Development of a LC-FTMS method to quantify natural sweeteners in red wines

Regenerative agriculture (RA) practices aim to minimize soil disturbance, keep soil covered, maintain living roots underground, and integrate livestock to improve soil health and sustainability.

In the last few decades, minor vine genotypes traditionally cultivated on the Mount Etna slopes, have attracted the interest of both researchers and vine growers, as they offer an interesting oenological profile.

The « Condado de Huelva » Registered Appellation Origin Mark (RAOM) is located in the Province of Huelva, in the southwest of Andalucía (Spain), being limited by the Atlantic Ocean and the Province of Sevilla. « Zalema », a white high productive grapevine plant is its major cultivar. The predominant rootstocks used are « Rupestris du Lot », « Castel 196-17 », « Couderc 161-49 », Couderc 33-09 », « Richter 110 » and « Millardet 41-B ». Traditionally, « Zalema » cv. has been dedicated to the elaboration of amber, bouquet-flavoured wines and in the last years mainly to young, fruit-flavoured white table wines.

Climate change is leading to an increase in average temperature and in the severity and occurrence of heatwaves, and is already disrupting grapevine phenology. In Australia, with the evolution of the weather of grape growing regions that are already warm and hot, berry composition including flavonoids, for which biosynthesis depends on bunch microclimate, are expected to be impacted [1]. These compounds, such as anthocyanins and tannins, contribute substantially to grape and wine quality. The goal of this research was to determine how flavonoid extraction is impacted when bunches are exposed to high (>35 °C) and extreme high (>45 °C) temperatures during berry development and maturity.

Cis-acting regulatory elements are DNA sequences that can be bound by transcription factors to regulate the expression of genes in a condition-dependent and tissue-specific way. It is nowadays possible to search for DNA motives and sequences that a given transcription factor is binding or at least can, but it is still hard to have a glance at all the transcription factors that are contemporaneously located at the same locus. Inspired by an existing technique that uses the CRISPR-Cas system in mammal cells, we are trying to develop a protocol to study such regulation in Vitis vinifera. Using the highly sequence-specific binding capacity of a catalytically inactive Cas9 protein (dCas9), our idea is to set up a system to target a desired sequence and precipitate all the crosslinked proteins and distantly interacting chromatin at this locus and analyze them.