Fast, and full microbiological wine analysis using triple cellular staining.
Abstract
We propose here a brand new large routine microbiological analysis method intended for oenology, in flow cytometry, using high performance equipment and triple selective cell staining, activated by fluorescence. The results and practical applications of the method are presented: Brettanomyces (Dekkera) Monitoring, fermentations monitoring, bottling and enological practices monitoring.The method allow a complete new microbiological tool for wine industry.The method has been accredited ISO 17025 in our laboratories.
DOI:
Issue: IVAS 2022
Type: Poster
Authors
Dubernet Matthieu1, Husset Mélanie1, Adler Sophie1, Lefebvre Cyril1, Guichard Perrine1, Hernandez Fanny1 and Paricaud Tatiana1
1Laboratoires Dubernet
Contact the author
Keywords
Wine microbiology, cytometry, brettanomyces, saccharomyces, bacteria
Tags
Citation
Related articles…
Exploring resilience and competitiveness of wine estates in Languedoc-Roussillon in the recent past: a multi-level perspective
Climate change projections to support the transition to climate-smart viticulture
The Earth’s system is undergoing major changes through a wide range of spatial and temporal scales as a response to growing anthropogenic radiative forcing, which is pushing the whole system far beyond its natural variability. Sources of greenhouse gases largely exceed their sinks, thus leading to a strengthened greenhouse effect. More energy is thereby being supplied to the system, with inevitable shifts in climatic patterns and weather regimes. Over the last decades, these modifications have been manifested in the full statistical distributions of the atmospheric variables, with dramatic changes in the frequency and intensity of extremes. Natural hazards, such as severe droughts, floods, forest fires, or heatwaves, are being triggered by extreme atmospheric events worldwide, thus threatening human activities. Viticultculture is not only exposed to changing climates but is also highly vulnerable, as grapevine phenology and physiological development are strongly controlled by atmospheric conditions. Therefore, the assessment of climate change projections for a given region is critical for climate change adaptation and risk reduction in viticulture. By adopting timely and suitable measures, the future sustainability and resiliency of the sector can be fostered. Climate-grapevine chain modelling is an essential tool for better planning and management. However, the accuracy of the resulting projections is limited by many uncertainties that must be duly taken into account when transferring knowledge to stakeholders and decision-makers. Climate-smart viticulture will comprise ensembles of locally tuned strategies, envisioning both adaptation and mitigation, assisted by emerging technologies and decision-support systems.
Sustaining wine identity through intra-varietal diversification
With contemporary climate change, cultivated Vitis vinifera L. is at risk as climate is a critical component in defining ecologically fitted plant materiel. While winegrowers can draw on the rich diversity among grapevine varieties to limit expected impacts (Morales-Castilla et al., 2020), replacing a signature variety that has created a sense of local distinctiveness may lead to several challenges. In order to sustain wine identity in uncertain climate outcomes, the study of intra-varietal diversity is important to reflect the adaptive and evolutionary potential of current cultivated varieties. The aim of this ongoing study is to understand to what extent can intra-varietal diversity be a climate change adaptation solution. With a focus on early (Sauvignon blanc, Riesling, Grolleau, Pinot noir) to moderate late (Chenin, Petit Verdot, Cabernet franc) ripening varieties, data was collected for flowering and veraison for the various studied accessions (from conservatory plots) and clones. For these phenological growing stages, heat requirements were established using nearby weather stations (adapted from the GFV model, Parker et al., 2013) and model performances were verified. Climate change projections were then integrated to predict the future behaviour of the intra-varietal diversity. Study findings highlight the strong phenotypic diversity of studied varieties and the importance of diversification to enhance climate change resilience. While model performances may require improvements, this study is the first step towards quantifying heat requirements of different clones and how they can provide adaptation solutions for winegrowers to sustain local wine identity in a global changing climate. As genetic diversity is an ongoing process through point mutations and epigenetic adaptations, perspective work is to explore clonal data from a wide variety of geographic locations.
Phenological characterization of a wide range of Vitis Vinifera varieties
In order to study the impact of climate change on Bordeaux grape varieties and to assess the adaptation capacities of candidates to the grape varieties of this wine region to the new climatic conditions, an experimental block design composed of 52 grape varieties was set up in 2009 at the INRAE Bordeaux Aquitaine center. Among the many parameters studied, the three main phenological stages of the vine (budburst, flowering and veraison) have been closely monitored since 2012. Observations for each year, stage and variety were carried out on four independent replicates. Precocity indices have been calculated from the data obtained over the 2012-2021 period (Barbeau et al. 1998). This work allowed to group the phenological behaviour of the grapevine varieties, not only based on the timing of the subsequent developmental stages, but also on the overall precocity of the cycle and the total length of the cycle between budburst and veraison. Results regarding the variability observed among the different grape varieties for these phenological stages are presented as heat maps.
Influence of grapevine rootstock/scion combination on rhizosphere and root endophytic microbiomes
Soil is a reservoir of microorganisms playing important roles in biogeochemical cycles and interacting with plants whether in the rhizosphere or in the root endosphere. The composition of the microbial communities thus impacts the plant health. Rhizodeposits (such as sugar, organic and amino acids, secondary metabolites, dead root cells …) are released by the roots and influence the communities of rhizospheric microorganisms, acting as signaling compounds or carbon sources for microbes. The composition of root exudates varies depending on several factors including genotypes. As most of the cultivated grapevines worldwide are grafted plants, the aim of this study was to explore the influence of rootstock and scion genotypes on the microbial communities of the rhizosphere and the root endosphere. The work was conducted in the GreffAdapt plot (55 rootstocks x 5 scions), in which the 275 combinations have been planted into 3 blocks designed according to the soil resistivity. Samples of roots and rhizosphere of 10 scion x rootstock combinations were first collected in May among the blocks 2 and 3. The quantities of bacteria, fungi and archaea have been assessed in the rhizosphere by quantitative PCR, and by cultivable methods for bacteria and fungi. The communities of bacteria, fungi and arbuscular mycorrhizal fungi (AMF) was analyzed by Illumina sequencing of 16S rRNA gene, ITS and 28S rRNA gene, respectively. The level of mycorrhization was also evaluated using black ink coloration of newly formed roots harvested in October. The level of bacteria, fungi and archaea was dependent on rootstock and scion genotypes. A block effect was observed, suggesting that the soil characteristics strongly influenced the microorganisms from the rhizosphere and root endosphere. High-throughput sequencing of the different target genes showed different communities of bacteria, fungi and AMF associated with the scion x rootstock combinations. Finally, all the combinations were naturally mycorrhized. The root mycorrhization intensity was influenced by the rootstock genotype, but not by the scion one. Altogether, these results suggest that both rootstock and scion genotypes influence the rhizosphere and root endophytic microbiomes. It would be interesting to analyze the biochemical composition of the rhizodeposition of these genotypes for a better understanding of the processes involved in the modulation of these microbiomes. Moreover, crossing our data with the plant agronomic characteristics could provide insights into their roles on plant fitness.