Brettanomyces bruxellensis wine isolates show high geographical dispersal and long persistence in cellars

Abstract

Brettanomyces bruxellensis is the main wine spoiler yeast all over the world, yet the structure of the populations associated with winemaking remains elusive. In this work, we considered 1411 wine isolates from 21 countries that were genotyped using twelve microsatellite markers. We confirmed that B. bruxellensis isolates from wine environments show high genetic diversity, with 58 and 42% of putative triploid and diploid individuals respectively distributed in 5 main genetic groups. The distribution in the genetic groups varied greatly depending on the country and/or the wine-producing region. However, the two possible triploid wine groups showing sulfite resistance/tolerance were identified in almost all regions/countries. Genetically identical isolates were also identified. The analysis of these clone groups revealed that a given genotype could be isolated repeatedly in the same winery over decades, demonstrating unsuspected persistence ability. Besides cellar residency, a great geographic dispersal was also evidenced, with some genotypes isolated in wines from different continents. Finally, the study of old isolates and/or isolates from old vintages revealed that only the diploid groups were identified prior 1990 vintages. The putative triploid groups were identified in subsequent vintages, and their proportion has increased steadily these last decades, suggesting adaptation to winemaking practices such as sulfite use. A possible evolutionary scenario explaining these results is discussed.

Assessing the Biofilm Formation Capacity of the Wine Spoilage Yeast Brettanomyces bruxellensis through FTIR Spectroscopy

“…The remarkable adaptation capacity of B. bruxellensis is reflected via its genomic plasticity and polymorphisms, which consequently lead to expanded strain variability, while inevitably categorizing the strains vis a vis their spoilage potential [ 25 , 56 ]. Nevertheless, the favorable growth environment of the wine cellar promotes the invasion and surface adhesion of microorganisms [ 48 ] and at the same time shapes the genetic partners of the well adapted species [ 57 , 58 ]. A similar scenario has been proposed for Saccharomyces cerevisiae , where essential genes for adherence (FLO genes) have been maintained in wild lineages, indicating that biofilm formation is important for yeast survival in the wild [ 59 , 60 , 61 ].…”

+Brettanomyces bruxellensis Displays Variable Susceptibility to Chitosan Treatment in Wine

“…Thirteen were diploid and belong to the CBS2499 genetic group, 13 were triploid and belong to the genetic group AWRI1499, and 14 were triploid and belong to the genetic group AWRI1608. These three groups are the most frequently encountered in wine (Cibrario et al, 2019). The other 13 strains were distributed into the L14165, L0308, and CBS5512 genetic clusters.…”

“…B. bruxellensis gathers triploid and diploid strains distributed in at least six main genetic groups ( Avramova et al, 2018a ; Gounot et al, 2020 ). Wine isolates are also highly diverse and belong to 5 of these genetic groups ( Cibrario et al, 2019 ). This genetic diversity may thus support distinct sensitivities to chitosan as previously described toward sulfites ( Curtin et al, 2012 ; Avramova et al, 2018a , b ).…”

Prediction of Genetic Groups within Brettanomyces bruxellensis through Cell Morphology Using a Deep Learning Tool

“…The spoilage yeast Brettanomyces bruxellensis presents many strain dependent characteristics, such as volatile phenol production that contributes to the famous "Brett character" [1][2][3], or capacities to withstand many stresses associated with wine related environments (nutritional requirements, resistance to low pH values, capacity to enter in Viable But Not Cultivable state and SO 2 resistance) [4][5][6][7][8]. In addition to this ability of B. bruxellensis to persist in wine [7,[9][10][11] and in cellars [12,13], the evolution of oenological practices, such as reducing oenological inputs (like lower SO 2 doses) and the impact of climate change on the physicochemical characteristics of wines (like higher pH levels), make the control of B. bruxellensis more challenging [14,15] and, therefore, leads to serious financial losses for winemakers [16]. Consequently, it is important to develop tools to further discriminate from the species level toward the strain level, and therefore, potentially, to predict spoilage-related phenotypes.…”

“…Recently, microsatellite analysis was applied for intraspecific discrimination among B. bruxellensis species [21,28]. This technique became a highly discriminating, robust and reproducible method adapted to the study of large populations, discriminating strains in different genetic groups and providing information in particular about their ploidy state and SO 2 resistance [5,12,29,30]. Thus, the determination of the genetic group of a given isolate could allow for predicting the risk of SO 2 resistance and, thus, help winemakers to adapt their antimicrobial techniques [5].…”