Vanadium compounds increased the content and release of distinct isoflavones in a Trifolium pratense suspension culture. Regarding transport-mechanism inhibitors, the process was mostly facilitated by ABC proteins and vesicular transport. The transport of isoflavones and other secondary metabolites is an important part of metabolism within plants and cultures in vitro regarding their role in defence against various abiotic and biotic stressors. This research focuses on the way how to increase production and exudation of isoflavones by application of chemical elicitor and the basic identification of their transport mechanisms across cell membranes. The release of five isoflavones (genistin, genistein, biochanin A, daidzein, and formononetin) into a nutrient medium was determined in a Trifolium pratense var. DO-8 suspension culture after two vanadium compound treatments and cultivation for 24 and 48 h. The NH4VO3 solution caused a higher concentration of isoflavones in the medium after 24 h. This increased content of secondary metabolites was subsequently suppressed by distinct transport-mechanism inhibitors. The transport of isoflavones in T. pratense was mostly affected by ABC inhibitors from the multidrug-resistance-associated protein subfamily, but the genistein concentration in the medium was lower after treatment with multidrug-resistance protein subfamily inhibitors. Brefeldin A, which blocks vesicular transport, also decreased the concentration of some isoflavones in the nutrient medium.

• Keywords
• Abiotic elicitation, Plasma-membrane transport, Red clover, Secondary metabolites,
• MeSH
• Biological Transport drug effects   MeSH
• Cell Membrane drug effects   metabolism   MeSH
• Isoflavones metabolism   MeSH
• Trifolium drug effects   metabolism   MeSH
• Vanadium pharmacology   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• Isoflavones MeSH
• Vanadium MeSH

The family Fabaceae traditionally serves as a food and herbal remedies source. Certain plants serve for treatment of menopausal symptoms based on a presence of typical secondary metabolites, isoflavones. Beside soybean and clovers, other plants or cultures in vitro can produce these molecules. A cultivation in vitro can be enhanced by elicitation that stimulates metabolites biosynthesis via stress reaction. Vanadium compounds have been already described as potential elicitors, and the aim of this study was to determine the impact of NH₄VO₃ and VOSO₄ solutions on isoflavones production in Genista tinctoria L. cell cultures. The significant increase of isoflavones content, such as genistin, genistein, or formononetin, was measured in a nutrient medium or dry mass after NH₄VO₃ treatment for 24 or 48 h. The possible transport mechanism of isoflavones release as a result of elicitation was further evaluated. An incubation with different transport inhibitors prior to elicitation took effect on isoflavones content in the medium. However, there was a non-ended result for particular metabolites such as genistein and daidzein, where ATP-binding cassette (ABC) or, alternatively, multidrug and toxin extrusion (MATE) proteins can participate. Possible elicitation by some inhibitors was discussed as a result of their pleiotropic effect. Despite this outcome, the determination of the transport mechanism is an important step for identification of the specific transporter.

• Keywords
• Dyer’s Greenweed, elicitation, heavy metals, plasma membrane transport,
• MeSH
• Cell Culture Techniques methods   MeSH
• Genista chemistry   cytology   drug effects   MeSH
• Isoflavones chemistry   MeSH
• Secondary Metabolism drug effects   MeSH
• Vanadium Compounds pharmacology   MeSH
• Vanadates pharmacology   MeSH
• Publication type
• Journal Article MeSH
• Names of Substances
• ammonium metavanadate MeSH Browser
• Isoflavones MeSH
• Vanadium Compounds MeSH
• Vanadates MeSH
• vanadyl sulfate MeSH Browser