caa a22 4500 467900701 CHVBK 20180406152817.0 cr unu---uuuuu 170328e20060301xx s 000 0 eng 10.1007/s00299-005-0025-1 doi (NATIONALLICENCE)springer-10.1007/s00299-005-0025-1 The use of green fluorescent protein (GFP) improves Agrobacterium -mediated transformation of ‘Spadona' pear ( Pyrus communis L.) [Elektronische Daten] [S. Yancheva, L. Shlizerman, S. Golubowicz, Z. Yabloviz, A. Perl, U. Hanania, M. Flaishman] An efficient and reproducible system for Agrobacterium-mediated transformation of the pear (Pyrus communis L.) cultivar Spadona was developed. Leaf explants of in vitro propagated plants were cocultivated with the disarmed Agrobacterium strain EHA105 harboring the plasmid pME504, carrying the uidA-intron and nptII genes. Under selective conditions, 5% of the plantlets regenerated and were positively stained for GUS. However, most of the GUS-positive plants re-callused and subsequently died, leaving only 0.3-0.8% of these plantlets to reach maturity. In order to identify transformed shoots at early stages of regeneration, we introduced the green fluorescent protein (GFP) into the pear cultivar Spadona using the plasmid PZP carrying the nuclear-targeted GFP and nptII genes. High expression levels of GFP were detected in transgenic cells as early as 7 days after transformation. GFP marked-callii and transformed plants were observed after 14 and 24 days, respectively. Fluorescence microscopy screening of transformed plant material, under the selection of kanamycin, increased the transformation frequency to 3.0-4.0%. We conclude that the introduction of GFP improves the selection of transformed plants of Spadona pear. Springer-Verlag, 2005 Pear nationallicence Pyrus communis nationallicence Regeneration nationallicence Agrobacterium , GUS nationallicence GFP nationallicence Transformation nationallicence BA : 6-benzylaminopurine nationallicence CaMV : cauliflower mosaic virus nationallicence GA : gibberellic acid A3 nationallicence GFP : green fluorescent protein nationallicence GUS : β-glucuronidase nationallicence HP1 : heterochromatin protein 1 nationallicence IBA : indole-3-butyric acid nationallicence NAA : α-naphthaleneacetic acid nationallicence npt II : neomycin phosphotransferase II nationallicence PCR : polymerase chain reaction nationallicence SDS : sodium dodecyl sulphate nationallicence TDZ : thidiazuron (N-phenyl-N′-1,2,3-thiadiazol-5-yl urea) nationallicence uid A : β-glucuronidase gene of Escherichia coli nationallicence Yancheva S. Plant Biotechnology Laboratory, Agricultural University, 12 Mendeleev Street, 4000, Plovdiv, Bulgaria aut Shlizerman L. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Golubowicz S. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Yabloviz Z. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Perl A. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Hanania U. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Flaishman M. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut Plant Cell Reports Springer-Verlag 25/3(2006-03-01), 183-189 0721-7714 25:3<183 2006 25 299 https://doi.org/10.1007/s00299-005-0025-1 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s00299-005-0025-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Yancheva S. Plant Biotechnology Laboratory, Agricultural University, 12 Mendeleev Street, 4000, Plovdiv, Bulgaria aut NATIONALLICENCE 700 1- Shlizerman L. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 700 1- Golubowicz S. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 700 1- Yabloviz Z. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 700 1- Perl A. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 700 1- Hanania U. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 700 1- Flaishman M. The Department of Fruit Trees Sciences, Institute of Horticulture, ARO, The Volcani Center, Bet Dagan, P.O. Box 6, 50250, Rehovot, Israel aut NATIONALLICENCE 773 0- Plant Cell Reports Springer-Verlag 25/3(2006-03-01), 183-189 0721-7714 25:3<183 2006 25 299 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer