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

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.

Publikationsverlauf

Eingereicht: 10. Januar 2020

Angenommen nach Revision: 05. Mai 2020

Artikel online veröffentlicht:
03. Juni 2020

• 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
• 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
• 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
• 4 Stępień A, Aebisher D, Bartusik-Aebisher D. Biological properties of Cistus species. Eur J Clin Exp Med 2018; 16: 127-132
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 17 Seger C, Sturm S. Analytical aspects of plant metabolite profiling platforms: current standings and future aims. J Proteome Res 2007; 6: 480-497
• 18 Kim HK, Choi YH, Verpoorte R. NMR-based metabolomic analysis of plants. Nat Protoc 2010; 5: 536-549
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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
• 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