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Breitler J. C., Dechamp E., Campa Claudine, Rodrigues L. A. Z., Guyot Romain, Marraccini P., Etienne H. (2018). CRISPR/Cas9-mediated efficient targeted mutagenesis has the potential to accelerate the domestication of Coffea canephora. Plant Cell Tissue and Organ Culture, 134 (3), 383-394. ISSN 0167-6857

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Lien direct chez l'éditeur doi:10.1007/s11240-018-1429-2

Titre
CRISPR/Cas9-mediated efficient targeted mutagenesis has the potential to accelerate the domestication of Coffea canephora
Année de publication2018
Type de documentArticle référencé dans le Web of Science WOS:000441559700004
AuteursBreitler J. C., Dechamp E., Campa Claudine, Rodrigues L. A. Z., Guyot Romain, Marraccini P., Etienne H.
SourcePlant Cell Tissue and Organ Culture, 2018, 134 (3), p. 383-394. ISSN 0167-6857
RésuméGenome editing, which is an unprecedented technological breakthrough, has provided a valuable means of creating targeted mutations in plant genomes. In this study, we developed a genomic web tool to identify all gRNA target sequences in the coffee genome, along with potential off-targets. In all, 8,145,748 CRISPR guides were identified in the draft genome of Coffea canephora corresponding to 5,338,568 different sequences and, of these, 4,655,458 were single, and 514,591 were covering exons. The proof of concept was established by targeting the phytoene desaturase gene (CcPDS) using the Agrobacterium tumefaciens transformation technique and somatic embryogenesis as the plant regeneration method. An analysis of the RNA-guided genome-editing events showed that 22.8% of the regenerated plants were heterozygous mutants and 7.6% were homozygous mutants. Mutation efficiency at the target site was estimated to be 30.4%. We demonstrated that genome editing by the CRISPR/Cas9 method is an efficient and reliable way of knocking out genes of agronomic interest in the coffee tree, opening up the way for coffee molecular breeding. Our results also showed that the use of somatic embryogenesis, as the method for regenerating genome-edited plants, could restrict the choice of targeted genes to those that are not essential to the embryo development and germination steps.
Plan de classementSciences du monde végétal [076] ; Biotechnologies [084]
LocalisationFonds IRD [F B010073769]
Identifiant IRDfdi:010073769
Lien permanenthttp://www.documentation.ird.fr/hor/fdi:010073769

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