Genetic transformation of Alstroemeria using particle bombardment View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

2000-08

AUTHORS

Hsueh-Shih Lin, Caroline van der Toorn, Krit J.J.M. Raemakers, Richard G.F. Visser, Marjo J. De Jeu, Evert Jacobsen

ABSTRACT

Transgenic plants were obtained after particle bombardment of embryogenic callus derived from stem segments of two tetraploid Alstroemeria genotypes with plasmids containing different selection/reporter genes. Firstly, a plasmid containing a firefly luciferase reporter gene driven by the maize ubiquitin promoter (Ubi1), was bombarded into both friable embryogenic callus and proembryos. Transient and stable expression of luciferase was visually detected by a luminometer. This selection method is non-destructive and can be applied over the whole developmental process from callus to embryo and plantlet. Molecular proof of transformation was obtained both by PCR analysis and Southern hybridization. Secondly, a plasmid containing the bar gene together with an uidA gene coding for β-glucuronidase both driven by the Ubi1 promoter was bombarded into proembryos. The transgenic callus was effectively selected from the callus clumps four months after bombardment on a medium containing 5 mg/l phosphinotricin (PPT). Selection by PPT was efficient and labour-saving. Stable expression of GUS was confirmed by the histochemical staining assay and molecular proof was obtained by PCR analysis. More... »

PAGES

369-377

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  • Journal

    TITLE

    Molecular Breeding

    ISSUE

    4

    VOLUME

    6

    Author Affiliations

    Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1023/a:1009671105610

    DOI

    http://dx.doi.org/10.1023/a:1009671105610

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1021739747


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