Xerophytic Lichens from Gypsiferous Outcrops of Arid Areas of Andalusia as a Source of Anti-Phytopathogenic Depsides
Abstract
:1. Introduction
2. Materials and Methods
2.1. General
2.2. Lichen Material
2.3. Extraction
2.4. LC-MS/MS and Feature-Based Molecular Networking Analysis
2.5. Crude Extract Fractionation
2.6. Purification of Compounds
2.7. Anti-Phytopathogen Bioassays
2.8. Antitumoral Activity Assays
3. Results
3.1. Antifungal Activity of Lichen Extracts
3.2. Identification of Main Active Peaks from the Raw Extracts
3.3. Feature-Based Molecular Networking of Lichen Crude Extracts
3.4. Structural Elucidation of 8′-Hydroxyfumarprotocetraric Acid
3.5. Anti-Phytopathogenic Activity Characterization of Pure Compounds
3.6. Antitumoral Activity of Pure Compounds
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Conflicts of Interest
References
- Rizzo, D.M.; Lichtveld, M.; Mazet, J.A.K.; Togami, E.; Miller, S.A. Plant Health and Its Effects on Food Safety and Security in a One Health Framework: Four Case Studies. One Health Outlook 2021, 3, 6. [Google Scholar] [CrossRef]
- Doehlemann, G.; Ökmen, B.; Zhu, W.; Sharon, A. Plant Pathogenic Fungi. Microbiol. Spectr. 2017, 5, FUNK-0023-2016. [Google Scholar] [CrossRef]
- Callaway, E. Devastating Wheat Fungus Appears in Asia for First Time. Nature 2016, 532, 421–422. [Google Scholar] [CrossRef]
- Pegg, K.G.; Coates, L.M.; O’Neill, W.T.; Turner, D.W. The Epidemiology of Fusarium Wilt of Banana. Front. Plant Sci. 2019, 10, 1395. [Google Scholar] [CrossRef]
- Fisher, M.C.; Gurr, S.J.; Cuomo, C.A.; Blehert, D.S.; Jin, H.; Stukenbrock, E.H.; Stajich, J.E.; Kahmann, R.; Boone, C.; Denning, D.W.; et al. Threats Posed by the Fungal Kingdom to Humans, Wildlife, and Agriculture. mBio 2020, 11, e00449-20. [Google Scholar] [CrossRef]
- Bhattacharya, S. Deadly New Wheat Disease Threatens Europe’s Crops. Nature 2017, 542, 145–146. [Google Scholar] [CrossRef]
- Armstrong, R.A.; Armstrong, R.A. Adaptation of Lichens to Extreme Conditions. In Plant Adaptation Strategies in Changing Environment; Springer: Berlin/Heidelberg, Germany, 2017; pp. 1–27. [Google Scholar] [CrossRef]
- Françoise, L.D.; Holger, T.; Marie-Laurence, A.; David, D.; Joël, B. Oxidative Stress Regulation in Lichens and Its Relevance for Survival in Coastal Habitats. Adv. Bot. Res. 2014, 71, 467–503. [Google Scholar] [CrossRef]
- Kranner, I.; Beckett, R.; Hochman, A.; Nash, T.H. Desiccation-Tolerance in Lichens: A Review. Bryologist 2008, 111, 576–593. [Google Scholar] [CrossRef]
- Shukla, V.; Pant Joshi, G.; M Rawat, M.S. Lichens as a Potential Natural Source of Bioactive Compounds: A Review. Phytochem. Rev. 2010, 9, 303–314. [Google Scholar] [CrossRef]
- Srivastava, P.; Upreti, D.K.; Dhole, T.N.; Srivastava, A.K.; Nayak, M.T. Antimicrobial Property of Extracts of Indian Lichen against Human Pathogenic Bacteria. Interdiscip. Perspect. Infect Dis. 2013, 2013, 1–6. [Google Scholar] [CrossRef]
- Kosanić, M.; Šeklić, D.; Marković, S.; Ranković, B. Evaluation of antioxidant, antimicrobial and anticancer properties of selected lichens from Serbia. Dig. J. Nanomater. Biostruct. 2014, 9, 273–287. [Google Scholar]
- Martínez-Sánchez, J.J.; Casares-Porcel, M.; Guerra, J.; Gutiérrez-Carretero, L.; Ros, R.M.; Hernández-Bastida, J.; Cano, M.J. A Special Habitat for Bryophytes and Lichens in the Arid Zones of Spain. Lindbergia 1994, 19, 116–121. [Google Scholar]
- Escudero, A.; Palacio, S.; Maestre, F.T.; Luzuriaga, A.L. Plant Life on Gypsum: A Review of Its Multiple Facets. Biol. Rev. Camb. Philos. Soc. 2015, 90, 1–18. [Google Scholar] [CrossRef]
- Llimona, X. Las Comunidades de Líquenes de Los Yesos de España. Ph.D. Thesis, Universidad de Barcelona, Barcelona, Spain, 1974. Available online: https://www.researchgate.net/publication/50338234_Las_comunidades_de_liquenes_de_los_yesos_de_Espana (accessed on 20 October 2022).
- Crespo, A.; Barreno, E. Sobre Las Comunidades Líquénicas Rupícolas de Acarospora hilaris (Duf.) Hue en la Península Ibérica. Anales Inst. Bot. Cavanilles 1976, 33, 189–205. [Google Scholar]
- Guerra, J.; Ros, R.M.; Cano, M.J.; Casares-Porcel, M. Gypsiferous Outcrops in SE Spain, Refuges of Rare, Vulnerable and Endangered Bryophytes and Lichens. Cryptogam. Bryol. Lichenol 1995, 16, 125–135. [Google Scholar]
- Gutiérrez-Carretero, L.; Casares-Porcel, M. Los Líquenes de Los Afloramientos de Yeso de La Península Ibérica. In Diversidad Vegetal de las Yeseras Ibéricas; Mota, J.F., Sánchez-Gómez, P., Guirado, J.S., Eds.; ADIF-Mediterráneo Asesores Consultores: Almería, Spain, 2011; pp. 549–567. [Google Scholar]
- Casares-Porcel, M.; Gutiérrez-Carretero, L. Síntesis de La Vegetación Liquénica Gipsícola Termo-y Mesomediterránea de La Península Ibérica. Cryptogamie. Bryol. Lichénologie 1993, 14, 361–388. [Google Scholar]
- Hidalgo, M.E.; Fernández, E.; Quilhot, W.; Lissi, E. Antioxidant Activity of Depsides and Depsidones. Phytochemistry 1994, 37, 1585–1587. [Google Scholar] [CrossRef]
- Luo, H.; Yamamoto, Y.; Kim, J.A.; Jung, J.S.; Koh, Y.J.; Hur, J.S. Lecanoric Acid, a Secondary Lichen Substance with Antioxidant Properties from Umbilicaria antarctica in Maritime Antarctica (King George Island). Polar Biol. 2009, 32, 1033–1040. [Google Scholar] [CrossRef]
- Nugraha, A.S.; Laksono, T.A.; Firli, L.N.; Sukrisno Putri, C.P.Z.; Pratoko, D.K.; Zulfikar, Z.; Untari, L.F.; Wongso, H.; Lambert, J.M.; Dillon, C.T.; et al. Anti-Cancer Evaluation of Depsides Isolated from Indonesian Folious Lichens: Physcia millegrana, Parmelia dilatata and Parmelia aurulenta. Biomolecules 2020, 10, 1420. [Google Scholar] [CrossRef]
- Mohammadi, M.; Bagheri, L.; Badreldin, A.; Fatehi, P.; Pakzad, L.; Suntres, Z.; van Wijnen, A.J. Biological Effects of Gyrophoric Acid and Other Lichen Derived Metabolites, on Cell Proliferation, Apoptosis and Cell Signaling Pathways. Chem. Biol. Interact 2022, 351, 109768. [Google Scholar] [CrossRef]
- Girardot, M.; Millot, M.; Hamion, G.; Billard, J.L.; Juin, C.; Ntoutoume, G.M.A.N.; Sol, V.; Mambu, L.; Imbert, C. Lichen Polyphenolic Compounds for the Eradication of Candida albicans Biofilms. Front. Cell Infect Microbiol. 2021, 11, 849. [Google Scholar] [CrossRef] [PubMed]
- Oh, J.M.; Kim, Y.J.; Gang, H.S.; Han, J.; Ha, H.H.; Kim, H. Antimicrobial Activity of Divaricatic Acid Isolated from the Lichen Evernia mesomorpha against Methicillin-Resistant Staphylococcus Aureus. Molecules 2018, 23, 3068. [Google Scholar] [CrossRef]
- Nugraha, A.S.; Dayli, I.R.; Sukrisno Putri, C.P.Z.; Firli, L.N.; Widhi Pratama, A.N.; Triatmoko, B.; Untari, L.F.; Wongso, H.; Keller, P.A.; Wangchuk, P. Isolation of Antibacterial Depside Constituents from Indonesian Folious Lichen, Candelaria fibrosa. JBAPN 2022, 12, 24–32. [Google Scholar] [CrossRef]
- White, P.A.S.; Oliveira, R.C.M.; Oliveira, A.P.; Serafini, M.R.; Araújo, A.A.S.; Gelain, D.P.; Moreira, J.C.F.; Almeida, J.R.G.S.; Quintans, J.S.S.; Quintans-Junior, L.J.; et al. Antioxidant Activity and Mechanisms of Action of Natural Compounds Isolated from Lichens: A Systematic Review. Molecules 2014, 19, 14496–14527. [Google Scholar] [CrossRef]
- Martín, J.; Crespo, G.; González-Menéndez, V.; Pérez-Moreno, G.; Sánchez-Carrasco, P.; Pérez-Victoria, I.; Ruiz-Pérez, L.M.; González-Pacanowska, D.; Vicente, F.; Genilloud, O.; et al. MDN-0104, an antiplasmodial betaine lipid from Heterospora chenopodii. J. Nat. Prod. 2014, 77, 2118–2123. [Google Scholar] [CrossRef]
- Pérez-Victoria, I.; Martín, J.; Reyes, F. Combined LC/UV/MS and NMR Strategies for the Dereplication of Marine Natural Products. Planta Med. 2016, 82, 857–871. [Google Scholar] [CrossRef]
- Nothias, L.-F.; Petras, D.; Schmid, R.; Dührkop, K.; Rainer, J.; Sarvepalli, A.; Protsyuk, I.; Ernst, M.; Tsugawa, H.; Fleischauer, M.; et al. Feature-Based Molecular Networking in the GNPS Analysis Environment. Nat. Methods 2020, 17, 905–908. [Google Scholar] [CrossRef]
- Monteiro, M.C.; de La Cruz, M.; Cantizani, J.; Moreno, C.; Tormo, J.R.; Mellado, E.; de Lucas, J.R.; Asensio, F.; Valiante, V.; Brakhage, A.A.; et al. A New Approach to Drug Discovery: High-Throughput Screening of Microbial Natural Extracts against Aspergillus fumigatus Using Resazurin. J. Biomol. Screen 2012, 17, 542–549. [Google Scholar] [CrossRef]
- Lohezic-Le Devehat, F.; Legouin, B.; Couteau, C.; Boustie, J.; Coiffard, L. Lichenic Extracts and Metabolites as UV Filters. J. Photochem. Photobiol. B 2013, 120, 17–28. [Google Scholar] [CrossRef]
- Culberson, C.F. Stenosporic Acid, a New Depside in Ramalina stenospora. Phytochemistry 1970, 9, 841–844. [Google Scholar] [CrossRef]
- Candan, M.; Yilmaz, M.; Tay, T.; Kivanç, M.; Türk, H. Antimicrobial Activity of Extracts of the Lichen Xanthoparmelia pokornyi and Its Gyrophoric and Stenosporic Acid Constituents. Z. Naturforsch C J. Biosci. 2006, 61, 319–323. [Google Scholar] [CrossRef] [PubMed]
- Marijana, K.; Branislav, R.; Slobodan, S. Antimicrobial Activity of the Lichen Lecanora frustulosa and Parmeliopsis hyperopta and Their Divaricatic Acid and Zeorin Constituents. Afr. J. Microbiol. Res. 2010, 4, 885–890. [Google Scholar]
