobacco production without selection. Plant Mol. Biol. 23: 203-211
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
Bock R (2001) Transgenic plastids in basic research and plant biotechnology. J. Mol. Biol. 312: 425-438
Buchholz F & Stewart AF (2001) Alteration of Cre recombinase site specificity by substrate-linked protein evolution. Nat. Biotechnol. 19: 1047-1052
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
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
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
Dale PJ, Clarke B & Fontes EMG (2002) Potential for the environmental impact of transgenic crops. Nat. Biotech. 20: 567-574
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
Daniell H (2002) Molecular strategies for gene containment in transgenic crops. Nat. Biotech. 20: 581-586
Daniell H, Khan MS & Allison L (2002) Milestones in chloroplast genetic engineering: an environmentally friendly era in biotechnology. Trends Plant Sci. 7: 84-91
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
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
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
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
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
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
Ebinuma H, Sugita K, Matsunaga E, Yamakado M & Komamine A (1997b) Principle of MAT vector. Plant Biotechnol. 14: 133-139
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
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
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
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
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
Gorbunova V & Levy AA (1999) How plants make ends meet: DNA double-strand break repair. Trends Plant Sci. 4: 263-269
Gorbunova V & Levy AA (2000) Analysis of extrachromosomal Ac/Ds transposableelements. Genetics 155: 349-359
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
Hajdukiewicz PT, Gilbertson L & Staub JM (2001) Multiple pathways for Cre/ lox-mediated recombination in plastids. Plant J. 27: 161-70
Hare P & Chua N-H (2002) Eviction of selectable marker genes from transgenic plants. Nat. Biotech. 20: 575-580
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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