Tissue specific expression and genomic organization of bitter sesquiterpene lactone biosynthesis in Cichorium intybus L. (Asteraceae)

Bogdanović﻿﻿﻿, Milica; Cankar, Katarina; Todorović, Slađana; Dragićević, Milan; Simonović, Ana; Houwelingen, Adèle van; Schijlen, Elio; Schipper, Bert; Gagneul, David; Hendriks, Théo; Quillet, Marie-Christine; Bouwmeester, Harro J.; Bosch, Dirk; Beekwilder, Jules

doi:10.1016/j.indcrop.2018.12.011

Cited by 19 publications

(33 citation statements)

References 32 publications

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“…In chicory, four different germacrene A synthase genes ( Ci GAS) are present, one copy of GAS‐ long ( Ci GAS‐L) and three physically linked copies of GAS‐ short (Ci GAS‐S1, Ci GAS‐S2 and Ci GAS‐S3) (Bogdanović et al ., 2019 ). Expression analysis of the Ci GAS genes has shown that both the Ci GAS‐L and Ci GAS‐ short genes are expressed in chicory roots, while predominantly only the Ci GAS‐L is expressed in chicory leaves (Bogdanović et al ., 2019 ; Bouwmeester et al ., 2002 ). Silencing of the Ci GAS genes by a microRNA approach resulted in reduced expression of both the long and short germacrene A synthases and a partial decrease of STL levels (Bogdanovic et al ., 2020 ).…”

Section: Introductionmentioning

confidence: 99%

Inactivation of the germacrene A synthase genes by CRISPR/Cas9 eliminates the biosynthesis of sesquiterpene lactones in Cichorium intybus L.

Cankar

Bundock

Sévenier

et al. 2021

Plant Biotechnology Journal

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Chicory (Cichorium intybus var. sativum) is an industrial crop species cultivated for the production of a fructose polymer inulin, which is used as a low-calorie sweetener and prebiotic. Besides, inulin chicory taproots also accumulate sesquiterpene lactones (STLs). These are bitter tasting compounds, which need to be removed during inulin extraction, resulting in additional costs. In this work, we describe chicory lines where STL accumulation is almost completely eliminated. Genome editing using the CRISPR/Cas9 system was used to inactivate four genes that encode the enzyme that performs the first dedicated step in STL synthesis, germacrene A synthase (CiGAS). Chicory lines were obtained that carried null mutations in all four CiGAS genes. Lines lacking functional CiGAS alleles showed a normal phenotype upon greenhouse cultivation and show nearly complete elimination of the STL synthesis in the roots. It was shown that the reduction in STLs could be attributed to mutations in genetically linked copies of the CiGAS-short gene and not the CiGAS-long gene, which is relevant for breeding the trait into other cultivars. The inactivation of the STL biosynthesis pathway led to increase in phenolic compounds as well as accumulation of squalene in the chicory taproot, presumably due to increased availability of farnesyl pyrophosphate (FFP). These results demonstrate that STLs are not essential for chicory growth and that the inhibition of the STL biosynthesis pathway reduced the STL levels chicory which will facilitate inulin extraction.

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Section: Introductionmentioning

confidence: 99%

Inactivation of the germacrene A synthase genes by CRISPR/Cas9 eliminates the biosynthesis of sesquiterpene lactones in Cichorium intybus L.

Cankar

Bundock

Sévenier

et al. 2021

Plant Biotechnology Journal

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“…Primers were designed according to Schwab et al 26 AmiRNA sequences specific for either CiGASlo (amiGASlo, TATCTAAGAT- Figure 1. STL biosynthesis in chicory and the most abundant guaianolide compounds, modified from Bogdanović et al 13 GASgermacrene A synthase, 12 GAOsgermacrene A oxidases, 14,15 COScostunolide synthase, 16,17 KLSkauniolide synthase. 18 Silencing target -GAS is marked by the red arrow.…”

Section: Vector Constructionmentioning

confidence: 99%

“…The most diversified and abundant group of STLs in chicory are the guaianolides, 11 all derived from a common precursorgermacrene A. 12 Present both in leaves and roots of the plant, the most common guaianolides in chicory are lactucin, lactucopicrin, and 8-deoxylactucin, as well as their modified formsoxalates, sulfates and glycosides, with oxalates being the most abundant form. 3 These compounds are mainly responsible for the bitter taste of chicory and lettuce.…”

Section: Introductionmentioning

confidence: 99%

“…6 The first step in guaianolide biosynthesis is the conversion of farnesyl diphosphate (FPP) to germacrene A (Figure 1), catalyzed by germacrene A synthase (GAS). 12 This enzyme is encoded in chicory by two genes, whose cDNAs -CiGASlo (long isoform) and CiGASsh (short isoform)have been isolated previously. 19 The two isozymes have 72% similarity at the amino acid level, and both convert FPP to germacrene A, as confirmed by heterologous expression.…”

Section: Introductionmentioning

confidence: 99%

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Silencing of germacrene A synthase genes reduces guaianolide oxalate content in Cichorium intybus L.

et al. 2019

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Chicory (Cichorium intybus L.) is a medicinal and industrial plant from the Asteraceae family that produces a variety of sesquiterpene lactones (STLs), most importantly bitter guaianolides: lactucin, lactucopicrin and 8-deoxylactucin as well as their modified forms such as oxalates. These compounds have medicinal properties; however, they also hamper the extraction of inulina very important food industry product from chicory roots. The first step in guaianolide biosynthesis is catalyzed by germacrene A synthase (GAS) which in chicory exists in two isoforms-GAS long (encoded by CiGASlo) and GAS short (encoded by CiGASsh). AmiRNA silencing was used to obtain plants with reduced GAS gene expression and level of downstream metabolites, guaianolide-15oxalates, as the major STLs in chicory. This approach could be beneficial for engineering new chicory varieties with varying STL content, and especially varieties with reduced bitter compounds more suitable for inulin production.

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“…The major STLs in chicory are derived from germacrene A. The most abundant are lactucin, lactucopicrin, 8-deoxylactucin and their oxalate and glucoside derivatives [ 8 , 9 ]. Lactucin and lactucopicrin are responsible for the antimalarial properties of chicory.…”

Section: Introductionmentioning

confidence: 99%

Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens

et al. 2021

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Chicory (Cichorium intybus L.) is an important industrial crop cultivated mainly to extract the dietary fiber inulin. However, chicory also contains bioactive compounds such as sesquiterpene lactones and certain polyphenols, which are currently discarded as waste. Plants are an important source of active pharmaceutical ingredients, including novel antimicrobials that are urgently needed due to the global spread of drug-resistant bacteria and fungi. Here, we tested different extracts of chicory for a range of bioactivities, including antimicrobial, antifungal and cytotoxicity assays. Antibacterial and antifungal activities were generally more potent in ethyl acetate extracts compared to water extracts, whereas supercritical fluid extracts showed the broadest range of bioactivities in our assays. Remarkably, the chicory supercritical fluid extract and a purified fraction thereof inhibited both methicillin-resistant Staphylococcus aureus (MRSA) and ampicillin-resistant Pseudomonas aeruginosa IBRS P001. Chicory extracts also showed higher antibiofilm activity against the yeast Candida albicans than standard sesquiterpene lactone compounds. The cytotoxicity of the extracts was generally low. Our results may thus lead to the development of novel antibacterial and antifungal preparations that are both effective and safe for human use.

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Tissue specific expression and genomic organization of bitter sesquiterpene lactone biosynthesis in Cichorium intybus L. (Asteraceae)

Cited by 19 publications

References 32 publications

Inactivation of the germacrene A synthase genes by CRISPR/Cas9 eliminates the biosynthesis of sesquiterpene lactones in Cichorium intybus L.

Inactivation of the germacrene A synthase genes by CRISPR/Cas9 eliminates the biosynthesis of sesquiterpene lactones in Cichorium intybus L.

Silencing of germacrene A synthase genes reduces guaianolide oxalate content in Cichorium intybus L.

Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens

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