Abstract
Different Eryngium species have been used with ornamental, agricultural and medicinal purposes, as a consequence of their chemical constituents. In the southwest Europe the endemic Eryngium viviparum, presents a high risk of extinction and ex situ strategies are high recommended for efficient conservation and re-introduction program. The objective of this study was to satisfy a dual objective: (i) to develop an ex situ conservation strategy through micropropagation and (ii) taking advantage of the extraordinary potential of plant tissue culture, produce a considerable amount of plant material to carry out a preliminary phytochemical study, based on the accumulation of phenolic compounds and their associated antioxidant activity. First a factorial design was conducted in order to study the effect of two cytokinins (6- benzylaminopurine, BAP, and kinetin, KIN), at three levels (0, 1 and 2 mg L−1), on shoot multiplication. Later another factorial design was applied, by using three levels of MS medium salt strength (full, half and quarter- strength) and four sucrose levels (0, 1, 2, and 3%) for improving shoot elongation and rooting. In parallel, a preliminary quantification of total phenolic and flavonoid contents from E. viviparum aerial parts was determined. The simple micropropagation protocol designed allowed obtaining a high rates of shoot multiplication (5.1–5.8 new shoots), rooting (100%) with healthy long roots (3.1–3.5 cm) and plantlet acclimatization (96%). Moderate antioxidant activity was recorded in hydromethanolic extracts from E. viviparum aerial parts. High correlation between total phenolic content and BAP levels in the culture media was found. In conclusion, the micropropagation procedure described here for the endangered E. viviparum can be used as new and very efficient ex situ conservation strategy, and as potential source of antioxidants, conferring an added-value to this plant.
Key message
In this work, we addresses the development of an efficient in vitro culture procedure of Eryngium viviparum as ex situ conservation methodology, which leads to a plant reintroduction programs, and as new source for secondary metabolites (mainly phenolic compounds), without ecological impact in their limited populations (either using seeds or wild plants as source materials).
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References
Agbo MO, Uzor PF, Akazie Nneji UN et al (2015) Antioxidant, total phenolic and flavonoid content of selected nigerian medicinal plants. Dhaka Univ J Pharm Sci 14:35–41. https://doi.org/10.3329/dujps.v14i1.23733
Agresti A (1996) An introduction to categorical data analysis, 1st edn. John Wiley & Sons, New York
Aguiar C (2003) O Eryngium viviparum Gay. afinal não está extinto em Portugal. In: Silva Lusitana. Estação Florestal Nacional, Bragança, pp 231–232
Ainsworth EA, Gillespie KM (2007) Estimation of total phenolic content and other oxidation substrates in plant tissues using Folin-Ciocalteu reagent. Nat Protoc 2:875–877. https://doi.org/10.1038/nprot.2007.102
Ali M, Abbasi BH, Ul-haq I (2013) Production of commercially important secondary metabolites and antioxidant activity in cell suspension cultures of Artemisia absinthium L. Ind Crops Prod 49:400–406. https://doi.org/10.1016/J.INDCROP.2013.05.033
Ayuso M, Ramil-Rego P, Landin M et al (2017) Computer-assisted recovery of threatened plants: keys for breaking seed dormancy of Eryngium viviparum. Front Plant Sci. https://doi.org/10.3389/fpls.2017.02092
Bañares A, Blanca G, Güemes J et al (2004) Atlas y libro rojo de la flora vascular amenazada de españa. Dirección General de Conservación de la Naturaleza, Madrid
