Abstract
The genus Brachystelma morphologically ranges from slender climbers to geophytes that mainly occur in Australia, Southeast Asia and sub-Saharan Africa. These species are well-known for their nutrition and medicinal values. The current study evaluated the effect of cytokinins (CKs) on the phenolic acid content and antioxidant activity of two in vitro-regenerated Brachystelma species. We quantified the phenolic acids (using Ultra-high performance liquid chromatography-tandem mass spectrometry, UHPLC-MS/MS) and antioxidant (Oxygen radical absorbance capacity, ORAC) potential of two (B. pulchellum and B. pygmaeum) species that were successfully micropropagated, with the use of three cytokinins (CKs): benzyladenine (BA), isopentenyladenine (iP) and meta-topolin riboside (mTR). The presence of CKs strongly influenced the concentrations of the 11 phenolic acids (consisting of seven hydroxybenzoic acids and four hydroxycinnamic acids) and ORAC of the in vitro-regenerated B. pulchellum and B. pygmaeum. Generally, the levels of hydroxycinnamic acids were higher than the hydroxybenzoic acids in both Brachystelma species. Ferulic (9–55 µg/g DW) and sinapic (14–97 µg/g DW) acids were the major phenolic acids in the micro-propagated plants. In both Brachystelma species, the majority of the CK-regenerated plants had higher ORAC relative to the control. The best antioxidant activity (> 70 μmol/g Trolox equivalents, TE) was obtained from 25 μM iP and mTR treatments for B. pulchellum and B. pygmaeum, respectively. Overall, the current study demonstrated the ability of CKs to serve as a suitable tool for the production of valuable therapeutic chemicals in the two Brachystelma species.
Key message
Application of cytokinins (CKs) had a significant effect on the quality and quantity of phenolic acids in in vitro cultured Brachystelma pulchellum and B. pygmaeum. We observed the accumulation of 11 phenolic acids (7 hydroxybenzoic acids and 4 hydroxycinnanmic acids) at varying concentrations. Ferulic and sinapic acids were the most abundant phenolic acids in the in vitro-regenerants and increased in response to the CK treatments. In addition, the presence of CKs successfully enhanced the ORAC of the in vitro-regenerated B. pulchellum.
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Abbreviations
- AAPH:
-
2, 2ʹ-Azobis (2-methylpropionamidine) dihydrochloride
- ANOVA:
-
One-way analysis of variance
- BA:
-
N6-benzyladenine
- CKs:
-
Cytokinins
- DMRT:
-
Duncan’s multiple range test
- DW:
-
Dry weight
- iP:
-
Isopentenyladenine
- mTR:
-
meta-Topolin riboside
- NAUC:
-
Net area under curve
- ORAC:
-
Oxygen radical absorbance capacity
- TE:
-
Trolox equivalents
- UHPLC-MS/MS:
-
Ultra-high performance liquid chromatography-tandem mass spectrometry
- UKZN:
-
University of KwaZulu-Natal
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Acknowledgements
We are grateful to Dr Adam Shuttleworth (UKZN) for providing the stock material used in the current study. The technical assistance rendered by Lucie Slobodianová (Czech Republic) and Mrs A. Young and her staff at UKZN Botanical Garden (South Africa) during the greenhouse stage of the study are greatly appreciated. We thank Mrs Lee Warren for assisting with the editing of the manuscript.
Funding
Financial support provided by the National Research Foundation (NRF), University of KwaZulu-Natal and North-West University is appreciated. This work was also supported by the Ministry of Education, Youth and Sport of the Czech Republic, ERDF project “Development of pre-applied research in nanotechnogy and biotechnology” (No. CZ.02.1.01/0.0/0.0/17_048/0007323). The funder had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.
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Conceptualization, N.P.H. and A.O.A; formal analysis, N.P.H., A.O.A., J.G.; investigation, N.P.H., J.G.; resources, J.V.S., J.F.F.; writing—original draft preparation, N.P.H.; writing—review and editing, A.O.A., J.G., J.V.S. J.F.F.; supervision, J.V.S., J.F.F.; project administration, J.V.S. J.F.F.; funding acquisition, J.G., J.V.S., J.F.F. All authors have read and approved the manuscript for submission.
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Communicated by Ewa Grzebelus.
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Hlophe, N.P., Aremu, A.O., Gruz, J. et al. Influence of different cytokinins on the phenolic acids and antioxidant activity of two Brachystelma species. Plant Cell Tiss Organ Cult 145, 689–699 (2021). https://doi.org/10.1007/s11240-021-02038-x
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DOI: https://doi.org/10.1007/s11240-021-02038-x