Journal Article

Transformation and Characterization of Transgenic Plants of <i>Solanum dulcamara</i> L.—Incidence of Transgene Silencing

I. S. Curtis, J. B. Power, P. Hedden, A. Phillips, K. C. Lowe, D. A. Ward and M. R. Davey

in Annals of Botany

Published on behalf of The Annals of Botany Company

Volume 86, issue 1, pages 63-71
Published in print July 2000 | ISSN: 0305-7364
Published online July 2000 | e-ISSN: 1095-8290 | DOI: http://dx.doi.org/10.1006/anbo.2000.1159
Transformation and Characterization of Transgenic Plants of Solanum dulcamara L.—Incidence of Transgene Silencing

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  • Ecology and Conservation
  • Evolutionary Biology
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A transformation system is described for Solanum dulcamara using the supervirulentAgrobacterium tumefaciens strain 1065, carrying both the β-glucuronidase (gus) and neomycin phosphotransferase II (npt II) genes adjacent to the right and left T-DNA borders, respectively. Leaf explants were more efficient for the production of transformed plants compared to stem explants on medium containing 50 mg l−1of kanamycin sulphate. A 1:10 (v:v) dilution of an overnight culture ofAgrobacterium gave optimal transformation in terms of transgenic plant regeneration. From a total of 174 kanamycin-resistant plants selected by their antibiotic resistance, 16 failed to exhibit GUS activity. Southern analysis revealed that these GUS-negative transformants originated from three independently transformed cell lines. Restriction enzyme analyses showed that the GUS-negative plants had both the gus and npt II genes integrated into their genome (one plant had a single copy of each gene; the other two plants had multiple copies), with major rearrangement of the gus gene occurring in plants with several copies of the transgene. GUS-negative plants showed leaf malformations, delayed flowering and a reduction in flower, fruit and seed production compared to GUS-positive and non-transformed (control) plants. Although gene silencing of the gus gene occurred, albeit at a low frequency (9.2%), the transformation system described generates large numbers of phenotypically normal, stably transformed plants. Copyright 2000 Annals of Botany Company

Keywords: Agrobacterium -mediated transformation, gene silencing, Solanum dulcamara L. (Bittersweet, Woody Nightshade), T-DNA truncation, transgene expression

Journal Article.  34 words. 

Subjects: Ecology and Conservation ; Evolutionary Biology ; Plant Sciences and Forestry

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