Macrowine 2021
IVES 9 IVES Conference Series 9 Characterization of Brettanomyces bruxellensis biofilm, a resistance strategy to persist in wine-related environments

Characterization of Brettanomyces bruxellensis biofilm, a resistance strategy to persist in wine-related environments

Abstract

AIM: Biofilm is a resistance mechanism deployed by microorganisms to adapt to stresses, leading to their persistence in the environment. In the case of Brettanomyces bruxellensis, a wine spoilage yeast, knowledge about its capacity to form biofilm remains limited although this potential strategy could explain its recurring presence in cellars. In this study, we propose to investigate the capacity of strains of B. bruxellensis to form biofilm according to different conditions and to characterize its structure.

METHODS: Sixty-five isolates of B. bruxellensis were sampled from a cellar and discriminated into genetic groups by microsatellite analysis. The capacity of selected strains to adhere and form biofilm has been investigated in different conditions of media and supports, related to wine environment. The structure of the biofilm and its components were explored using several microscopic techniques as confocal laser scanning microscopy, electronic microscopy and epifluorescence microscopy.

RESULTS: Twelve strains among the 65 isolates were selected and have showed the capacity to form biofilm on polystyrene surface. Microscopic observations of the biofilm revealed microcolonies, filamentous cells and extracellular polymeric substances despite a small thickness. Then, the study of the impact of wine on B. bruxellensis biofilm revealed biofilm cell released and growth of these released biofilm cells, probable contamination source of the wine. Finally, for both planktonic and biofilm lifestyles in wine, a new chlamydospore-like element was observed for B. bruxellensis, described as a resistance form in other fungi.

CONCLUSIONS: This study validates the capacity of B. bruxellensis to form biofilm and highlights structural element of this mode of life. Moreover, an additional resistance strategy was suggested through the description of the chlamydospore-like elements. New insights into the persistence of B. bruxellensis during the winemaking process and in wineries have emerged.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Manon Lebleux, Hany ABDO, Louise BASMACIYAN, Julie LAURENT, Chloé ROULLIER-GALL, Hervé ALEXANDRE, Michèle GUILLOUX-BENATIER, Stéphanie WEIDMANN, Sandrine ROUSSEAUX

Univ. Bourgogne Franche-Comté, AgroSup Dijon, PAM UMR A 02.102, VAlMiS Laboratory-IUVV, Dijon, France

Contact the author

Keywords

Brettanomyces bruxellensis, biofilm, wine spoilage

Citation

Related articles…

The concept of terroir: what place for microbiota?

Microbes play key roles on crop nutrient availability via biogeochemical cycles, rhizosphere interactions with roots as well as on plant growth and health. Recent advances in technologies, such as High Throughput Sequencing Techniques, allowed to gain deeper insight on the structure of bacterial and fungal communities associated with soil, rhizosphere and plant phyllosphere. Over the past 10 years, numerous scientific studies have been carried out on the microbial component of the vineyard. Whether the soil or grape compartments have been taken into account, many studies agree on the evidence of regional delineations of microbial communities, that may contribute to regional wine characteristics and typicity. Some authors proposed the term “microbial terroir” including “yeast terroir” for grapes to describe the connection between microbial biogeography and regional wine characteristics. Many factors are involved in terroir including climate, soil, cultivar and human practices as well as their interactions. Studies considering “microbial terroir” greatly contributed to improve our knowledge on factors that shape the vineyard microbial structure and diversity. However, the potential impact of “microbial terroir” on wine composition has yet not received strong scientific evidence and many questions remain to be addressed, related to the functional characterization of the microbial community and its impact on plant physiology and grape composition, the origins and interannual stability of vineyard microbiota, as well as their impact on wine sensorial attributes. The presentation will give an overview on the role of microbiota as a terroir component and will highlight future perspectives and challenges on this key subject for the wine industry.

Modeling the suitability of Pinot Noir in Oregon’s Willamette Valley in a changing climate

Air temperature is the key driver of grapevine phenology and a significant environmental factor impacting yield and quality for a winegrape growing region. In this study the optimal downscaled CMIP5 ensemble for computing thegrowing season average temperature (GST) viticulture climate classification index was determined to spatially compute on a decadal basis predictions of the GST climate index and the grapevine sugar ripeness (GSR) model for Pinot Noir throughout the Willamette Valley (WV) American Viticultural Area (AVA). Forecasts for average temperature and a 220 g/L target sugar concentration level were computed using daily Localized Constructed Analogs (LOCA) downscaled CMIP5 historic and Representative Concentration Pathways (RCP) future climate projections of minimum and maximum daily temperature. We explore spatiotemporal trends of the GST climate classification index and Pinot Noir specific applications of the GSR phenology model for the WV AVA. Spatiotemporal computations of the GST climate index and Pinot Noir specific applications of the GSR model enable the opportunity to explore relationships between their computed values with one intent being to provide updated GST ranges that better align with current temperature-based modeling understanding of Pinot Noir grapevine phenology and the viticultural application of LOCA CMIP5 climate projections for the WV AVA. The Pinot Noir specific applications of the GSR model or the GST index with updated bounds indicate that the percent of the WV AVA area suitable for Pinot Noir production is currently at or near its peak value in the upper 80s to lower 90s of this century.

