IMR Press / FBL / Volume 28 / Issue 9 / DOI: 10.31083/j.fbl2809201
Open Access Original Research
CRISPR/Cas9-Targeted Mutagenesis of CiGAS and CiGAO to Reduce Bitterness in Chicory (Cichorium intybus L.)
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1 UMR Transfrontalière BioEcoAgro N° 1158, Univ. Lille, INRAE, Univ. Liège, UPJV, ISA, Univ. Artois, Univ. Littoral Côte d’Opale, Institut CharlesViollette, Joint Laboratory CHIC41H, University of Lille-Florimond Desprez, Cité scientifique, 59655 Villeneuve d’ascq, France
*Correspondence: (Caroline Rambaud)
Front. Biosci. (Landmark Ed) 2023, 28(9), 201;
Submitted: 28 November 2022 | Revised: 18 January 2023 | Accepted: 13 March 2023 | Published: 14 September 2023
Copyright: © 2023 The Author(s). Published by IMR Press.
This is an open access article under the CC BY 4.0 license.

Background: Chicory (Cichorium intybus L.), a member of the Asteraceae family, is known for its numerous health benefits, including its prebiotic, digestive, antioxidant or anti-inflammatory effects. Used as a coffee substitute, chicory roots is also appreciated for its bitterness, which can prove to be a disadvantage for other uses in food. The bitterness of chicory is largely linked to the presence of sesquiterpene lactones (STLs) in the roots. Methods: In order to create less bitter industrial chicory varieties, CRISPR/Cas9 technology was used to inhibit the first two genes of the STL biosynthetic pathway: germacrene A synthase (CiGAS), short form, and germacrene A oxidase (CiGAO). To determine the impact of these reductions on the perception of bitterness, a sensory analysis of 13 field-grown chicories genotypes, contrasting for their STL composition, allowed the construction of obtain a bitterness scale by correlating STL content with perceived bitterness. The edited chicories were positioned on this scale according to their STL content. Results: Biallelic mutations in two of the copies of CiGAS-short form or in the CiGAO gene led to a reduction in STL content of edited chicories and a reduction in bitterness, or even an absence of perception, was obtained for some mutants. Conclusions: The use of the CRISPR/Cas9 tool as well as the choice of targets therefore makes it possible to modulate the bitterness of chicory.

Cichorium intybus L.
sesquiterpene lactones
hairy roots
germacrene A synthase
germacrene A oxidase
Région des Hauts-de-France
Fig. 1.
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