Belkaid A, Currie J-C, Desgagnés J, Annabi B (2006) The chemopreventive properties of chlorogenic acid reveal a potential new role for the microsomal glucose-6-phosphate translocase in brain tumor progression. Cancer Cell Int 6:7. https://doi.org/10.1186/1475-2867-6-7
Brand-Williams W, Cuvelier ME, Berset C (1995) Use of a free radical method to evaluate antioxidant activity. LWT-Food Sci Technol 28:25–30. https://doi.org/10.1016/S0023-6438(95)80008-5
Chandrika R, Vyshali P, Saraswathi K, Kaliwal B (2011) Rapid multiplication of mature flowering plant of Eryngium foetidum L. by in vitro technique. Int J Biotechnol Appl 3:114–117
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
de la Cádiz-Gurrea ML, Fernández-Arroyo S, Joven J, Segura-Carretero A (2013) Comprehensive characterization by UHPLC-ESI-Q-TOF-MS from an Eryngium bourgatii extract and their antioxidant and anti-inflammatory activities. Food Res Int 50:197–204. https://doi.org/10.1016/J.FOODRES.2012.09.038
Debergh P, Aitken-Christie J, Cohen D et al (1992) Reconsideration of the term ‘vitrification’ as used in micropropagation. Plant Cell, Tissue Organ Cult 30:135–140. https://doi.org/10.1007/BF00034307
Dias MI, Sousa MJ, Alves RC, Ferreira ICFR (2016) Exploring plant tissue culture to improve the production of phenolic compounds: a review. Ind Crops Prod 82:9–22. https://doi.org/10.1016/J.INDCROP.2015.12.016
Engelmann F (2011) Use of biotechnologies for the conservation of plant biodiversity. Vitr Cell Dev Biol 47:5–16. https://doi.org/10.1007/s11627-010-9327-2
Erdem SA, Nabavi SF, Orhan IE et al (2015) Blessings in disguise: a review of phytochemical composition and antimicrobial activity of plants belonging to the genus Eryngium. DARU J Pharm Sci 23:53. https://doi.org/10.1186/s40199-015-0136-3
Fay M (1992) Conservation of rare and endangered plants using in vitro methods. Vitr Cell Dev Biol 28:1–4
Gaspar T, Kevers C, Penel C et al (1996) Plant hormones and plant growth regulators in plant tissue culture. Vitr Cell Dev Biol 32:272–289. https://doi.org/10.1007/BF02822700
Gayatri M, Madhu M, Kavyashree R, Dhananjaya S (2006) A protocol for in vitro regeneration of Eryngium foetidum L. Indian J Botechnol 5:249–251
George EF, Hall MA, De Klerk G-J (eds) (2008) Plant propagation by tissue culture. Springer the Netherlands, New Delhi
González-Benito M, Martín C (2011) In vitro preservation of Spanish biodiversity. Vitr Cell Dev Biol 47:46–54
Gugliucci A, Bastos DHM (2009) Chlorogenic acid protects paraoxonase 1 activity in high density lipoprotein from inactivation caused by physiological concentrations of hypochlorite. Fitoterapia 80:138–142. https://doi.org/10.1016/J.FITOTE.2009.01.001
Hu J, Gao S, Liu S et al (2019) An aseptic rapid propagation system for obtaining plumbagin of Ceratostigma willmottianum Stapf. Plant Cell, Tissue Organ Cult 137:369–377. https://doi.org/10.1007/s11240-019-01577-8
Isah T, Umar S, Mujib A et al (2018) Secondary metabolism of pharmaceuticals in the plant in vitro cultures: strategies, approaches, and limitations to achieving higher yield. Plant Cell, Tissue Organ Cult 132:239–265. https://doi.org/10.1007/s11240-017-1332-2
Karuppusamy S (2009) A review on trends in production of secondary metabolites from higher plants by in vitro tissue, organ and cell cultures. J Med plants Res 3:1222–1239
Kikowska M, Budzianowski J, Krawczyk A, Thiem B (2012) Accumulation of rosmarinic, chlorogenic and caffeic acids in in vitro cultures of Eryngium planum L. Acta Physiol Plant 34:2425–2433. https://doi.org/10.1007/s11738-012-1011-1
