Abstract
Plants are important sources of chemical compounds with various applications. These are usually associated with the secondary metabolism of plants which is tightly linked to the interactions with the surrounding environment. These compounds usually accumulate in low amounts, sometimes leading to the overexploitation of the source plant. Hence, development of alternative platforms for the production of these compounds represents a more sustainable solution. Among such platforms, hairy roots (HR) cultures are often used for their ability to produce the same secondary metabolites as the original plant. Cynara cardunculus L. or cardoon has been used in folk medicine to treat various diseases. Distinct parts of the plant have been studied and found to produce secondary metabolites with health promoting properties. In this work, HR cultures from cardoon were characterized regarding their secondary metabolites production and potential biological activities. Methanolic extracts contained compounds that belong, almost exclusively, to the hydroxycinnamic acid group. These extracts showed maximum total phenolic content and antioxidant activity of 46 mg GAE/g DW and 98 μmolTE/g DW, respectively, at day 35 of the growth curve. This extract also showed antiproliferative activity on colorectal cancer cells (EC50 1.16 ± 0.07 mg/mL). This work demonstrates the potential of cardoon HR as alternative sources for valuable hydroxycinnamic acid compounds with important biological activities.
Key message
Hairy root cultures from cardoon produced mainly hydroxycinnamic acid compounds, which showed antioxidant activity and antiproliferative properties on colorectal cancer cell lines.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- 1,5-DCQA:
-
1,5-O-Dicaffeoylquinic acid
- AUC:
-
Area under the curve
- CGA:
-
Chlorogenic acid
- DW:
-
Dry weight
- FC:
-
Folin-Ciocalteu reagent
- GAE:
-
Gallic acid equivalent
- HPLC-MS:
-
High Performance Liquid Chromatography-Mass Spectrometry
- HR:
-
Hairy roots
- ORAC:
-
Oxygen radical absorbance capacity
- TE:
-
Trolox equivalent
- TPC:
-
Total phenolic content
References
Abu-Reidah IM, Arráez-Román D, Segura-Carretero A, Fernández-Gutiérrez A (2013) Extensive characterisation of bioactive phenolic constituents from globe artichoke (Cynara scolymus L.) by HPLC–DAD-ESI-QTOF-MS. Food Chem 141:2269–2277. https://doi.org/10.1016/j.foodchem.2013.04.066
Amselem J, Tepfer M (1992) Molecular basis for novel root phenotypes induced by Agrobacterium rhizogenes A4 on cucumber. Plant Mol Biol 19:421–432. https://doi.org/10.1007/BF00023390
Bakkali AT, Jaziri M, Foriers A et al (1997) Lawsone accumulation in normal and transformed cultures of henna, Lawsonia inermis. Plant Cell Tissue Organ Cult 51:83–87. https://doi.org/10.1023/A:1005926228301
Ben Salem M, Affes H, Athmouni K et al (2017) Chemicals compositions, antioxidant and anti-inflammatory activity of Cynara scolymus leaves extracts, and analysis of major bioactive polyphenols by HPLC. Evid Based Complement Altern Med 2017:1–14. https://doi.org/10.1155/2017/4951937
Bulgakov VP, Veremeichik GN, Grigorchuk VP et al (2016) The rolB gene activates secondary metabolism in arabidopsis calli via selective activation of genes encoding MYB and bHLH transcription factors. Plant Physiol Biochem 102:70–79. https://doi.org/10.1016/j.plaphy.2016.02.015
Chen R, Liu X, Zou J et al (2013) Qualitative and quantitative analysis of phenylpropanoids in cell culture, regenerated plantlets and herbs of Saussurea involucrata. J Pharm Biomed Anal 74:39–46. https://doi.org/10.1016/j.jpba.2012.10.010
Ciau-Uitz R, Miranda-Ham ML, Coello-Coello J et al (1994) Indole alkaloid production by transformed and non-transformed root cultures of Catharanthus roseus. Vitr Cell Dev Biol Plant 30P:84–88
Clifford MN, Knight S, Kuhnert N (2005) Discriminating between the six isomers of dicaffeoylquinic acid by LC-MS n. J Agric Food Chem 53:3821–3832. https://doi.org/10.1021/jf050046h
Dai J, Mumper RJ (2010) Plant phenolics: extraction, analysis and their antioxidant and anticancer properties. Molecules 15:7313–7352. https://doi.org/10.3390/molecules15107313
