TY  - JOUR
AU  - Aničić, Neda
AU  - Matekalo, Dragana
AU  - Skorić, Marijana
AU  - Gašić, Uroš M.
AU  - Nestorović Živković, Jasmina
AU  - Dmitrović, Slavica
AU  - Božunović, Jelena
AU  - Milutinović, Milica
AU  - Petrović, Luka
AU  - Dimitrijević, Milena
AU  - Anđelković, Boban D.
AU  - Mišić, Danijela
PY  - 2024
UR  - http://cherry.chem.bg.ac.rs/handle/123456789/6454
AB  - Iridoids, a class of atypical monoterpenes, exhibit exceptional diversity within the Nepeta genus (subfam. Nepetoidae, fam. Lamiaceae).The majority of these plants produce iridoids of the unique stereochemistry, with nepetalactones (NLs) predominating; however, a few Nepeta species lack these compounds. By comparatively analyzing metabolomics, transcriptomics, gene co-expression, and phylogenetic data of the iridoid-producing N. rtanjensis Diklić & Milojević and iridoid-lacking N. nervosa Royle & Bentham, we presumed that one of the factors responsible for the absence of these compounds in N. nervosa is iridoid synthase (ISY). Two orthologues of ISY were mined from leaves transcriptome of N. rtanjensis (NrPRISE1 and NrPRISE2), while in N. nervosa only one (NnPRISE) was identified, and it was phylogenetically closer to the representatives of the Family 1 isoforms, designated as P5βRs. Organ-specific and MeJA-elicited profiling of iridoid content and co-expression analysis of IBG candidates, highlighted NrPRISE2 and NnPRISE as promising candidates for ISY orthologues, and their function was confirmed using in vitro assays with recombinant proteins, after heterologous expression of recombinant proteins in E. coli and their His-tag affinity purification. NrPRISE2 demonstrated ISY activity both in vitro and likely in planta, which was supported by the 3D modeling and molecular docking analysis, thus reclassification of NrPRISE2 to NrISY is accordingly recommended. NnPRISE also displays in vitro ISY-like activity, while its role under in vivo conditions was not here unambiguously confirmed. Most probably under in vivo conditions the NnPRISE lacks substrates to act upon, as a result of the loss of function of some of the upstream enzymes of the iridoid pathway. Our ongoing work is conducted towards re-establishing the biosynthesis of iridoids in N. nervosa.
PB  - Frontiers Media
T2  - Frontiers in Plant Science
T1  - Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae)
VL  - 14
DO  - 10.3389/fpls.2023.1211453
ER  - 

@article{
author = "Aničić, Neda and Matekalo, Dragana and Skorić, Marijana and Gašić, Uroš M. and Nestorović Živković, Jasmina and Dmitrović, Slavica and Božunović, Jelena and Milutinović, Milica and Petrović, Luka and Dimitrijević, Milena and Anđelković, Boban D. and Mišić, Danijela",
year = "2024",
abstract = "Iridoids, a class of atypical monoterpenes, exhibit exceptional diversity within the Nepeta genus (subfam. Nepetoidae, fam. Lamiaceae).The majority of these plants produce iridoids of the unique stereochemistry, with nepetalactones (NLs) predominating; however, a few Nepeta species lack these compounds. By comparatively analyzing metabolomics, transcriptomics, gene co-expression, and phylogenetic data of the iridoid-producing N. rtanjensis Diklić & Milojević and iridoid-lacking N. nervosa Royle & Bentham, we presumed that one of the factors responsible for the absence of these compounds in N. nervosa is iridoid synthase (ISY). Two orthologues of ISY were mined from leaves transcriptome of N. rtanjensis (NrPRISE1 and NrPRISE2), while in N. nervosa only one (NnPRISE) was identified, and it was phylogenetically closer to the representatives of the Family 1 isoforms, designated as P5βRs. Organ-specific and MeJA-elicited profiling of iridoid content and co-expression analysis of IBG candidates, highlighted NrPRISE2 and NnPRISE as promising candidates for ISY orthologues, and their function was confirmed using in vitro assays with recombinant proteins, after heterologous expression of recombinant proteins in E. coli and their His-tag affinity purification. NrPRISE2 demonstrated ISY activity both in vitro and likely in planta, which was supported by the 3D modeling and molecular docking analysis, thus reclassification of NrPRISE2 to NrISY is accordingly recommended. NnPRISE also displays in vitro ISY-like activity, while its role under in vivo conditions was not here unambiguously confirmed. Most probably under in vivo conditions the NnPRISE lacks substrates to act upon, as a result of the loss of function of some of the upstream enzymes of the iridoid pathway. Our ongoing work is conducted towards re-establishing the biosynthesis of iridoids in N. nervosa.",
publisher = "Frontiers Media",
journal = "Frontiers in Plant Science",
title = "Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae)",
volume = "14",
doi = "10.3389/fpls.2023.1211453"
}

