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 431484
Title One-Step Agrobacterium Mediated Transformation of Eight Genes Essential for Rhizobium Symbiotic Signaling Using the Novel Binary Vector System pHUGE
Author(s) Untergasser, A.; Bijl, G.J.M.; Liu, W.; Bisseling, T.; Schaart, J.G.; Geurts, R.
Source PLoS One 7 (2012)10. - ISSN 1932-6203
DOI https://doi.org/10.1371/journal.pone.0047885
Department(s) Laboratory of Molecular Biology
WUR Plant Breeding
Publication type Refereed Article in a scientific journal
Publication year 2012
Keyword(s) site-specific recombination - root-nodule organogenesis - selectable marker gene - genomic dna fragments - medicago-truncatula - plant transformation - nodulation factors - lotus-japonicus - host-range - cytokinin
Abstract Advancement in plant research is becoming impaired by the fact that the transfer of multiple genes is difficult to achieve. Here we present a new binary vector for Agrobacterium tumefaciens mediated transformation, pHUGE-Red, in concert with a cloning strategy suited for the transfer of up to nine genes at once. This vector enables modular cloning of large DNA fragments by employing Gateway technology and contains DsRED1 as visual selection marker. Furthermore, an R/Rs inducible recombination system was included allowing subsequent removal of the selection markers in the newly generated transgenic plants. We show the successful use of pHUGE-Red by transferring eight genes essential for Medicago truncatula to establish a symbiosis with rhizobia bacteria as one 74 kb T-DNA into four non-leguminous species; strawberry, poplar, tomato and tobacco. We provide evidence that all transgenes are expressed in the root tissue of the non-legumes. Visual control during the transformation process and subsequent marker gene removal makes the pHUGE-Red vector an excellent tool for the efficient transfer of multiple genes.
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