Silene latifolia serves as a model species to study dioecy, the evolution of sex chromosomes, dosage compensation and sex-determination systems in plants. Currently, no protocol for genetic transformation is available for this species, mainly because S. latifolia is considered recalcitrant to in vitro regeneration and infection with Agrobacterium tumefaciens. Using cytokinins and their synthetic derivatives, we markedly improved the efficiency of regeneration. Several agrobacterial strains were tested for their ability to deliver DNA into S. latifolia tissues leading to transient and stable expression of the GUS reporter. The use of Agrobacterium rhizogenes strains resulted in the highest transformation efficiency (up to 4.7% of stable transformants) in hairy root cultures. Phenotypic and genotypic analyses of the T1 generation suggested that the majority of transformation events contain a small number of independent T-DNA insertions and the transgenes are transmitted to the progeny in a Mendelian pattern of inheritance. In short, we report an efficient and reproducible protocol for leaf disc transformation and subsequent plant regeneration in S. latifolia, based on the unique combination of infection with A. rhizogenes and plant regeneration from hairy root cultures using synthetic cytokinins. A protocol for the transient transformation of S.latifolia protoplasts was also developed and applied to demonstrate the possibility of targeted mutagenesis of the sex linked gene SlAP3 by TALENs and CRISPR/Cas9.

• MeSH
• Agrobacterium genetika   MeSH
• chromozomy rostlin genetika   MeSH
• CRISPR-Cas systémy MeSH
• DNA bakterií genetika   MeSH
• exprese genu MeSH
• genetické techniky MeSH
• geneticky modifikované rostliny MeSH
• modely genetické MeSH
• molekulární evoluce MeSH
• regenerace genetika   MeSH
• reportérové geny MeSH
• Silene genetika   mikrobiologie   fyziologie   MeSH
• TALENs MeSH
• transformace genetická * MeSH
• Publikační typ
• časopisecké články MeSH