Production of phytochemicals by using habituated and long-term cultured cells

Cultured plant cell lines that grew and produced phytochemicals in the absence of exogenous plant hormones were established and then examined for their culture characteristics and the features of their produced phytochemicals. Cell lines that produce the following phytochemicals were established by selection and passage: anthocyanin-producing cell lines of cherry, peach, and Asiatic dayflower (Commelina communis L.); a carotenoid-producing cell line of Asiatic dayflower; a betacyanin-producing cell line of an Amaranthus species; and an isoflavone-producing cell line of soybean. The strains of cherry, peach, and Asiatic dayflower were established by passage in hormone-free media, whereas those of the Amaranthus sp. and soybean required passage with concentrations of auxins and cytokinins that were progressively halved. The anthocyanin-producing cell line of pear required low levels of phytohormones (0.156 mu g l(-1) of 2,4-dichlorophenoxyacetic acid and 0.0156 mu g l(-1) of 6-benzylaminopurine), and no cell line that grew in the absence of exogenous phytohormones could be obtained from pear. The anthocyanins malonylawobanin and flavocommelin of blue Asiatic dayflower petal were not found in extracts from the blue-violet anthocyanin-producing cultured cells. The anthocyanin-producing cherry cell line that requires no phytohormone is amenable to long-term culture over 300 passages. In suspension culture, the strain produces anthocyanin at a higher concentration in modified B5 medium than in Murashige and Skoog medium.