caa a22 4500 467899991 CHVBK 20180406152815.0 cr unu---uuuuu 170328e20060601xx s 000 0 eng 10.1007/s00299-005-0087-0 doi (NATIONALLICENCE)springer-10.1007/s00299-005-0087-0 Molecular characterization of grapevine plants transformed with GFLV resistance genes: II [Elektronische Daten] [Fatemeh Maghuly, Stephan Leopold, Artur da Câmara Machado, Eduviges Borroto Fernandez, Mahmood Ali Khan, Giorgio Gambino, Ivana Gribaudo, Angelika Schartl, Margit Laimer] A collection of 127 putatively transgenic individuals of Vitis vinifera cv. Russalka was characterized by PCR and Southern hybridization. Six different constructs containing the neomycin phosphotransferase (nptII) marker gene and sequences of the Grapevine Fanleaf Virus Coat Protein (GFLV CP) gene including non-translatable and truncated forms were transferred via Agrobacterium-mediated transformation. Detection of transgenic sequences by PCR was positive in all lines. Southern blot analysis revealed that the number of inserted T-DNA copies ranged from 1 to 6. More than 46% of the tested transgenic lines contain one copy of the inserted T-DNA, qualifying them as interesting candidates for further breeding programs. Southern data of one line indicate the presence of an incomplete copy of the T-DNA, thus confirming previous PCR results. Since many putative transgenic lines shared identical hybridization patterns, they were clustered into 39 lines and considered as having originated from independent transformation events. The detection of the tetracycline (TET) resistance genes in 15% of the lines shows that an integration of plasmid backbone sequences beyond the T-DNA borders occurred. Enzyme-linked immunosorbent assay (ELISA) performed on leaf tissue did not show any accumulation of the GFLV CP in the 39 transgenic lines analyzed. Reverse transcription polymerase chain reaction (RT-PCR) and Northern blot were carried out; RT-PCR analyses showed that the GFLV CP mRNA was expressed at variable levels. Springer-Verlag, 2005 Nepovirus nationallicence Coat protein-mediated virus resistance nationallicence Pathogen-derived resistance nationallicence Somatic embryogenesis nationallicence Vitis vinifera nationallicence Virus resistance nationallicence Grapevine Fanleaf disease nationallicence Maghuly Fatemeh Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut Leopold Stephan Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut da Câmara Machado Artur Centro Biotecnologia dos Açores, Departamento Ciencias Agrarias, Universidade dos Açores, Terra Cha, 9700, Angra do Heroismo, Azores aut Borroto Fernandez Eduviges Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut Ali Khan Mahmood Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut Gambino Giorgio Viticulture Unit, Plant Virology Institute CNR, Via L. da Vinci 44, I-10095, Grugliasco (TO), Italy aut Gribaudo Ivana Viticulture Unit, Plant Virology Institute CNR, Via L. da Vinci 44, I-10095, Grugliasco (TO), Italy aut Schartl Angelika Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut Laimer Margit Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut Plant Cell Reports Springer-Verlag 25/6(2006-06-01), 546-553 0721-7714 25:6<546 2006 25 299 https://doi.org/10.1007/s00299-005-0087-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0087-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Maghuly Fatemeh Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 700 1- Leopold Stephan Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 700 1- da Câmara Machado Artur Centro Biotecnologia dos Açores, Departamento Ciencias Agrarias, Universidade dos Açores, Terra Cha, 9700, Angra do Heroismo, Azores aut NATIONALLICENCE 700 1- Borroto Fernandez Eduviges Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 700 1- Ali Khan Mahmood Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 700 1- Gambino Giorgio Viticulture Unit, Plant Virology Institute CNR, Via L. da Vinci 44, I-10095, Grugliasco (TO), Italy aut NATIONALLICENCE 700 1- Gribaudo Ivana Viticulture Unit, Plant Virology Institute CNR, Via L. da Vinci 44, I-10095, Grugliasco (TO), Italy aut NATIONALLICENCE 700 1- Schartl Angelika Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 700 1- Laimer Margit Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer Lände 11, A-1190, Vienna, Austria aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/6(2006-06-01), 546-553 0721-7714 25:6<546 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer