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…

Soil quality in Beaujolais vineyard. Importance of pedology and cultural practices

A pedological study was carried out from 2009 to 2017 in Beaujolais vineyard, to improve physical and chemical knowledge of soils. It was completed in 2016 and 2017 by the current study, dealing with microbial aspects, in order to build a reference frame for improved advice in soil management. Microbial biomass was measured on representative plots of the six most common soil types identified in Beaujolais and, for each soil type, on plots with different levels of the main impacting parameters: total organic carbon, pH, cation exchange capacity, extractable copper. A total of 59 soil samples were collected. Confirming the results of various trials carried out in Beaujolais over the past 20 years, the results of the present study showed that the soils were still alive, but exhibited a large variability of biological parameters, which appeared dependant on both pedological and anthropic factors. Therefore, a good interpretation of biological parameters and advice for vine growers must rely on a pedologically-based referential with differentiated main driving factors. For example, the control of pH is of primary importance in granitic soils and in no way organic matter addition can improve soil quality if pH is too low. Conversely, in calcareous soils, biological parameters are more directly affected by direct or indirect (cover crops for example) inputs of organic matter. The use of biological parameters, such as microbial biomass, is of great potential value to improve advice on agro-viticultural practices (soil management, fertilization, liming, etc.), basis of a sustainable wine production on fragile soils.

Traditional agroforestry vineyards, sources of inspiration for the agroecological transition of viticulture

A unique “terroir” can be found in southern Bolivia, which combines the specific features of climate, topography and altitude of high valleys, with the management of grapevines staked on trees. It is one of the rare remnants of agroforestry viticulture. A survey was carried out among 29 grapegrowers in three valleys, to characterize the structure and management of these vineyards, and identify the services they expect from trees. Farms were small (2.2 ha on average) and 85% of vineyards were less than 1 ha. Viticulture was associated with vegetable, fruit and fodder production, sometimes in the same fields. Molle trees were found in all plots, together with one or two other native tree species. Traditional grapevine varieties such as Negra Criolla, Moscatel de Alejandría and Vicchoqueña were grown with a large range of densities from 1550 to 9500 vines ha-1. From 18 to 30% of them were staked on trees, with 1.2 to 4.9 vines per tree. The management of these vineyards (irrigation, fertilization and grapevine protection) was described, the most particular technical operation being the coordinated pruning of trees and grapevines. Three types of management could be identified in the three valleys. Grapegrowers had a clear idea of the ecosystem services they expected from trees in their vineyards. The main one was protection against climate hazards (hail, frost, flood). Then they expected benefits in terms of pest and disease control, improvement of soil fertility and resulting yield. At last, some producers claimed that tree-staking was quicker and cheaper than conventional trellising. It can be hypothesized then that agroforestry is a promising technique for the agroecological transition of viticulture. Its contribution to the “terroir” of the high valleys of southern Bolivia and its link with the specificities of the wines and spirits produced there remain to be explored.

Metabolomic discrimination of grapevine water status for Chardonnay and Pinot noir

Water status impact in viticulture has been widely explored, as it strongly affects grapevine physiology and grape chemical composition. It is considered as a key component of vitivinicultural terroir. Most of the studies concerning grapevine water status have focused on either physiological traits, or berry compounds, or traits involved in wine quality. Here, the response of grapevine to water availability during the ripening period is assessed through non-targeted metabolomics analysis of grape berries by ultra-high resolution mass spectrometry. The grapevine water status has been assessed during 2 consecutive years (2019 & 2020), through carbon isotope discrimination on juices from berries collected at maturity (21.5 brix approx.) for 2 Vitis vinifera cv. Pinot noir (PN) and Chardonnay (CH). A total of 220 grape juices were collected from 5 countries worldwide (Italy; Argentina; France; Germany; Portugal). Measured δ13C (‰) varied from -28.73 to -22.6 for PN, and from -28.79 to -21.67 for CH. These results also clearly revealed higher water stress for the 2020 vintage. The same grape juices have been analysed by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-qTOF-MS), leading to the detection of up to 4500 CHONS containing elemental compositions, and thus likely tens of thousands of individual compounds, which include fatty acids, organic acids, peptides, phenolics, also with high levels of glycosylation. Multivariate statistical analysis revealed that up to 160 elemental compositions, covering the whole range of detected masses (100 –1000 m/z), were significantly correlated to the observed gradients of water status. Examples of chemical markers, which are representative of these complex fingerprints, include various derivatives of the known abscisic acid (ABA), such as phaesic acid or abscisic acid glucose ester, which are significantly correlated with higher water stress, regardless of the variety. Cultivar-specific behaviours could also be identified from these fingerprints. Our results provide an unprecedented representation of the metabolic diversity, which is involved in the water status regulation at the grape level, and which could contribute to a better knowledge of the grapevine mitigation strategy in a climate change context.

