EFFECTS OF SUPPLIED CINNAMIC-ACIDS AND BIOSYNTHETIC INTERMEDIATES ON THE ANTHOCYANINS ACCUMULATED BY WILD CARROT SUSPENSION-CULTURES

Anthocyanins isolated and characterized from the wild carrot suspensio n cultures used here were yranosyl-(1-->2)-]beta-D<-galactopyranosylcy anidin (1), pyranosyl-(1-->2)-beta-D-galactopyranosyl]cyanidin (2), et a-D-xylopyranosyl-(1-->2)-]beta-D-galactopyranos ylcyanidin (3), pyran osyl-(1-->2)-]beta-D-galactopyranosylcyanidin (4), beta-D-xylopyranosy l-(1-->2)-]beta-D-galactopyrano sylcyanidin (5), pyranosyl-(1-->2)-]be ta-D-galactopyranosylcyanidin (6), anosyl-(1-->6)-[beta-D-xylopyranosy l-(1-->2)-]beta D-galactopyranosylcyanidin (7), pyranosyl-(1-->6)-beta -D-galactopyranosyl]cyanidin (8), and 3-O-(beta-D-galactopyranosyl)cya nidin (9). Except when cinnamic acids were provided in the culture med ium, the major anthocyanin present in the two clones examined was 2. W hen the naturally occurring and some non-naturally occurring cinnamic acids were provided individually in the medium, 1 and 2 were minor com ponents and the anthocyanin acylated with the supplied cinnamic acid, namely 3, 4, 5, 6, or 7 was the major anthocyanin present in the tissu e. When caffeic acid was provided the major anthocyanin in the tissue was 4, thereby suggesting that the caffeic acid was methylated before its use in anthocyanin biosynthesis. Other cinnamic acids supplied had limited effects on the anthocyanins accumulated and appeared not to r esult in the accumulation of new anthocyanins by the tissue. Thus the tissue can use some but not all analogues of sinapic acid to acylate a nthocyanins. Additional anthocyanins were detected in extracts of the wild carrot tissue cultures using mass spectrometry (both MSIMS and HP LC/MS). The additional compounds detected have also been found in cult ures of black carrot, an Afghan cultivar of Daucus carota ssp. sativa and the flowers of wild carrot giving no evidence for qualitative diff erences in the anthocyanins synthesized by subspecies, cell cultures f rom subspecies, or clones from cell cultures. There are major differen ces in the amounts of individual anthocyanins found in cultures from d ifferent subspecies and in different clones from cell cultures. Here a nthocyanins without acyl groups were usually found in the tissues and their accumulation is discussed. On the basis of the structures of the isolated anthocyanins, a likely pathway from cyanidin to the accumula ted anthocyanins is proposed and discussed.