caa a22 4500 463160258 CHVBK 20180406164745.0 cr unu---uuuuu 170326e20071201xx s 000 0 eng 10.1007/s10535-007-0148-z doi (NATIONALLICENCE)springer-10.1007/s10535-007-0148-z High efficiency of stable genetic transformation in Dendrobium via microprojectile bombardment [Elektronische Daten] [C. Suwanaketchanatit, J. Piluek, S. Peyachoknagul, P. Huehne] Several parameters affecting transient β-glucuronidase (GUS) expression in protocorms of Dendrobium cv. Jaquelyn Thomas were examined using bombardment technique with pActin-1D plasmid. The GUS activity in Dendrobium protocorm was not significantly affected by size of the target, type of particles, and helium gas pressure. However, the numbers of surviving tissues after bombardment were different. Transgenic orchids were established by bombardment of actively dividing protocorms with gold particles coated with pMNK1005 plasmid containing hygromycin phosphotransferase (hgh) and green fluorescent protein (gfp) genes driven by a ubiquitin promoter. A high efficiency of orchid transformation was established using three selection steps. The bombarded protocorms were screened on medium supplemented with 5 mg dm−3 and 25 mg dm−3 hygromycin and surviving protocorms were stringently selected on medium containing 30 mg dm−3 hygromycin. The transformation efficiency was 19.87 % and GFP expressing protocorms were not chimeras. The integration of the transgene into genomic DNA of transgenic plantlets was confirmed by PCR and Southern blot hybridization. All but one of the transgenic lines contained multiple copies of the transgene. Institute of Experimental Botany, ASCR, 2007 β-glucuronidase nationallicence green fluorescent protein nationallicence orchid nationallicence PCR nationallicence Southern blot nationallicence GFP : green fluorescent protein nationallicence GUS : β-glucuronidase nationallicence VW : Vacin and Went medium nationallicence Suwanaketchanatit C. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut Piluek J. Department of Horticulture, Faculty of Agriculture, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut Peyachoknagul S. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut Huehne P. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut Biologia Plantarum Springer Netherlands 51/4(2007-12-01), 720-727 0006-3134 51:4<720 2007 51 10535 https://doi.org/10.1007/s10535-007-0148-z text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10535-007-0148-z text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Suwanaketchanatit C. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut NATIONALLICENCE 700 1- Piluek J. Department of Horticulture, Faculty of Agriculture, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut NATIONALLICENCE 700 1- Peyachoknagul S. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut NATIONALLICENCE 700 1- Huehne P. Department of Genetics, Faculty of Science, Kasetsart University, 10900, Jatuchak, Bangkok, Thailand aut NATIONALLICENCE 773 0- Biologia Plantarum Springer Netherlands 51/4(2007-12-01), 720-727 0006-3134 51:4<720 2007 51 10535 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer