Agrobacterium rhizogenes-Mediated Genetic Transformation and Establishment of CRISPR/Cas9 Genome-Editing Technology in Limonium bicolor

Limonium bicolor is a perennial herbaceous plant belonging to the Plumbaginaceae family. It can be used as dried flower or in cut arrangements and serves model recretohalophyte. Its genome sequencing has been recently completed. However, research on L. limited by absence of highly efficient genetic transformation system. In this study, we established Agrobacterium rhizogenes-mediated method. The transgenic hairy roots were induced from hypocotyl using A. rhizogenes strain K599 harboring pRdGa4Cas9 plasmid (which carries an expression cassette 35S::DsRed2). shoots regenerated root segments (~0.1 cm diameter), induction efficiency was achieved at 100%. with 4–5 rosette leaves directly planted into soil induce roots. Therefore, plantlets produced. DsRed2 reliable reporter gene screening plantlets. Furthermore, also CRISPR/Cas9 system employing approach. method CRIPSP/Cas9 will provide valuable tool for functional genomics investigation trait improvement bicolor.