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Planta Med 2020; 86(15): 1148-1155
DOI: 10.1055/a-1176-1937
Natural Product Chemistry and Analytical Studies
Original Papers

Differentiation between Cistus L. (Sub-) Species (Cistaceae) Using NMR Metabolic Fingerprinting[#]

Simon Moosmang
1   University of Innsbruck, Institute for Pharmacy/Department of Pharmacognosy, Center for Molecular Biosciences Innsbruck (CMBI), Innsbruck, Austria
,
Sonja Sturm
1   University of Innsbruck, Institute for Pharmacy/Department of Pharmacognosy, Center for Molecular Biosciences Innsbruck (CMBI), Innsbruck, Austria
,
Johannes Novak
2   University of Veterinary Medicine Vienna, Institute for Animal Nutrition and Functional Plant Compounds, Vienna, Austria
,
Brigitte Lukas
2   University of Veterinary Medicine Vienna, Institute for Animal Nutrition and Functional Plant Compounds, Vienna, Austria
,
Hermann Stuppner
1   University of Innsbruck, Institute for Pharmacy/Department of Pharmacognosy, Center for Molecular Biosciences Innsbruck (CMBI), Innsbruck, Austria
› Author AffiliationsSupported by: Austrian Science Fund P 29305
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Abstract

The genus Cistus is taxonomically complex, as taxonomic classification of individual species based on morphological criteria is often difficult and ambiguous. However, specific species contain valuable natural products, especially terpenoids and polyphenols, which exert various biological effects and might therefore be used for treatment of a broad array of disorders. Hence, a fast and reliable method for clear identification of different Cistus (sub-) species is required. Approaches for analysis of secondary metabolite profiles, e.g., with NMR, might remedy the challenging classification of Cistus (sub-) species and help to identify specific markers for differentiation between them. In the present study, 678 samples from wild-growing Cistus populations, including 7 species and 6 subspecies/varieties thereof, were collected in 3 years from populations in 11 countries all over the Mediterranean basin. Samples were extracted with buffered aqueous methanol and analysed with NMR. From the resulting 1D-1H-NOESY and J-Res profile spectra, marker signals or spectral regions for the individual (sub-) species were identified with multivariate statistical tools. By examining the NMR profiles of these extracts, we were able to identify discriminators and specific markers for the investigated Cistus (sub-) species. Various influencing factors, like (sub-) species, wild harvestings of different populations from several countries, numerous collection sites, different years, and cultivation in greenhouses have been considered in this work. As the here identified markers are independent from these influencing factors, the results can be considered a robust model and might be used for future differentiation between Cistus (sub-) species.

Key words

Cistus L. - Cistaceae - NMR-secondary metabolites - metabolic fingerprinting - chemotaxonomic classification

# Dedicated to Prof. Wolfgang Kubelka on the occasion of his 85th birthday.


Supporting Information

    Information of NMR pulse programs, country-specific floras used for taxonomic classification, a breakdown of sample numbers by country and species, PCA and OPLS-DA scores and loadings plots, results of permutation tests used for model validation, boxplots comparing signal intensities between different Cistus species, and plots with mean bucket integrals of different Cistus (sub-) species are available as Supporting Information.

  • Supporting Information


Publication History

Received: 10 January 2020

Accepted after revision: 05 May 2020

Article published online:
03 June 2020

© 2020. Thieme. All rights reserved.

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  • References

  • 1 Guzmán B, Vargas P. Systematics, character evolution, and biogeography of Cistus L. (Cistaceae) based on ITS, trnL-trnF, and matK sequences. Mol Phylogenetics Evol 2005; 37: 644-660
    CrossrefPubMedSearch in Google Scholar
  • 2 Papaefthimiou D, Papanikolaou A, Falara V, Givanoudi S, Kostas S, Kanellis AK. Genus Cistus: a model for exploring labdane-type diterpenesʼ biosynthesis and a natural source of high value products with biological, aromatic, and pharmacological properties. Front Chem 2014; 2: 35
    CrossrefPubMedSearch in Google Scholar
  • 3 Frazão DF, Raimundo JR, Domingues JL, Quintela-Sabarís C, Gonçalves JC, Delgado F. Cistus ladanifer (Cistaceae): a natural resource in Mediterranean-type ecosystems. Planta 2018; 247: 289-300
    CrossrefPubMedSearch in Google Scholar
  • 4 Stępień A, Aebisher D, Bartusik-Aebisher D. Biological properties of Cistus species. Eur J Clin Exp Med 2018; 16: 127-132
    CrossrefPubMedSearch in Google Scholar
  • 5 Ehrhardt C, Hrincius ER, Korte V, Mazur I, Droebner K, Poetter A, Dreschers S, Schmolke M, Planz O, Ludwig S. A polyphenol rich plant extract, CYSTUS052, exerts anti influenza virus activity in cell culture without toxic side effects or the tendency to induce viral resistance. Antiviral Res 2007; 76: 38-47
    CrossrefPubMedSearch in Google Scholar
  • 6 Rebensburg S, Helfer M, Schneider M, Koppensteiner H, Eberle J, Schindler M, Gürtler L, Brack-Werner R. Potent in vitro antiviral activity of Cistus incanus extract against HIV and Filoviruses targets viral envelope proteins. Sci Rep 2016; 6: 20394
    CrossrefPubMedSearch in Google Scholar
  • 7 Raab-Straube E. Cistaceae. Euro+Med Plantbase – the information resource for Euro-Mediterranean plant diversity. Available at (Accessed January 7, 2020): http://ww2.bgbm.org/EuroPlusMed/PTaxonDetail.asp?NameCache=Cistus
  • 8 Lo Bianco M, Grillo O, Cañadas E, Venora G, Bacchetta G. Inter- and intraspecific diversity in Cistus L. (Cistaceae) seeds, analysed with computer vision techniques. Plant Biol 2017; 19: 183-190
    CrossrefPubMedSearch in Google Scholar
  • 9 Anastasaki T, Demetzos C, Perdetzoglou D, Gazouli M, Loukis A, Harvala C. Analysis of labdane-type diterpenes from Cistus creticus (subsp. creticus and subsp. eriocephalus), by GC and GC-MS. Planta Med 1999; 65: 735-739
    Thieme ConnectPubMedSearch in Google Scholar
  • 10 Paolini J, Falchi A, Quilichini Y, Desjobert JM, De Cian MC, Varesi L, Costa J. Morphological, chemical and genetic differentiation of two subspecies of Cistus creticus L. (C. creticus subsp. eriocephalus and C. creticus subsp. corsicus). Phytochem 2009; 70: 1146-1160
    CrossrefPubMedSearch in Google Scholar
  • 11 Barrajón-Catalán E, Fernández-Arroyo S, Roldán C, Guillén E, Saura D, Segura-Carretero A, Micol V. A systematic study of the polyphenolic composition of aqueous extracts deriving from several Cistus genus species: evolutionary relationship. Phytochem Anal 2011; 22: 303-312
    CrossrefPubMedSearch in Google Scholar
  • 12 Demoly JP, Montserrat P. Cistus . In: Castroviejo S, Aedo C, Cirujano S, Laínz M, Montserrat P, Morales R, Muñoz-Garmendia F, Navarro C, Paiva J, Soriano C. eds. Flora Ibérica, Vol. 3. Madrid: Consejo de Investigaciones Cientificas; 1993: 319-337
    Search in Google Scholar
