A transformation method for obtaining marker-free plants of a cross-pollinating and vegetatively propagated crop View Full Text


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Article Info

DATE

2003-03-10

AUTHORS

Nick de Vetten, Anne-Marie Wolters, Krit Raemakers, Ingrid van der Meer, Renaldo ter Stege, Els Heeres, Paul Heeres, Richard Visser

ABSTRACT

It is generally thought that transformation of plant cells using Agrobacterium tumefaciens occurs at a very low frequency. Therefore, selection marker genes are used to identify the rare plants that have taken up foreign DNA. Genes encoding antibiotic and herbicide resistance are widely used for this purpose in plant transformation1,2. Over the past several years, consumer and environmental groups have expressed concern about the use of antibiotic- and herbicide-resistance genes from an ecological and food safety perspective. Although no scientific basis has been determined for these concerns, generating marker-free plants would certainly contribute to the public acceptance of transgenic crops. Several methods have been reported to create marker gene–free transformed plants, for example co-transformation, transposable elements, site-specific recombination, or intrachromosomal recombination3,4,5,6,7,8,9. Not only are most of these systems time-consuming and inefficient, but they are also employed on the assumption that isolation of transformants without a selective marker gene is not feasible10. Here we present a method that permits the identification of transgenic plants without the use of selectable markers. This strategy relies on the transformation of tissue explants or cells with a virulent A. tumefaciens strain and selection of transformed cells or shoots after PCR analysis. Incubation of potato explants with A. tumefaciens strain AGL0 resulted in transformed shoots at an efficiency of 1–5% of the harvested shoots, depending on the potato genotype used. Because this system does not require genetic segregation or site-specific DNA-deletion systems to remove marker genes, it may provide a reliable and efficient tool for generating transgenic plants for commercial use, especially in vegetatively propagated species like potato and cassava. More... »

PAGES

439-442

References to SciGraph publications

  • 2001-02. Chemical-regulated, site-specific DNA excision in transgenic plants in NATURE BIOTECHNOLOGY
  • 2001-06. Efficient production of transgenic plants by Agrobacterium-mediated transformation of cassava (Manihot esculenta Crantz) in EUPHYTICA
  • 1993-11. Transposition Mediated Re–positioning and Subsequent Elimination of Marker Genes from Transgenic Tomato in NATURE BIOTECHNOLOGY
  • 1996-12. Production of transgenic cassava (Manihot esculenta Crantz) plants by particle bombardment using luciferase activity as selection marker in MOLECULAR BREEDING
  • 1995-11. Factors affecting the inhibition by antisense RNA of granule-bound starch synthase gene expression in potato in MOLECULAR GENETICS AND GENOMICS
  • 2001-02. The right chemistry for marker gene removal? in NATURE BIOTECHNOLOGY
  • 1999-05. Selectable marker-free transgenic plants without sexual crossing: transient expression of cre recombinase and use of a conditional lethal dominant gene in PLANT MOLECULAR BIOLOGY
  • 1983-05. A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid in NATURE
  • 1991-10. A DNA Transformation–Competent Arabidopsis Genomic Library in Agrobacterium in BIO/TECHNOLOGY
  • 2000-04. Intrachromosomal recombination between attP regions as a tool to remove selectable marker genes from tobacco transgenes in NATURE BIOTECHNOLOGY
  • 2002. Transformation of a large number of potato varieties: genotype-dependent variation in efficiency and somaclonal variability in EUPHYTICA
  • 1983-07. A chimaeric antibiotic resistance gene as a selectable marker for plant cell transformation in NATURE
  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/nbt801

    DOI

    http://dx.doi.org/10.1038/nbt801

    DIMENSIONS

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    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/12627169


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