Modulation of Phenolic Profiles and Antioxidant Capacity by Culture Medium and Tissue Type in Echinacea purpurea and Echinacea pallida

Year 2025, Volume: 9 Issue: 3, 768 - 777, 27.09.2025

Münüre Tanur Erkoyuncu

https://doi.org/10.31015/2025.3.15

Abstract

This study aimed to compare the accumulation of secondary metabolites in root and stem tissues of Echinacea purpurea and Echinacea pallida grown in vitro on different basal media (MS, WPM, B5). Five major caffeic acid derivatives (CADs: caftaric, chlorogenic, caffeic, echinacoside, and cichoric acid), total phenolic content (TPC), total flavonoid content (TFC), and total antioxidant capacity (TAC) were measured. In E. pallida, the highest levels of caftaric acid (11.29 ± 0.06 mg/g DW) and cichoric acid (11.79 ± 0.01 mg/g DW) were found in root tissues on B5 medium, while echinacoside reached 8.29 ± 0.26 mg/g DW on WPM. Chlorogenic acid was most abundant in roots on MS (1.57 ± 0.06 mg/g DW). Caffeic acid was found in low amounts in WPM and B5 media only. In E. purpurea, cichoric acid reached the highest value overall in B5-grown root tissue (70.64 ± 0.41 mg/g DW). The same tissue also showed high caftaric (11.05 ± 0.03 68 mg/g DW) and caffeic acid (0.35 ± 0.0168 mg/g DW) levels. Chlorogenic acid peaked in WPM roots (5.25 ± 0.68 mg/g DW). Echinacoside was not detected in this species. Antioxidant parameters also differed significantly by species, tissue, and medium. In E. purpurea root tissues, the highest TPC (69.12 ± 1.98 mg GAE/g DW) and TFC (15.74 ± 0.93 mg QE/g DW) were found in B5 medium, while the highest TAC (%85.90) was recorded in WPM. E. pallida generally had lower antioxidant values. Overall, secondary metabolite accumulation was strongly influenced by species, tissue type, and medium. B5 was most effective for CADs, and WPM supported greater antioxidant capacity. E. purpurea outperformed E. pallida in both cichoric acid content and antioxidant potential, indicating its promising potential for targeted metabolite production in plant tissue culture.

Keywords

Echinacea , Caffeic acid derivatives , TPC , TFC , Antioxidant activity , In vitro culture , Basal media

