Agrobacterium rhizogenes-mediated transformation of a dioecious plant model Silene latifolia
Language English Country Netherlands Media print-electronic
Document type Journal Article
PubMed
29656128
DOI
10.1016/j.nbt.2018.04.001
PII: S1871-6784(17)30515-0
Knihovny.cz E-resources
- Keywords
- CRISPR/Cas9, Genetic transformation, In vitro regeneration, Protoplast assay, Silene latifolia,
- MeSH
- Agrobacterium genetics MeSH
- Chromosomes, Plant genetics MeSH
- CRISPR-Cas Systems MeSH
- DNA, Bacterial genetics MeSH
- Gene Expression MeSH
- Genetic Techniques MeSH
- Plants, Genetically Modified MeSH
- Models, Genetic MeSH
- Evolution, Molecular MeSH
- Regeneration genetics MeSH
- Genes, Reporter MeSH
- Silene genetics microbiology physiology MeSH
- Transcription Activator-Like Effector Nucleases MeSH
- Transformation, Genetic * MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- DNA, Bacterial MeSH
- T-DNA MeSH Browser
- Transcription Activator-Like Effector Nucleases MeSH
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.
References provided by Crossref.org
Sexy ways: approaches to studying plant sex chromosomes
How to use CRISPR/Cas9 in plants: from target site selection to DNA repair
Sex and the flower - developmental aspects of sex chromosome evolution
