Effect of salicylic acid on the gene transcript and protein accumulation of flavonoid biosynthesis-related enzymes in Vitis vinifera cell suspension cultures

Abstract

Salicylic acid (SA) as a plant signal molecule plays an important regulatory role in stimulating the accumulation of plants’ defensive compounds. Numerous studies have shown its regulating effects on the biosynthesis of flavonoids. However, the controlling mechanism needs to be made clear further. Here, Vitis vinifera L. ‘Cabernet Sauvignon’ cell suspension cultures were treated with SA and its synthesis inhibitor paclobutrazol (Pac) to investigate their effects on flavonoid biosynthesis. The accumulation of gene transcripts and proteins of flavonoid biosynthesis pathway enzymes, including chalcone synthase (VvCHS), chalcone isomerase (VvCHI), dihydroflavonol 4-reductase (VvDFR), and anthocyanidin synthase (VvANS), were detected using real-time polymerase chain reaction (RT-PCR), quantitative polymerase chain reaction (qPCR), and westernblotting techniques. The results showed that treatment with exogenous SA at the proper concentration enhanced the production of flavonoids such as anthocyanins and proanthocyanidins (PAs) inside the suspension-cultured cells. The induction of anthocyanins and PAs was found to be time course-dependent. The mRNA and protein accumulation of VvCHS, VvCHI, VvDFR, and VvANS in the treatment system were enhanced too, and the changes were observed concomitantly. Treatment of Pac inhibited the induction of the accumulation of gene transcripts and proteins in accordance with the decreased accumulation of flavonoids. These results suggest that exogenous SA could induce both transcript and protein accumulation of flavonoid biosynthesis-related enzymes and in turn enhance the accumulation of flavonoid compounds such as anthocyanins and PAs in suspension-cultured grape cells.