Kikowska M, Thiem B, Sliwinska E et al (2014) The effect of nutritional factors and plant growth regulators on micropropagation and production of phenolic acids and saponins from plantlets and adventitious root cultures of Eryngium maritimum L. J Plant Growth Regul 33:809–819. https://doi.org/10.1007/s00344-014-9428-y
Kikowska M, Thiem B, Sliwinska E et al (2016) Micropropagation of Eryngium campestre L. via shoot culture provides valuable uniform plant material with enhanced content of phenolic acids and antimicrobial activity. Acta Biol Crac Bot 58:43–56. https://doi.org/10.1515/abcsb-2016-0009
Küpeli E, Kartal M, Aslan S, Yesilada E (2006) Comparative evaluation of the anti-inflammatory and antinociceptive activity of Turkish Eryngium species. J Ethnopharmacol 107:32–37. https://doi.org/10.1016/J.JEP.2006.02.005
Lansdown R (2011) Eryngium viviparum. In: Walter K, Gillett H (eds) The IUCN red list of threatened species 2011, 1st edn. The World Conservation Monitoring Center, IUCN-The World Conservation Union, Cambridge, p 862
Le Claire E, Schwaiger S, Banaigs B et al (2005) Distribution of a new rosmarinic acid derivative in Eryngium alpinum L. and other Apiaceae. J Agric Food Chem 53:4367–4372. https://doi.org/10.1021/JF050024V
Makunga NP, Jäger AK, van Staden J (2003) Micropropagation of Thapsia garganica—a medicinal plant. Plant Cell Rep 21:967–973. https://doi.org/10.1007/s00299-003-0623-8
Marín JA, Gella R (1987) Acclimatization of the micropropagated cherry rootstock Masto de montaña (Prunus cerasus L.). Acta Hortic 212:603–606. https://doi.org/10.17660/actahortic.1987.212.99
Matkowski A (2008) Plant in vitro culture for the production of antioxidants—a review. Biotechnol Adv 26:548–560. https://doi.org/10.1016/J.BIOTECHADV.2008.07.001
Máximo WPF, Santos PAA, Martins GS et al (2018) In vitro multiplication of Eucalyptus hybrid via temporary immersion bioreactor: culture media and cytokinin effects. Crop Breed Appl Biotechnol 18:131–138. https://doi.org/10.1590/1984-70332018v18n2a19
Miliauskas G, Venskutonis PR, van Beek TA (2004) Screening of radical scavenging activity of some medicinal and aromatic plant extracts. Food Chem 85:231–237. https://doi.org/10.1016/J.FOODCHEM.2003.05.007
Mize CW, Koehler KJ, Compton ME (1999) Statistical considerations for in vitro research: II—data to presentation. Vitr Cell Dev Biol 35:122–126. https://doi.org/10.1007/s11627-999-0021-1
Mongkolsilp S, Pongbupakit I, Sae-Lee N, Sitthithaworn W (2004) Radical scavenging activity and total phenolic content of medicinal plants used in primary health care. SWU J Pharm Sci 9:32–35
Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497. https://doi.org/10.1111/j.1399-3054.1962.tb08052.x
Nicole Cotelle BSP (2001) Role of flavonoids in oxidative stress. Curr Top Med Chem 1:569–590. https://doi.org/10.2174/1568026013394750
Pękal A, Pyrzynska K (2014) Evaluation of aluminium complexation reaction for flavonoid content assay. Food Anal Methods 7:1776–1782. https://doi.org/10.1007/s12161-014-9814-x
Petersen M, Simmonds MS (2003) Rosmarinic acid. Phytochemistry 62:121–125. https://doi.org/10.1016/S0031-9422(02)00513-7
Rice-Evans CA, Miller NJ, Bolwell PG et al (1995) The relative antioxidant activities of plant-derived polyphenolic flavonoids. Free Radic Res 22:375–383. https://doi.org/10.3109/10715769509145649
Rice-Evans CA, Miller NJ, Paganga G (1996) Structure-antioxidant activity relationships of flavonoids and phenolic acids. Free Radic Biol Med 20:933–956. https://doi.org/10.1016/0891-5849(95)02227-9
Romero M, Ramil-Rego P, Rubinos M (2004) Conservation status of Eryngium viviparum Gay. Acta Bot Gall 151:55–64
Sarasan V, Cripps R, Ramsay MM et al (2006) Conservation in vitro of threatened plants—progress in the past decade. Vitr Cell Dev Biol 42:206–214. https://doi.org/10.1079/IVP2006769