Aničić, N., Matekalo, D., Skorić, M., Gašić, U. M., Nestorović Živković, J., Dmitrović, S., Božunović, J., Milutinović, M., Petrović, L., Dimitrijević, M., Anđelković, B. D.,& Mišić, D.. (2024). Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae). in Frontiers in Plant Science
Frontiers Media., 14.
https://doi.org/10.3389/fpls.2023.1211453

Aničić N, Matekalo D, Skorić M, Gašić UM, Nestorović Živković J, Dmitrović S, Božunović J, Milutinović M, Petrović L, Dimitrijević M, Anđelković BD, Mišić D. Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae). in Frontiers in Plant Science. 2024;14.
doi:10.3389/fpls.2023.1211453 .

Aničić, Neda, Matekalo, Dragana, Skorić, Marijana, Gašić, Uroš M., Nestorović Živković, Jasmina, Dmitrović, Slavica, Božunović, Jelena, Milutinović, Milica, Petrović, Luka, Dimitrijević, Milena, Anđelković, Boban D., Mišić, Danijela, "Functional iridoid synthases from iridoid producing and non-producing Nepeta species (subfam. Nepetoidae, fam. Lamiaceae)" in Frontiers in Plant Science, 14 (2024),
https://doi.org/10.3389/fpls.2023.1211453 . .

TY  - JOUR
AU  - Banjanac, Tijana
AU  - Dragičević, Milan
AU  - Šiler, Branislav
AU  - Gašić, Uroš M.
AU  - Bohanec, Borut
AU  - Nestorović Živković, Jasmina
AU  - Trifunović, Snežana S.
AU  - Mišić, Danijela
PY  - 2017
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/2477
AB  - Species within the genus Centaurium readily hybridize and polyploid complexes are often seen in natural populations. We describe phytochemical profiles of newly discovered allohexaploid hybrid, here named Centaurium pannonicum, and its parental tetraploid species C. erythraea and rare C. littorale ssp. compressum. Our aim was to examine chemodiversity of these taxa in the area of Vojvodina (North Serbia) and to perform metabolomics search for chemical classifiers which would provide high resolution discrimination of parental and hybrid individuals. In sum, UHPLC-MS/MS Orbitrap metabolomics fingerprinting revealed seventy compounds in methanol extracts. Despite the lack of qualitative chemical novelty in hybrid plants, UHPLC-qqqMS targeted metabolomics approach, aimed at three secoiridoid compounds and seventeen phenolics, pointed to considerable differences in quantitative composition of these dominant compounds among the plant taxa studied. In addition to the difference in the ploidy levels, the hybrid taxon was well distinguished from both parental species based on metabolite profiles, and, for most individuals, positioned intermediately to the parental taxa in both PCA and hierarchical clustering. After optimizing and comparing several statistical learning methods, it was possible to narrow the number of taxonomic classifiers to five (three xanthones, one secoiridoid glycoside, and one phenolic acid), while increasing the differentiation resolution. The presented metabolomics approach will certainly, along with morphometrics and molecular genetics studies, have high impact on further elucidation of complex relationships among taxa within the genus Centaurium.
PB  - Pergamon-Elsevier Science Ltd, Oxford
T2  - Phytochemistry
T1  - Chemodiversity of two closely related tetraploid Centaurium species and their hexaploid hybrid: Metabolomic search for high-resolution taxonomic classifiers
VL  - 140
SP  - 27
EP  - 44
DO  - 10.1016/j.phytochem.2017.04.005
ER  - 

