caa a22 4500 463235835 CHVBK 20180405153256.0 cr unu---uuuuu 170326e20070201xx s 000 0 eng 10.1007/s11240-006-9190-3 doi (NATIONALLICENCE)springer-10.1007/s11240-006-9190-3 Highly efficient Agrobacterium tumefaciens -mediated transformation of celery ( Apium graveolens L.) through somatic embryogenesis [Elektronische Daten] [Guo-Qing Song, A. Loskutov, K. Sink] A protocol was developed for rapid and efficient production of transgenic celery plants via somatic embryo regeneration from Agrobacterium tumefaciens- inoculated leaf sections, cotyledons and hypocotyls. These explants were excised from in vitro seedlings of the cvs. XP166 and XP85 and inoculated with A. tumefaciens strain EHA105 containing the binary vector pBISN1. PBISN1 has the neomycin phosphotransferase gene (nptII) and an intron interrupted β-glucuronidase (GUS) reporter gene (gusA). Co-cultivation was carried out for 4d in the dark on callus induction medium (CIM): Gamborg B5+2.79μM kinetin+2.26μM 2,4-dichlorophenoxyacetic acid (2,4-D) supplemented with 100μM acetosyringone. Embryogenic calluses resistant to kanamycin (Km) were then recovered on CIM+25mgl−1 Km+250mgl−1 timentin after 12weeks. Subsequently, a large number of Km-resistant and GUS-positive transformants, tens to hundreds per explant were regenerated via somatic embryogenesis on Gamborg B5+4.92μM 6 (γ,γ-dimethylallylamino)-purine (2iP)+1.93μM α-naphthaleneacetic acid (NAA)+25mgl−1 Km+250mgl−1 timentin after 8weeks. Using this protocol, the transformation frequency was 5.0% and 5.0% for leaf sections, 17.8% and 18.3% for cotyledons, and 15.9% and 16.7% for hypocotyl explants of cvs. XP85 and XP166, respectively. Stable integration of the model transgenes with 1-3 copy numbers was confirmed in all ten randomly selected transgenic events by Southern blot analysis of gusA. Progeny analysis by histochemical GUS assay showed stable Mendelian inheritance of the transgenes. Thus, A. tumefaciens-mediated transformation of cotyledons or hypocotyls provides an effective and reproducible protocol for large-scale production of transgenic celery plants. Springer Science+Business Media B.V., 2006 β-glucuronidase nationallicence Plant regeneration nationallicence Somatic embryo nationallicence Transgenic plant nationallicence Song Guo-Qing Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut Loskutov A. Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut Sink K. Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/2(2007-02-01), 193-200 0167-6857 88:2<193 2007 88 11240 https://doi.org/10.1007/s11240-006-9190-3 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11240-006-9190-3 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Song Guo-Qing Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Loskutov A. Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Sink K. Plant Transformation Center, Department of Horticulture, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 773 0- Plant Cell, Tissue and Organ Culture Kluwer Academic Publishers 88/2(2007-02-01), 193-200 0167-6857 88:2<193 2007 88 11240 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer