Terroir 2004 banner
IVES 9 IVES Conference Series 9 The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

The use of epifluorescence versus plating to monitor the effect of different parameters on microorganisms in wine

Abstract

The monitoring of the number of micro-orgranisms in wine is crucial for the wine producer. Traditional counting methods include microscopic enumeration and plating on selective media, which measures the culturability of the cells. The use of epifluorescence microscopy is, however, a method, which can measure both culturability and viability in wine. This method distinguishes between live and dead cells. Research showed that little difference existed between plating and epifluorescence numbers to enumerate lactic acid bacteria in wine. However, a difference exists between these two methods to distinguish between acetic acid bacteria numbers in wine. Plating counting numbers were lower than plate numbers for Acetobacter pasteurianus in wine under anaerobic conditions. This difference was, however, negated by the addition of oxygen to the wine. SO2 additions lowered the culturability of A. pasteurianus at dosages higher than 0.35mg/L molecular SO2, but higher dosages were required to lower epifluorescence intensity, which is an indication of viability. Brettanomyces bruxellensis culturability was inhibited at lower dosages, but total cell numbers according to epifluorescence microscopy were affected at higher molecular SO2 dosages. Epifluorescence microscopy and plating also showed that B. bruxellensis was drastically affected after 120 min after molecular SO2 addition and it’s culturability after only 30 min. An exposure time of 5 min to molecular SO2 reduced the cell’s viability drastically and 45 min completely inhibiting the viability after two days. The bonded form of sulphur dioxide did not affect both micro-organisms. Epifluorescence microscopy can thus be used as a quick alternative to assess micro-organisms numbers and culturability in wine. This technique has both advantages and disadvantages over traditional enumeration methods, which will also be discussed.

DOI:

Publication date: January 12, 2022

Issue: Terroir 2004

Type: Article

Authors

W.J. du Toit (1), I.S. Pretorius (1,2) and A. Lonvaud-Funel (3)

(1) Department of Viticulture and Oenology, Institute for Wine Biotechnology, Stellenbosch University, Victoria Street, ZA 7600 Stellenbosch, South Africa
(2)The Australian Wine Research Institute, Waite Road, Urrbrae, SA 5064 Adealide, Australia
(3) Faculté d’Oenologie, 351, Cours de la Libération, 33405 Talence, France

Contact the author

Tags

IVES Conference Series | Terroir 2004

Citation

Related articles…

Frost risk projections in a changing climate are highly sensitive in time and space to frost modelling approaches

Late spring frost is a major challenge for various winegrowing regions across the world, its occurrence often leading to important yield losses and/or plant failure. Despite a significant increase in minimum temperatures worldwide, the spatial and temporal evolution of spring frost risk under a warmer climate remains largely uncertain. Recent projections of spring frost risk for viticulture in Europe throughout the 21st century show that its evolution strongly depends on the model approach used to simulate budburst. Furthermore, the frost damage modelling methods used in these projections are usually not assessed through comparison to field observations and/or frost damage reports.
The present study aims at comparing frost risk projections simulated using six spring frost models based on two approaches: a) models considering a fixed damage threshold after the predicted budburst date (e.g BRIN, Smoothed-Utah, Growing Degree Days, Fenovitis) and b) models considering a dynamic frost sensitivity threshold based on the predicted grapevine winter/spring dehardening process (e.g. Ferguson model). The capability of each model to simulate an actual frost event for the Vitis vinifera cv. Chadonnay B was previously assessed by comparing simulated cold thermal stress to reports of events with frost damage in Chablis, the northernmost winegrowing region of Burgundy. Models exhibited scores of κ > 0.65 when reproducing the frost/non-frost damage years and an accuracy ranging from 0.82 to 0.90.
Spring frost risk projections throughout the 21st century were performed for all winegrowing subregions of Bourgogne-Franche-Comté under two CMIP5 concentration pathways (4.5 and 8.5) using statistically downscaled 8×8 km daily air temperature and humidity of 13 climate models. Contrasting results with region-specific spring frost risk trends were observed. Three out of five models show a decrease in the frequency of frost years across the whole study area while the other two show an increase that is more or less pronounced depending on winegrowing subregion. Our findings indicate that the lack of accuracy in grapevine budburst and dehardening models makes climate projections of spring frost risk highly uncertain for grapevine cultivation regions.

De novo Vitis champinii whole genome assembly allows rootstock-specific identification of potential candidate genes for drought and salt tolerance

Vitis champinii cultivars Ramsey and Dog-ridge are main choices for rootstocks to adapt viticulture in semi-arid and arid regions thanks to their distinctive tolerance to drought and salinity. However, genetic studies on non-vinifera rootstocks have heavily relied on the grapevine (Vitis vinifera) reference genome, which difficulted the assessment of the genetic variation between rootstock species and grapevines. In the present study, this limitation is addressed by introducing a novo phased genome assembly and annotation of Vitis champinii. This new Vitis champinii genome was employed as reference for mapping RNA-seq reads from the same species under drought and salt stresses, and for comparison the same reads were also mapped to the Vitis vinifera PN40024.V4 reference genome. A significant increase in alignment rate was gained when mapping Vitis champinii RNA-seq reads to its own genome, compared to the Vitis vinifera PN40024.V4 reference genome, thus revealing the expression levels of genes specific to Vitis champinii. Moreover, differences in coding sequences were observed in ortholog genes between Vitis champinii and Vitis vinifera, which therefore challenges previous differential expression analyses performed between contrasting Vitis genotypes on the same gene from the Vitis vinifera genome. Genes with possible implications in drought and salt tolerance have been identified across the genome of Vitis champinii, and the same genomic data can potentially guide the discovery of candidate genes specific from Vitis champinii for other traits of interest, therefore becoming a valuable resource for rootstock breeding designs, specially towards increased drought and salinity due to climate change.

VineyardFACE: Investigation of a moderate (+20%) increase of ambient CO2 level on berry ripening dynamics and fruit composition

Climate change and rising atmospheric carbon dioxide concentration is a concern for agriculture, including viticulture. Studies on elevated carbon dioxide have already been on grapevines, mainly taking place in greenhouses using potted plants or using field grown vines under higher CO2 enrichment, i.e. >650 ppm. The VineyardFACE, located at Hochschule Geisenheim University, is an open field Free Air CO2 Enrichment (FACE) experimental set-up designed to study the effects of elevated carbon dioxide using field grown vines (Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon). As the carbon dioxide fumigation started in 2014, the long term effects of elevated carbon dioxide treatment can be investigated on berry ripening parameters and fruit metabolic composition.
The present study aims to investigate the effect on fruit composition under a moderate increase (+20%; eCO2) of carbon dioxide concentration, as predicted for 2050 on both Riesling and Cabernet Sauvignon. Berry composition was determined for primary (sugars, organic acids, amino acids) and secondary metabolites (anthocyanins). Special focus was given on monitoring of berry diameter and ripening rates throughout three growing seasons. Compared to previous results of the early adaptative phase of the vines [1], our results show little effect of eCO2 treatment on primary metabolites composition in berries. However, total anthocyanins concentration in berry skin was lower for eCO2 treatment in 2020, although the ratio between anthocyanins derivatives did not differ.
[1] Wohlfahrt Y., Tittmann S., Schmidt D., Rauhut D., Honermeier B., Stoll M. (2020) The effect of elevated CO2 on berry development and bunch structure of Vitis vinifera L. cvs. Riesling and Cabernet Sauvignon. Applied Science Basel 10: 2486

Aromatic maturity is a cornerstone of terroir expression in red wine

Harvesting grapes at adequate maturity is key to the production of high-quality red wines. Enologists and wine makers define several types of maturity, including technical maturity, phenolic maturity and aromatic maturity. Technical maturity and phenolic maturity are relatively well documented in the scientific literature, while articles on aromatic maturity are scarcer. This is surprising, because aromatic maturity is, without a doubt, the most important of the three in determining wine quality and typicity (including terroir expression). Optimal terroir expression can be obtained when the different types of maturity are reached at the same time, or within a short time frame. This is more likely to occur when the ripening takes place under mild temperatures, neither too cool, nor too hot. Aromatic expression in wine can be driven, from low to high maturity, by green, herbal, fresh fruit, ripe fruit, jammy fruit, candied fruit or cooked fruit aromas. Green and cooked fruit aromas are not desirable in red wines, while the levels of other aromatic compounds contribute to the typicity of the wine in relation to its origin. Wines produced in cool climates, or on cool soils in temperate climates, are likely to express herbal or fresh fruit aromas; while wines produced under warm climates, or on warm soils in temperate climates, may express ripe fruit, jammy fruit or candied fruit aromas. Growers can optimize terroir expression through their choice of grapevine variety. Early ripening varieties perform better in cool climates and late ripening varieties in warm climates. Additionally, maturity can be advanced or delayed by different canopy management practices or training systems.

Effect of one-year cover crop and arbuscular mycorrhiza inocululation in the microbial soil community of a vineyard

The microbial composition of the soil is an important factor to consider in viticulture, since its influence on the “terroir” and on the organoleptic properties of the wine have been demonstrated. Different agronomic techniques have the potential to modify the composition and functionality of the soil microbial community. Maintaining green covers is known to increase soil microbial diversity. The direct application of inoculum of beneficial microorganisms to the soil has also been used to increase their abundance. However, the environmental conditions of each site seem to have a determining weight in the result of these practices. In this study, we compared the effect on the microbial community of a cover crop with legumes in autumn and the inoculation of grapevines with commercial inoculum bases on Rhizophagus irregularis and Funeliformis mosseae in the previous spring. The study has been carried out in a vineyard in Binissalem, Mallorca, Spain. After applying the treatments, we will analyze the soil microbial communities using the data obtained from Illumina amplification of soil DNA from the 16S and ITS regions to analyze bacteria and fungi community, respectively. In addition, we will record the physicochemical characteristics of the soil at each sampling point. The result showed that agronomic management, in the short term, has less influence than soil characteristics on the composition of the soil microbiome. With these results, we can conclude that in a vineyard, agricultural techniques should focus on improving the characteristics of the soil to improve the biodiversity of the soil microbiota.