caa a22 4500 46323684X CHVBK 20180405153300.0 cr unu---uuuuu 170326e20071101xx s 000 0 eng 10.1007/s11240-007-9228-1 doi (NATIONALLICENCE)springer-10.1007/s11240-007-9228-1 Agrobacterium -mediated transformation of sorghum ( Sorghum bicolor (L.) Moench) using an improved invitro regeneration system [Elektronische Daten] [Tuong-Van Nguyen, Tran Thanh Thu, Martine Claeys, Geert Angenon] An improved regeneration protocol suitable for transformation of sorghum was developed. The improvements focused on limiting the production of phenolic compounds and the use of suitable culture vessels for each developmental stage in plant regeneration from immature embryo derived calli. The addition of activated charcoal in the callus induction medium reduced the production of black pigments, however it also inhibited the callus formation on immature embryo explants. Cold pre-treatment of the immature seeds from which embryo explants were excised had a positive effect on both explant survival and callus formation. A one-day 4°C treatment of immature seeds significantly improved the callus formation from immature embryos and reduced the need for frequent subculture. Petri dishes with ventilation were suitable for the callus induction phase, but not for plant regeneration. Regeneration of plants could be improved by using disposal plastic boxes (250ml volume) instead of Petri dishes. Agrobacterium-mediated transformation using the improved regeneration protocol and the hygromycin phosphotransferase gene as selectable marker resulted in the recovery of 15 transgenic plants from 300 initial immature embryos (5% efficiency). The transgenic nature of the obtained plants was demonstrated by Southern hybridisation and progeny analysis. The transgenes were inherited in a Mendelian fashion and were integrated at a single locus in the majority of the analysed lines. Springer Science+Business Media B.V., 2007 Cereals nationallicence Cold pre-treatment nationallicence Culture vessels nationallicence Phenolic compounds nationallicence Transgenic plant nationallicence AC : Activated charcoal nationallicence CH : Casein hydrolysate nationallicence SCIM : Sorghum callus induction medium nationallicence SRM : Sorghum regeneration medium nationallicence Nguyen Tuong-Van Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut Thanh Thu Tran Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut Claeys Martine Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut Angenon Geert Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut Plant Cell, Tissue and Organ Culture Springer Netherlands 91/2(2007-11-01), 155-164 0167-6857 91:2<155 2007 91 11240 https://doi.org/10.1007/s11240-007-9228-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-007-9228-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Nguyen Tuong-Van Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Thanh Thu Tran Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Claeys Martine Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut NATIONALLICENCE 700 1- Angenon Geert Laboratory of Plant Genetics, Institute for Molecular Biology and Biotechnology, Vrije Universiteit Brussel, Pleinlaan 2, 1050, Brussels, Belgium aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Springer Netherlands 91/2(2007-11-01), 155-164 0167-6857 91:2<155 2007 91 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer