TY  - CONF
AU  - Bogdanović, Milica
AU  - Todorović, Slađana
AU  - Ćuković, Katarina
AU  - van Arkel, Jeroen
AU  - Bosch, Dirk
AU  - Cankar, Katarina
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/6155
AB  - Root chicory (Cichorium intybus var. sativum), an industrial crop species used for the production of a fructose polymer inulin, has been shown to contain a mixture of bitter tasting sesquiterpene lactones (STLs), that are currently discarded as waste. For several STLs found in plants of the Asteraceae family including chicory, interesting bioactivities have been demonstrated, including potent anti-cancer, anti-malarial, anti-inflammatory, anti-fungal and anti-bacterial activity. This activity is mainly attributed to guaianolide STLs; in chicory the most abundant STLs are lactucin, lactucopicrin, and 8-deoxylactucin, found predominantly in their oxalated forms in the latex of the plant. Several steps in the biosynthetic pathway of these compounds have been unraveled recently. However, the enzymes involved in the formation of STL oxalates, the most abundant form of STLs in chicory, have not yet been identified. Candidate genes for the chicory oxalate-CoA ligase (CiOxL) and chicory STL oxalyl transferases (CiOxT) putatively involved in the STL-oxalate formation were identified. Next, introduction of CRISPR/Cas reagents into chicory by Agrobacterium tumefaciens-mediated stable transformation was used to inactivate gene candidates putatively involved in STL-oxalate formation, and several chicory lines with edited genes were successfully regenerated. Detailed genotyping of mutant lines revealed the presence of indels leading to frame-shift predominantly, varying from 1 to 44 base pairs in length. Detailed genotyping also confirmed previous observations that plants transformed via Agrobacterium often showed chimerism, and a mixture of different on-target edits in one plant was observed. Leaves of plants carrying mutations in CiOxL or CiOxT were characterized by LC-MS to determine changes in terpene profile. The analysis showed that the production of STLs was reduced or eliminated in leaves of several CiOxL and CiOxT4 plants. Surprisingly, not only the oxalated terpenes were reduced but also the non-oxalated STLs, perhaps due to feedback regulation or toxicity of non-oxalated forms. These results contribute to further elucidation of the STL pathway in chicory and show that Agrobacterium-mediated plant transformation with CRISPR/Cas reagents requires detailed genotyping for characterization of genome edited plants.
PB  - COST Action CA18111
C3  - Book of abstracts: 4th PlantEd Conference; 2023 Sep 18-20; Porto, Portugal
T1  - CRISPR/Cas targeted inactivation of guaianolide oxalate formation in chicory
SP  - 74
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_6155
ER  - 

@conference{
author = "Bogdanović, Milica and Todorović, Slađana and Ćuković, Katarina and van Arkel, Jeroen and Bosch, Dirk and Cankar, Katarina",
year = "2023",
abstract = "Root chicory (Cichorium intybus var. sativum), an industrial crop species used for the production of a fructose polymer inulin, has been shown to contain a mixture of bitter tasting sesquiterpene lactones (STLs), that are currently discarded as waste. For several STLs found in plants of the Asteraceae family including chicory, interesting bioactivities have been demonstrated, including potent anti-cancer, anti-malarial, anti-inflammatory, anti-fungal and anti-bacterial activity. This activity is mainly attributed to guaianolide STLs; in chicory the most abundant STLs are lactucin, lactucopicrin, and 8-deoxylactucin, found predominantly in their oxalated forms in the latex of the plant. Several steps in the biosynthetic pathway of these compounds have been unraveled recently. However, the enzymes involved in the formation of STL oxalates, the most abundant form of STLs in chicory, have not yet been identified. Candidate genes for the chicory oxalate-CoA ligase (CiOxL) and chicory STL oxalyl transferases (CiOxT) putatively involved in the STL-oxalate formation were identified. Next, introduction of CRISPR/Cas reagents into chicory by Agrobacterium tumefaciens-mediated stable transformation was used to inactivate gene candidates putatively involved in STL-oxalate formation, and several chicory lines with edited genes were successfully regenerated. Detailed genotyping of mutant lines revealed the presence of indels leading to frame-shift predominantly, varying from 1 to 44 base pairs in length. Detailed genotyping also confirmed previous observations that plants transformed via Agrobacterium often showed chimerism, and a mixture of different on-target edits in one plant was observed. Leaves of plants carrying mutations in CiOxL or CiOxT were characterized by LC-MS to determine changes in terpene profile. The analysis showed that the production of STLs was reduced or eliminated in leaves of several CiOxL and CiOxT4 plants. Surprisingly, not only the oxalated terpenes were reduced but also the non-oxalated STLs, perhaps due to feedback regulation or toxicity of non-oxalated forms. These results contribute to further elucidation of the STL pathway in chicory and show that Agrobacterium-mediated plant transformation with CRISPR/Cas reagents requires detailed genotyping for characterization of genome edited plants.",
publisher = "COST Action CA18111",
journal = "Book of abstracts: 4th PlantEd Conference; 2023 Sep 18-20; Porto, Portugal",
title = "CRISPR/Cas targeted inactivation of guaianolide oxalate formation in chicory",
pages = "74",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6155"
}

Bogdanović, M., Todorović, S., Ćuković, K., van Arkel, J., Bosch, D.,& Cankar, K.. (2023). CRISPR/Cas targeted inactivation of guaianolide oxalate formation in chicory. in Book of abstracts: 4th PlantEd Conference; 2023 Sep 18-20; Porto, Portugal
COST Action CA18111., 74.
https://hdl.handle.net/21.15107/rcub_ibiss_6155

Bogdanović M, Todorović S, Ćuković K, van Arkel J, Bosch D, Cankar K. CRISPR/Cas targeted inactivation of guaianolide oxalate formation in chicory. in Book of abstracts: 4th PlantEd Conference; 2023 Sep 18-20; Porto, Portugal. 2023;:74.
https://hdl.handle.net/21.15107/rcub_ibiss_6155 .

Bogdanović, Milica, Todorović, Slađana, Ćuković, Katarina, van Arkel, Jeroen, Bosch, Dirk, Cankar, Katarina, "CRISPR/Cas targeted inactivation of guaianolide oxalate formation in chicory" in Book of abstracts: 4th PlantEd Conference; 2023 Sep 18-20; Porto, Portugal (2023):74,
https://hdl.handle.net/21.15107/rcub_ibiss_6155 .

TY  - JOUR
AU  - Salvagnin, Umberto
AU  - Unkel, Katharina
AU  - Sprink, Thorben
AU  - Bundock, Paul
AU  - Sevenier, Robert
AU  - Bogdanović, Milica
AU  - Todorović, Slađana
AU  - Cankar, Katarina
AU  - Hakkert, Johanna Christina
AU  - Schijlen, Elio
AU  - Nieuwenhuis, Ronald
AU  - Hingsamer, Maria
AU  - Kulmer, Veronika
AU  - Kernitzkyi, Michael
AU  - Bosch, Dirk
AU  - Martens, Stefan
AU  - Malnoy, Mickael
PY  - 2023
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5794
AB  - Root chicory (Cichorium intybus L. var. sativum) is used to extract inulin, a fructose polymer used as a natural sweetener and prebiotic. However, bitter tasting sesquiterpene lactones, giving chicory its known flavour, need to be removed during inulin extraction. To avoid this extraction and associated costs, recently chicory variants with a lower sesquiterpene lactone content were created by inactivating the four copies of the germacrene A synthase gene (CiGAS-S1, -S2, -S3, -L) which encode the enzyme initiating bitter sesquiterpene lactone biosynthesis in chicory. In this study, different delivery methods for CRISPR/Cas9 reagents have been compared regarding their efficiency to induce mutations in the CiGAS genes, the frequency of off-target mutations as well as their environmental and economic impacts. CRISPR/Cas9 reagents were delivered by Agrobacterium-mediated stable transformation or transient delivery by plasmid or preassembled ribonucleic complexes (RNPs) using the same sgRNA. All methods used lead to a high number of INDEL mutations within the CiGAS-S1 and CiGAS-S2 genes, which match the used sgRNA perfectly; additionally, the CiGAS-S3 and CiGAS-L genes, which have a single mismatch with the sgRNA, were mutated but with a lower mutation efficiency. While using both RNPs and plasmids delivery resulted in biallelic, heterozygous or homozygous mutations, plasmid delivery resulted in 30% of unwanted integration of plasmid fragments in the genome. Plants transformed via Agrobacteria often showed chimerism and a mixture of CiGAS genotypes. This genetic mosaic becomes more diverse when plants were grown over a prolonged period. While the genotype of the on-targets varied between the transient and stable delivery methods, no off-target activity in six identified potential off-targets with two to four mismatches was found. The environmental impacts (greenhouse gas (GHG) emissions and primary energy demand) of the methods are highly dependent on their individual electricity demand. From an economic view - like for most research and development activities - employment and value-added multiplier effects are high; particularly when compared to industrial or manufacturing processes. Considering all aspects, we conclude that using RNPs is the most suitable method for genome editing in chicory since it led to a high efficiency of editing, no off-target mutations, non-transgenic plants with no risk of unwanted integration of plasmid DNA and without needed segregation of transgenes.
PB  - Lausanne: Frontiers Media SA
T2  - Frontiers in Plant Science
T1  - A comparison of three different delivery methods for achieving CRISPR/Cas9 mediated genome editing in Cichorium intybus L
VL  - 14
DO  - 10.3389/fpls.2023.1111110
SP  - 1111110
ER  - 

@article{
author = "Salvagnin, Umberto and Unkel, Katharina and Sprink, Thorben and Bundock, Paul and Sevenier, Robert and Bogdanović, Milica and Todorović, Slađana and Cankar, Katarina and Hakkert, Johanna Christina and Schijlen, Elio and Nieuwenhuis, Ronald and Hingsamer, Maria and Kulmer, Veronika and Kernitzkyi, Michael and Bosch, Dirk and Martens, Stefan and Malnoy, Mickael",
year = "2023",
abstract = "Root chicory (Cichorium intybus L. var. sativum) is used to extract inulin, a fructose polymer used as a natural sweetener and prebiotic. However, bitter tasting sesquiterpene lactones, giving chicory its known flavour, need to be removed during inulin extraction. To avoid this extraction and associated costs, recently chicory variants with a lower sesquiterpene lactone content were created by inactivating the four copies of the germacrene A synthase gene (CiGAS-S1, -S2, -S3, -L) which encode the enzyme initiating bitter sesquiterpene lactone biosynthesis in chicory. In this study, different delivery methods for CRISPR/Cas9 reagents have been compared regarding their efficiency to induce mutations in the CiGAS genes, the frequency of off-target mutations as well as their environmental and economic impacts. CRISPR/Cas9 reagents were delivered by Agrobacterium-mediated stable transformation or transient delivery by plasmid or preassembled ribonucleic complexes (RNPs) using the same sgRNA. All methods used lead to a high number of INDEL mutations within the CiGAS-S1 and CiGAS-S2 genes, which match the used sgRNA perfectly; additionally, the CiGAS-S3 and CiGAS-L genes, which have a single mismatch with the sgRNA, were mutated but with a lower mutation efficiency. While using both RNPs and plasmids delivery resulted in biallelic, heterozygous or homozygous mutations, plasmid delivery resulted in 30% of unwanted integration of plasmid fragments in the genome. Plants transformed via Agrobacteria often showed chimerism and a mixture of CiGAS genotypes. This genetic mosaic becomes more diverse when plants were grown over a prolonged period. While the genotype of the on-targets varied between the transient and stable delivery methods, no off-target activity in six identified potential off-targets with two to four mismatches was found. The environmental impacts (greenhouse gas (GHG) emissions and primary energy demand) of the methods are highly dependent on their individual electricity demand. From an economic view - like for most research and development activities - employment and value-added multiplier effects are high; particularly when compared to industrial or manufacturing processes. Considering all aspects, we conclude that using RNPs is the most suitable method for genome editing in chicory since it led to a high efficiency of editing, no off-target mutations, non-transgenic plants with no risk of unwanted integration of plasmid DNA and without needed segregation of transgenes.",
publisher = "Lausanne: Frontiers Media SA",
journal = "Frontiers in Plant Science",
title = "A comparison of three different delivery methods for achieving CRISPR/Cas9 mediated genome editing in Cichorium intybus L",
volume = "14",
doi = "10.3389/fpls.2023.1111110",
pages = "1111110"
}

Salvagnin, U., Unkel, K., Sprink, T., Bundock, P., Sevenier, R., Bogdanović, M., Todorović, S., Cankar, K., Hakkert, J. C., Schijlen, E., Nieuwenhuis, R., Hingsamer, M., Kulmer, V., Kernitzkyi, M., Bosch, D., Martens, S.,& Malnoy, M.. (2023). A comparison of three different delivery methods for achieving CRISPR/Cas9 mediated genome editing in Cichorium intybus L. in Frontiers in Plant Science
Lausanne: Frontiers Media SA., 14, 1111110.
https://doi.org/10.3389/fpls.2023.1111110

Salvagnin U, Unkel K, Sprink T, Bundock P, Sevenier R, Bogdanović M, Todorović S, Cankar K, Hakkert JC, Schijlen E, Nieuwenhuis R, Hingsamer M, Kulmer V, Kernitzkyi M, Bosch D, Martens S, Malnoy M. A comparison of three different delivery methods for achieving CRISPR/Cas9 mediated genome editing in Cichorium intybus L. in Frontiers in Plant Science. 2023;14:1111110.
doi:10.3389/fpls.2023.1111110 .

