Effect of 2,5-Dicarbonyl-3-Isobutyl-Piperazine on 3-Isobutyl-2-Methoxypyrazine Biosynthesis in Wine Grape.

The metabolic pathway of 3-alkyl-2-methoxypyrazines (MPs) in grape remains largely unclear except for the final step. In this study, the 2,5-dicarbonyl-3-isobutyl-piperazine (DCIP), which is proposed as the key intermediate of 3-isobutyl-2-methoxypyrazine (IBMP) biosynthesis, was incorporated into Cabernet Sauvignon clusters in situ using a soaking method. The IBMP concentration of grape and the expression patterns of VvOMTs in berry skin were monitored over two consecutive years. The results showed that the IBMP concentration of grape treated with DCIP was significantly increased at maturity in both years. The relative expression levels of VvOMT1 and VvOMT3 in berry skin were positively correlated with the IBMP accumulation. After DCIP incorporation, the relative expression level of VvOMT1 and particularly that of VvOMT3 were obviously up-regulated and closely mirrored the IBMP accumulation pattern in two consecutive years. Therefore, we speculate that DCIP may be a key intermediate involved in the biosynthesis of IBMP and plays an important role in regulating IBMP accumulation.

R2R3-MYB Transcription Factors Regulate Anthocyanin Biosynthesis in Grapevine Vegetative Tissues.

Anthocyanins with important physiological functions mainly accumulate in grape berry, but teinturier grape cultivars can accumulate anthocyanins in both reproductive and vegetative tissues. The molecular regulatory mechanisms of anthocyanin biosynthesis in grapevine reproductive and vegetative tissues are different. Therefore, teinturier grapevine cultivar provides opportunities to investigate transcriptional regulation of vegetative anthocyanins, and to compare with mechanisms that regulate grape berry anthocyanins. Yan73 is a teinturier Vitis vinifera variety with vegetative tissues able to accumulate anthocyanins, but the anthocyanin pattern and the molecular mechanism regulating anthocyanin biosynthesis in these tissues remain uncharacterized. We analyzed the anthocyanin metabolic and transcriptome profiles of the vegetative tissues of Yan73 and its male parent with HPLC-ESI-MS/MS and RNA-sequencing technologies. Yan73 vegetative tissues had relatively high 3'-OH, acylated, and methoxylated anthocyanins. Furthermore, peonidin-3-O-(trans-6-coumaryl)-glucoside is the most abundant anthocyanin in Yan73 grapevine vegetative tissues. A total of 30,17 and 10 anthocyanin biosynthesis genes showed up-regulated expression in Yan73 leaf, stem and tendril, respectively, indicating anthocyanin biosynthesis in Yan73 vegetative tissues is regulated by transcription factors. The up-regulated expression of VvMYBA1 on chromosome 2 and VvMYBA5, VvMYBA6, and VvMYBA7 on chromosome 14 are responsible for the anthocyanin patterns of Yan73 vegetative tissues. The expression of a set of R2R3-MYB C2 repressor genes is activated and may negatively regulate anthocyanin biosynthesis in Yan73 vegetative tissues. These findings enhance our understanding of anthocyanin biosynthesis in grapevine.

Foliar-sprayed manganese sulfate improves flavonoid content in grape berry skin of Cabernet Sauvignon (Vitis vinifera L.) growing on alkaline soil and wine chromatic characteristics.

Flavonoids are key determinants of grape quality and wine color. Grapevines growing in alkaline soil are prone to manganese deficiency, which can decrease the contents of secondary metabolites, including flavonoids. We determined the effects of a foliar Mn treatment (MnSO4·H2O) of Cabernet Sauvignon grapevines (V. vinifera L.) growing in alkaline soil on the flavonoid contents in grape skin, and the quality of wine. The Mn treatments were applied in 2017 and 2018, and tended to increase the grape sugars, berry weight, and the contents of phenolic compounds from veraison until harvest. The Mn treatments increased the amounts of acetylated, methylated, and total anthocyanins, as well as the total flavonol contents in grape berry skin at harvest. The wines prepared from these grapes had a higher color intensity than those prepared from grapes from control vines. Foliar-applied MnSO4·H2O can promote flavonoid biosynthesis in grape berries, and improve the color of wine.

Behavior of 3-isobutyl-2-methoxypyrazine biosynthesis related to proposed precursor and intermediate in wine grape.

Although 3-alkyl-2-methoxypyrazines (MPs) are very important flavor compounds in grape and wine, the metabolic pathway of these compounds has not been fully elucidated, and only a few studies have focused on it to date. In this study, we performed in situ incorporation of the proposed precursor (l-Leu) and the key intermediate (2-amino-4-methylpentanamide; AMPA) of 3-isobutyl-2-methoxypyrazine (IBMP) biosynthesis into Cabernet Sauvignon clusters. The IBMP content and the expression levels of key related genes were monitored. IBMP content decreased shortly after l-Leu or AMPA treatment, but subsequently increased significantly, with raised levels of O-methyltransferase genes (VvOMT1 and VvOMT3) in the berries between veraison and maturation. We speculate that l-Leu may be a precursor of IBMP biosynthesis and that AMPA may be a key intermediate.

Methoxypyrazines biosynthesis and metabolism in grape: A review.

This review summarizes research on the discovery, biosynthesis, accumulation, transport, and metabolism of 3-alkyl-2-methoxypyrazines (MPs) in grape. The MPs are a family of potent volatile compounds distributed throughout biological kingdoms. These compounds impart herbaceous/green/vegetal sensory attributes to certain varieties of wine. Generally, high levels of MPs in wine are derived mainly from the corresponding grapes. Although two pathways for MPs biosynthesis have been proposed, only the final step and the enzymes that catalyze it has been confirmed in grape, and the metabolic intermediates and key enzymes involved in other steps are still unknown. The limited understanding of MPs metabolism has restricted research on these compounds, and some empirical results cannot be explained by the current knowledge of MPs metabolism. This review provides insights into research on MPs biosynthesis and metabolism, and proposes directions for further research on this important class of flavour/odour compounds.