Flavonol Synthase Activity and the Regulation of Flavonol and Anthocyanin Biosynthesis during Flower Development in Dianthus caryophyllus L. (Carnation)

Abstract

Flavonol synthase (FLS) was demonstrated in crude extracts from flower buds of Dianthus caryophyllus (carnation). The enzyme catalyzed the conversion of dihydrokaempferol and ihydroquercetin to kaempferol and quercetin, respectively. The reaction required 2-oxoglutarate, ferrous ion and ascorbate as co-factors and had a pH optimum at about 7.4. The demonstration of FLS activity allowed comparative studies on flavonol and anthocyanin biosynthesis during bud and flower development. Besides FLS the flavonoid enzymes chalcone synthase (CHS), flavanone 3-hydroxylase (FH T) and dihydroflavonol 4-reductase (DFR) were measured. DFR is specifically involved in anthocyanin synthesis, while CHS and FHT provide dihydroflavonol, the common substrate for both FLS and DFR . Maximum expression of CHS, FHT and FLS activity was already observed in small buds, whereas DFR activity started to increase much later and reached its highest level in opened flowers. A substantial correlation was observed between the time courses of FLS and DFR activity and the accumulation of flavonols and anthocyanins, respectively. The competition of FLS and DFR for dihydroflavonols was found to be largely circumvented by different substrate specificities and by the sequential expression of the two enzymes. Both flavonols and anthocyanins are obviously not, or only to some extent, subject to degradation.