IVAS 2022 banner
IVES 9 IVES Conference Series 9 IVAS 9 IVAS 2022 9 Profiling the Metschnikowia yeast populations in spontaneous fermentation of Amarone della Valpolicella

Profiling the Metschnikowia yeast populations in spontaneous fermentation of Amarone della Valpolicella

Abstract

The microbial diversity during spontaneous grape must fermentation has a determinant influence on the chemical composition and sensory properties of wine. Therefore, yeast diversity is an important target to better understand wine regionality. Hence, the aim of this study was to isolate, identify, and characterize the yeast core microbiota in grape must during the early stage of lab-scale spontaneous fermentation of withered grapes to produce Amarone della Valpolicella wine (Verona, Italy). At the end of the withering process, chosen grape bunches, mainly of Corvina and Corvinone varieties, were pressed and transferred to glass bottles in the laboratory for spontaneous fermentation (SF). To investigate the impact of grape washing on microbial removal, one batch of grapes was submitted to a washing step in aqueous solution (1% w/v citric acid) at the winery before pressing. Microbial composition was investigated during the first five days of fermentation, a stage known for greater variability of microorganisms, isolating yeast colonies from WL agar plates. Overall, 67 colonies were purified and the partial 26S rRNA gene sequencing allowed the identification of six different species, among which Metschnikowia spp. was prevalent. Indeed, 42 isolates of this genus were obtained, deriving from musts of washed grapes (22) and from non-washed grapes (20). Interestingly, the washing step did not impact on the presence of pulcherrimin-producing isolates. A more in-depth characterization was carried out on those 42 isolates, as Metschnikowia spp. are acknowledged to contribute to the diversity and complexity of wine taste. A strain level analysis was performed by means of fingerprinting profiles (primer (GTG)5) and phenotypic characterization (sulfite reductase, β-glucosidase, and esterase enzymatic activities). 11 genotypic profiles and 6 different phenotypic combinations were observed among the 42 isolates. Considering both approaches, it was possible to define the presence of 19 strains of Metschnikowia spp., most of them isolated only once, but some present in both washed and non-washed grapes, throughout the whole sampling period. Results obtained in this study shed light on the native Metschnikowia yeast community of washed and non-washed withered grapes, that is composed by diverse strains, and highlight that this biodiversity can be underestimated if only genotypic or phenotypic properties are investigated. This diversity represents a reservoir of strains with enological/pro technological significance that could be applied and combined to improve the sensory characteristics of wine and fermented beverages. 

DOI:

Publication date: June 27, 2022

Issue: IVAS 2022

Type: Poster

Authors

Martelli Francesco1, Checchia Ilaria1, Troiano Eleonora1, Gatto Veronica1, Leal Binati Renato1, Torriani Sandra1 and Felis Giovanna E.1

1Department of Biotechnology, University of Verona, Italy

Contact the author

Keywords

non-Saccharomyces; microbial diversity; spontaneous fermentation; wine quality; grape washing

Tags

IVAS 2022 | IVES Conference Series

Citation

Related articles…

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.

Leaf vine content in nutrients and trace elements in La Mancha (Spain) soils: influence of the rootstock

The use of rootstock of American origin has been the classic method of fighting against Phylloxera for more than 100 years. For this reason, it is interesting to establish if different rootstock modifies nutrient composition as well as trace elements content that could be important for determining the traceability of the vine products. A survey of four classic rootstocks (110-Richter, SO4, FERCAL and 1103-Paulsen) and four new ones (M1, M2, M3 and M4) provided by Agromillora Iberia. S.L.U., all of them grafted with the Tempranillo variety, has been carried out during 2019. The eight rootstocks were planted in pots of 500 cc, on three soils with very different characteristics from Castilla-La Mancha (Spain). In the month of July, the leaves were collected and dried in a forced air oven for seven days at 40ºC. Then, the samples were prepared for the analysis determination, carried out by X-Ray fluorescence spectrometry. The results obtained showed that in the case of content in mineral elements in leaf, separated by soil type, we can report the importance of few elements such as Si, Fe, Pb and, especially, Sr. The rootstock does not influence the composition of the vine leaf for the studied elements that are the most important in determining the geochemical footprint of the soil. The influence of the soil can be discriminated according to some elements such as Fe, Pb, Si and, especially, Sr.

Different soil types and relief influence the quality of Merlot grapes in a relatively small area in the Vipava Valley (Slovenia) in relation to the vine water status

Besides location and microclimatic conditions, soil plays an important role in the quality of grapes and wine. Soil properties influence…

Use of a new, miniaturized, low-cost spectral sensor to estimate and map the vineyard water status from a mobile 

Optimizing the use of water and improving irrigation strategies has become increasingly important in most winegrowing countries due to the consequences of climate change, which are leading to more frequent droughts, heat waves, or alteration of precipitation patterns. Optimized irrigation scheduling can only be based on a reliable knowledge of the vineyard water status.

In this context, this work aims at the development of a novel methodology, using a contactless, miniaturized, low-cost NIR spectral tool to monitor (on-the-go) the vineyard water status variability. On-the-go spectral measurements were acquired in the vineyard using a NIR micro spectrometer, operating in the 900–1900 nm spectral range, from a ground vehicle moving at 3 km/h. Spectral measurements were collected on the northeast side of the canopy across four different dates (July 8th, 14th, 21st and August 12th) during 2021 season in a commercial vineyard (3 ha). Grapevines of Vitis vinifera L. Graciano planted on a VSP trellis were monitored at solar noon using stem water potential (Ψs) as reference indicators of plant water status. In total, 108 measurements of Ψs were taken (27 vines per date).

Calibration and prediction models were performed using Partial Least Squares (PLS) regression. The best prediction models for grapevine water status yielded a determination coefficient of cross-validation (r2cv) of 0.67 and a root mean square error of cross-validation (RMSEcv) of 0.131 MPa. This predictive model was employed to map the spatial variability of the vineyard water status and provided useful, practical information towards the implementation of appropriate irrigation strategies. The outcomes presented in this work show the great potential of this low-cost methodology to assess the vineyard stem water potential and its spatial variability in a commercial vineyard.

1H-NMR-based Metabolomics to assess the impact of soil type on the chemical composition of Mediterranean red wines

The aim of this study was to evaluate the effects of different soil types on the chemical composition of Mediterranean red wines, through untargeted and targeted 1H-NMR metabolomics. One milliliter of raw wine was analyzed by means of a Bruker Avance II 400 spectrometer operating at 400.15 MHz. The spectra were recorded by applying the NOESYGPPS1D pulse sequency, to achieve water and ethanol signals suppression. No modification of the pH was performed to avoid any chemical alteration of the matrix. The generation of input variables for untargeted analysis was done via bucketing the spectra. The resulting dataset was preprocessed prior to perform unsupervised PCA, by means of MetaboAnalyst web-based tool suite. The identification of compounds for the targeted analysis was performed by comparison to pure compounds spectra by means of SMA plug-in of MNova 14.2.3 software. The dataset containing the concentrations (%) of identified compounds was subjected to one-way analysis of variance (ANOVA) to highlight significant differences among the wines. The untargeted analysis, carried out through the PCA, revealed a clear differentiation among the wines. The fragments of the spectra contributing mostly to the separation were attributed to flavonoids, aroma compounds and amino acids. The targeted analysis leaded to the identification of 68 compounds, whose concentrations were significant different among the wines. The results were related to soils physical-chemical analysis and showed that: 1) high concentrations of flavan-3-ols and flavonols are correlated with high clay content in soils; 2) high concentrations of anthocyanins, amino acids, and aroma compounds are correlated with neutral and moderately alkaline soil pH; 3) low concentrations of flavonoids and aroma compounds are correlated with high soil organic matter content and acidic pH. The 1H-NMR metabolomic analysis proved to be an excellent tool to discriminate between wines originating from grapes grown on different soil types and revealed that soils in the Mediterranean area exert a strong impact on the chemical composition of the wines.