@article{
author = "Banjanac, Tijana and Dragičević, Milan and Šiler, Branislav and Gašić, Uroš M. and Bohanec, Borut and Nestorović Živković, Jasmina and Trifunović, Snežana S. and Mišić, Danijela",
year = "2017",
abstract = "Species within the genus Centaurium readily hybridize and polyploid complexes are often seen in natural populations. We describe phytochemical profiles of newly discovered allohexaploid hybrid, here named Centaurium pannonicum, and its parental tetraploid species C. erythraea and rare C. littorale ssp. compressum. Our aim was to examine chemodiversity of these taxa in the area of Vojvodina (North Serbia) and to perform metabolomics search for chemical classifiers which would provide high resolution discrimination of parental and hybrid individuals. In sum, UHPLC-MS/MS Orbitrap metabolomics fingerprinting revealed seventy compounds in methanol extracts. Despite the lack of qualitative chemical novelty in hybrid plants, UHPLC-qqqMS targeted metabolomics approach, aimed at three secoiridoid compounds and seventeen phenolics, pointed to considerable differences in quantitative composition of these dominant compounds among the plant taxa studied. In addition to the difference in the ploidy levels, the hybrid taxon was well distinguished from both parental species based on metabolite profiles, and, for most individuals, positioned intermediately to the parental taxa in both PCA and hierarchical clustering. After optimizing and comparing several statistical learning methods, it was possible to narrow the number of taxonomic classifiers to five (three xanthones, one secoiridoid glycoside, and one phenolic acid), while increasing the differentiation resolution. The presented metabolomics approach will certainly, along with morphometrics and molecular genetics studies, have high impact on further elucidation of complex relationships among taxa within the genus Centaurium.",
publisher = "Pergamon-Elsevier Science Ltd, Oxford",
journal = "Phytochemistry",
title = "Chemodiversity of two closely related tetraploid Centaurium species and their hexaploid hybrid: Metabolomic search for high-resolution taxonomic classifiers",
volume = "140",
pages = "27-44",
doi = "10.1016/j.phytochem.2017.04.005"
}

Banjanac, T., Dragičević, M., Šiler, B., Gašić, U. M., Bohanec, B., Nestorović Živković, J., Trifunović, S. S.,& Mišić, D.. (2017). Chemodiversity of two closely related tetraploid Centaurium species and their hexaploid hybrid: Metabolomic search for high-resolution taxonomic classifiers. in Phytochemistry
Pergamon-Elsevier Science Ltd, Oxford., 140, 27-44.
https://doi.org/10.1016/j.phytochem.2017.04.005

Banjanac T, Dragičević M, Šiler B, Gašić UM, Bohanec B, Nestorović Živković J, Trifunović SS, Mišić D. Chemodiversity of two closely related tetraploid Centaurium species and their hexaploid hybrid: Metabolomic search for high-resolution taxonomic classifiers. in Phytochemistry. 2017;140:27-44.
doi:10.1016/j.phytochem.2017.04.005 .

Banjanac, Tijana, Dragičević, Milan, Šiler, Branislav, Gašić, Uroš M., Bohanec, Borut, Nestorović Živković, Jasmina, Trifunović, Snežana S., Mišić, Danijela, "Chemodiversity of two closely related tetraploid Centaurium species and their hexaploid hybrid: Metabolomic search for high-resolution taxonomic classifiers" in Phytochemistry, 140 (2017):27-44,
https://doi.org/10.1016/j.phytochem.2017.04.005 . .

TY  - JOUR
AU  - Mišić, Danijela
AU  - Šiler, Branislav
AU  - Gašić, Uroš M.
AU  - Avramov, Stevan N.
AU  - Živković, Suzana
AU  - Nestorović Živković, Jasmina
AU  - Milutinovic, Milica
AU  - Tešić, Živoslav Lj.
PY  - 2015
UR  - https://cherry.chem.bg.ac.rs/handle/123456789/1663
AB  - Introduction - Nepeta species contain a variety of secondary metabolites, including iridoid monoterpenes - nepetalactones and phenolic acids - that are considered the main bioactive constituents. This work represents the first attempt to comparatively explore variations in these two major groups of secondary metabolites within the genus. Objective - To develop an efficient analytical methodology for simultaneous analysis of nepetalactones and phenolic acids in methanol extracts of selected Nepeta species, and to evaluate its potential application in chemotaxonomic studies. Material and methods - A UHPLC combined with linear-trap quadrupole (LTQ) orbitrap MS method was used to characterise chemical diversity and complexity of phenolics among 12 selected Nepeta species. A targeted metabolomic approach using UHPLC coupled to a diode array detector (DAD) and combined with (+/-) heated electrospray ionisation (HESI) MS/MS was developed and validated for quantitative analysis of six hydroxycinnamic acid derivatives and four nepetalactones. Results - Phenolic profiling provided a valuable database of bioactive compounds in the plant group studied, including phenolic acids (hydroxybenzoic and hydroxycinnamic acids) and flavonoids (flavones, flavonols and flavanones). Principal component analysis and cluster analysis suggested the applicability of 10 targeted compounds as chemomarkers for chemotaxonomic studies. Pearson's correlation analysis revealed significant positive correlations between metabolites involved in different biosynthetic pathways (phenylpropanoid or monoterpenoid). Conclusion - The described targeted metabolomic approach proved to be highly beneficial in designing a phytochemical overview of the genus Nepeta, and might have applications in further clarification of phylogenetic relations. Furthermore, it has the potential to be implemented in a routine quality control of plant material and herbal preparations.
PB  - Wiley-Blackwell, Hoboken
T2  - Phytochemical Analysis
T1  - Simultaneous UHPLC/DAD/( plus /-)HESI-MS/MS Analysis of Phenolic Acids and Nepetalactones in Methanol Extracts of Nepeta Species: A Possible Application in Chemotaxonomic Studies
VL  - 26
IS  - 1
SP  - 72
EP  - 85
DO  - 10.1002/pca.2538
ER  - 

