Transgenic poplars: expression of chimeric genes using four different constructs |
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Authors: | Jean Charles Leple Ana Cristina Miranda Brasileiro Marie France Michel Francis Delmotte Lise Jouanin |
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Affiliation: | (1) Station d'Amélioration des Arbres Forestiers, Institut National de la Recherche Agronomique, F-45160 Ardon, Olivet, France;(2) Laboratoire de Biologie Cellulaire, Institut National de la Recherche Agronomique, Route de Saint-Cyr, F-78026 Versailles Cedex, France;(3) Centre de Biophysique Moléculaire, Département de Biochimie des Glycoconjugués et Lectines Endogènes, Centre National de la Recherche Scientifique et Université d'Orléans, F-45071 Orléans Cedex 2, France |
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Abstract: | Summary Leaf or stem explants of a hybrid poplar clone (Populus tremula X Populus alba), sensitive to Agrobacterium tumefaciens, were co-cultivated either by an octopine or a nopaline disarmed A. tumefaciens modified strain. Transformed poplar shoots were readily regenerated from explants. The protocol was improved using the nopaline disarmed strain C58/pMP90 with the binary vector pBI121. This protocol was then used to test three other vectors. The first one, possessing a nptII gene fused to the CaMV 19S promoter, permitted regeneration of transformed shoots in presence of 50 to 100 mg/l kanamycin. The two other vectors carried an additional nptII gene under the control of the CaMV 35S or CaMV 35S promoter with a double enhancer sequence (CaMV 70). CaMV 70 promoter provided consistently higher level of gene expression than the other promoters in both callus and leaf tissues.Abbreviations CaMV Cauliflower Mosaïc Virus - 2iP 2-isopentenyladenine - GUS and gus ß-glucuronidase - NAA 1-naphthaleneacetic acid - NPTII and nptII neomycin phosphotransferase II - NOS Nopaline synthase, X-Gluc: 5-bromo-4-chloro-3-indolyl ß-D glucuronide - Ap ampicillin - Gn gentamycin - Km kanamycin - Rf rifampicin - St streptomycinThis work is dedicated to the late Marie France Michel who initiated the poplar biotechnology project at INRA. |
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