RESEARCHERS DEVELOP CRISPR SYSTEM FOR SALSIFIES
Tragopogon or salsifies from the Asteraceae family is an excellent model for polyploidy studies. An efficient CRISPR-Cas9-based genome editing technique for salsifies will greatly aid studies on the genetics of polyploidy. University of Florida researchers led by Shengchen Shan aimed at developing a CRISPR-Cas9 system for salsifies.
The team established a feasible tissue culture and transformation method for Tragopogon. Through protoplast transient assays, the TragCRISPR system (the CRISPR-Cas9 system adapted for Tragopogon) was able to introduce site-specific mutations in Tragopogonprotoplasts. Agrobacterium‐mediated transformation with Cas9‐sgRNA constructs targeting the phytoene desaturase gene (TraPDS) was implemented in this model polyploid system.
Analysis of the albino shoots of the transformed individuals indicated simultaneous mutations of two TraPDS alleles from T. porrifolius, a diploid. In T. mirus, a tetraploid, four TraPDS alleles were also found to have mutations. The frequencies of the successfully transformed calli with the albino phenotype were 87% and 78% in the diploid and polyploid, respectively.
This study reveals that the application of CRISPR-Cas9 system can enable investigations and permit unique studies in polyploids.
For more information, read the article in Molecular Ecology Resources.