Unraveling the biosynthetic pathway of sesquiterpene lactones in industrial chicory and witloof using CRISPR/Cas9 genome editing

Industrial chicory (Cichorium intybus L. var. sativum) and witloof (Cichorium intybus L. var.
foliosum) are economically important crops in Flanders. Industrial chicory is mainly produced for
inulin, while witloof is a well-known vegetable. However, today the bitter taste of these crops
limits their use. Reducing the bitterness will create new economic opportunities. Industrial chicory
can then be used as a flour for the bakery industry and for the production of novel food products,
opening a complete new market. For witloof, changing bitterness can lead to product
differentiation by creating a more diverse range of flavors, maximizing consumers acceptance. The
bitterness is caused by sesquiterpene lactones (SL). The goal of our project is to gain more
knowledge about the regulatory and biosynthetic genes involved in these bitter compounds.
Therefor we will use a new breeding technique CRISPR/Cas9 which allows to mutate plants on very
specific places in the DNA. To do this we will work with an innovative strategy based on
protoplasts, which are cells without a cell wall, that enables a more efficient DNA uptake. Since
CRISPR/Cas9 has not yet been performed in Cichorium, protocols will be optimized. Obtained
plants will be screened to find the right mutants. In the end the project will lead to the creation of
new plants with changed metabolite profiles in a faster and more precise way, compared to the
traditional breeding techniques used up till now.