References

• Abbasi, B. H., Stiles, A. R., Saxena, P. K., & Liu, C.-Z. (2012). Gibberellic acid increases secondary metabolite production in Echinacea purpurea hairy roots. Applied biochemistry and biotechnology, 168(7), 2057-2066. https://doi.org/DOI 10.1007/s12010-012-9917-z
• Abbasi, B. H., Tian, C.-L., Murch, S. J., Saxena, P. K., & Liu, C.-Z. (2007). Light-enhanced caffeic acid derivatives biosynthesis in hairy root cultures of Echinacea purpurea. Plant Cell Reports, 26(8), 1367-1372. https://doi.org/DOI 10.1007/s00299-007-0344-5
• An, D., Wu, C.-H., Wang, M., Wang, M., Chang, G.-N., Chang, X.-J., & Lian, M.-L. (2022). Methyl jasmonate elicits enhancement of bioactive compound synthesis in adventitious root co-culture of Echinacea purpurea and Echinacea pallida. In Vitro Cellular & Developmental Biology-Plant, 58(1), 181-187. https://doi.org/https://doi.org/10.1007/s11627-021-10195-z
• Barnes, J., Anderson, L. A., Gibbons, S., & Phillipson, J. D. (2005). Echinacea species (Echinacea angustifolia (DC.) Hell., Echinacea pallida (Nutt.) Nutt., Echinacea purpurea (L.) Moench): a review of their chemistry, pharmacology and clinical properties. Journal of Pharmacy and Pharmacology, 57(8), 929-954. https://doi.org/https://doi.org/10.1211/0022357056127
• Chang, C.-C., Yang, M.-H., Wen, H.-M., & Chern, J.-C. (2002). Estimation of total flavonoid content in propolis by two complementary colorimetric methods. Journal of food and drug analysis, 10(3).
• Cui, H.-Y., Abdullahil Baque, M., Lee, E.-J., & Paek, K.-Y. (2013). Scale-up of adventitious root cultures of Echinacea angustifolia in a pilot-scale bioreactor for the production of biomass and caffeic acid derivatives. Plant biotechnology reports, 7(3), 297-308. https://doi.org/10.1007/s11816-012-0263-y
• Dalby-Brown, L., Barsett, H., Landbo, A.-K. R., Meyer, A. S., & Mølgaard, P. (2005). Synergistic antioxidative effects of alkamides, caffeic acid derivatives, and polysaccharide fractions from Echinacea purpurea on in vitro oxidation of human low-density lipoproteins. Journal of Agricultural and Food Chemistry, 53(24), 9413-9423. https://doi.org/https://doi.org/10.1021/jf0502395
• Demirci, T. (2022). Determination of secondary metabolite production efficiency in Echinacea purpurea callus, shoot, and root in vitro cultures with methyl jasmonate applications. Acta Physiologiae Plantarum, 44(12), 128. https://doi.org/doi.org/10.1007/s11738-022-03468-6
• Demirci, T., Çelikkol Akçay, U., & Göktürk Baydar, N. (2020). Effects of 24-epibrassinolide and l-phenylalanine on growth and caffeic acid derivative production in hairy root culture of Echinacea purpurea L. Moench. Acta Physiologiae Plantarum, 42(4), 66. https://doi.org/doi.org/10.1007/s11738-020-03055-7
• Demirci, T., Özmen, S., Yılmaz, E. G., Aşcı, Ö. A., & Baydar, N. G. (2017). The influence of methyl jasmonate on growth and caffeic acid derivative contents of in vitro shoot and roots in echinaceae (Echinacea purpurea). Indian J Pharm Educ Res, 51(3), 513-517. https://doi.org/10.5530/ijper.51.3s.77
• Erenler, R., Telci, I., Ulutas, M., Demirtas, I., Gul, F., Elmastas, M., & Kayir, O. (2015). Chemical Constituents, Quantitative Analysis and Antioxidant Activities of E chinacea purpurea (L.) M oench and E chinacea pallida (N utt.) N utt. Journal of food biochemistry, 39(5), 622-630.
• Erkoyuncu, M. T., & Yorgancilar, M. (2021). Optimization of callus cultures at Echinacea purpurea L. for the amount of caffeic acid derivatives. Electronic Journal of Biotechnology, 51, 17-27. https://doi.org/https://doi.org/10.1016/j.ejbt.2021.02.003
• Fan, M.-z., Wu, X.-h., Li, X.-f., Piao, X.-c., Jiang, J., & Lian, M.-l. (2021). Co-cultured adventitious roots of Echinacea pallida and Echinacea purpurea inhibit lipopolysaccharide-induced inflammation via MAPK pathway in mouse peritoneal macrophages. Chinese Herbal Medicines, 13(2), 228-234. https://doi.org/https://doi.org/10.1016/j.chmed.2021.01.001
