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

Leroy Thierry, Marraccini Pierre, Dufour Magali, Montagnon Christophe, Lashermes Philippe, Sabau Xavier, Ferreira Lucia Pires, Jourdan Isabelle, Pot David, Andrade Alan Carvalho, Glaszmann Jean-Christophe, Vieira Luiz Gonzaga Esteves, Piffanelli Pietro. 2005. Construction and characterization of a Coffea canephora BAC library to study the organization of sucrose biosynthesis genes. Theoretical and Applied Genetics, 111 (6) : 1032-1041.

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

Article de revue ; Article de revue à facteur d'impact

Version publiée - Anglais
Accès réservé aux personnels Cirad
Utilisation soumise à autorisation de l'auteur ou du Cirad.
document_529101.pdf
Télécharger (544kB)

Résumé : 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.

Mots-clés Agrovoc : Coffea canephora, banque de gènes, génome, pool de gènes, RFLP, clone, biotechnologie végétale, caractère agronomique, génie génétique, chromosome

Mots-clés complémentaires : Bac, Chromosome artificiel

Classification Agris : F30 - Génétique et amélioration des plantes

Champ stratégique Cirad : Axe 1 (2005-2013) - Intensification écologique

Auteurs et affiliations