PROBING GRAPEVINE-BOTRYTIS CINEREA INTERACTION THROUGH MASS SPECTROMETRY IMAGING

The interest in indigenous non-Saccharomyces yeast for use in wine production has increased in recent years because they contribute to the complex character of the wine. The aim of this work was to investigate the fermentation products of ten indigenous strains selected from a collection of native yeasts established at the Institute for Adriatic Crops and Karst Reclamation in 2021, previously isolated from Croatian Maraština grapes, belonging to Hypopichia pseudoburtonii, Metschnikowia pulcherrima, Metschnikowia sinensis, Metschnikowia chrysoperlae, Lachancea thermotolerans, Pichia kluyveri, Hanseniaspora uvarum, Hanseniaspora guillermondii, Hanseniaspora pseudoguillermondii, and Starmerella apicola species, and compare it with commercial non-Saccharomyces and Saccharomyces strains.

Infection of grapes (Vitis vinifera) by Botrytis cinerea (grey mould) is a frequent occurrence in vineyards and during prolonged wet and humid conditions can lead to significant detrimental impact on yield and overall quality. Growth of B. cinerea causes oxidisation of phenolic compounds resulting in a loss of colour and formation of a suite of off-flavours and odours in wine made from excessively infected fruit. Apart from wine grapes, developing post-harvest B. cinerea infection in high-value horticultural products during storage, shipment and marketing may cause significant loss in fresh fruits, vegetables and other crops. A rapid and sensitive assessment method to detect, screen and quantify fungal infection would greatly assist viticultural growers and winemakers in determining fruit quality.

Brettanomyces bruxellensis is considered as the main spoilage yeast in oenology. Its presence in red wine leads to off-flavour due to the production of volatile phenols such as 4-vinylphenol, 4-vinylguaiacol, 4-ethylphenol and 4-ethylguaiacol, whose aromatic notes are unpleasant (e.g. animal, leather, horse or pharmaceutical). Beside wine, B. bruxellensis is commonly isolated from beer, kombucha and bioethanol production, where its role can be described as negative or positive. Recent genomic studies unveiled the existence of various populations.

Enzymatic browning (BE) of must is caused by polyphenol oxidases (PPOs), tyrosinase and laccase. Both PPOs can oxidize diphenols such as hydroxycinnamic acids (HA) to quinones, which can later polymerize to form melanins [1], which are responsible of BE in white wines and of oxidasic haze in red wines. SO₂ is the main tool used to protect must from BE thanks to its capacity to inhibit PPOs [2]. However, the current trend in winemaking is to reduce and even eliminate this unfriendly additive. Among the different possible alternatives for protecting must against BE, the inoculation with a selected Metschnikowia pulcherrima MP1 is without any doubt one of the most promising ones.

Oenococcus oeni is the main Lactic Acid Bacteria responsible for malolactic fermentation, converting malic acid into lactic acid and carbon dioxide in wines. Following the alcoholic fermentation, this second fermentation ensures a deacidification and remains essential for the release of aromatic notes and the improvement of microbial stability in many wines. Nevertheless, wine is a harsh environment for microbial growth, especially because of its low pH (between 2.9 and 3.6 depending on the type of wine) and nutrient deficiency. In order to maintain homeostasis and ensure viability, O. oeni possesses different cellular mechanisms including organic acid metabolisms which represent also the major pathway to synthetize energy in wine.