Project
Introducing Phytophthora infestans resistance in potato using CRISPR (CRISPR-SOLANUM)
Can we use a CRISPR based approach to introduce Phytophthora infestans (P.i.) resistance in potato? Can we design a transfection and a screening system for this? Can we knock out plant susceptibility genes to avoid infection, while limiting pleiotropic effects on other plant processes? Can we use a phenotypical test to evaluate the effect of these mutations? These are the main research questions in the CRISPR-SOLANUM project.
Research approach
First, we develop an efficient protoplast-based transfection and regeneration protocol starting from 5-6 commercial cultivars, and select 1-2 for the rest of the project. Second, we identify candidate S-genes, design guide RNAs, construct vectors, transfect protoplasts and molecularly characterize mutation events. Finally, we phenotype regenerated material by testing disease resistance and screening for pleiotropic effects.
Relevance/Valorisation
Ultimately, 2 potato cultivars were chosen for this project, ‘Bintje’ and ‘Spunta’. For both genotypes protoplasts could be successfully transfected and mutated but only ‘Spunta’ protoplasts could regenerate into plants. Transfection and screening systems were developed and optimized and after that susceptibility genes (S genes) were targeted with CRISPR/Cas. Several mutants were regenerated. In a later phase the phenotypic consequences of these mutations (mainly whether the susceptibility for Phytophthora late blight has been reduced or not) will be tested; therefore, a biotest was selected during this PhD. The knowledge developed in this project is particularly relevant for potato breeders and growers and molecular breeders.