CRISPR/Cas9-mediated efficient targeted mutagenesis has the potential to accelerate the domestication of Coffea canephora
Autor: | Jean-Christophe Breitler, Eveline Dechamp, Claudine Campa, Hervé Etienne, Romain Guyot, Leonardo Augusto Zebral Rodrigues, Pierre Marraccini |
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Přispěvatelé: | UMR - Interactions Plantes Microorganismes Environnement (UMR IPME), Institut de Recherche pour le Développement (IRD [France-Sud])-Université de Montpellier (UM)-Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Département Systèmes Biologiques (Cirad-BIOS), Centre de Coopération Internationale en Recherche Agronomique pour le Développement (Cirad), Federal University of Lavras, ANR-10-LABX-0001,AGRO,Agricultural Sciences for sustainable Development(2010) |
Jazyk: | angličtina |
Rok vydání: | 2018 |
Předmět: |
0106 biological sciences
0301 basic medicine Phytoene desaturase Targeted mutagenesis Agrobacterium Mutagénèse F62 - Physiologie végétale - Croissance et développement Computational biology Horticulture Biology Coffea canephora 01 natural sciences Genome F30 - Génétique et amélioration des plantes 03 medical and health sciences Genome editing Embryogénèse somatique CRISPR Gene 2. Zero hunger Physiologie du développement Génome Cas9 Domestication des plantes Biologie moléculaire CRISPR/CA9 CRISPRCas9 Somatic embryogenesis biology.organism_classification [SDV.BV.AP]Life Sciences [q-bio]/Vegetal Biology/Plant breeding 030104 developmental biology Phytoene Desaturase 010606 plant biology & botany |
Zdroj: | Plant Cell, Tissue and Organ Culture Plant Cell, Tissue and Organ Culture, Springer Verlag, 2018, 134 (3), pp.383-394. ⟨10.1007/s11240-018-1429-2⟩ 27th Biennial ASIC Conference, Portland, 16-20 September 2018. Book of abstracts 2019 |
ISSN: | 0167-6857 1573-5044 |
Popis: | International audience; Genome 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 |
Databáze: | OpenAIRE |
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