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An alternative approach for gene transfer in trees using wild-typeAgrobacterium strains

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Abstract

Micropropagated shoots of three forest tree species, poplar (Populus tremula × P.alba), wild cherry (Prunus avium L.) and walnut (Juglans nigra × J. regia), were inoculated each with six different wild-typeAgrobacterium strains. Poplar and wild cherry developed tumors that grew hormone-independently, whereas on walnut, gall formation was weak. On poplar and wild cherry, tumors induced by nopaline strains developed spontaneously shoots that had a normal phenotype and did not carry oncogenic T-DNA. From these observations, we have established a co-inoculation method to transform plants, using poplar as an experimental model. The method is based on inoculation of stem internodes with anAgrobacterium suspension containing both an oncogenic strain that induces shoot differentiation and a disarmed strain that provides the suitable genes in a binary vector. We used the vector pBI121 carryingneo (kanamycin resistance) anduidA (β-glucuronidase) genes to facilitate early selection and screening. Poplar plants derived from kanamycin-resistant shoots that did not carry oncogenic T-DNA, were shown to contain and to expressneo anduidA genes. These results suggest that wild-typeAgrobacterium strains that induce shoot formation directly from tumors can be used as a general tool for gene transfer, avoiding difficult regeneration procedures.

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Author notes

  1. Joris Muzzin

    Present address: Piccoplant Mikrovermehrungen, Brockhauser Weg 75, D-2900, Oldenburg, Germany

Authors and Affiliations

  1. Laboratoire de Biologie Cellulaire, INRA, route de Saint-Cyr, F-78026, Versailles Cedex, France

    Ana Cristina Miranda Brasileiro & Lise Jouanin

  2. Station d'Amélioration des Arbres Forestiers, INRA, Ardon, F-45160, Olivet, France

    Jean-Charles Leplé, Joris Muzzin, Dalila Ounnoughi & Marie-France Michel

Authors
  1. Ana Cristina Miranda Brasileiro
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  2. Jean-Charles Leplé
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  3. Joris Muzzin
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  4. Dalila Ounnoughi
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  5. Marie-France Michel
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  6. Lise Jouanin
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Additional information

This work is dedicated to the late Marie-France Michel who initiated the poplar biotechnology project at INRA.

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Brasileiro, A.C.M., Leplé, JC., Muzzin, J. et al. An alternative approach for gene transfer in trees using wild-typeAgrobacterium strains. Plant Mol Biol 17, 441–452 (1991). https://doi.org/10.1007/BF00040638

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  • DOI: https://doi.org/10.1007/BF00040638

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