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Transgenic coffee fruits from Coffea arabica genetically modified by bombardment

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Abstract

The genetic modification of Coffea arabica fruits is an important tool for the investigation of physiological characteristics and functional validation of genes related to coffee bean quality traits. In this work, plants of C. arabica cultivar Catuaí Vermelho were successfully genetically modified by bombardment of embryogenic calli. Calli were obtained from 90% of the leaf explants cultivated in a callogenesis-inducing medium modified with 20 μM 2,4-dichlorophenoxyacetic acid (2,4-D). The resulting calli were bombarded with the pBI426 vector containing a uidA and nptII gene fusion that was driven by the double CaMV35s promoter. Kanamycin-selected embryos were positive for β-glucuronidase (GUS) activity in histochemical assays and for target gene amplification by polymerase chain reaction. Integration of the nptII gene was confirmed by Southern blot and showed a low copy number (one to three) of insertions. Transformed plants showed normal development and settled fruits. GUS expression was assessed in the flower and fruit organs demonstrating the capacity of the double CaMV35s promoter to drive long-term stable expression of uidA in C. arabica fruit tissues. Moreover, we obtained a T1 progeny presenting 3:1 Mendelian segregation of the uidA gene. This investigation is the first to report exogenous gene expression in coffee fruits and transgenic inheritance in C. arabica plants.

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Acknowledgments

C. arabica seeds were kindly provided by Dr. Antonio A. Pereira from EPAMIG. This research was supported by the “Consórcio Brasileiro de Pesquisa e Desenvolvimento do Café” and Embrapa.

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Authors and Affiliations

  1. Embrapa Recursos Genéticos e Biotecnologia, Laboratório de Interação Molecular Planta-Praga, PqEB Av. W5-Norte (final), Brasilia, 707770-900, DF, Brazil

    Erika V. S. Albuquerque, Welcimar G. Cunha, Aulus E. A. D. Barbosa, Poliene M. Costa, João B. Teixeira, Giovanni R. Vianna, Glaucia B. Cabral & Maria F. Grossi-de-Sa

  2. Institut de Recherche pour le Développement, Résistance des Plantes aux Bioagresseurs IRD-UMR-186 IRD-Cirad-UM2, BP 64501, 34394, Montpellier, Cedex 5, France

    Diana Fernandez

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  1. Erika V. S. Albuquerque
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  2. Welcimar G. Cunha
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  3. Aulus E. A. D. Barbosa
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  4. Poliene M. Costa
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  5. João B. Teixeira
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  6. Giovanni R. Vianna
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  7. Glaucia B. Cabral
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  8. Diana Fernandez
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  9. Maria F. Grossi-de-Sa
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Correspondence to Erika V. S. Albuquerque.

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Editor: N. J. Taylor

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Albuquerque, E.V.S., Cunha, W.G., Barbosa, A.E.A.D. et al. Transgenic coffee fruits from Coffea arabica genetically modified by bombardment. In Vitro Cell.Dev.Biol.-Plant 45, 532–539 (2009). https://doi.org/10.1007/s11627-009-9254-2

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  • DOI: https://doi.org/10.1007/s11627-009-9254-2

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