- Poncet, R.; le Dévéhat, F.L.; Ferron, S.; Hivert, J.; Fontaine, C.; Picot, F.; Bidault, E.; Kervran, L. The Genus Ramalina (Ascomycota, Lecanoromycetes, Ramalinaceae) from the Scattered Islands (French Southern and Antarctic Lands), with Descriptions of Three New Species. Plant Fungal Syst. 2021, 66, 211–224. [Google Scholar] [CrossRef]
Lichen | Place | GPS Coordinate | Voucher Code |
---|---|---|---|
Diploschistes ocellatus (Fr.) Norman var. almeriensis Llimona | Sorbas, Almeria | 37°5′17″ N/2°5′15″ O | GDA-Lichen 3966 |
Seirophora lacunosa (Rupr.) Fröden | Sorbas, Almeria | 37°5′22″ N/2°5′16″ O | GDA-Lichen 3970 |
Cladonia foliacea (Huds.) Willd. f. convoluta (Lam.) J. Steiner | Venta de los yesos, Tabernas, Almeria | 37°4′60″ N/2°17′37″ O | GDA-Lichen 3965 |
Acarospora placodiiformis (Nyl.) H. Olivier | Venta de los yesos, Tabernas, Almeria | 37°4′60″ N/2°17′38″ O | GDA-Lichen 3964 |
Xanthoparmelia pokornyi (Körb.) O. Blanco, A. Crespo, Elix, D. Hawksw. & Lumbsch | Venta de los yesos, Tabernas, Almeria | 37°4′60″ N/2°17′37″ O | GDA-Lichen 3969 |
Squamarina lentigera (Weber) Poelt | Venta de los yesos, Tabernas, Almeria | 37°4′60″ N/2°17′38″ O | GDA-Lichen 3968 |
Cluster | tR (min) | Present in Sample | m/z | Ion Type | Molecular Formula | Identified Compound * |
---|---|---|---|---|---|---|
C1 | 4.65 | S. lentigera | 345.098 | [M + H]+ | C18H16O7 | Usnic acid |
5.41 | X. pokornyi | 389.160 | [M + H]+ | C21H24O7 | Divaricatic acid | |
5.53 | S. lentigera | 419.377 | [M + H]+ | C22H26O8 | Sekikaic acid | |
5.96 | X. pokornyi | 417.192 | [M + H]+ | C23H28O7 | Stenosporic acid | |
6.06 | S. lentigera | 447.202 | [M + H]+ | C24H30O8 | 4′-O-Methylpaludosic acid | |
C2 | 3.03 | C. foliacea | 375.072 | [M + H]+ | C18H14O9 | Protocetraric acid |
3.13 | C. foliacea | 492.115 | [M + NH4]+ | C22H18O12 | 8′-Hydroxyfumarprotocetraric acid | |
4.91 | C. foliacea | 490.099 | [M + NH4]+ | C22H16O12 | Fumarprotocetraric acid | |
C3 | 2.76 | X. pokornyi | 197.178 | [M + H]+ | C10H12O4 | Divaric acid |
3.90 | X. pokornyi | 211.204 | [M + H]+ | C11H14O4 | Ethyl everninate |
CD3OD | HMBC | ||
1H NMR | 13C NMR | ||
N° | d in ppm, mult, J in Hz | d in ppm | |
1 | 111.7 | ||
2 | 161.3 | ||
3 | 115.6 | ||
4 | 161.1 | ||
5 | 6.64, s | 115.1 | 3, 4, 6, 8 and 9 |
6 | 144.6 | ||
7 | 162.5 | ||
8 | 4.96, s | 52.9 | 3, 4 and 5 |
9 | 2.44, s | 19.3 | 1, 5, 6 and 7 |
1′ | 110.3 | ||
2′ | 158.6 | ||
3′ | 112.2 | ||
4′ | 147.7 | ||
5′ | 142.4 | ||
6′ | 134.8 | ||
7′ | 172.3 | ||
8′ | 5.41, s | 56.1 | 2′, 3′, 4′ and 1″ |
9′ | 2.84, s | 14.1 | 4′, 5′, 6′ and 7′ |