Salvagnin, Umberto, Unkel, Katharina, Sprink, Thorben, Bundock, Paul, Sevenier, Robert, Bogdanović, Milica, Todorović, Slađana, Cankar, Katarina, Hakkert, Johanna Christina, Schijlen, Elio, Nieuwenhuis, Ronald, Hingsamer, Maria, Kulmer, Veronika, Kernitzkyi, Michael, Bosch, Dirk, Martens, Stefan, Malnoy, Mickael, "A comparison of three different delivery methods for achieving CRISPR/Cas9 mediated genome editing in Cichorium intybus L" in Frontiers in Plant Science, 14 (2023):1111110,
https://doi.org/10.3389/fpls.2023.1111110 . .

TY  - CONF
AU  - Stojković, Dejan
AU  - Ivanov, Marija
AU  - Ćirić, Ana
AU  - Petrović, Jovana
AU  - Kostić, Marina
AU  - Glamočlija, Jasmina
AU  - Soković, Marina
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5158
AB  - Chicory (Cichorium intybus L.) is an important industrial crop cultivated mainly to extract the
dietary fiber inulin. This study aimed to investigate antimicrobial, antibiofilm and wound healing
potentials of the extract obtained by supercritical fluid extraction (SFE). Antibacterial activity
of extract ranged from 0.25 to 4.00 mg/mL regarding minimal inhibitory concentra-tions (MICs),
while minimal bactericidal concentrations (MBCs) were noted within the range of 0.5 to 8.00 mg/
mL. The extract has shown promising antifungal potential with MICs rang-ing 0.125- 0.75 mg/
mL and MFCs 0.25-1.0 mg/mL against Candida species. Incubation with the extract has reduced
the ability of both C. albicans and P. aeruginosa to attach to the sur-face and begin the process
of biofilm formation. Concentration equal to previously deter-mined MIC has almost completely
(>90% inhibition) reduced the biofilm biomass formed by P. aeruginosa. Even twice as lower concentrations
(0.5 MIC) has limited the biofilm forming ability and induce more than 80% inhibition
in P. aeruginosa as bacterial biofilm model or-ganism. Impact on fungal biofilm was less profound
and MIC concentration was able to re-duce the biofilm formation for less than 60%. HaCaT (human
immortalized keratinocytes cell line) cells were treated with IC20 (24 h) concentration of chicory
SFE extract. It is estimated that SFE extract has slight effect on wound closure in keratinocyte
monolayer, when com-pared to untreated control. Since, Chicory SFE fraction possessed excellent
antibacterial po-tential against E. coli and K. pneumoniae and antifungal activity against Candida
species these results are considered very important in the future inventions
PB  - Belgrade: Serbian Plant Physiology Society
C3  - 4th International Conference on Plant Biology and 23rd SPPS Meeting; 2022 6-8; Belgrade, Serbia
T1  - Supercritical fluid extraction of Chicory reveals its antimicrobial, antibiofilm and wound healing potentials
SP  - 96
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5158
ER  - 

@conference{
author = "Stojković, Dejan and Ivanov, Marija and Ćirić, Ana and Petrović, Jovana and Kostić, Marina and Glamočlija, Jasmina and Soković, Marina",
year = "2022",
abstract = "Chicory (Cichorium intybus L.) is an important industrial crop cultivated mainly to extract the
dietary fiber inulin. This study aimed to investigate antimicrobial, antibiofilm and wound healing
potentials of the extract obtained by supercritical fluid extraction (SFE). Antibacterial activity
of extract ranged from 0.25 to 4.00 mg/mL regarding minimal inhibitory concentra-tions (MICs),
while minimal bactericidal concentrations (MBCs) were noted within the range of 0.5 to 8.00 mg/
mL. The extract has shown promising antifungal potential with MICs rang-ing 0.125- 0.75 mg/
mL and MFCs 0.25-1.0 mg/mL against Candida species. Incubation with the extract has reduced
the ability of both C. albicans and P. aeruginosa to attach to the sur-face and begin the process
of biofilm formation. Concentration equal to previously deter-mined MIC has almost completely
(>90% inhibition) reduced the biofilm biomass formed by P. aeruginosa. Even twice as lower concentrations
(0.5 MIC) has limited the biofilm forming ability and induce more than 80% inhibition
in P. aeruginosa as bacterial biofilm model or-ganism. Impact on fungal biofilm was less profound
and MIC concentration was able to re-duce the biofilm formation for less than 60%. HaCaT (human
immortalized keratinocytes cell line) cells were treated with IC20 (24 h) concentration of chicory
SFE extract. It is estimated that SFE extract has slight effect on wound closure in keratinocyte
monolayer, when com-pared to untreated control. Since, Chicory SFE fraction possessed excellent
antibacterial po-tential against E. coli and K. pneumoniae and antifungal activity against Candida
species these results are considered very important in the future inventions",
publisher = "Belgrade: Serbian Plant Physiology Society",
journal = "4th International Conference on Plant Biology and 23rd SPPS Meeting; 2022 6-8; Belgrade, Serbia",
title = "Supercritical fluid extraction of Chicory reveals its antimicrobial, antibiofilm and wound healing potentials",
pages = "96",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5158"
}

Stojković, D., Ivanov, M., Ćirić, A., Petrović, J., Kostić, M., Glamočlija, J.,& Soković, M.. (2022). Supercritical fluid extraction of Chicory reveals its antimicrobial, antibiofilm and wound healing potentials. in 4th International Conference on Plant Biology and 23rd SPPS Meeting; 2022 6-8; Belgrade, Serbia
Belgrade: Serbian Plant Physiology Society., 96.
https://hdl.handle.net/21.15107/rcub_ibiss_5158

Stojković D, Ivanov M, Ćirić A, Petrović J, Kostić M, Glamočlija J, Soković M. Supercritical fluid extraction of Chicory reveals its antimicrobial, antibiofilm and wound healing potentials. in 4th International Conference on Plant Biology and 23rd SPPS Meeting; 2022 6-8; Belgrade, Serbia. 2022;:96.
https://hdl.handle.net/21.15107/rcub_ibiss_5158 .

Stojković, Dejan, Ivanov, Marija, Ćirić, Ana, Petrović, Jovana, Kostić, Marina, Glamočlija, Jasmina, Soković, Marina, "Supercritical fluid extraction of Chicory reveals its antimicrobial, antibiofilm and wound healing potentials" in 4th International Conference on Plant Biology and 23rd SPPS Meeting; 2022 6-8; Belgrade, Serbia (2022):96,
https://hdl.handle.net/21.15107/rcub_ibiss_5158 .

TY  - CONF
AU  - Ćirić, Ana
AU  - Ivanov, Marija
AU  - Stojković, Dejan
AU  - Glamočlija, Jasmina
AU  - Kostić, Marina
AU  - Petrović, Jovana
AU  - Soković, Marina
PY  - 2022
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5581
AB  - Cichorium intybus L. (Asteraceae), is an important medicinal plant, rich in sesquiterpene
lactones (STLs), inulin, biologically relevant elements, and vitamins which exert potent
pro-health effects on the human organism. The aim of this study was to screen two
extracts of chicory containing STLs, the optimized extract (Ci_SFEopt) and the
supercritical fluid extract (Ci_SFEpur), for their antimicrobial activities against a range
of pathogenic microorganisms. Both extracts showed antibacterial activity, with
Ci_SFEpur being more potent than Ci_SFEopt. The most sensitive species was
Pseudomonas aeruginosa (minimal inhibitory concentration, MIC 0.06 mg/mL).
Ci_SFEpur and Ci_SFEopt extracts demonstrated good antifungal activity against
Candida species (MIC 0.03-1.00 mg/mL). Thin layer chromatography (TLC)
bioautography was used to screen the extracts and provide basic information for the
isolation of bioactive compounds in the future. Ci_SFEopt was active against all tested
microorganisms (Rf values were 0.18-0.88). The Ci_SFEpur extract efficiently inhibited
(Rf 0.76) Staphylococcus aureus, P. aeruginosa and Candida krusei. The identification
of powerful antimicrobial activity of Ci_SFEpur against P. aeruginosa and Candida
species is an important step towards the development of novel antimicrobial therapeutics
and efficient utilization of this widely distributed plant.
PB  - Belgrade: Serbian Biological Society
C3  - Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
T1  - Evaluation of antimicrobial properties of chicory extracts
SP  - 257
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5581
ER  - 

@conference{
author = "Ćirić, Ana and Ivanov, Marija and Stojković, Dejan and Glamočlija, Jasmina and Kostić, Marina and Petrović, Jovana and Soković, Marina",
year = "2022",
abstract = "Cichorium intybus L. (Asteraceae), is an important medicinal plant, rich in sesquiterpene
lactones (STLs), inulin, biologically relevant elements, and vitamins which exert potent
pro-health effects on the human organism. The aim of this study was to screen two
extracts of chicory containing STLs, the optimized extract (Ci_SFEopt) and the
supercritical fluid extract (Ci_SFEpur), for their antimicrobial activities against a range
of pathogenic microorganisms. Both extracts showed antibacterial activity, with
Ci_SFEpur being more potent than Ci_SFEopt. The most sensitive species was
Pseudomonas aeruginosa (minimal inhibitory concentration, MIC 0.06 mg/mL).
Ci_SFEpur and Ci_SFEopt extracts demonstrated good antifungal activity against
Candida species (MIC 0.03-1.00 mg/mL). Thin layer chromatography (TLC)
bioautography was used to screen the extracts and provide basic information for the
isolation of bioactive compounds in the future. Ci_SFEopt was active against all tested
microorganisms (Rf values were 0.18-0.88). The Ci_SFEpur extract efficiently inhibited
(Rf 0.76) Staphylococcus aureus, P. aeruginosa and Candida krusei. The identification
of powerful antimicrobial activity of Ci_SFEpur against P. aeruginosa and Candida
species is an important step towards the development of novel antimicrobial therapeutics
and efficient utilization of this widely distributed plant.",
publisher = "Belgrade: Serbian Biological Society",
journal = "Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia",
title = "Evaluation of antimicrobial properties of chicory extracts",
pages = "257",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5581"
}

Ćirić, A., Ivanov, M., Stojković, D., Glamočlija, J., Kostić, M., Petrović, J.,& Soković, M.. (2022). Evaluation of antimicrobial properties of chicory extracts. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia
Belgrade: Serbian Biological Society., 257.
https://hdl.handle.net/21.15107/rcub_ibiss_5581

Ćirić A, Ivanov M, Stojković D, Glamočlija J, Kostić M, Petrović J, Soković M. Evaluation of antimicrobial properties of chicory extracts. in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia. 2022;:257.
https://hdl.handle.net/21.15107/rcub_ibiss_5581 .

Ćirić, Ana, Ivanov, Marija, Stojković, Dejan, Glamočlija, Jasmina, Kostić, Marina, Petrović, Jovana, Soković, Marina, "Evaluation of antimicrobial properties of chicory extracts" in Knjiga sažetaka: Treći Kongres biologa Srbije: Osnovna i primenjena istraživanja: Metodika nastave; 2022 Sep 21-25; Zlatibor, Serbia (2022):257,
https://hdl.handle.net/21.15107/rcub_ibiss_5581 .