Dias MI, Barros L, Barreira JCM et al (2018) Phenolic profile and bioactivity of cardoon (Cynara cardunculus L.) inflorescence parts: selecting the best genotype for food applications. Food Chem 268:196–202. https://doi.org/10.1016/j.foodchem.2018.06.081
Folgado A, Pires AS, Figueiredo AC et al (2019) Toward alternative sources of milk coagulants for cheese manufacturing: establishment of hairy roots culture and protease characterization from Cynara cardunculus L. Plant Cell Rep 39:89–100. https://doi.org/10.1007/s00299-019-02475-1
Habibi P, Soccol CR, Grossi-de-Sa MF (2018) Hairy root-mediated biotransformation recent advances and exciting prospects. Hairy roots. Springer Singapore, Singapore, pp 185–211
Häkkinen ST, Moyano E, Cusidó RM, Oksman-Caldentey K-M (2016) Exploring the metabolic stability of engineered hairy roots after 16 years maintenance. Front Plant Sci 7:1486. https://doi.org/10.3389/fpls.2016.01486
Huang D, Ou B, Hampsch-Woodill M et al (2002) High-throughput assay of oxygen radical absorbance capacity (ORAC) using a multichannel liquid handling system coupled with a microplate fluorescence reader in 96-well format. J Agric Food Chem 50:4437–4444. https://doi.org/10.1021/jf0201529
Jeong G-T, Park D-H (2006) Enhanced secondary metabolite biosynthesis by elicitation in transformed plant root system effect of abiotic elicitors. Appl Biochem Biotechnol 130:436–446. https://doi.org/10.1385/ABAB:130:1:436
Kim YJ, Weathers PJ, Wyslouzil BE (2002) Growth of Artemisia annua hairy roots in liquid- and gas-phase reactors. Biotechnol Bioeng 80:454–464. https://doi.org/10.1002/bit.10389
Kukić J, Popović V, Petrović S et al (2008) Antioxidant and antimicrobial activity of Cynara cardunculus extracts. Food Chem 107:861–868. https://doi.org/10.1016/j.foodchem.2007.09.005
Lin L-Z, Harnly JM (2008) Identification of hydroxycinnamoylquinic acids of arnica flowers and burdock roots using a standardized LC-DAD-ESI/MS profiling method. J Agric Food Chem 56:10105–10114. https://doi.org/10.1021/jf802412m
Liu CZ, Wang YC, Ouyang F et al (1997) Production of artemisinin by hairy root cultures of Artemisia annua L. Biotechnol Lett 19:927–929. https://doi.org/10.1023/A:1018362309677
Lodise O, Patil K, Karshenboym I et al (2019) Inhibition of prostate cancer cells by 4,5-dicaffeoylquinic acid through cell cycle arrest. Prostate Cancer 2019:4520645. https://doi.org/10.1155/2019/4520645
Menin B, Moglia A, Comino C et al (2013) In vitro callus-induction in globe artichoke (Cynara cardunculus L. var. scolymus ) as a system for the production of caffeoylquinic acids. J Hortic Sci Biotechnol 88:537–542. https://doi.org/10.1080/14620316.2013.11513003
Mukherjee C, Samanta T, Mitra A (2016) Redirection of metabolite biosynthesis from hydroxybenzoates to volatile terpenoids in green hairy roots of Daucus carota. Planta 243:305–320. https://doi.org/10.1007/s00425-015-2403-4
Nemoto K, Hara M, Suzuki M et al (2009) Function of the aux and rol genes of the Ri plasmid in plant cell division in vitro. Plant Signal Behav 4:1145–1147. https://doi.org/10.4161/psb.4.12.9904
Oliveira-Alves SC, Pereira RS, Pereira AB et al (2020) Identification of functional compounds in baru (Dipteryx alata Vog.) nuts: nutritional value, volatile and phenolic composition, antioxidant activity and antiproliferative effect. Food Res Int 131:109026. https://doi.org/10.1016/j.foodres.2020.109026
Pandino G, Lombardo S, Mauromicale G, Williamson G (2011) Phenolic acids and flavonoids in leaf and floral stem of cultivated and wild Cynara cardunculus L. genotypes. Food Chem 126:417–422. https://doi.org/10.1016/j.foodchem.2010.11.001
Pandino G, Lombardo S, Mauromicale G (2013) Globe artichoke leaves and floral stems as a source of bioactive compounds. Ind Crops Prod 44:44–49. https://doi.org/10.1016/j.indcrop.2012.10.022
Pereira L, Silva P, Duarte M et al (2017) Targeting colorectal cancer proliferation, stemness and metastatic potential using brassicaceae extracts enriched in isothiocyanates: a 3D cell model-based study. Nutrients 9:368. https://doi.org/10.3390/nu9040368