  • 13 Barrajón-Catalán E, Tomás-Menor L, Morales-Soto A, Martí Bruñá N, Saura López D, Segura-Carretero A, Micol V. Rockroses (Cistus sp.) Oils. In: Preedy VR. ed. Essential Oils in Food Preservation, Flavor and Safety. Cambridge (MA): Academic Press; 2016: 649-658
    CrossrefSearch in Google Scholar
  • 14 Guzmán B, Vargas P. Historical biogeography and character evolution of Cistaceae (Malvales) based on analysis of plastid rbcL and trnL-trnF sequences. Org Divers Evol 2009; 9: 83-99
    CrossrefPubMedSearch in Google Scholar
  • 15 Civeyrel L, Leclercq J, Demoly JP, Agnan Y, Quèbre N, Pélissier C, Otto T. Molecular systematics, character evolution, and pollen morphology of Cistus and Halimium (Cistaceae). Plant Syst Evol 2011; 295: 23-54
    CrossrefPubMedSearch in Google Scholar
  • 16 Kim HK, Choi YH, Verpoorte R. NMR-based plant metabolomics: where do we stand, where do we go?. Trends Biotechnol 2011; 29: 267-275
    CrossrefPubMedSearch in Google Scholar
  • 17 Seger C, Sturm S. Analytical aspects of plant metabolite profiling platforms: current standings and future aims. J Proteome Res 2007; 6: 480-497
    CrossrefPubMedSearch in Google Scholar
  • 18 Kim HK, Choi YH, Verpoorte R. NMR-based metabolomic analysis of plants. Nat Protoc 2010; 5: 536-549
    CrossrefPubMedSearch in Google Scholar
  • 19 Kim HK, Choi YH, Erkelens C, Lefeber AWM, Verpoorte R. Metabolic fingerprinting of Ephedra species using 1H-NMR spectroscopy and principal component analysis. Chem Pharm Bull 2005; 53: 105-109
    CrossrefPubMedSearch in Google Scholar
  • 20 Kim HK, Khan S, Wilson EG, Kricun SDP, Meissner A, Goraler S, Deelder AM, Choi YH, Verpoorte R. Metabolic classification of South American Ilex species by NMR-based metabolomics. Phytochem 2010; 71: 773-784
    CrossrefPubMedSearch in Google Scholar
  • 21 Safer S, Cicek SS, Pieri V, Schwaiger S, Schneider P, Wissemann V, Stuppner H. Metabolic fingerprinting of Leontopodium species (Asteraceae) by means of 1H NMR and HPLC-ESI-MS. Phytochem 2011; 72: 1379-1389
    CrossrefPubMedSearch in Google Scholar
  • 22 Lund JA, Brown PN, Shipley PR. Differentiation of Crataegus spp. guided by nuclear magnetic resonance spectrometry with chemometric analyses. Phytochem 2017; 141: 11-19
    CrossrefPubMedSearch in Google Scholar
  • 23 Triba MN, Le Moyec L, Amathieu R, Goossens C, Bouchemal N, Nahon P, Rutledge DN, Savarin P. PLS/OPLS models in metabolomics: the impact of permutation of dataset rows on the K-fold cross-validation quality parameters. Mol Biosyst 2015; 11: 13-19
    CrossrefPubMedSearch in Google Scholar
  • 24 Politeo O, Maravić A, Burčul F, Carev I, Kamenjarin J. Phytochemical composition and antimicrobial activity of essential oils of wild growing Cistus species in Croatia. Nat Prod Commun 2018; 13: 771-774
    PubMedSearch in Google Scholar
  • 25 Bechlaghem K, Allali H, Benmehdi H, Aissaoui N, Flamini G. Chemical analysis of the essential oils of three Cistus species growing in North-West of Algeria. Agric Conspec Sci 2019; 84: 283-293
    PubMedSearch in Google Scholar
  • 26 Lukas B, Bragagna L, Starzyk K, Labedz K, Stolze K, Novak J. Polyphenol-Variabilität von zypriotischem Cistus creticus L. Julius-Kuhn-Archiv 2018; 460: 121-125
    PubMedSearch in Google Scholar
  • 27 Maggi F, Lucarini D, Papa F, Peron G, DallʼAcqua S. Phytochemical analysis of the labdanum-poor Cistus creticus subsp. eriocephalus (Viv.) Greuter et Burdet growing in central Italy. Biochem Syst Ecol 2016; 66: 50-57
    CrossrefPubMedSearch in Google Scholar