TY  - CONF
AU  - Ćirić, Ana
AU  - Ivanov, Marija
AU  - Stojković, Dejan
AU  - Glamočlija, Jasmina
AU  - Kostić, Marina
AU  - Soković, Marina
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4381
AB  - Chicory (Cichorium intybus L.) is a plant widely cultivated for the inulin production. The 
plant is underutilized regarding sesquiterpene lactones that might express potent 
pharmacological activities, and are currently discharged as a byproduct. Our study aimed to 
investigate both fraction and lactone antimicrobial potential and ability to reduce viability of 
P. aeruginosa cells in biofilms (MTT assay), as well as their ability to produce 
exopolysaccharides (Congo red binding assay), extracellular DNA and form biofilms on 
urinary catheters. Chicory fraction and 8-deoxylactucin exhibited promising antibacterial 
activity towards antibiotic resistant strain of P. aeruginosa (MIC 0.5 and 0.06 mg/mL, 
respectively). Chicory fraction has reduced drastically (>50% inhibition with MIC of agent) 
viability of P. aeruginosa cells in biofilm, while the activity of 8-deoxylactucin was less 
profound (<30% inhibition). Likewise, application of chicory fraction in MIC concentration 
has significantly limited the ability of P. aeruginosa cells to produce exopolysaccharides 
(>40% inhibition) and eDNA (>60% inhibition), known biofilm resistance factors. Biofilms 
formed on urinary catheters are challenging threat in hospital environments. Though, ability 
of both chicory fraction and 8-deoxylactucin to reduce ability of P. aeruginosa to attach to 
catheter surface should be further explored as promising novel antibacterial treatment since 
application of both agents at their MIC concentrations has induced more than 50% inhibition.
PB  - International Association of Food Nutrition and Safety
C3  - 5th International Symposium on Phytochemicals in Medicine and Food; 2021 Aug 25- Sep 1; Nanchang, China
T1  - Chicory fraction and 8-deoxylactucin inhibit growth and virulence traits of resistant Pseudomonas aeruginosa
SP  - 394
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4381
ER  - 

@conference{
author = "Ćirić, Ana and Ivanov, Marija and Stojković, Dejan and Glamočlija, Jasmina and Kostić, Marina and Soković, Marina",
year = "2021",
abstract = "Chicory (Cichorium intybus L.) is a plant widely cultivated for the inulin production. The 
plant is underutilized regarding sesquiterpene lactones that might express potent 
pharmacological activities, and are currently discharged as a byproduct. Our study aimed to 
investigate both fraction and lactone antimicrobial potential and ability to reduce viability of 
P. aeruginosa cells in biofilms (MTT assay), as well as their ability to produce 
exopolysaccharides (Congo red binding assay), extracellular DNA and form biofilms on 
urinary catheters. Chicory fraction and 8-deoxylactucin exhibited promising antibacterial 
activity towards antibiotic resistant strain of P. aeruginosa (MIC 0.5 and 0.06 mg/mL, 
respectively). Chicory fraction has reduced drastically (>50% inhibition with MIC of agent) 
viability of P. aeruginosa cells in biofilm, while the activity of 8-deoxylactucin was less 
profound (<30% inhibition). Likewise, application of chicory fraction in MIC concentration 
has significantly limited the ability of P. aeruginosa cells to produce exopolysaccharides 
(>40% inhibition) and eDNA (>60% inhibition), known biofilm resistance factors. Biofilms 
formed on urinary catheters are challenging threat in hospital environments. Though, ability 
of both chicory fraction and 8-deoxylactucin to reduce ability of P. aeruginosa to attach to 
catheter surface should be further explored as promising novel antibacterial treatment since 
application of both agents at their MIC concentrations has induced more than 50% inhibition.",
publisher = "International Association of Food Nutrition and Safety",
journal = "5th International Symposium on Phytochemicals in Medicine and Food; 2021 Aug 25- Sep 1; Nanchang, China",
title = "Chicory fraction and 8-deoxylactucin inhibit growth and virulence traits of resistant Pseudomonas aeruginosa",
pages = "394",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4381"
}

Ćirić, A., Ivanov, M., Stojković, D., Glamočlija, J., Kostić, M.,& Soković, M.. (2021). Chicory fraction and 8-deoxylactucin inhibit growth and virulence traits of resistant Pseudomonas aeruginosa. in 5th International Symposium on Phytochemicals in Medicine and Food; 2021 Aug 25- Sep 1; Nanchang, China
International Association of Food Nutrition and Safety., 394.
https://hdl.handle.net/21.15107/rcub_ibiss_4381

Ćirić A, Ivanov M, Stojković D, Glamočlija J, Kostić M, Soković M. Chicory fraction and 8-deoxylactucin inhibit growth and virulence traits of resistant Pseudomonas aeruginosa. in 5th International Symposium on Phytochemicals in Medicine and Food; 2021 Aug 25- Sep 1; Nanchang, China. 2021;:394.
https://hdl.handle.net/21.15107/rcub_ibiss_4381 .

Ćirić, Ana, Ivanov, Marija, Stojković, Dejan, Glamočlija, Jasmina, Kostić, Marina, Soković, Marina, "Chicory fraction and 8-deoxylactucin inhibit growth and virulence traits of resistant Pseudomonas aeruginosa" in 5th International Symposium on Phytochemicals in Medicine and Food; 2021 Aug 25- Sep 1; Nanchang, China (2021):394,
https://hdl.handle.net/21.15107/rcub_ibiss_4381 .

TY  - JOUR
AU  - Häkkinen, Suvi T.
AU  - Soković, Marina
AU  - Nohynek, Liisa
AU  - Ćirić, Ana
AU  - Ivanov, Marija
AU  - Stojković, Dejan
AU  - Tsitko, Irina
AU  - Matos, Melanie
AU  - Baixinho, João P.
AU  - Ivasiv, Viktoriya
AU  - Fernández, Naiara
AU  - Nunes Dos Santos, Claudia
AU  - Oksman-Caldentey, Kirsi-Marja
PY  - 2021
UR  - https://www.mdpi.com/1424-8247/14/9/941
UR  - http://www.pubmedcentral.nih.gov/articlerender.fcgi?artid=PMC8469098
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4493
AB  - 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.
PB  - Basel: MDPI
T2  - Pharmaceuticals (Basel, Switzerland)
T1  - Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens.
IS  - 9
VL  - 14
DO  - 10.3390/ph14090941
SP  - 941
ER  - 

@article{
author = "Häkkinen, Suvi T. and Soković, Marina and Nohynek, Liisa and Ćirić, Ana and Ivanov, Marija and Stojković, Dejan and Tsitko, Irina and Matos, Melanie and Baixinho, João P. and Ivasiv, Viktoriya and Fernández, Naiara and Nunes Dos Santos, Claudia and Oksman-Caldentey, Kirsi-Marja",
year = "2021",
abstract = "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.",
publisher = "Basel: MDPI",
journal = "Pharmaceuticals (Basel, Switzerland)",
title = "Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens.",
number = "9",
volume = "14",
doi = "10.3390/ph14090941",
pages = "941"
}

