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Construction and characterization of a Coffea canephora BAC library to study the organization of sucrose biosynthesis genes

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Abstract

The first bacterial artificial chromosome (BAC) library of Robusta coffee (Coffea canephora) was constructed, with the aim of developing molecular resources to study the genome structure and evolution of this perennial crop. Clone 126, which is highly productive and confers good technological and organoleptic qualities of beverage, was chosen for development of this library. The BAC library contains 55,296 clones, with an average insert size of 135 Kb per plasmid, therefore representing theoretically nine haploid genome equivalents of C. canephora. Its validation was achieved with a set of 13 genetically anchored single-copy and 4 duplicated RFLP probes and yielded on average 9 BAC clones per probe. Screening of this BAC library was also carried out with partial cDNA probes coding for enzymes of sugar metabolism like invertases and sucrose synthase, with the aim of characterizing the organization and promoter structure of this important class of genes. It was shown that genes for both cell wall and vacuolar forms of invertases were probably unique in the Robusta genome whereas sucrose synthase was encoded by at least two genes. One of them (CcSUS1) was cloned and sequenced, showing that our BAC library is a valuable tool to rapidly identify genes of agronomic interest or linked to cup quality in C. canephora.

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Acknowledgments

We thank the robotic team of CIRAD for the management of the BAC library. We are also very grateful to K. Avia for his help concerning gene mapping. The work was supported by the French Embassy in Brazil (DCSUR-BRE-4C5-008) to P. Marraccini. T. Leroy should be contacted for further information about the access to the BAC library.

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Authors and Affiliations

  1. Centre de Coopération Internationale en Recherche Agronomique pour le Développement (CIRAD), TA 80/03, Avenue d’Agropolis, 34398, Montpellier Cedex 5, France

    T. Leroy, P. Marraccini, M. Dufour, C. Montagnon, X. Sabau, I. Jourdan, D. Pot, J. C. Glaszmann & P. Piffanelli

  2. Instituto Agronômico do Paraná (IAPAR), LBI-AMG, CP 481, 86001-970, Londrina (PR), Brazil

    P. Marraccini, L. P. Ferreira & L. G. E. Vieira

  3. Institut de Recherche pour le Développement (IRD), 911 Avenue Agropolis, BP 64501, 34394, Montpellier Cedex 5, France

    P. Lashermes

  4. Parque Estação Biológica, Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) -Cenargen, CP 02372, 70770-900, Brasilia (DF), Brazil

    A. C. Andrade

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  1. T. Leroy
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  2. P. Marraccini
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  3. M. Dufour
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  4. C. Montagnon
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  5. P. Lashermes
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  6. X. Sabau
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  7. L. P. Ferreira
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  8. I. Jourdan
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  9. D. Pot
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  10. A. C. Andrade
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  11. J. C. Glaszmann
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  12. L. G. E. Vieira
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  13. P. Piffanelli
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Corresponding author

Correspondence to T. Leroy.

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Communicated by H. Nybom

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Leroy, T., Marraccini, P., Dufour, M. et al. Construction and characterization of a Coffea canephora BAC library to study the organization of sucrose biosynthesis genes. Theor Appl Genet 111, 1032–1041 (2005). https://doi.org/10.1007/s00122-005-0018-z

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  • DOI: https://doi.org/10.1007/s00122-005-0018-z

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