How can historical cultivars mitigate the effects of climate change?

IFV, INRAe and the national network “Partenaires de la Sélection Vigne” representing 37 organizations from the different wine regions, have been working increasingly closely over the last 2 decades towards the preservation of the French varietal patrimony. There are approximately 600 patrimonial varieties according to INRAe and SupAgro Montpellier experts, including ancient cultivars (400) and intravarietal crossbreeds obtained since the 19th century. In the context of a drastic reduction in such varieties from the mid 1980’s in favor of mainstream varieties, it was essential to carry out an inventory of old vines and vineyards. INRAe Vassal collection plays a key role here as it holds the largest diversity available, along with a rich bibliography and herbariums, offering us the opportunity to document and double check the identity of a cultivar, consolidating the expertise of ampelographers. The work is carried out in several stages, from verifying the existence of a variety in a small region, through to rehabilitation. During this session, the authors present the process that leads to the official registration of a variety. After this, IFV selection center takes over to initiate the process of selection and propagation. A specific focus within regions such as the Alps, Champagne and the South-West will provide details of the full procedure. Bia, Bouysselet, Chardonnay rose, Mecle and the aptly named Tardif, are some of the cultivars that have followed this procedure. Furthermore, a recent regulation established by INAO on “varieties of interest for adaptation purposes” might boost uptake by growers. Since 2006, 36 historical cultivars have been registered. Most of these have been neglected in the past due to late maturity, lack of sugar and high titratable acidity at harvest time. Such characteristics are today considered as positive qualities, not only in mitigation of the effects of climate change, but also as an opportunity for restoring diversity…

Better understand the soil wet bulb formation with subsurface or aerial drip irrigation in viticulture

The gradual change in rainfall patterns experienced in the south of France vineyards, especially around the Mediterranean sea, means that the vines are increasingly subject to summer drought. The winegrowers developped the use of irrigation techniques to ensure the maintenance of competitive yields in the production of wines under Protected Geographical Indication label. In practice, drip irrigation pipes can be installed above the ground or buried into the soil as well as at different distances from the vine row. The objective of this study was to examine the profiles of the wet bulbs of the soil obtained from two drip irrigation systems : aerial drip located under the vine row and subsurface drip placed in the middle of the inter-row. This experiment took place over two consecutive seasons (2020-2021) on a 3.4 ha Viognier plot in the Mediterranean region (PGI Oc, France) on sandy clay soil. The annual rainfalls were less than 400 mm. Soil water content probes were installed at different depths (20 – 40 – 60 – 80 cm) and at different lateralities from the vine row (30 – 60 – 90 – 120 cm) to control the formation of the soil wet bulb during irrigation. The mapping and the analysis of the data allowed a better understanding and differentiation of the water percolation when irrigating with subsurface or aerial drip. For the same amount of water and without differences of vine water status, it is shown that in a subsurface drip irrigation situation, the size of the wet bulb formed is larger than in aerial drip irrigation system.

Adaptability of grapevines to climate change: characterization of phenology and sugar accumulation of 50 varieties, under hot climate conditions

Climate is the major factor influencing the dynamics of the vegetative cycle and can determine the timing of phenological periods. Knowledge of the phenology of varieties, their chronological duration, and thermal requirements, allows not only for the better management of interventions in the vineyard, but also to predict the varieties’ behaviour in a scenario of climate change, giving the wine producer the possibility of selecting the grape varieties that are best adapted to the climatic conditions of a certain terroir. In 2014, Symington Family Estates, Vinhos, established two grape variety libraries in two different places with distinctive climate conditions (Douro Superior, and Cima Corgo), with the commitment of contributing to a deeper agronomic and oenological understanding of some grape varieties, in hot climate conditions. In these research vineyards are represented local varieties that are important in the regional and national viticulture, but also others that have over time been forgotten — as well as five international reference cultivars. From 2017 to 2021, phenological observations have been made three times a week, following a defined protocol, to determine the average dates of budbreak, flowering and veraison. With the climate data of each location, the thermal requirements of each variety and the chronological duration of each phase have been calculated. During maturation, berry samples have been gathered weekly to study the dynamics of sugar accumulation, between other parameters. The data was analysed applying phenological and sugar accumulation models available in literature. The results obtained show significant differences between the varieties over several parameters, from the chronological duration and thermal requirements to complete the various stages of development, to the differences between the two locations, confirming the influence of the climate on phenology and the stages of maturation, in these specific conditions.