• Fu, R., Zhang, P., Deng, Z., Jin, G., Guo, Y., & Zhang, Y. (2021). Diversity of antioxidant ingredients among Echinacea species. Industrial Crops and Products, 170, 113699. https://doi.org/https://doi.org/10.1016/j.indcrop.2021.113699
• Gamborg, O. L. c., Miller, R. A., & Ojima, K. (1968). Nutrient requirements of suspension cultures of soybean root cells. Experimental cell research, 50(1), 151-158.
• Gao, Y., Wu, C., Piao, X., Han, L., Gao, R., & Lian, M. (2018). Optimization of culture medium components and culture period for production of adventitious roots of Echinacea pallida (Nutt.) Nutt. Plant Cell, Tissue and Organ Culture (PCTOC), 135(2), 299-307. https://doi.org/doi.org/10.1007/s11240-018-1464-z
• Gurav, S. S., Gurav, N. S., Patil, A. T., & Duragkar, N. J. (2020). Effect of explant source, culture media, and growth regulators on callogenesis and expression of secondary metabolites of Curcuma longa. Journal of Herbs, Spices & Medicinal Plants, 26(2), 172-190. https://doi.org/https://doi.org/10.1080/10496475.2019.1689542
• H Naghdi, B. (2013). Influence of KNO [3], CaCl [2] and MgSO [4] concentrations on growth and cichoric acid accumulation in hairy root culture of purple coneflower [echinacea purpurea L.].
• Hazrati, R., Zare, N., Asghari-Zakaria, R., Sheikhzadeh, P., & Johari-Ahar, M. (2022). Factors affecting the growth, antioxidant potential, and secondary metabolites production in hazel callus cultures. AMB Express, 12(1), 109. https://doi.org/doi.org/10.1186/s13568-022-01449-z
• Khalili, S., Moieni, A., & Abdoli, M. (2014). Influence of different strains of Agrobacterium rhizogenes, culture medium, age and type of explant on hairy root induction in Echinacea angustifolia. Iranian Journal of Genetics and Plant Breeding, 3(1), 56-49.
• Lee, S. O., Liu, Y., Murphy, P., & Hendrich, S. (2006). Bioavailability of Echinacea root powder phenolics in humans. In: Wiley Online Library.
• Liu, J., Yang, L., Dong, Y., Zhang, B., & Ma, X. (2018). Echinacoside, an inestimable natural product in treatment of neurological and other disorders. Molecules, 23(5), 1213. https://doi.org/https://doi.org/10.3390/molecules23051213
• Lloyd, G., & McCown, B. (1980). Commercially-feasible micropropagation of mountain laurel, Kalmia latifolia, by use of shoot-tip culture.
• Lopez, T., Corbin, C., Falguieres, A., Doussot, J., Montguillon, J., Hagège, D., Hano, C., & Lainé, É. (2016). Secondary metabolite accumulation, antibacterial and antioxidant properties of in vitro propagated Clidemia hirta L. extracts are influenced by the basal culture medium. Comptes Rendus. Chimie, 19(9), 1071-1076. https://doi.org/https://doi.org/10.1016/j.crci.2016.03.012
• Mobin, M., Wu, C.-H., Tewari, R. K., & Paek, K.-Y. (2015). Studies on the glyphosate-induced amino acid starvation and addition of precursors on caffeic acid accumulation and profiles in adventitious roots of Echinacea purpurea (L.) Moench. Plant Cell, Tissue and Organ Culture (PCTOC), 120(1), 291-301. https://doi.org/10.1007/s11240-014-0606-1
• Monfort, L. E. F., Bertolucci, S. K. V., Lima, A. F., de Carvalho, A. A., Mohammed, A., Blank, A. F., & Pinto, J. E. B. P. (2018). Effects of plant growth regulators, different culture media and strength MS on production of volatile fraction composition in shoot cultures of Ocimum basilicum. Industrial Crops and Products, 116, 231-239. https://doi.org/https://doi.org/10.1016/j.indcrop.2018.02.075
• Murashige, T., & Skoog, F. (1962). A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiologia plantarum, 15(3).
• Murch, S. J., Peiris, S. E., Shi, W. L., Zobayed, S., & Saxena, P. K. (2006). Genetic diversity in seed populations of Echinacea purpurea controls the capacity for regeneration, route of morphogenesis and phytochemical composition. Plant Cell Reports, 25(6), 522-532. https://doi.org/DOI 10.1007/s00299-006-0118-5