Häkkinen, S. T., Soković, M., Nohynek, L., Ćirić, A., Ivanov, M., Stojković, D., Tsitko, I., Matos, M., Baixinho, J. P., Ivasiv, V., Fernández, N., Nunes Dos Santos, C.,& Oksman-Caldentey, K.. (2021). Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens.. in Pharmaceuticals (Basel, Switzerland)
Basel: MDPI., 14(9), 941.
https://doi.org/10.3390/ph14090941

Häkkinen ST, Soković M, Nohynek L, Ćirić A, Ivanov M, Stojković D, Tsitko I, Matos M, Baixinho JP, Ivasiv V, Fernández N, Nunes Dos Santos C, Oksman-Caldentey K. Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens.. in Pharmaceuticals (Basel, Switzerland). 2021;14(9):941.
doi:10.3390/ph14090941 .

Häkkinen, Suvi T., Soković, Marina, Nohynek, Liisa, Ćirić, Ana, Ivanov, Marija, Stojković, Dejan, Tsitko, Irina, Matos, Melanie, Baixinho, João P., Ivasiv, Viktoriya, Fernández, Naiara, Nunes Dos Santos, Claudia, Oksman-Caldentey, Kirsi-Marja, "Chicory Extracts and Sesquiterpene Lactones Show Potent Activity against Bacterial and Fungal Pathogens." in Pharmaceuticals (Basel, Switzerland), 14, no. 9 (2021):941,
https://doi.org/10.3390/ph14090941 . .

TY  - CONF
AU  - Stojković, Dejan
AU  - Ivanov, Marija
AU  - Petrović, Jovana
AU  - Ćirić, Ana
AU  - Soković, Marina
PY  - 2021
UR  - https://radar.ibiss.bg.ac.rs/handle/123456789/4377
AB  - Cichorium intybus L. (Asteraceae) or chicory is a leafy biennial or perennial plant, with a strong fleshy 
taproot, that can grow up to 75 cm in length. Chicory has a long history being used as medicinal plant. The 
major sesquiterpene lactones (STLs) of chicory belong to the class of guaianolide sesquiterpene lactones 
and are commonly derived from a single sesquiterpene, germacrene A. The main STLs in chicory are 
lactucin, lactucopicrin, 8-deoxylactucin and their oxalate and glucoside derivatives. 
The aim of this study was to screen the different types of chicory extracts and terpene compounds for their 
bioactivities and to understand better how the valuable currently unused part of chicory could be taken to 
use. 
Antimicrobial activity of terpene samples was tested via analysing the type of growth inhibition (biostatic 
or biocidic) using modified microdilution method. For the determination of the antimicrobial activity of 
terpene samples, a modified bioautographic method was used as well. Impact of extracts and compounds 
on biofilm formation were determined on Candida albicans 475/15 and Pseudomonas aeruginosa (AMPR). 
Cristal violet assay was used for the determination of antiproliferative effect, on human gingival fibroblasts 
cells HGF-1 (ATCC® CRL-2014™). 
It was observed that EtOAc extract of chicory root showed good antimicrobial potential against all tested 
bacteria (MIC 0.75-3 mg/mL) except for P. aeruginosa (AMPR) strain, while water root extract did not 
display antimicrobial effect with tested concentrations. Costunolide, lactucopicrin, 11β,13-dihydrolactucin, 
compounds found in chicory roots, showed the most prominent effect against resistant strain of P. 
aeruginosa. All tested standard compounds demonstrated very good antifungal activity against Candida 
species (MIC 0.03-1.00 mg/mL). By using bioautographic method we have shown that none of the extract 
fractions were active against Escherichia coli, while two fractions of EtOAc extract were active against P. 
aeruginosa. EtOAc extract of chicory showed the most promising activity in inhibiting C. albicans biofilm 
formation, while slightly lower activity was observed for compound parthenolide. Water extract did not 
exhibit any cytotoxicity up to 1 mg/mL. On the other hand, EtOAc extract was cytotoxic to HGF-1 cell at 
GI50(%) concentration of 0.750 mg/mL. As for the tested compounds, 11β-13-dihydrolactucin was the less 
toxic to HGF-1 cells. All other compounds were toxic at the lowest concentration used
PB  - Bragança, Portugal: Instituto Politécnico de Bragança (IPB)
C3  - 1st Natural products application: Health, Cosmetic and Food: book of abstracts
T1  - Biological activity of chicory extracts and compounds
SP  - 118
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_4377
ER  - 

@conference{
author = "Stojković, Dejan and Ivanov, Marija and Petrović, Jovana and Ćirić, Ana and Soković, Marina",
year = "2021",
abstract = "Cichorium intybus L. (Asteraceae) or chicory is a leafy biennial or perennial plant, with a strong fleshy 
taproot, that can grow up to 75 cm in length. Chicory has a long history being used as medicinal plant. The 
major sesquiterpene lactones (STLs) of chicory belong to the class of guaianolide sesquiterpene lactones 
and are commonly derived from a single sesquiterpene, germacrene A. The main STLs in chicory are 
lactucin, lactucopicrin, 8-deoxylactucin and their oxalate and glucoside derivatives. 
The aim of this study was to screen the different types of chicory extracts and terpene compounds for their 
bioactivities and to understand better how the valuable currently unused part of chicory could be taken to 
use. 
Antimicrobial activity of terpene samples was tested via analysing the type of growth inhibition (biostatic 
or biocidic) using modified microdilution method. For the determination of the antimicrobial activity of 
terpene samples, a modified bioautographic method was used as well. Impact of extracts and compounds 
on biofilm formation were determined on Candida albicans 475/15 and Pseudomonas aeruginosa (AMPR). 
Cristal violet assay was used for the determination of antiproliferative effect, on human gingival fibroblasts 
cells HGF-1 (ATCC® CRL-2014™). 
It was observed that EtOAc extract of chicory root showed good antimicrobial potential against all tested 
bacteria (MIC 0.75-3 mg/mL) except for P. aeruginosa (AMPR) strain, while water root extract did not 
display antimicrobial effect with tested concentrations. Costunolide, lactucopicrin, 11β,13-dihydrolactucin, 
compounds found in chicory roots, showed the most prominent effect against resistant strain of P. 
aeruginosa. All tested standard compounds demonstrated very good antifungal activity against Candida 
species (MIC 0.03-1.00 mg/mL). By using bioautographic method we have shown that none of the extract 
fractions were active against Escherichia coli, while two fractions of EtOAc extract were active against P. 
aeruginosa. EtOAc extract of chicory showed the most promising activity in inhibiting C. albicans biofilm 
formation, while slightly lower activity was observed for compound parthenolide. Water extract did not 
exhibit any cytotoxicity up to 1 mg/mL. On the other hand, EtOAc extract was cytotoxic to HGF-1 cell at 
GI50(%) concentration of 0.750 mg/mL. As for the tested compounds, 11β-13-dihydrolactucin was the less 
toxic to HGF-1 cells. All other compounds were toxic at the lowest concentration used",
publisher = "Bragança, Portugal: Instituto Politécnico de Bragança (IPB)",
journal = "1st Natural products application: Health, Cosmetic and Food: book of abstracts",
title = "Biological activity of chicory extracts and compounds",
pages = "118",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_4377"
}

