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Plant Cell Rep ; 25(3): 183-9, 2006 Mar.
Article de Anglais | MEDLINE | ID: mdl-16328389

RÉSUMÉ

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.


Sujet(s)
Protéines à fluorescence verte/génétique , Protéines à fluorescence verte/métabolisme , Pyrus/génétique , Rhizobium/génétique , Transformation génétique , Acclimatation , Feuilles de plante/cytologie , Racines de plante/croissance et développement , Pousses de plante/cytologie , Végétaux génétiquement modifiés , Pyrus/physiologie , Régénération
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