• Murthy, H. N., Lee, E.-J., & Paek, K.-Y. (2014). Production of secondary metabolites from cell and organ cultures: strategies and approaches for biomass improvement and metabolite accumulation. Plant Cell, Tissue and Organ Culture (PCTOC), 118(1), 1-16. https://doi.org/10.1007/s11240-014-0467-7
• Osman, N. I., Sidik, N. J., & Awal, A. (2016). Effects of variations in culture media and hormonal treatments upon callus induction potential in endosperm explant of Barringtonia racemosa L. Asian Pacific Journal of Tropical Biomedicine, 6(2), 143-147. https://doi.org/https://doi.org/10.1016/j.apjtb.2015.10.007
• Ozyigit, I. I., Dogan, I., Hocaoglu-Ozyigit, A., Yalcin, B., Erdogan, A., Yalcin, I. E., Cabi, E., & Kaya, Y. (2023). Production of secondary metabolites using tissue culture-based biotechnological applications. Frontiers in Plant Science, 14, 1132555.
• Pellati, F., Benvenuti, S., Magro, L., Melegari, M., & Soragni, F. (2004). Analysis of phenolic compounds and radical scavenging activity of Echinacea spp. Journal of Pharmaceutical and Biomedical Analysis, 35(2), 289-301. https://doi.org/https://doi.org/10.1016/S0731-7085(03)00645-9
• Romero, F. R., Delate, K., Kraus, G. A., Solco, A. K., Murphy, P. A., & Hannapel, D. J. (2009). Alkamide production from hairy root cultures of Echinacea. In Vitro Cellular & Developmental Biology-Plant, 45(5), 599-609. https://doi.org/10.1007/s11627-008-9187-1
• Roy, A., & Bharadvaja, N. (2017). Effect of different culture medias on shoot multiplication and stigmasterol content in accessions of Centella asiatica. International Journal of Ayurvedic and Herbal Medicine, 7, 2643-2650. https://doi.org/ http://www.interscience.org.uk
• Sun, C., Shen, X., Zhang, Y., Song, T., Xu, L., & Xiao, J. (2023). Molecular Defensive Mechanism of Echinacea purpurea (L.) Moench against PAH Contaminations. International Journal of Molecular Sciences, 24(13), 11020. https://doi.org/https://doi.org/10.3390/ijms241311020
• Thomsen, M. O., Fretté, X. C., Christensen, K. B., Christensen, L. P., & Grevsen, K. (2012). Seasonal variations in the concentrations of lipophilic compounds and phenolic acids in the roots of Echinacea purpurea and Echinacea pallida. Journal of Agricultural and Food Chemistry, 60(49), 12131-12141. https://doi.org/https://doi.org/10.1021/jf303292t
• Vanisree, M., Lee, C.-Y., Lo, S.-F., Nalawade, S. M., Lin, C. Y., & Tsay, H.-S. (2004). Studies on the production of some important secondary metabolites from medicinal plants by plant tissue cultures. Bot. Bull. Acad. Sin, 45(1), 1-22.
• Wu, C., Wang, M., Song, H., & Cui, X. (2013). Medium salt strength and sucrose concentration affect root growth and secondary metabolite contents in adventitious root cultures of Echinacea pallida. Nat Prod Res Dev, 25, 1167-1171.
• Wu, C.-H., Dewir, Y. H., Hahn, E.-J., & Paek, K.-Y. (2006). Optimization of culturing conditions for the production of biomass and phenolics from adventitious roots of Echinacea angustifolia. Journal of Plant Biology, 49(3), 193-199.
• Wu, C.-H., Huang, T., Cui, X.-H., & Paek, K. (2012). Induction of adventitious roots of Echinacea pallida and accumulation of caffeic acid derivatives. China Journal of Chinese Materia Medica, 37(24), 3768-3772.
• Wu, C.-H., Murthy, H. N., Hahn, E.-J., & Paek, K.-Y. (2007). Large-scale cultivation of adventitious roots of Echinacea purpurea in airlift bioreactors for the production of chichoric acid, chlorogenic acid and caftaric acid. Biotechnology letters, 29(8), 1179-1182. https://doi.org/10.1007/s10529-007-9399-1
• Xu, C.-g., Tang, T.-x., Chen, R., Liang, C.-h., Liu, X.-y., Wu, C.-l., Yang, Y.-s., Yang, D.-p., & Wu, H. (2014). A comparative study of bioactive secondary metabolite production in diploid and tetraploid Echinacea purpurea (L.) Moench. Plant Cell, Tissue and Organ Culture (PCTOC), 116(3), 323-332. https://doi.org/10.1007/s11240-013-0406-z