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Agrobacterium rhizogenes-mediated DNA transfer to Aesculus hippocastanum L. and the regeneration of transformed plants

  • Genetic Transformation and Hybridization
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Abstract

Hairy roots were induced from androgenic embryos of horse chestnut (Aesculus hippocastanum L.) by infection with Agrobacterium rhizogenes strain A4GUS. Single roots were selected according to their morphology in the absence of antibiotic or herbicide resistance markers. Seventy-one putative transformed hairy root lines from independent transformation events were established. Regeneration was induced in MS liquid medium supplemented with 30 μΜ 6-benzylaminopurine (BA), and the regenerants were multiplied on MS solid medium containing 10 μM BA. Following elongation on MS medium supplemented with 1 μM BA and 500 mg/l polyvinylpyrrolidone, the shoots were subjected to a root-inducing treatment. Stable integration of TL-DNA within the horse chestnut genome was confirmed by Southern hybridization. The copy number of transgenes was estimated to be from two to four.

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Acknowledgements

The authors are grateful to Dr. M. Nešković, University of Belgrade, and to Dr. B. Watillon,Centre Wallon de Recherches Agronomiques, Gembloux, Belgium, for their critical suggestions during this research. We would like to thank Dr. P. Landré, Université Pierre et Marie Curie VI, Paris, for kindly providing us with an Agrobacterium rhizogenes culture, Ms. S. Michiels, CRAW, Gembloux, Belgium, for excellent assistance in molecular analysis and Dr. Tatjana Savić, Institute for Biological Research, Belgrade, for helping with the statistical analysis. The Ministry of Science, Technology and Development of Serbia, Contract no. 1573, supported this research.

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Authors and Affiliations

  1. Institute for Biological Research “S. Stanković” 29, Novembar 142, Belgrade, Serbia and Montenegro

    S. Zdravković-Korać, D. Ćalić & L. Radojević

  2. CRAW-Department Biotechnology, Chaussée de Charleroi 234, 5030, Gembloux, Belgium

    Y. Muhovski & P. Druart

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  1. S. Zdravković-Korać
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  2. Y. Muhovski
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  3. P. Druart
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  4. D. Ćalić
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  5. L. Radojević
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Correspondence to S. Zdravković-Korać.

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Communicated by E.D. Earle

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Zdravković-Korać, S., Muhovski, Y., Druart, P. et al. Agrobacterium rhizogenes-mediated DNA transfer to Aesculus hippocastanum L. and the regeneration of transformed plants. Plant Cell Rep 22, 698–704 (2004). https://doi.org/10.1007/s00299-004-0756-4

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  • DOI: https://doi.org/10.1007/s00299-004-0756-4

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