Inheritance of two foreign genes co-introduced intoPetunia hybrida by direct gene transfer |
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Authors: | Denis Tagu Catherine Bergounioux Claudette Perennes Pierre Gadal |
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Institution: | (1) Laboratoire de Physiologie Végétale Moléculaire, UA CNRS 1128, bât. 430, Université Paris-Sud, Centre d'Orsay, 91.405 Orsay Cedex, France;(2) Present address: Departamento de Genetica Molecular, Centro de Investigacion y Desarollo, CSIC, Jordi Girona Salgado 18-26, 08034 Barcelona, Spain |
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Abstract: | In previous work, transformedPetunia hybrida plants were obtained by direct gene transfer, using two different genes on separate plasmids (NPT II gene and a cDNA of PEPC from green sorghum leaves). In this study, we have analysed the sexual transmission of the acquired genes by genetic crossing analysis of 2 of the transgenic petunias. The ploïdies of the two clones were determined by flow cytometric analysis showing that one was 2n and the other 4n. Self and back crosses show that the kanamycin character was inherited as a single dominant trait, and that the two clones were heterozygotes for this character. Therefore, the 4n clone probably arises from an endoploidization followed by a transformation event. Southern blot analyses show that all of the resistant progenies which were analysed harboured the kanamycin gene, and expressed the phosphorylation activity in vitro. The DNA of several progenies were also tested for the presence of co-transformed PEPC cDNA sequence. All of the kanamycin-resistant progenies tested contained the second coding sequence, indicating that the two foreign genes might be genetically co-inherited in the transgenic plants. The way in which the two genes are integrated into the genome is discussed.Abbreviations NPT
neomycin phosphotransferase
- PEG
polyethyleneglycol
- PEPC
phosphoenolpyruvate carboxylase |
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Keywords: | Petunia hybrida Sorghum vulgare co-transformation electroporation inheritance phosphoenolpyruvate carboxylase protoplast |
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