Sharma RK, Wakhlu AK, Boleria M (2004) Micropropagation of Anethum graveolens L through axillary shoot proliferation. J Plant Biochem Biotechnol 13:157–159. https://doi.org/10.1007/BF03263214
Silveira FAO, Negreiros D, Barbosa NPU et al (2016) Ecology and evolution of plant diversity in the endangered campo rupestre: a neglected conservation priority. Plant Soil 403:129–152. https://doi.org/10.1007/s11104-015-2637-8
Sutter E, Langhans RW (1982) Formation of epicuticular wax and its effect on water loss in cabbage plants regenerated from shoot-tip culture. Can J Bot 60:2896–2902. https://doi.org/10.1139/b82-350
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
Thiem B, Kikowska M, Krawczyk A et al (2013) Phenolic acid and DNA contents of micropropagated Eryngium planum L. Plant Cell, Tissue Organ Cult 114:197–206. https://doi.org/10.1007/s11240-013-0315-1
Treutter D (2010) Managing phenol contents in crop plants by phytochemical farming and breeding—visions and constraints. Int J Mol Sci 11:807–857. https://doi.org/10.3390/ijms11030807
Tusevski O, Vinterhalter B, Krstić Milošević D et al (2017) Production of phenolic compounds, antioxidant and antimicrobial activities in hairy root and shoot cultures of Hypericum perforatum L. Plant Cell, Tissue Organ Cult 128:589–605. https://doi.org/10.1007/s11240-016-1136-9
Vieitez AM, Vieitez ML (1980) Culture of chestnut shoots from buds in vitro. J Hortic Sci 55:83–84. https://doi.org/10.1080/00221589.1980.11514906
Villaño D, Fernández-Pachón MS, Moyá ML et al (2007) Radical scavenging ability of polyphenolic compounds towards DPPH free radical. Talanta 71:230–235. https://doi.org/10.1016/J.TALANTA.2006.03.050
Walter K, Gillett H (1998) 1997 IUCN Red List of threatened plants, 1st edn. The World Conservation Monitoring Center. IUCN-The World Conservation Union, Gland and Cambridge
Wang P, Su Z, Yuan W et al (2012) Phytochemical constituents and pharmacological activities of Eryngium L. (Apiaceae). Pharm Crop 3:99–120
Yip ECH, Chan ASL, Pang H et al (2006) Protocatechuic acid induces cell death in HepG2 hepatocellular carcinoma cells through a c-Jun N-terminal kinase-dependent mechanism. Cell Biol Toxicol 22:293–302. https://doi.org/10.1007/s10565-006-0082-4
Funding
This research was supported by TREMEDAL-Inland wetlands of Northern Iberian Peninsula: management and restoration of mires and wet environments European Union (LIFE11 NAT/ES/000707, 2012-2015). This work was funded by Xunta de Galicia, Spain (CITACA Strategic Partnership, Reference: ED431E 2018/07) and “Red de Uso Sostenible de Recursos y Residuos” (ED431D 2017/18). The authors acknowledge the FPU grant from the Spanish Ministry of Education, Culture and Sport (reference FPU15/04849) to P. García-Pérez.
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MA: Performed micropropagation experiments; MA and PGP: Performed phenolic compound determination and evaluation of RSA; PR-R: Contributed with plant seeds, reagents and materials; MA, PPG and MB: Conceived and designed the experiments. All authors contributed to the writing of the manuscript.
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Communicated by Ali R. Alan.
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Ayuso, M., García-Pérez, P., Ramil-Rego, P. et al. In vitro culture of the endangered plant Eryngium viviparum as dual strategy for its ex situ conservation and source of bioactive compounds. Plant Cell Tiss Organ Cult 138, 427–435 (2019). https://doi.org/10.1007/s11240-019-01638-y
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DOI: https://doi.org/10.1007/s11240-019-01638-y