Functional characterization of grapevine MLO genes to define their roles in Powdery mildew susceptibility by CRISPR/Cas9 genome editing

Malolactic fermentation (MLF)¹, the decarboxylation of L-malic acid into L-lactic acid, is performed by lactic acid bacteria (LAB). MLF has a deacidifying effect that may compromise freshness or microbiological stability in wines² and can be inhibited by fumaric acid [E297] (FA). In wine, can be added at a maximum allowable dose of 0.6 g/L³. Its inhibition with FA is being studied as an alternative strategy to minimize added doses of SO₂⁴. In addition, wine yeasts are capable of metabolizing and storing small amounts of FA and during alcoholic fermentation (AF).

In the current context of market competition and consumption evolution, it is necessary to produce wines of a genuine typicity. The Terroir represents an unique and irreproducible inheritance that can be valorized through the origin and the sensory characteristics of the wines.

The South Western Cape of South Africa is exposed to strong southerly and south easterly synoptic winds during the growth period of the grapevine. The development of sea breezes in the afternoon is also a phenomenon associated with the ripening period of grapes cultivated in this coastal area. Wind is one of the environmental variables having the greatest spatial variation but the implications of regular exposure to wind for the performance of the grapevine has not yet been determined for vineyards in the South Western Cape. This study was initiated to meet this need.
The study was conducted in a hedge-trellised vineyard of Vitis vinifera L. cv Merlot with north east – south west row direction. Thirty experimental sites, each consisting of 14 vines, were identified as being exposed to wind or sheltered based on hand-held anemometer readings during the 2001/2002 season. Four stationary anemometers were strategically positioned between the thirty sites. Stomatal conductance and leaf temperature were measured with a PP systems porometer. Vegetative and yield measurements were performed during the 2002/2003 season. The t-test of equal variance was used to determine significant differences in measured parameters between exposed and sheltered grapevines.
Stomatal conductance and leaf area were significantly reduced by exposure to wind. This was associated with a significant reduction in the leaf area of primary shoots, related to shorter shoots, but a significant augmentation of secondary shoot leaf number and area. The number of bunches per vine and yield were also reduced for exposed vines. The berry potassium content was significantly increased for exposed grapevines.
This demonstrates that exposure to wind can result in significant within-vineyard, and potentially between-vineyard, variability in grapevine physiology, vegetative growth, yield and berry composition, with implications for wine style and quality.

Wine aroma represents one of the most important quality parameter and it is influenced by various factors (viticulture and vinification techniques, climate or storage conditions etc.). Wines produced from conventionally and ecologically grown grapes of same variety have different chemical composition and aroma profile [1]. Aroma profile of wine can be also influenced by additional treatment of wine, such as concentration of wine by reverse osmosis (RO). Reverse osmosis represents a pressure-driven membrane separation technique that separates the initial wine on the retentate or concentrate that is retained on the membrane, and permeate that passes through it [2]. Wine permeate usually containes water, ethanol, acetic acid and several low molecular weight compounds that can pass through the membrane. This property enables the use of reverse osmosis membranes for wine concentration, partial dealcoholization, acetic acid or aroma correction [3,4].

Most of the issues surrounding trade in wine and spirits focus on the fight against non-tariff measures.