Agrobacterium-mediated transformation of sweet orange and regeneration of transgenic plants |
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Authors: | Leandro Peña Magdalena Cervera José Juárez Antonio Navarro José A Pina Nuria Durán-Vila Luis Navarro |
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Institution: | (1) Dpto. de protección vegetal y biotecnología, Institute Valenciano de Investigaciones Agrarias, Apartado oficial, E-46113 Moncada, Spain |
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Abstract: | Summary Transgenic sweet orange (Citrus sinensis L. Osbeck) plants have been obtained by Agrobacterium tumefaciens-mediated gene transfer. An hypervirulent A. tumefaciens strain harboring a binary vector that contains the chimeric neomycin phosphotransferase II (NPT II) and ß-glucuronidase (GUS) genes was cocultivated with stem segments from in vivo grown seedlings. Shoots regenerated under kanamycin selection were harvested from the stem segments within 12 weeks. Shoot basal portions were assayed for GUS activity and the remaining portions were shoot tip grafted in vitro for production of plants. Integration of the GUS gene was confirmed by Southern analysis. This transformation procedure showed the highest transgenic plant production efficiency reported for Citrus.Abbreviations BA
benzyladenine
- CaMV
cauliflowermosaic virus
- GUS
ß-glucuronidase
- LB
Luria Broth
- MS
Murashige and Skoog
- NAA
naphthalenacetic acid
- NOS
nopaline synthase
- NPT II
neomycin phosphotransferase II
- PEG
polyethylene glycol
- RM
rooting medium
- SRM
shoot regeneration medium |
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Keywords: | Citrus sinensis L Osbeck cv Pineapple Sweet orange Agrobacterium tumefaciens Genetic transformation |
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