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Establishment of callus-cultures of the Argentinean mistletoe, Ligaria cuneifolia (R. et P.) Tiegh (Loranthaceae) and screening of their polyphenolic content

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Abstract

Ligaria cuneifolia (R. et P.) Tiegh (Loranthaceae), known as liga, muérdago criollo, or Argentinean mistletoe, is a hemiparasitic plant with a broad distribution in central and northern Argentina. Pharmacological studies showed that L. cuneifolia extracts have hypolipemic, antioxidant, antibacterial, and immunomodulatory effects. We have established callus cultures from embryo and haustoria fragments. The highest frequency of callus formation from embryos (85%) was obtained on White medium with 4% (w/v) sucrose and 2.5 µM 1-naphtalene acetic acid and 9.2 µM kinetin as plant growth regulators (PGRs). From haustoria, the best result (35%) was obtained on Gamborg medium with 3% (w/v) sucrose and 0.45 µM 2,4-dichlorephenoxyacetic acid and 0.47 µM zeatin as PGRs. Thin layer chromatography showed that callus methanolic extract (2.5% w/v) had a lower content of flavonoids and proanthocyanins as compared to the wild plant (5% w/v for leaves, stems, and flowers), but a higher content of hydroxycinnamic acids. High performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS) showed the presence of quercetin glycosides and phenolic acids in the methanolic extracts both from the parent plant and the callus obtained from embryo.

Key message

Callus cultures were established from embryo and haustorium explants of Ligaria cuneifolia. Leaves, stems, and meristems were recalcitrant to in vitro culture. Callus tissues contained quercetin glycosides and phenolic acids.

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Acknowledgements

We wish to thank Dr. Sabrina Flor for her assistance in the mass spectrometry analysis (Pharmaceutical Technology Department of the Faculty of Pharmacy and Biochemistry from the University of Buenos Aires), Dr. Javier Calcagno (CONICET/CEBBAD) for his advice regarding the statistical analysis, Dr. Chana Pilberg (Universidad Maimónides) for kindly providing us from plant material from Merlo, and M. Julian Schecter for his advice and careful revision of English. This work was supported by Fondo Nacional de Ciencia y Tecnología (FONCyT), Ministerio de Ciencia, Tecnología e Innovación Productiva from Argentina (PICT2015-2024), Universidad de Buenos Aires, and Universidad Maimónides. M A Alvarez and M Laguia-Becher are researchers from CONICET, MVR has a scholarship from CONICET-Universidad Maimónides, and MLB has a scholarship from FONCyT.

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Authors and Affiliations

  1. Carreras de Farmacia y Bioquímica, Cátedra de Farmacobotánica y Farmacognosia, Facultad de Ciencias de la Salud, Universidad Maimónides, Hidalgo 775, Ciudad de Buenos Aires, Argentina

    M. V. Ricco, M. L. Bari, F. Bagnato, C. Cornacchioli, L. U. Spairani & M. A. Álvarez

  2. Consejo Nacional de Investigaciones Científicas y Tecnicas, Godoy Cruz 2290, Ciudad de Buenos Aires, Argentina

    M. V. Ricco, M. Laguia-Becher & M. A. Álvarez

  3. Departamento de Farmacología, Cátedra de Farmacobotánica, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, Ciudad de Buenos Aires, Argentina

    C. Dobrecky, R. A. Ricco & M. L. Wagner

  4. Facultad de Turismo y Urbanismo, Universidad Nacional de San Luis, Av del Libertador s/n, Barranca Colorada, Villa de Merlo, San Luis, Argentina

    A. Posadaz

  5. Centro de Estudios Biotecnológicos, Biológicos, Ambientales y Diagnóstico (CEBBAD), Universidad Maimónides, Hidalgo 775, Ciudad de Buenos Aires, Argentina

    M. Laguia-Becher

  6. Instituto Antártico Argentino, Dirección Nacional del Antártico, Av. 25 de Mayo 1143, San Martín, Buenos Aires, Argentina

    L. U. Spairani

  7. Cátedra de Tecnología Farmacéutica I, Facultad de Farmacia y Bioquímica, Universidad de Buenos Aires, Junín 956, Ciudad de Buenos Aires, Argentina

    C. Dobrecky

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  1. M. V. Ricco
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  2. M. L. Bari
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  5. M. Laguia-Becher
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  6. L. U. Spairani
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  7. A. Posadaz
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  8. C. Dobrecky
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  9. R. A. Ricco
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  10. M. L. Wagner
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  11. M. A. Álvarez
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Contributions

MVR and MLB carried out the experiments and participated in drafting the manuscript; CC and FB carried out experiments; ML-B and CD participated in the analysis of the results; AP participated in selecting, collecting, and classifying the plant material; LUS performed statistical analysis; MLW, RAR and MAA initiated the project and supervised the work throughout, MAA also drafted the manuscript. All authors read and approved the final manuscript.

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Correspondence to M. A. Álvarez.

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Communicated by Sergio J. Ochatt.

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11240_2019_1615_MOESM1_ESM.docx

Supplementary material 1 (DOCX 19 kb). On line resource 1: Antioxidant treatments used for avoiding browning in Ligaria cuneifolia leaves in vitro cultures. Explants (n = 10) were washed with the antioxidants during 5 min, 15 min and 30 min. Then, explants were transferred to Petri dishes with White medium plus 2,4-d (4.5 µM).The treatments were performed in dark or light, with similar results in both cases. l-CysHCl: l-Cysteine hydrochloride monohydrate

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Ricco, M.V., Bari, M.L., Bagnato, F. et al. Establishment of callus-cultures of the Argentinean mistletoe, Ligaria cuneifolia (R. et P.) Tiegh (Loranthaceae) and screening of their polyphenolic content. Plant Cell Tiss Organ Cult 138, 167–180 (2019). https://doi.org/10.1007/s11240-019-01615-5

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