Investigating the impact of grape exposure and UV radiations on rotundone in Vitis vinifera L. Tardif grapes under field trial conditions

Rotundone is the main aroma compound responsible for peppery notes in wines whose biosynthesis is negatively affected by heat and drought. Through the alteration of precipitation regime and the increase in temperature during maturation, climate change is expected to affect wine peppery typicality. In this context there is a demand for developing sustainable viticultural strategies to enhance rotundone accumulation or limit its degradation. It was recently proposed that ultraviolet (UV) radiations could stimulate rotundone production. The aim of this study was to investigate under field trial conditions the impact of grape exposure and UV treatments on rotundone in Vitis vinifera L. Tardif, an almost extinct grape variety from south-west France that can express particularly high rotundone levels. Four different treatments were compared in 2021 to a control treatment using a randomised complete block design with three replications per treatment. Grape exposure was manipulated through early or late defoliation. Leaf and laterals shoots were removed at Eichorn Lorenz growth stages 32 or 34 on the morning-sun side of the canopy. During grape maturation, UV radiations were either reduced by 99% by installing UV radiation-shielding sheets, or applied four times using the Boxilumix™ non thermal device (Asclepios Tech, Tournefeuille) with the aim of activating plant signalling pathway. Loggers displayed in solar radiation shields were used to assess the effect of such shielding sheets on air temperature within the bunch zone. The composition of grapes subjected to these treatments will be soon analysed for their rotundone content and basic classical laboratory analyses. Grapes will be harvested to elaborate wines under standardized small-scale vinification conditions (60kg) that will be assessed by a trained sensory panel.

Ecophysiological performance of Vitis rootstocks under water stress

The use of rootstocks tolerant to soil water deficit is an interesting strategy to cope with limited water availability. Currently, several nurseries are breeding new genotypes, but the physiological basis of its responses under water stress are largely unknown. To this end, an ecophysiological assessment of the conventional 110-Richter (110R) and SO4, and the new M1 and M4 rootstocks was carried out in potted ungrafted plants. During one season, these Vitis genotypes were grown under greenhouse conditions and subjected to two water regimes, well-watered and water deficit. Water potentials of plants under water deficit down to < -1.4 MPa, and net photosynthesis (AN) <5 μmol m-2 s-1 did not cause leaf oxidative stress damage compared to well-watered conditions in any of the genotypes. The antioxidant capacity was sufficient to neutralize the mild oxidative stress suffered. Under both treatments, gravimetric differences in daily water use were observed among genotypes, leading to differences in the biomass of root, shoot and leaf. Under well-watered conditions, SO4 and 110R were the most vigorous and M1 and M4 the least. However, under water stress, SO4 exhibited the greatest reduction in biomass while M4 showed the lowest. Remarkably, under these conditions, SO4 reached the least negative stem water potential (Ψstem), while M1 reduced stomatal conductance (gs) and AN the most. In addition, SO4 and M1 genotypes also showed the highest and lowest hydraulic conductance values, respectively. Our results suggest that there are differences in water use regulation among genotypes, not only attributed to differences in stomatal regulation or intrinsic water use efficiency at the leaf level. Therefore, because no differences in canopy-to-root ratio were achieved, it is hypothesized that xylem vessel anatomical differences may be driving the reported differences among rootstocks performance. Results demonstrate that each Vitis rootstock differs in its ecophysiological responses under water stress.