Synthesis of flavonols and expression of flavonol synthase genes in the developing grape berries of Shiraz and Chardonnay (Vitis vinifera L.)

Flavonols were determined in Shiraz and Chardonnay grapes throughout berry development. The predominant flavonols were quercetin-3-glycosides with trace amounts of kaempferol-3-glycosides detected in Shiraz flowers but not in developing berries. Flavonols were present in the skin of ripening grapes but were not detected in seeds or flesh. Flavonols were also present in buds, tendrils, inflorescences, anthers and leaves. The concentration of flavonols in flowers (mg/g fresh weight) was high and decreased between flowering and berry set then remained relatively constant through berry development. The total amount of flavonols in berries (mg/berry) was low until pre-veraison then increased during berry development, particularly before veraison, the onset of ripening, in Chardonnay and during ripening in Shiraz. Two cDNA fragments with homology to genes encoding the enzyme flavonol synthase (FLS) were isolated from Shiraz flowers. In the overlapping region of the two cDNAs, they had 80% sequence identity at the nucleotide level and both had high homology to FLS genes from other plants. VvFLS1 was expressed in leaves, tendrils, pedicels, buds and inflorescences as well as in developing grapes. Expression was highest between flowering and fruit set then declined, increasing again during ripening coincident with the increase in flavonols per berry. Expression of VvFLS2 was much lower than for VvFLS1 and did not change during berry development. The results indicate that two distinct periods of flavonol synthesis occur in grapes, the first around flowering and the second during ripening of the developing berries.