Vanadium elicitation of Trifolium pratense L. cell culture and possible pathways of produced isoflavones transport across the plasma membrane.

KEY MESSAGE: 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.

Isoflavones Production and Possible Mechanism of Their Exudation in Genista tinctoria L. Suspension Culture after Treatment with Vanadium Compounds.

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.

Effect of Gibberellic Acid on Growing-Point Development of Non-Vernalized Wheat Plants under Long-Day Conditions.

The goal of this study was to determine whether the application of gibberellic acid (GA3) to seeds of common wheat varieties with different vernalization and photoperiod requirements affects the transition from vegetative to generative stage. Three varieties of wheat with different photoperiod sensitivities and vernalization were selected for the experiment-the winter varieties, Mironovskaya and Bezostaya, and the spring variety, Sirael. Seeds were treated with different concentrations of GA3 and plants were grown under long-day conditions with monitoring of their photosynthetic activity (Fv/Fm, Pn, E, gs). We monitored the activity of the photosynthetic apparatus by checking the plants to see if they were growing properly. The phenological stages of the wheat species were checked for indications of a transition from the vegetative to the generative stage. Selected concentrations of GA3 had no effect on the compensation of the vernalization process (transition to the generative phase). Chlorophyll fluorescence was one of the factors for monitoring stress. The variety, Bezostaya, is similar to the spring variety, Sirael, in its trends and values. The growth conditions of Bezostaya and Sirael were not affected by the activity of the photosynthetic apparatus. The development of growing points in winter varieties occurred at the prolonged single ridge stage. The spring variety reached the stage of head emergence after sixty days of growth (changes to the flowering phase did not appear in winter wheat). Application of GA3 to the seeds had no effect on the transition of the growing point to the double-ridge generative stage. The present study highlights the priming effect of GA3 on seeds of common wheat varieties with different vernalization and photoperiod requirements as it affected the transition from vegetative to generative stage.