Asexual production of selectable marker-free transgenic woody plants, vegetatively propagated species View Full Text


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Article Info

DATE

2002-09

AUTHORS

Etsuko Matsunaga, Koichi Sugita, Hiroyasu Ebinuma

ABSTRACT

We describe here a practical system for generating selectable marker-free transgenic woody plants independent of sexual crossing. We previously reported that the GST-MAT vector system could produce marker-free transgenic tobacco plants containing a single-copy transgene at high frequency. The GST-MAT vector system consists of a DNA excision cassette of the R/RS site-specific recombination system from Zygosaccharomyces rouxii into which the isopentenyltransferase gene from Agrobacterium tumefaciens has been inserted. In this study, we applied this new GST-MAT vector to hybrid aspen (Populus Sieboldii X Populus grandidentata), a model of vegetatively propagated plant species, to produce selectable marker-free transgenic woody plants. In the new GST-MAT vector, the chimeric ipt gene fused with a light-inducible rbcS promoter efficiently produced transgenic ipt-shooty with GUS activity from 38.0% of infected stems. Upon excision of the R and ipt genes between RS sites, regulated by the inducible promoter of the maize glutathione-S-transferase (GST-II-27) gene, 3 (21.4%) transgenic hybrid aspen plants with marker-free and normal phenotype were generated from 14 ipt-shooty lines within 2 months after cutting induction. These results suggest that the MAT-vector system might be useful for removing a selectable marker gene independent of sexual crossing in vegetatively propagated species. More... »

PAGES

95-106

References to SciGraph publications

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  • 1996-12. A novel principle for selection of transgenic plant cells: positive selection in PLANT CELL REPORTS
  • 2002-03. A new GST-MAT vector containing both ipt and iaaM/H genes can produce marker-free transgenic tobacco plants with high frequency in PLANT CELL REPORTS
  • 1998-11. Positive selection: a plant selection principle based on xylose isomerase, an enzyme used in the food industry in PLANT CELL REPORTS
  • 2001-03. Mat (Multi-Auto-Transformation) vector system. The oncogenes of Agrobacterium as positive markers for regeneration and selection of marker-free transgenic plants in IN VITRO CELLULAR & DEVELOPMENTAL BIOLOGY - PLANT
  • 2001-02. Chemical-regulated, site-specific DNA excision in transgenic plants in NATURE BIOTECHNOLOGY
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  • 2002. GST-MAT Vector for the Efficient and Practical Removal of Marker Genes from Transgenic Plants in TESTING FOR GENETIC MANIPULATION IN PLANTS
  • 1987-09. Genomic organization, sequence analysis and expression of all five genes encoding the small subunit of ribulose-1,5-bisphosphate carboxylase/oxygenase from tomato in MOLECULAR GENETICS AND GENOMICS
  • 2000. Selection of Marker-Free Transgenic Plants Using the Oncogenes (ipt, rol A, B, C) of Agrobacterium as Selectable Markers in MOLECULAR BIOLOGY OF WOODY PLANTS
  • 1986-09. Benzyladenine modulation of the expression of two genes for nuclear-encoded chloroplast proteins in Lemna gibba: Apparent post-transcriptional regulation in PLANTA
  • Journal

    TITLE

    Molecular Breeding

    ISSUE

    1-2

    VOLUME

    10

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1020308216747

    DOI

    http://dx.doi.org/10.1023/a:1020308216747

    DIMENSIONS

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    38 schema:description We describe here a practical system for generating selectable marker-free transgenic woody plants independent of sexual crossing. We previously reported that the GST-MAT vector system could produce marker-free transgenic tobacco plants containing a single-copy transgene at high frequency. The GST-MAT vector system consists of a DNA excision cassette of the R/RS site-specific recombination system from Zygosaccharomyces rouxii into which the isopentenyltransferase gene from Agrobacterium tumefaciens has been inserted. In this study, we applied this new GST-MAT vector to hybrid aspen (Populus Sieboldii X Populus grandidentata), a model of vegetatively propagated plant species, to produce selectable marker-free transgenic woody plants. In the new GST-MAT vector, the chimeric ipt gene fused with a light-inducible rbcS promoter efficiently produced transgenic ipt-shooty with GUS activity from 38.0% of infected stems. Upon excision of the R and ipt genes between RS sites, regulated by the inducible promoter of the maize glutathione-S-transferase (GST-II-27) gene, 3 (21.4%) transgenic hybrid aspen plants with marker-free and normal phenotype were generated from 14 ipt-shooty lines within 2 months after cutting induction. These results suggest that the MAT-vector system might be useful for removing a selectable marker gene independent of sexual crossing in vegetatively propagated species.
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