Stojković, D., Ivanov, M., Petrović, J., Ćirić, A.,& Soković, M.. (2021). Biological activity of chicory extracts and compounds. in 1st Natural products application: Health, Cosmetic and Food: book of abstracts
Bragança, Portugal: Instituto Politécnico de Bragança (IPB)., 118.
https://hdl.handle.net/21.15107/rcub_ibiss_4377

Stojković D, Ivanov M, Petrović J, Ćirić A, Soković M. Biological activity of chicory extracts and compounds. in 1st Natural products application: Health, Cosmetic and Food: book of abstracts. 2021;:118.
https://hdl.handle.net/21.15107/rcub_ibiss_4377 .

Stojković, Dejan, Ivanov, Marija, Petrović, Jovana, Ćirić, Ana, Soković, Marina, "Biological activity of chicory extracts and compounds" in 1st Natural products application: Health, Cosmetic and Food: book of abstracts (2021):118,
https://hdl.handle.net/21.15107/rcub_ibiss_4377 .

TY  - CONF
AU  - Bogdanović, Milica
AU  - Todorović, Slađana
AU  - Ćuković, Katarina
AU  - Dragićević, Milan
AU  - Beekwilder, Jules
AU  - Cankar, Katarina
AU  - Simonović, Ana
PY  - 2020
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5438
AB  - Chicory (Cichorium intybus L.) is a valuable crop grown mostly in northern France, Belgium and the Netherlands, whose roots, leaves and etiolated shoots are used as vegetable, coffee replacement and remedy for several conditions including inflammation. Roots of chicory are an important source of inulin, a prebiotic fiber and a sweetener. Chicory also contains bioactive terpene compounds that protect the plant against herbivores, but are also responsible for medicinal properties of the plant, as they have anti-microbial, anti-cancer and anti-inflammatory activity. Biosynthesis of plant bioactive metabolites can be modified using new plant breeding techniques, especially new gene editing techniques like CRISPR/Cas9 to improve their health-promoting qualities. Chicory was transformed with Agrobacterium tumefaciens carrying CRISPR/Cas9 constructs to knock-out expression of key genes in the sesquiterpene biosynthesis pathway. Regenerated plants were selected on media with kanamycin and further tested by PCR for the presence of transgenes. Obtained plants were also tested for mutations in target genes, by PCR with primers surrounding the mutation site detecting larger indels and by Sanger sequencing detecting smaller indels. Further analysis was performed detecting heteroduplex DNA fragments as a result of either hetero- or homozygous mutation events.
PB  - East Sarajevo: Faculty of Agriculture
C3  - Book of abstracts: XI International Scientific Agriculture Symposium: Agrosym 2020; 2020 Oct 8-9; Jahorina, Republika Srpska, Bosnia and Herzegovina
T1  - Metabolic engeneering in chicory by CRISPR/Cas9 editing
SP  - 152
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5438
ER  - 

@conference{
author = "Bogdanović, Milica and Todorović, Slađana and Ćuković, Katarina and Dragićević, Milan and Beekwilder, Jules and Cankar, Katarina and Simonović, Ana",
year = "2020",
abstract = "Chicory (Cichorium intybus L.) is a valuable crop grown mostly in northern France, Belgium and the Netherlands, whose roots, leaves and etiolated shoots are used as vegetable, coffee replacement and remedy for several conditions including inflammation. Roots of chicory are an important source of inulin, a prebiotic fiber and a sweetener. Chicory also contains bioactive terpene compounds that protect the plant against herbivores, but are also responsible for medicinal properties of the plant, as they have anti-microbial, anti-cancer and anti-inflammatory activity. Biosynthesis of plant bioactive metabolites can be modified using new plant breeding techniques, especially new gene editing techniques like CRISPR/Cas9 to improve their health-promoting qualities. Chicory was transformed with Agrobacterium tumefaciens carrying CRISPR/Cas9 constructs to knock-out expression of key genes in the sesquiterpene biosynthesis pathway. Regenerated plants were selected on media with kanamycin and further tested by PCR for the presence of transgenes. Obtained plants were also tested for mutations in target genes, by PCR with primers surrounding the mutation site detecting larger indels and by Sanger sequencing detecting smaller indels. Further analysis was performed detecting heteroduplex DNA fragments as a result of either hetero- or homozygous mutation events.",
publisher = "East Sarajevo: Faculty of Agriculture",
journal = "Book of abstracts: XI International Scientific Agriculture Symposium: Agrosym 2020; 2020 Oct 8-9; Jahorina, Republika Srpska, Bosnia and Herzegovina",
title = "Metabolic engeneering in chicory by CRISPR/Cas9 editing",
pages = "152",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5438"
}

Bogdanović, M., Todorović, S., Ćuković, K., Dragićević, M., Beekwilder, J., Cankar, K.,& Simonović, A.. (2020). Metabolic engeneering in chicory by CRISPR/Cas9 editing. in Book of abstracts: XI International Scientific Agriculture Symposium: Agrosym 2020; 2020 Oct 8-9; Jahorina, Republika Srpska, Bosnia and Herzegovina
East Sarajevo: Faculty of Agriculture., 152.
https://hdl.handle.net/21.15107/rcub_ibiss_5438

Bogdanović M, Todorović S, Ćuković K, Dragićević M, Beekwilder J, Cankar K, Simonović A. Metabolic engeneering in chicory by CRISPR/Cas9 editing. in Book of abstracts: XI International Scientific Agriculture Symposium: Agrosym 2020; 2020 Oct 8-9; Jahorina, Republika Srpska, Bosnia and Herzegovina. 2020;:152.
https://hdl.handle.net/21.15107/rcub_ibiss_5438 .

