use of green fluorescent protein (GFP) improves Agrobacterium-mediated transformation of 'Spadona' pear (Pyrus communis L.)

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-i...

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Bibliographic Details
Published in:	Plant cell reports Vol. 25; no. 3; pp. 183 - 189
Main Authors:	Yancheva, S.D, Shlizerman, L.A, Golubowicz, S, Yabloviz, Z, Perl, A, Hanania, U, Flaishman, M.A
Format:	Journal Article
Language:	English
Published:	Berlin Springer 01-03-2006 Springer Nature B.V
Subjects:	acclimation Acclimatization adventitious shoots Agrobacterium tumefaciens beta-glucuronidase Biological and medical sciences biomarkers Biotechnology callus culture Cultivars Eukaryotic cell cultures explants Fluorescence microscopy fruit trees Fruits Fundamental and applied biological sciences. Psychology gene expression regulation genetic transformation genetic vectors green fluorescent protein Green Fluorescent Proteins - genetics Green Fluorescent Proteins - metabolism histology kanamycin kinase leaves Methods. Procedures. Technologies micropropagation Miscellaneous molecular sequence data nucleotide sequences pears Plant cells and fungal cells plant genetics Plant Leaves - cytology plant morphology plant physiology plant regeneration Plant Roots - growth & development Plant Shoots - cytology planting Plants, Genetically Modified Proteins Pyrus - genetics Pyrus - physiology Pyrus communis Regeneration reporter genes Rhizobium - genetics rooting Transformation, Genetic transgenes transgenic plants Pyrus communis Vegetals Transformation Genetic transformation Pear GUS Rosaceae Optimization GFP Regeneration Agrobacterium Dicotyledones Angiospermae Green fluorescent protein Bacteria Spermatophyta Rhizobiaceae
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Summary:	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.
Bibliography:	http://dx.doi.org/10.1007/s00299-005-0025-1 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0721-7714 1432-203X
DOI:	10.1007/s00299-005-0025-1