Planta Med 2010; 76 - P029
DOI: 10.1055/s-0030-1264327

In vitro shoot cultures of endemic Cyclopia genistoides (Honeybush) as a source of valuable polyphenolic compounds

M Luczkiewicz 1, A Kokotkiewicz 1, A Hering 2, K Gorynski 3, A Bucinski 3, R Ochocka 1
  • 1Department of Pharmacognosy, Medical University of Gdansk, Pharmacognosy with Medicinal Plant Garden, Al. gen. J. Hallera107, 80–416 Gdansk, Poland
  • 2Department of Biology and Pharmaceutical Botany, Medical University of Gdansk, Biology and Pharmaceutical Botany, Al. gen. J. Hallera 107, 80–416 Gdansk, Poland
  • 3Department of Biopharmacy, Nicolaus Copernicus University in Torun, Collegium Medicum in Bydgoszcz, Biopharmacy, Sklodowskiej-Curie st 9, 85–095 Bydgoszcz, Poland

The plants from Cyclopia genus (Honeybush) are endemic, South-African shrubs characteristic for the fynbos plant formation of the Cape Floristic Region. Aerial parts of several Cyclopia species, including C. genistoides, are used to manufacture the honeybush herbal tea, recognized by its dinstictive sweet, honey-like aroma, low level of tannins and lack of caffeine [1,2]. Extracts obtained from Cyclopia plants are rich in polyphenols, including xanthones (mangiferin and isomangiferin), flavanones (hesperidin), flavones and isoflavones, and as such exhibit substantial antioxidative and antimutagenic activity [1,2,3]. In vitro propagation protocol for C. genistoides has been established. Solid Schenk-Hildebrandt (SH) medium supplemented with 2.0mg l-1 6-(γ,γ-dimethylallylamino)purine (2iP) and 0.22mg l-1 thidiazuron (TDZ) have been shown to be the best for shoot culture initiation, as well as for the development of high number of microshoots. Shoot elongation procedure involved explant cultivation on the full strength SH medium supplemented with 1.0mg l-1 indole-3-butyric acid (IBA), followed by 30-day long growth on SH medium with reduced sucrose level, supplemented with the same amount of IBA. As a result, elongated shoots representing intact plant morphology were obtained. Root induction was achieved on solid SH medium with reduced sucrose and nitrate concentrations, supplemented with 5.0mg l-1 indole-3-acetic acid (IAA) and 50.0mg l-1 citric acid. The regenerated plants were acclimatized in the glasshouse, with the use of peat/gravel/perlite subtrate (1:1:1). Both regenerated plants and in vitro microshoots of C. genistoides were evaluated for the production of selected polyphenolic compounds (LC-ESI-MS and LC-DAD analysis).

Acknowledgements: the study was supported by grant no N N302 041936 from Polish Ministry of Education and Science.

References: 1. Joubert E. et al. (2008)J. Ethnopharmacol. 19:376–412.

2. Kokotkiewicz A., Luczkiewicz M. (2009) Fitoterapia 80:3–11.

3. Joubert E. et al. (2008)J. Agric. Food Chem. 56:954–963.