Amb motiu del tancament d'estiu, la validació de documents es reprendrà a partir del 28 d'agost de 2026. Disculpeu les molèsties.
Con motivo del cierre de verano, la validación de documentos se reanudará a partir del 28 de agosto de 2026. Disculpad las molestias
Due to the summer closure, document validation will resume starting August 28, 2026. We apologize for any inconvenience.

Files

690717.pdf (2.19 MB)

Document type

Article

Version

Published version

Publication date

2019-10-20

Publication license

cc-by (c) Hooghvorst, Isidre et al., 2019
Please use this identifier to cite or link to this item: https://hdl.handle.net/2445/150815

Efficient knockout of Phytoene desaturase gene using CRISPR/Cas9 in melon

Journal Title

Authors

Director/Tutor

Journal ISSN

Volume Title

Related resource

https://doi.org/10.1038/s41598-019-53710-4

Abstract

CRISPR/Cas9 system has been widely applied in many plant species to induce mutations in the genome for studying gene function and improving crops. However, to our knowledge, there is no report of CRISPR/Cas9-mediated genome editing in melon (Cucumis melo). In our study, phytoene desaturase gene of melon (CmPDS) was selected as target for the CRISPR/Cas9 system with two designed gRNAs, targeting exons 1 and 2. A construct (pHSE-CmPDS) carrying both gRNAs and the Cas9 protein was delivered by PEG-mediated transformation in protoplasts. Mutations were detected in protoplasts for both gRNAs. Subsequently, Agrobacterium-mediated transformation of cotyledonary explants was carried out, and fully albino and chimeric albino plants were successfully regenerated. A regeneration efficiency of 71% of transformed plants was achieved from cotyledonary explants, a 39% of genetic transformed plants were successful gene edited, and finally, a 42-45% of mutation rate was detected by Sanger analysis. In melon protoplasts and plants most mutations were substitutions (91%), followed by insertions (7%) and deletions (2%). We set up a CRISPR/Cas9-mediated genome editing protocol which is efficient and feasible in melon, generating multi-allelic mutations in both genomic target sites of the CmPDS gene showing an albino phenotype easily detectable after only few weeks after Agrobacteriummediated transformation.

Subject

Mutació (Biologia), Fruita

Subject (English)

Mutation (Biology), Fruit

Citation

Collections

Articles publicats en revistes (Biologia Evolutiva, Ecologia i Ciències Ambientals)

Citation

HOOGHVORST, Isidre, LÓPEZ CRISTOFFANINI, Camilo Alonso and NOGUÉS MESTRES, Salvador. Efficient knockout of Phytoene desaturase gene using CRISPR/Cas9 in melon. Scientific Reports. 2019. Vol. 9, num. 17077. ISSN 2045-2322. [consulted: 14 of August of 2026]. Available at: https://hdl.handle.net/2445/150815

Export metadata

JSON - METS

Share record