Ramos PAB, Santos SAO, Guerra ÂR et al (2014) Phenolic composition and antioxidant activity of different morphological parts of Cynara cardunculus L. var. altilis (DC). Ind Crops Prod 61:460–471. https://doi.org/10.1016/j.indcrop.2014.07.042
Ramos PAB, Guerra ÂR, Guerreiro O et al (2017) Antiproliferative effects of Cynara cardunculus L. Var. altilis (DC) lipophilic extracts. Int J Mol Sci. https://doi.org/10.3390/ijms18010063
Rency AS, Pandian S, Kasinathan R et al (2019) Hairy root cultures as an alternative source for the production of high-value secondary metabolites. Natural bio-active compounds. Springer Singapore, Singapore, pp 237–264
Rodrigues L, Silva I, Poejo J et al (2016) Recovery of antioxidant and antiproliferative compounds from watercress using pressurized fluid extraction. RSC Adv 6:30905–30918
Sapio L, Salzillo A, Illiano M et al (2020) Chlorogenic acid activates ERK1/2 and inhibits proliferation of osteosarcoma cells. J Cell Physiol 235:3741–3752. https://doi.org/10.1002/jcp.29269
Skala E, Kicel A, Olszewska MA et al (2015) Establishment of hairy root cultures of Rhaponticum carthamoides (Willd.) Iljin for the production of biomass and caffeic acid derivatives. Biomed Res Int 2015:1–11. https://doi.org/10.1155/2015/181098
Sonnante G, D’Amore R, Blanco E et al (2010) Novel hydroxycinnamoyl-coenzyme A quinate transferase genes from artichoke are involved in the synthesis of chlorogenic acid. Plant Physiol 153:1224–1238. https://doi.org/10.1104/pp.109.150144
Thaipong K, Boonprakob U, Crosby K et al (2006) Comparison of ABTS, DPPH, FRAP, and ORAC assays for estimating antioxidant activity from guava fruit extracts. J Food Compos Anal 19:669–675. https://doi.org/10.1016/j.jfca.2006.01.003
Thakore D, Srivastava AK (2017) Production of biopesticide azadirachtin using plant cell and hairy root cultures. Eng Life Sci 17:997–1005. https://doi.org/10.1002/elsc.201700012
Vereshchagina YV, Bulgakov VP, Grigorchuk VP et al (2014) The rolC gene increases caffeoylquinic acid production in transformed artichoke cells. Appl Microbiol Biotechnol 98:7773–7780. https://doi.org/10.1007/s00253-014-5869-2
Verpoorte R (1998) Exploration of nature’s chemodiversity: the role of secondary metabolites as leads in drug development. Drug Discov Today 3:232–238. https://doi.org/10.1016/S1359-6446(97)01167-7
Weremczuk-Jeżyna I, Grzegorczyk-Karolak I, Frydrych B et al (2013) Hairy roots of Dracocephalum moldavica: rosmarinic acid content and antioxidant potential. Acta Physiol Plant 35:2095–2103. https://doi.org/10.1007/s11738-013-1244-7
Wetterauer B, Wildi E, Wink M (2018) Production of the anticancer compound camptothecin in root and hairy root cultures of Ophiorrhiza mungos L. In: Kumar N (ed) BT-biotechnological approaches for medicinal and aromatic plants conservation genetic improvement and utilization. Springer Singapore, Singapore, pp 303–341
Wink M (1988) Plant breeding: importance of plant secondary metabolites for protection against pathogens and herbivores. Theor Appl Genet 75:225–233. https://doi.org/10.1007/BF00303957
Acknowledgements
This work was supported by Fundação para a Ciência e Tecnologia (FCT, Portugal) through project ref. PTDC/BAA-AGR/30447/2017, Research Unit ref. UIDB/04551/2020 and PhD Fellowship to AF (ref. PD/BD/114488/2016, Plants for Life PhD Program), Portuguese Mass Spectrometry Network (Rede Nacional de Espectrometria de Massa—RNEM; Lisboa-01-0145-FEDER-402-022125), iNOVA4Health-UIDB/04462/2020 and INTERFACE Program (Innovation, Technology and Circular Economy Fund (FITEC). The authors wish to thank Ana Bárbara Pereira for technical support in the preparation of cell extracts.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Conflict of interest
The authors declare they have no conflict of interest.
Additional information
Communicated by Ali R. Alan.
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Folgado, A., Serra, A.T., Prazeres, I. et al. Hairy root cultures of Cynara cardunculus L. as a valuable source of hydroxycinnamic acid compounds. Plant Cell Tiss Organ Cult 147, 37–47 (2021). https://doi.org/10.1007/s11240-021-02102-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11240-021-02102-6