Exogenous ethylene stimulates the long-term expression of genes related to anthocyanin biosynthesis in grape berries

The treatment of grape berries (Vitis vinifera L. cv. Cabernet Sauvignon) with the ethylene-releasing compound, 2-chloroethylphosphonic acid (2-CEPA), at veraison is a method known to enhance grape skin colour. We observed that it produced a 6-fold increase, up to 30 pmol g(-1) FW, of the cluster internal ethylene compared to untreated controls within the 24 h following treatment. This ethylene upsurge was associated with increased levels of chalcone synthase (CHS) and flavanone 3-hydroxylase (F3H) transcripts, which persisted over the following 20 days. Transcript levels of leucoanthocyanidin dioxygenase (LDOX) and UDP glucose-flavonoid 3-O-glucosyl transferase (UFGT) were similarly enhanced by 2-CEPA, although to a lesser extent. The effect on UFGT was confirmed at the protein level by an immunoblot analysis. The transcript accumulation of dihydroflavonol 4-reductase (DFR) was unaffected by 2-CEPA treatment. Examination of the levels of CHS, F3H and UFGT mRNAs in berries during bunch exposure to ethylene, revealed elevated levels of each transcript within the first 6 h of treatment when compared to nonethylene-treated controls. HPLC analyses of berry skin extracts showed that levels of each of the anthocyanins analysed ( delphinidin, cyanidin, petunidin, peonidin and malvidin) increased over the 10 days following the ethylene burst, and decreased thereafter. However, anthocyanin levels at harvest were still higher in ethylene treated grapes than in controls. This data is the first evidence that ethylene triggers gene expression related to anthocyanin synthesis in grapes, and in addition, our results also confirm the existence of other regulatory modes in the anthocyanin biosynthetic pathway.