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An Efficient Procedure to Stably Introduce Genes into an Economically Important Pulp Tree (Eucalyptus grandis × Eucalyptus urophylla)

Transgenic Research volume 12, pages 403–411 (2003)Cite this article

Abstract

Regeneration problems are one of the main limitations preventing the wider application of genetic engineering strategies to the genus Eucalyptus. Seedlings from Eucalyptus grandis × Eucalyptus urophylla were selected according to their regeneration (adventitious organogenesis) and transformation capacity. After in vitro cloning, the best genotype of 250 tested was transformed via Agrobacterium tumefaciens. A cinnamyl alcohol dehydrogenase (CAD) antisense cDNA from Eucalyptus gunnii was transferred, under the control of the 35S CaMV promoter with a double enhancer sequence, into a selected genotype. According to kanamycin resistance and PCR verification, 120 transformants were generated. 58% were significantly inhibited for CAD activity, and nine exhibited the highest down-regulation, ranging from 69 to 78% (22% residual activity). Southern blot hybridisation showed a low transgene copy number, ranging from 1 to 4, depending on the transgenic line. Northern analyses on the 5–16 and 3–23 lines (respectively one and two insertion sites) demonstrated the antisense origin of CAD gene inhibition. With respectively 26 and 22% of residual CAD activity, these two lines were considered as the most interesting and transferred to the greenhouse for further analyses.

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Author notes

  1. Gisele de Andrade

    Present address: University of Georgia, School of Forest Resources, Athens, GA, 30602, USA

Affiliations

  1. UMR 5546, Pôle de Biotechnologie Végétale, 24 Chemin de Borde Rouge, BP17 Auzeville, 31326, Castanet-Tolosan, France

    Vincent Tournier, Sabine Grat, Christiane Marque, Walid El Kayal, Alain-Michel Boudet & Chantal Teulières

  2. AracruCelulose S.A. Centro de Pesquisa e Tecnologia, Rodovia ES-257, km 25, Aracruz, ES 29197-000, Brazil

    Ricardo Penchel & Gisele de Andrade

Authors
  1. Vincent Tournier
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  2. Sabine Grat
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  3. Christiane Marque
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  4. Walid El Kayal
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  5. Ricardo Penchel
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  6. Gisele de Andrade
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  7. Alain-Michel Boudet
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  8. Chantal Teulières
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Correspondence to Chantal Teulières.

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Tournier, V., Grat, S., Marque, C. et al. An Efficient Procedure to Stably Introduce Genes into an Economically Important Pulp Tree (Eucalyptus grandis × Eucalyptus urophylla). Transgenic Res 12, 403–411 (2003). https://doi.org/10.1023/A:1024217910354

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  • DOI: https://doi.org/10.1023/A:1024217910354

  • antisense
  • cinnamyl alcohol dehydrogenase
  • eucalyptus
  • genetic engineering
  • juvenile clones