@article{
author = "Mišić, Danijela and Šiler, Branislav and Gašić, Uroš M. and Avramov, Stevan N. and Živković, Suzana and Nestorović Živković, Jasmina and Milutinovic, Milica and Tešić, Živoslav Lj.",
year = "2015",
abstract = "Introduction - Nepeta species contain a variety of secondary metabolites, including iridoid monoterpenes - nepetalactones and phenolic acids - that are considered the main bioactive constituents. This work represents the first attempt to comparatively explore variations in these two major groups of secondary metabolites within the genus. Objective - To develop an efficient analytical methodology for simultaneous analysis of nepetalactones and phenolic acids in methanol extracts of selected Nepeta species, and to evaluate its potential application in chemotaxonomic studies. Material and methods - A UHPLC combined with linear-trap quadrupole (LTQ) orbitrap MS method was used to characterise chemical diversity and complexity of phenolics among 12 selected Nepeta species. A targeted metabolomic approach using UHPLC coupled to a diode array detector (DAD) and combined with (+/-) heated electrospray ionisation (HESI) MS/MS was developed and validated for quantitative analysis of six hydroxycinnamic acid derivatives and four nepetalactones. Results - Phenolic profiling provided a valuable database of bioactive compounds in the plant group studied, including phenolic acids (hydroxybenzoic and hydroxycinnamic acids) and flavonoids (flavones, flavonols and flavanones). Principal component analysis and cluster analysis suggested the applicability of 10 targeted compounds as chemomarkers for chemotaxonomic studies. Pearson's correlation analysis revealed significant positive correlations between metabolites involved in different biosynthetic pathways (phenylpropanoid or monoterpenoid). Conclusion - The described targeted metabolomic approach proved to be highly beneficial in designing a phytochemical overview of the genus Nepeta, and might have applications in further clarification of phylogenetic relations. Furthermore, it has the potential to be implemented in a routine quality control of plant material and herbal preparations.",
publisher = "Wiley-Blackwell, Hoboken",
journal = "Phytochemical Analysis",
title = "Simultaneous UHPLC/DAD/( plus /-)HESI-MS/MS Analysis of Phenolic Acids and Nepetalactones in Methanol Extracts of Nepeta Species: A Possible Application in Chemotaxonomic Studies",
volume = "26",
number = "1",
pages = "72-85",
doi = "10.1002/pca.2538"
}

Mišić, D., Šiler, B., Gašić, U. M., Avramov, S. N., Živković, S., Nestorović Živković, J., Milutinovic, M.,& Tešić, Ž. Lj.. (2015). Simultaneous UHPLC/DAD/( plus /-)HESI-MS/MS Analysis of Phenolic Acids and Nepetalactones in Methanol Extracts of Nepeta Species: A Possible Application in Chemotaxonomic Studies. in Phytochemical Analysis
Wiley-Blackwell, Hoboken., 26(1), 72-85.
https://doi.org/10.1002/pca.2538

Mišić D, Šiler B, Gašić UM, Avramov SN, Živković S, Nestorović Živković J, Milutinovic M, Tešić ŽL. Simultaneous UHPLC/DAD/( plus /-)HESI-MS/MS Analysis of Phenolic Acids and Nepetalactones in Methanol Extracts of Nepeta Species: A Possible Application in Chemotaxonomic Studies. in Phytochemical Analysis. 2015;26(1):72-85.
doi:10.1002/pca.2538 .

Mišić, Danijela, Šiler, Branislav, Gašić, Uroš M., Avramov, Stevan N., Živković, Suzana, Nestorović Živković, Jasmina, Milutinovic, Milica, Tešić, Živoslav Lj., "Simultaneous UHPLC/DAD/( plus /-)HESI-MS/MS Analysis of Phenolic Acids and Nepetalactones in Methanol Extracts of Nepeta Species: A Possible Application in Chemotaxonomic Studies" in Phytochemical Analysis, 26, no. 1 (2015):72-85,
https://doi.org/10.1002/pca.2538 . .