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oai:riuma.uma.es:10630/161992026-02-03T12:24:43Zcom_10630_2254col_10630_37959

Targeted mutagenesis of FaTM6 in the Octoploid Strawberry (Fragaria x ananassa) using the CRISPR/Cas9 System Martín-Pizarro, Carmen Duarte-Conde, José Antonio Posé-Padilla, David Fresas Strawberry Fragaria x ananassa CRISPR/Cas9 Flower Stamen Petal TM6 AP3 The B-class of MADS-box transcription factors has been studied in many plant species, but remain functionally uncharacterized in the Rosaceae family. APETALA3 (AP3), a member of this class, controls the identity of petals and stamens in Arabidopsis thaliana. In this work, we identified two members of the AP3 lineage in the cultivated strawberry (Fragaria × ananassa): FaAP3 and FaTM6. Interestingly, FaTM6, and not FaAP3, shows an expression pattern equivalent to that of AP3 in Arabidopsis. Genome editing using Cluster Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 system is becoming a robust tool for targeted and stable mutagenesis of DNA. However, whether it can be efficiently used in an octoploid species such as F. × ananassa is not known. In our study, we report the application of the CRISPR/Cas9 in F. × ananassa to characterize the function of FaTM6 in flower development. An exhaustive analysis by high-throughput sequencing of the FaTM6 locus spanning the target sites showed a high efficiency genome editing already in the T0 generation. The phenotypic characterization of the mutant lines indicates that FaTM6 plays a key role in petal and especially in anther development in strawberry. Our results validate the CRISPR/Cas9 strategy for gene functional analysis in an octoploid species such as F. × ananassa, and offer new opportunities for engineering strawberry to improve traits of interest in breeding programs. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech. 2018-07-12T09:07:44Z 2018-07-12T09:07:44Z 2018 2018-07-12 conference output https://hdl.handle.net/10630/16199 eng XIV Reunión de Biología Molecular de Plantas Salamanca Julio 2018 Attribution-NonCommercial-NoDerivatives 4.0 Internacional Attribution-NonCommercial-NoDerivatives 4.0 Internacional http://creativecommons.org/licenses/by-nc-nd/4.0/ open access application/pdf