Enhanced production of the pharmaceutically important polyphenolic compounds in L. shoot cultures by precursor feeding strategy.

Agitated L. shoot cultures were established to analyse the content of selected pharmaceutically important flavonoids and phenolic acids. Two variants (selected from nine ones) of MS medium were prepared: A (BAP 1 mg/L; NAA 0.5 mg/L; GA 0.25 mg/L) and B (BAP 2 mg/L; NAA 0.5 mg/L). The biomass was harvested after 1, 2, 3,4, 5 and 6 weeks. Four-week cultures (variant A) were selected to perform the precursor feeding experiment. The L-phenylalanine dose of 1.6 g/L appears to be the most advantageous. Compared to the control cultures, the content of the individual compounds increased in a range from 1.4 to 17.3-fold (e.g. -coumaric acid - 17.3 fold; casticin - 4.8-fold). The biomass from in vitro cultures is richer in neochlorogenic acid (16-fold), p-coumaric acid (5.3-fold), rutin (2.8-fold), caffeic acid (1.5-fold) and cinaroside (1.5-fold) than the leaves of its parent greenhouse-cultivated plants. Extracts contained 30 mg/100 g DW of casticin, but after the hydrolysis its amount increased up to 200 mg/100 g DW and twice exceeded the content in the greenhouse leaves. The results indicate that agitated shoot cultures might be considered as a potential biotechnological source of some pharmaceutically important compounds, especially casticin, rutin, neochlorogenic and -coumaric acids.