Functional Analysis of TM6 MADS box gene in the Octoploid Strawberry by CRISPR/Cas9 directed mutagenesis

Martin-Pizarro, C.; Trivino, J. C.; Pose, D.

doi:10.1101/351296

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Functional Analysis of TM6 MADS box gene in the Octoploid Strawberry by CRISPR/Cas9 directed mutagenesis

Martin-Pizarro, C.·Trivino, J. C.·Pose, D.

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Abstract

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 x 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. x ananassa is not known. Here we report for the first time the application of CRISPR/Cas9 in F. x 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. in an octoploid species such as F. x ananassa, and offer new opportunities for engineering strawberry to improve traits of interest in breeding programs.

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Martin-Pizarro, C., Trivino, J. C., Pose, D.. 2018-06-20. Functional Analysis of TM6 MADS box gene in the Octoploid Strawberry by CRISPR/Cas9 directed mutagenesis. https://doi.org/10.1101/351296

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