Proceedings of OENO IVAS 2019

Grapes from warm(ing) climates often contain excessive sugars but lack acidity. This can lead to highly alcoholic wines with compromised stability and balance. The yeast Lachancea thermotolerans can ameliorate such wines due to its metabolic peculiarity – partial fermentation of sugars to lactic acid. This study aimed to elucidate the population-wide diversity in L. thermotolerans, whilst selecting superior strains for wine sector. An extensive collection of isolates (~200) sourced from different habitats worldwide was first genotyped on 14 microsatellite loci. This revealed differentiation of L. thermotolerans genetic groups based on the isolation substrate and geography. The 94 genotyped strains were then characterised in Vitis vinifera cv. Chardonnay fermentations.

Over the last decade, the use of non-Saccharomyces yeasts in the winemaking process has been re-assessed and accepted by winemakers. These yeasts can be used to achieve specific objectives such as lowering the ethanol content, preventing wine spoilage and increasing the production of specific aroma compounds. Since these species are unable to complete alcoholic fermentation, strategies of co- and sequential inoculation of non-Saccharomyces and Saccharomyces cerevisiae have been developed. However, when mixed starter cultures are used, several parameters (e.g. strain yeast, inoculation timing and nutrient competitions) impact the growth of the individual yeasts, the fermentation kinetics and the metabolites/aroma production. In particular, competition for nitrogen compounds could have a major impact, potentially leading to sluggish fermentation when the yeast assimilable nitrogen (YAN) availability is low. Moreover, many aroma compounds produced by the yeasts are directly produced and influenced by nitrogen metabolism such as higher alcohols, acetate esters and ethyl esters which participate in the organoleptic complexity of wine.

Alcoholic fermentation is the main step for winemaking, mainly performed by the yeast Saccharomyces cerevisiae. But other wine yeasts called non-Saccharomyces may contribute to alcoholic fermentation and modulate the wine aroma complexity. The recurrent problem with the use of these non-Saccharomyces yeasts is their trend to die off prematurely during alcoholic fermentation, leading to a lack of their interesting aromatic properties searched in the desired wine. This phenomenon appears to be mainly due to interactions with S. cerevisiae. These interactions are often negatives but remain unclear because of the species and strain specific response. Among the non-Saccharomyces yeasts, Lachancea thermotolerans is a wine yeast naturally found in grape must and well known as a great L-lactic acid producer and an aromatic molecules enhancer, but its behavior during alcoholic fermentation can be completely different in co-fermentation with S. cerevisiae in function of strain used.

Although there are detailed studies on the microbiota of Vitis vinifera L. grapes, little is known about the diversity of yeast communities present in non-vinifera Vitis ecosystems (i.e., grapes and spontaneously fermenting grape musts). Potentially scientific and/or enological valuable yeast strains from these non-vinifera Vitis ecosystems might never be isolated from V. vinifera L. Using a standard culture-dependent strategy, we studied the population of yeast species during initial stages of spontaneous fermentation of V. labrusca L. (Isabella) grape musts. Rare non-Saccharomyces yeast species were recognized in Isabella, including Candida azymoides, Pichia cecembensis, Candida californica, Candida bentonensis, Issatchenkia hanoiensis and Candida apicola.