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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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