Staff Publications

Staff Publications

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    'Staff publications' is the digital repository of Wageningen University & Research

    'Staff publications' contains references to publications authored by Wageningen University staff from 1976 onward.

    Publications authored by the staff of the Research Institutes are available from 1995 onwards.

    Full text documents are added when available. The database is updated daily and currently holds about 240,000 items, of which 72,000 in open access.

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Record number 346265
Title Seed-expressed fluorescent proteins as versatile tools for easy (co)transformation and high-throughput funtional genomics in Arabidopsis
Author(s) Stuitje, A.R.; Verbree, E.C.; Linden, K.H. van der; Mietkiewska, E.M.; Nap, J.P.H.; Kneppers, T.J.A.
Source Plant Biotechnology Journal 1 (2003)4. - ISSN 1467-7644 - p. 301 - 309.
Department(s) PRI Bioscience
Publication type Refereed Article in a scientific journal
Publication year 2003
Keyword(s) agrobacterium-mediated transformation - t-dna integration - vacuum infiltration - transgenic plants - brassica-napus - thaliana - recombination - tumefaciens - gene
Abstract We demonstrate that fluorescent proteins can be used as visual selection markers for the transformation of Arabidopsis thaliana by the floral dip method. Seed-specific expression of green fluorescent protein (GFP) variants, as well as DsRed, permits the identification of mature transformed seeds in a large background of untransformed seeds by fluorescence microscopy. In planta visualization of transformed seeds in siliques shows that susceptibility to floral dip transformation is limited to a small, defined window in flower development. In the competent stage, the random transformation of up to 25% of the seeds within a single silique may occur. The use of fluorescent proteins with different spectral characteristics allows a rapid identification and genetic analysis of seeds that have received multiple genes-of-interest in co-transformation experiments. The data reveal that co-transformation does not occur at random, since the co-transformed genes are integrated at a single genetic locus in ¿ 70% of the cases. This genetic linkage of the co-transformed genes greatly simplifies metabolic pathway engineering by reverse genetics in Arabidopsis. Additional advantages of using visual selection instead of antibiotic resistance include a rapid identification of the effect of the T-DNA insertion or the transgene on seed development and/or germination. This technology, of tagging and identifying transformed seeds by fluorescence provides a novel high-throughput screening system with many potential applications in plant biotechnology.
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