Bogdanović, Milica, Todorović, Slađana, Ćuković, Katarina, Dragićević, Milan, Beekwilder, Jules, Cankar, Katarina, Simonović, Ana, "Metabolic engeneering in chicory by CRISPR/Cas9 editing" in Book of abstracts: XI International Scientific Agriculture Symposium: Agrosym 2020; 2020 Oct 8-9; Jahorina, Republika Srpska, Bosnia and Herzegovina (2020):152,
https://hdl.handle.net/21.15107/rcub_ibiss_5438 .

TY  - CONF
AU  - Cankar, Katarina
AU  - Bogdanović, Milica
AU  - Todorović, Slađana
AU  - Dragićević, Milan
AU  - Simonović, Ana
AU  - van Houwelingen, Adele
AU  - Schijlen, Elio
AU  - van de Geest, Henri
AU  - Hendriks, Theo
AU  - Quillet, MC
AU  - Bouwmeester, Harro
AU  - Bosch, Dirk
AU  - Beekwilder, Jules
PY  - 2016
UR  - http://radar.ibiss.bg.ac.rs/handle/123456789/5441
AB  - Sesquiterpene lactones (STLs) give chicory its characteristic bitter taste and are highly concentrated in the latex channels of chicory taproot. The study of STLs is interesting to control the bitterness of industrial and consumption chicory, including witloof. The major STLs in chicory are lactucin, deoxylactucin, and lactucopicrin and oxalate and sulphate conjugates thereof. Formation of chicory STLs requires the biosynthesis of costunolide. Two germacrene A synthases, two germacrene A oxidases and a costunolide synthase have been described from chicory. In this work, the genetic organization of the genes involved in STL biosynthesis in chicory linkage groups is described. A number of genomic regions hosting STL biosynthetic genes have been addressed by NGS sequencing. Gene expression in different tissues is addressed by following their expression
using an RNAseq approach and by promoter activity studies using GFP. Finally, the downregulation of STL gene expression in chicory is achieved in hairy root culture regenerants expressing amiRNAs, and the consequences for STL content in different tissues is tested. These results will contribute to successful breeding strategies for chicory germplasm with well-defined STL content and bitterness.
PB  - Ghent : Phytochemical Society of Europe
C3  - Book of abstracts: Plant omics and biotechnology for human health: POBHH; 2016 Nov 21-24; Gent, Belgium
T1  - Tissue specific expression and genomic organization of sesquiterpene lactone biosynthetic genes in chicory
SP  - 65
UR  - https://hdl.handle.net/21.15107/rcub_ibiss_5441
ER  - 

@conference{
author = "Cankar, Katarina and Bogdanović, Milica and Todorović, Slađana and Dragićević, Milan and Simonović, Ana and van Houwelingen, Adele and Schijlen, Elio and van de Geest, Henri and Hendriks, Theo and Quillet, MC and Bouwmeester, Harro and Bosch, Dirk and Beekwilder, Jules",
year = "2016",
abstract = "Sesquiterpene lactones (STLs) give chicory its characteristic bitter taste and are highly concentrated in the latex channels of chicory taproot. The study of STLs is interesting to control the bitterness of industrial and consumption chicory, including witloof. The major STLs in chicory are lactucin, deoxylactucin, and lactucopicrin and oxalate and sulphate conjugates thereof. Formation of chicory STLs requires the biosynthesis of costunolide. Two germacrene A synthases, two germacrene A oxidases and a costunolide synthase have been described from chicory. In this work, the genetic organization of the genes involved in STL biosynthesis in chicory linkage groups is described. A number of genomic regions hosting STL biosynthetic genes have been addressed by NGS sequencing. Gene expression in different tissues is addressed by following their expression
using an RNAseq approach and by promoter activity studies using GFP. Finally, the downregulation of STL gene expression in chicory is achieved in hairy root culture regenerants expressing amiRNAs, and the consequences for STL content in different tissues is tested. These results will contribute to successful breeding strategies for chicory germplasm with well-defined STL content and bitterness.",
publisher = "Ghent : Phytochemical Society of Europe",
journal = "Book of abstracts: Plant omics and biotechnology for human health: POBHH; 2016 Nov 21-24; Gent, Belgium",
title = "Tissue specific expression and genomic organization of sesquiterpene lactone biosynthetic genes in chicory",
pages = "65",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_5441"
}

Cankar, K., Bogdanović, M., Todorović, S., Dragićević, M., Simonović, A., van Houwelingen, A., Schijlen, E., van de Geest, H., Hendriks, T., Quillet, M., Bouwmeester, H., Bosch, D.,& Beekwilder, J.. (2016). Tissue specific expression and genomic organization of sesquiterpene lactone biosynthetic genes in chicory. in Book of abstracts: Plant omics and biotechnology for human health: POBHH; 2016 Nov 21-24; Gent, Belgium
Ghent : Phytochemical Society of Europe., 65.
https://hdl.handle.net/21.15107/rcub_ibiss_5441

Cankar K, Bogdanović M, Todorović S, Dragićević M, Simonović A, van Houwelingen A, Schijlen E, van de Geest H, Hendriks T, Quillet M, Bouwmeester H, Bosch D, Beekwilder J. Tissue specific expression and genomic organization of sesquiterpene lactone biosynthetic genes in chicory. in Book of abstracts: Plant omics and biotechnology for human health: POBHH; 2016 Nov 21-24; Gent, Belgium. 2016;:65.
https://hdl.handle.net/21.15107/rcub_ibiss_5441 .

Cankar, Katarina, Bogdanović, Milica, Todorović, Slađana, Dragićević, Milan, Simonović, Ana, van Houwelingen, Adele, Schijlen, Elio, van de Geest, Henri, Hendriks, Theo, Quillet, MC, Bouwmeester, Harro, Bosch, Dirk, Beekwilder, Jules, "Tissue specific expression and genomic organization of sesquiterpene lactone biosynthetic genes in chicory" in Book of abstracts: Plant omics and biotechnology for human health: POBHH; 2016 Nov 21-24; Gent, Belgium (2016):65,
https://hdl.handle.net/21.15107/rcub_ibiss_5441 .