Posters – Grape and wine microorganisms: diversity and adaptation

Oenococcus oeni is the main lactic acid bacteria (LAB) species which conducts the malolactic fermentation (MLF) in wine. During MLF, O. oeni converts malic acid into lactic acid, which modulates wine aroma composition leading to better balanced organoleptic properties. O. oeni is a highly specialized species only detected in environments containing alcohol such as wine, cider or kombucha. Genome analysis of more than 240 strains showed that they form at least 4 main phylogenetic lineages and several sublineages, which are associated with different beverages or types of wines.

Oenococcus oeni is the predominant lactic acid bacteria species in wine and cider, where it performs the malolactic fermentation (MLF) (Lonvaud-Funel, 1999). The O. oeni strains analyzed to date form four major genetic lineages named phylogroups A, B, C and D (Lorentzen et al., 2019). Most of the strains isolated from wine, cider, or kombucha belong to phylogroups A, B+C, and D, respectively, although B and C strains were also detected in wine (Campbell-Sills et al., 2015; Coton et al., 2017; Lorentzen et al., 2019;

Hanseniaspora vineae (Hv) is a fermenting non-Saccharomyces yeast that compared to Saccharomyces cerevisiae (Sc) present some peculiar features on its metabolism that make it attractive for its use in wine production. Among them, it has been reported a faster yeast lysis and release of polysaccharides, as well as increased ß-glucosidase activity. Hv also produces distinctive aroma compounds, including elevated levels of fermentative compounds such as ß-phenylethyl acetate and norisoprenoids like safranal. However, it is known for its high nutritional requirements, resulting in prolonged and sluggish fermentations, even when complemented with Sc strain and nutrients.

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Wine production is a complex biochemical process that involves a heterogeneous microbiota consisting of different microorganisms such as yeasts, bacteria, and filamentous fungi. Among these microorganisms, yeasts play a predominant role in the chemistry of wine, as they actively participate in alcoholic fermentation, a biochemical process that transforms the sugars in grapes into ethanol and carbon dioxide while producing additional by-products. The quality of the final product is greatly influenced by the microbiota present in the grape berry, and the demand for indigenous yeast starters adapted to specific grape must and reflecting the biodiversity of a particular region is increasing. This supports the concept that indigenous yeast strains can be associated with a "terroir".

Assyrtiko is a rare ancient grape variety that constitutes one of the most popular in Greece. The objective of the current research was to evaluate indigenous Saccharomyces cerevisiae isolates as fermentation starters and also test the possible strain impact on volatile profile of Assyrtiko wine. 163 S. cerevisiae isolates, which were previously selected from spontaneous alcoholic fermentation, were identified at strain level by interdelta-PCR genomic fingerprinting. Yeasts strains were examined for their fermentative capacity in laboratory scale fermentation on pasteurized Assyrtiko grape must.

Climate change leads to even more hostile and stressful for the wine microorganism conditions and consequently issues with fermentation rate progression and off-character formation are frequently observed. The objective of the current research was to classify a great collection of yeast isolates from Greek wines based on their technological properties with oenological interest. Towards this direction, fourteen spontaneously fermented wines from different regions of Greece were collected for further yeast typing. The yeast isolates were subjected in molecular analyses and identification at species level.

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.

Flow cytometry (FCM) is a powerful technique allowing the detection, characterization and quantification of microbial populations in different fields of application (medical environment, food industry, enology, etc.). Depending on the fluorescent markers and specific probes used, FCM provides information on the physiological state of the cell and allows the quantification of a microorganism of interest within a mixed population. For 15 years, the enological sector has shown growing interest in this technique, which is now used to determine the populations present (of interest or spoilage) and the physiological state of microorganisms at the different stages of winemaking.

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Non-Saccharomyces yeasts not only increase microbial diversity during wine fermentation, but also have a positive effect on improving wine aroma. Among these non-Saccharomyces yeast species, Metschnikowia pulcherrima is often studied and used in winemaking in recent years, but its application in icewine has been rarely reported. In this study, indigenous M. pulcherrima strains and Saccharomyces cerevisiae strains (commercial and indigenous strains) were sequentially inoculated for icewine fermentations; meanwhile, pure S. cerevisiae fermentations were used as the control; indigenous strains used above were screened from spontaneous fermentations of Vidal blanc icewine.

Brettanomyces bruxellensis is an ubiquitous yeast associated with different fermentation media such as beer and kombucha, where its presence is beneficial to bring an aromatic typicity. However, it is a main spoilage yeast in wines, in which it produces volatile phenols responsible for organoleptic deviations causing significant economic losses (Chatonnet et al., 1992). Cellar and winery equipment’s are considered as the first source of contamination, during fermentation and wine ageing process (Connel et al., 2002). Indeed, it is possible to find B. bruxellensis in the air, on walls and floors of the cellars, on small materials, vats and barrels.

During alcoholic fermentation, nitrogen is one of essential nutrient for yeast as it plays a key role in sugar transport and biosynthesis of and wine aromatic compounds (thiols, esters, higher alcohols). The main issue of a lack in yeast assimilable nitrogen (YAN) in winemaking is sluggish or stuck fermentations promoting the growth of alteration species and leads to economic losses. Currently, grape musts are often characterized by low YAN concentration and an increase of sugars concentration due to global warming, making alcoholic fermentations even more difficult. YAN depletion can be corrected by addition of inorganic (ammonia) or organic (yeast derivatives products) nitrogen during alcoholic fermentation.

Vitis vinifera L. cv. Pinot noir can be challenging to manage in the winery as its thin skins require careful handling to ensure sufficient extraction of wine colour to promote colour stability during ageing.1 Literature has shown that fermentation with flocculent yeasts can increase red wine colour density.2 As consumers prefer greater colour density in red wines,3 the development of tools to increase colour density would be useful for the wine industry. This research explored the impact of interspecies sequential inoculation and co-flocculation of commercial yeast on Pinot noir wine colour.