caa a22 4500 463236106 CHVBK 20180405153257.0 cr unu---uuuuu 170326e20070901xx s 000 0 eng 10.1007/s11240-007-9268-6 doi (NATIONALLICENCE)springer-10.1007/s11240-007-9268-6 Use of fused gfp and gus reporters for the recovery of transformed Medicago truncatula somatic embryos without selective pressure [Elektronische Daten] [Ana Duque, Susana de Sousa Araújo, Matilde Cordeiro, Dulce Santos, Manuel Fevereiro] We developed an alternative methodology for invitro selection of transgenic Medicago truncatula cv. Jemalong plants using a bifunctional construct in which the coding sequences for the green fluorescent protein (GFP) and the β-glucuronidase protein (GUS) are fused. An Agrobacterium-mediated transformation protocol was used followed by regeneration via somatic embryogenesis in the dark, to avoid the synthesis and the consequent autofluorescence of chlorophyll. This method is a clear advantage over antibiotic and herbicide selection in which survival of non-transformed tissue is commonly reported, with the reassurance that all the somatic embryos selected as GFP positive are transformed. This was subsequently corroborated by the detection of GUS activity in leaves, stems and roots of the regenerated plants. Without antibiotic selection, and performing the embryo induction in the dark, it was possible to attest the advantage of using GFP as an invivo detectable reporter for early embryo selection. The fusion with the GUS coding sequence provided additional evidence for the transformation of the previously selected embryos. Springer Science+Business Media B.V., 2007 Agrobacterium -mediated transformation nationallicence Antibiotic-free nationallicence Dark embryo induction nationallicence GFP : Green fluorescent protein nationallicence GUS : β-Glucuronidase nationallicence EIM : Embryo induction medium nationallicence EPM : Embryo proliferation medium nationallicence ECM : Embryo conversion medium nationallicence Duque Ana Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut de Sousa Araújo Susana Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut Cordeiro Matilde Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut Santos Dulce Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut Fevereiro Manuel Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut Plant Cell, Tissue and Organ Culture Springer Netherlands 90/3(2007-09-01), 325-330 0167-6857 90:3<325 2007 90 11240 https://doi.org/10.1007/s11240-007-9268-6 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-007-9268-6 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Duque Ana Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- de Sousa Araújo Susana Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Cordeiro Matilde Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Santos Dulce Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 700 1- Fevereiro Manuel Laboratory of Plant Cell Biotechnology, Instituto de Tecnologia Química e Biológica (ITQB), Apt. 127, 2781-901, Oeiras, Portugal aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Springer Netherlands 90/3(2007-09-01), 325-330 0167-6857 90:3<325 2007 90 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer