Transgene behaviour in populations of rice plants transformed using a new dual binary vector system: pGreen/pSoup View Full Text


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Article Info

DATE

2003-04-03

AUTHORS

P. Vain, A. S. Afolabi, B. Worland, J. W. Snape

ABSTRACT

Transgene integration, expression level and stability have been studied, across two generations, in a population of rice plants transformed using a new dual binary vector system: pGreen/pSoup. pGreen is a small Ti binary vector unable to replicate in Agrobacterium without the presence of another binary plasmid, pSoup, in the same strain. We engineered both pGreen and pSoup to contain each a different T-DNA. Transformation experiments were conducted using a pGreen vector containing the bar and gusA expression units (no transgene in pSoup) or with a pSoup vector containing an aphIV and gfp expression units (no transgene in pGreen). High plant transformation frequencies (up to 40%) were obtained using herbicide resistance (bar) or antibiotic resistance (aphIV) genes. Around 80% of the independently transformed plants expressed unselected reporter genes (gusA or gfp) present in the vectors. Backbone sequences transfer was frequent (45% of lines) and occurred often in multicopy lines. Around 15–20% of the rice plant lines contained a single T-DNA integration without backbone. Integration of additional transgene copies did not improve expression levels in either T0 plants or T1 progenies. Nearly all multicopy lines contained transgenes integrated at several loci in the plant genome, showing that T-DNAs from either pGreen or pSoup frequently integrated at unlinked loci. Precise determination of loci number required the analysis of transgene presence in progeny. Segregation of transgene phenotype was generally misleading and tended to underestimate the real number of transgenic loci. The contribution of this new dual-binary vector system to the development of high-throughput rice transformation systems and to the production of marker-free transgenic rice plants is discussed. More... »

PAGES

210-217

References to SciGraph publications

  • 2000-04. pGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation in PLANT MOLECULAR BIOLOGY
  • 2000-02. Evidence of multiple complex patterns of T-DNA integration into the rice genome in THEORETICAL AND APPLIED GENETICS
  • 1996-03. Intron-mediated enhancement of gene expression in maize (Zea mays L.) and bluegrass (Poa pratensis L.) in PLANT CELL REPORTS
  • 1998-02. Expression of an engineered cysteine proteinase inhibitor (Oryzacystatin-IΔD86) for nematode resistance in transgenic rice plants in THEORETICAL AND APPLIED GENETICS
  • 1996-06. High efficiency transformation of maize (Zea mays L.) mediated by Agrobacterium tumefaciens in NATURE BIOTECHNOLOGY
  • 2000-10. T-DNA vector backbone sequences are frequently integrated into the genome of transgenic plants obtained by Agrobacterium-mediated transformation in MOLECULAR BREEDING
  • 1996-08. Agrobacterium tumefaciens-mediated transformation of japonica and indica rice varieties in PLANTA
  • 2002-03. Identifying the chromosomes of the A- and C-genome diploid Brassica species B. rapa (syn. campestris) and B. oleracea in their amphidiploid B. napus in THEORETICAL AND APPLIED GENETICS
  • 2001-09. Characterization of rice transformed via an Agrobacterium-mediated inflorescence approach in MOLECULAR BREEDING
  • 2001-04. Efficient Co-Transformation of Nicotiana Tabacum by two Independent T-DNAs, the Effect of T-DNA Size and Implications for Genetic Separation in TRANSGENIC RESEARCH
  • 2002-08-22. Transgene behaviour across two generations in a large random population of transgenic rice plants produced by particle bombardment in THEORETICAL AND APPLIED GENETICS
  • 1997-09. Transformation of rice mediated by Agrobacterium tumefaciens in PLANT MOLECULAR BIOLOGY
  • 1996-01. Selection of large quantities of embryogenic calli from indica rice seeds for production of fertile transgenic plants using the biolistic method in PLANT CELL REPORTS
  • 1996-09. Agrobacterium-mediated transformation of Javanica rice in MOLECULAR BREEDING
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    http://scigraph.springernature.com/pub.10.1007/s00122-003-1255-7

    DOI

    http://dx.doi.org/10.1007/s00122-003-1255-7

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1050258094

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/12677408


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