1″ | 164.7 | ||
2″ | 6.73, d, 15.5 | 132.8 | 1″, 3″ and 4″ |
3″ | 6.72, d, 15.5 | 133.7 | 1″, 2″ and 4″ |
4″ | 166.1 |
Compound | IC50 (µg/mL) | |||||||||||||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Z. tritici | B. cinerea | C. acutatum | F. oxysporum TR4 | M. grisea | V. dahliae | F. proliferatum | ||||||||
Abs | Flu | Abs | Flu | Abs | Flu | Abs | Flu | Abs | Flu | Abs | Flu | Abs | Flu | |
8′-hydroxy fumarprotocetraric acid | >160 | >160 | >160 | >160 | >160 | >160 | >107 | >107 | >160 | >160 | >160 | >160 | >160 | >160 |
divaricatic acid | 12.4 | 10.67 | 36.87 | 30.29 | 56.84 | 75.75 | 79.00 | 83.28 | 48.85 | 51.85 | 49.38 | 32.11 | 83.13 | 88.07 |
stenosporic acid | 7.53 | 6.59 | 38.03 | 38.94 | 69.48 | 98.49 | 77.02 | 93.47 | 36.06 | 41.88 | 51.75 | 27.03 | 94.17 | 103.30 |
sekikaic acid | >160 | 134.6 | >160 | >160 | >160 | >160 | >107 | >107 | >160 | >160 | >160 | >160 | >160 | >160 |
4′-O-methylpaludosic acid | 38.87 | 24.88 | >160 | >160 | 157.7 | 106.8 | >107 | >107 | >160 | >160 | >160 | 89.78 | >160 | >160 |
Compound | ED 50 (µg/mL) | ||||
---|---|---|---|---|---|
A2058 | A549 | HepG2 | MCF7 | MIA PaCa-2 | |
8′-hydroxyfumarprotocetraric acid | >40.00 | >40.00 | >40.00 | >40.00 | >40.00 |
divaricatic acid | 23.75 | 26.99 | 28.41 | 15.12 | >40.00 |
stenosporic acid | >40.00 | >40.00 | >40.00 | 30.13 | >40.00 |
sekikaic acid | >40.00 | >40.00 | >40.00 | >40.00 | >40.00 |
4′-O-methylpaludosic acid | >40.00 | >40.00 | >40.00 | >40.00 | >40.00 |
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Fernández-Pastor, I.; González-Menéndez, V.; Martínez Andrade, K.; Serrano, R.; Mackenzie, T.A.; Benítez, G.; Casares-Porcel, M.; Genilloud, O.; Reyes, F. Xerophytic Lichens from Gypsiferous Outcrops of Arid Areas of Andalusia as a Source of Anti-Phytopathogenic Depsides. J. Fungi 2023, 9, 887. https://doi.org/10.3390/jof9090887
Fernández-Pastor I, González-Menéndez V, Martínez Andrade K, Serrano R, Mackenzie TA, Benítez G, Casares-Porcel M, Genilloud O, Reyes F. Xerophytic Lichens from Gypsiferous Outcrops of Arid Areas of Andalusia as a Source of Anti-Phytopathogenic Depsides. Journal of Fungi. 2023; 9(9):887. https://doi.org/10.3390/jof9090887
Chicago/Turabian StyleFernández-Pastor, Ignacio, Victor González-Menéndez, Kevin Martínez Andrade, Rachel Serrano, Thomas A. Mackenzie, Guillermo Benítez, Manuel Casares-Porcel, Olga Genilloud, and Fernando Reyes. 2023. "Xerophytic Lichens from Gypsiferous Outcrops of Arid Areas of Andalusia as a Source of Anti-Phytopathogenic Depsides" Journal of Fungi 9, no. 9: 887. https://doi.org/10.3390/jof9090887