caa a22 4500 378902024 CHVBK 20180305123520.0 cr unu---uuuuu 161128e20040701xx s 000 0 eng 10.2298/hel0440025m doi (NATIONALLICENCE)gruyter-10.2298/hel0440025m Agrobacterium-MEDIATED TRANSFORMATION OF TWO HIGH OLEIC SUNFLOWER (Helianthus annuus L.) GENOTYPES: ASSESSMENT AND OPTIMIZATION OF IMPORTANT PARAMETERS [Elektronische Daten] DETERMINACIÓN Y OPTIMIZACIÓN DE PARÁMETROS QUE APOYAN LA TRANSFORMACIÓN DE DOS GENOTIPOS DE GIRASOL (Helianthus annuus L.), MEDIANTE Agrobacterium / TRANSFORMATION DE DEUX TOURNESOLS OLÉICS (Helianthus annuus L.) GENOTYPES VIA Agrobacterium tumefaciens: ÉVALUATION ET PARAMÈTRES OPTIMUM / [Sh. Mohamed, R. Boehm, P.C. Binsfeld, H. Schnabl] The establishment of an efficient Agrobacterium-mediated transformation protocol for a recalcitrant crop like high oleic sunflower genotypes, cv. Capella and SWSR2 inbred line, is presented in this paper. The protocol requires the identification and optimization of parameters affecting T-DNA delivery and plant regeneration. The basis for identifying the most appropriate conditions for transformation was the fluorometric, histochemical GUS activity coupled with plant cell vitality. Agrobacterium tumefaciens-mediated transformation was performed using strain LBA4404 for cv. Capella and strain GV3101 for the inbred line. Both strains harboring the pBI121 plasmid with the gus gene were under the control of 35S promoter. The flourometric GUS activity was increased in cv. Capella and SWSR2 inbred line 1.9 and 1.6 fold, respectively, relative to transformation without inducers, by optimizing the bacterial density to OD600=1.0, by splitting shoot apices and by using MS medium as a co-cultivation medium supplemented with 200 μM acetosyringone. Pre-culturing the explants for three days on SIM2 medium before the bacterial inoculation followed by co-cultivation for 72 h increased the percentage of the GUS expression to 40% and 30% in cv. Capella and SWSR2 inbred line, respectively. It also improved the regeneration percentage as well as the vitality of cells. This article is distributed under the terms of the Creative Commons Attribution Non-Commercial License, which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Agrobacterium tumefaciens nationallicence β-glucuronidase nationallicence genetic transformation nationallicence Helianthus annuus L nationallicence shoot apices nationallicence Mohamed Sh Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut Boehm R. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut Binsfeld P.C. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut Schnabl H. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany, Phone.: +49-228-732832, Fax: +49-731696 aut HELIA De Gruyter 27/40(2004-07-01), 25-40 1018-1806 27:40<25 2004 27 helia https://doi.org/10.2298/hel0440025m text/html Onlinezugriff via DOI 1 research article jats NATIONALLICENCE 856 40 https://doi.org/10.2298/hel0440025m text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Mohamed Sh Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut NATIONALLICENCE 700 1- Boehm R. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut NATIONALLICENCE 700 1- Binsfeld P.C. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany aut NATIONALLICENCE 700 1- Schnabl H. Institute of Plant Molecular Physiology and Biotechnology, University of Bonn, Karlrobert-Kreiten-Str. 13, 53115 Bonn, Germany, Phone.: +49-228-732832, Fax: +49-731696 aut NATIONALLICENCE 773 0- HELIA De Gruyter 27/40(2004-07-01), 25-40 1018-1806 27:40<25 2004 27 helia CC0 http://creativecommons.org/publicdomain/zero/1.0 nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-gruyter