Transformation: FxaRT0001

Transformation Information

Transformation Name: FxaRT0001
Description: transformation FxaRT0001 to Hawaii 4
Genetic background: Hawaii 4
Project Name: CRISPR/Cas9?mediated mutagenesis of phytoene desaturase in diploid and octoploid strawberry
Description: Background: Gene editing using CRISPR/Cas9 is a simple and powerful tool for elucidating genetic controls and for crop improvement and its use has been reported in a growing number of important food crops, including recently Fragaria. In order to inform application of the technology in Fragaria, we targeted the visible endogenous marker gene PDS (phytoene desaturase) in diploid Fragaria vesca ssp. vesca Hawaii 4 and octoploid F. x ananassa Calypso. Results: Agrobacterium-mediated transformation of leaf and petiole explants was used for efficient stable integration of constructs expressing plant codon-optimised Cas9 and single guide sequences under control of the Arabidopsis U6-26 consensus promoter and terminator or Fragaria vesca U6III regulatory sequences. More than 80% (Hawaii 4) and 50% (Calypso) putative transgenic shoot lines (multiple shoots derived from a single callus) exhibited mutant phenotypes. Of mutant shoot line. Of mutant shoot lines selected for molecular analysis, approximately 75% (¿Hawaii 4¿) and 55% (¿Calypso¿) included albino regenerants with bi-allelic target sequence variants. Our results indicate the PDS gene is functionally diploid in ¿Calypso¿. Conclusion: We demonstrate that CRISPR/Cas9 may be used to generate biallelic mutants at high frequency within the genomes of diploid and octoploid strawberry. The methodology, observations and comprehensive data set presented will facilitate routine application of this technology in Fragaria to single and multiple gene copy targets where mutant phenotypes cannot be identified visually.

Construct for Transformation (FxaRT0001)


Plants for Transformation (FxaRT0001)

Note: The tree nodes can be clicked to show the generations of plants. T0 represents the 1st generation (Progenitor of T1), T1 represents the 2nd generation (Progeny of T0), and so on. Clicking the plant name will be directed to the plant information page.

This database is supported by NSF (IOS-1546625) and hosted by BTI.