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…

Postveraison shoot trimming in Tannat and Merlot: preliminary results on yield components, plant balance and berry composition

There is currently a trend towards the production of wines with low alcohol content. To achieve this, grapes with low sugar content must be used. There are techniques at the vineyard level that can delay ripening and avoid excessive sugar accumulation without, a priori, affecting the final polyphenol content. Postveraison shoot trimming (PVST) is experimentally evaluated for these purposes, but its impact under Uruguayan climatic conditions with high interannual variability is not known. The aim of this work is to assess the PVST in Tannat and Merlot cultivars and their impact on yield components, plant balance and berry primary composition. In this study, two commercial vineyards of 10 years old Tannat and Merlot (grafted on SO4) at Canelones Department were selected. During the 2020-201 growing season, grapevines were submitted to PVST when grapes reached 15º Brix. In a randomized block, trimmed (T) and control (C) plants were evaluated with three repetitions each cultivar. Evaluation of the evolution of primary berry composition during ripening, measurement of yield components and plant balance were performed. For both cultivars, PVST did not affect yield components. Merlot reached 5.4 kg per plant and Tannat 7.1 kg, with not statistical significance between treatments. However, statistical differences were observed in terms of plant balance. In Merlot Ravaz Index reached a difference of 5.3 (12.0 in T and 6.7 in C) meanwhile Tannat reached 3.5 of statistical difference (13.7 in T and 10.2 in C). The tendency to imbalance for the treated plants had an impact on the final grape composition. Merlot grapes showed statistical difference in final total acidity (0.3 g of difference between treatments) while treatments impact final sugar content on Tannat grapes (10.0 g of difference between treatments). Further studies are needed to assess the impact of different canopy management techniques in our conditions.

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.

Climate ethnography and wine environmental futures

Globalisation and climate change have radically transformed world wine production upsetting the established order of wine ecologies. Ecological risks and the future of traditional agricultural systems are widely debated in anthropology, but very little is understood of the particular challenges posed by climate change to viticulture which is seen by many as the canary in the coalmine of global agriculture. Moreover, wine as a globalised embedded commodity provides a particularly telling example for the study of climate change having already attracted early scientific attention. Studies of climate change in viticulture have focused primarily on the production of systematic models of adaptation and vulnerability, while the human and cultural factors, which are key to adaptation and sustainable futures, are largely missing. Climate experts have been unanimous in recognising the urgent need for a better understanding of the complex dynamics that shape how climate change is experienced and responded to by human systems. Yet this call has not yet been addressed. Climate ethnography, coined by the anthropologist Susan Crate (2011), aims to bridge this growing disjuncture between climate science and everyday life through the exploration of the social meaning of climate change. It seeks to investigate the confrontation of its social salience in different locations and under different environmental guises (Goodman 2018: 340). By understanding how wine producers make sense of the world (and the environment) and act in it, it proposes to focus on the co-production of interdisciplinary knowledge by identifying and foreshadowing problems (Goodman 2018: 342; Goodman & Marshall 2018). It seeks to offer an original, transformative and contrasted perspective to climate change scenarios by investigating human agency -individual or collective- in all its social, political and cultural diversity. An anthropological approach founded on detailed ethnographies of wine production is ideally placed to address economic, social and cultural disruptions caused by the emergence of these new environmental challenges. Indeed, the community of experts in environmental change have recently called for research that will encompass the human dimension and for more broad-based, integrated through interdisciplinarity, useful knowledge (Castree & al 2014). My paper seeks to engage with climate ethnography and discuss what it brings to the study of wine environmental futures while exploring the limitations of the anthropological environmental approach.

Impact on leaf morphology of Vitis vinifera L. cvs Riesling and Cabernet Sauvignon under Free Air Carbon dioxide Enrichment (FACE)

Atmospheric carbon dioxide (CO2) concentration has continuously increased since pre-industrial times from 280 ppm in 1750, and is predicted to exceed 700 ppm by the end of 21st century. For most of C3 plant species elevated CO2 (eCO2) improve photosynthetic apparatus results in an increased plant biomass production. To investigate the effects of eCO2 on morphological leaf characteristics the two Vitis vinifera L. cultivars, Riesling and Cabernet Sauvignon, grown in the Geisenheim VineyardFACE (Free Air Carbon dioxide Enrichment) system were used. The FACE site is located at Geisenheim University (49° 59′ N, 7° 57′ E, 94 m above sea level), Germany and was implemented in 2014 comparing future atmospheric CO2-concentrations (eCO2, predicted for the mid-21st century) with current ambient CO2-conditions (aCO2). Experiments were conducted under rain-fed conditions for two consecutive years (2015 and 2016). Six leaves per repetition of the CO2 treatment were sampled in the field and immediately fixed in a FAA solution (ethanol, H2O, formaldehyde and glacial acetic acid). After 24 h leaf samples were transferred and stored in an ethanol solution. Subsequently, leaf tissue was dehydrated using ethanol series and embedded in paraffin. By using a rotary microtomesections of 5 µm were prepared and fixed on microscopic slides. Subsequent the samples were stained using consecutive staining and washing solutions. Afterwards pictures of the leaf cross-sections were taken using a light microscope and consecutive measurements were conducted with an open source image software. Differences found in leaf cross-sections of the two CO2 treatments were detected for the palisade parenchyma. Leaf thickness, upper and lower epidermis and spongy parenchyma remained less affected under eCO2 conditions. The observed results within grapevine leaf tissues can provide first insights to seasonal adaptation strategies of grapevines under future elevated CO2 concentrations.

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.