Macrowine 2021
IVES 9 IVES Conference Series 9 Expanding the biotechnological potential of M. pulcherrima/fructicola clade for wine-related applications

Expanding the biotechnological potential of M. pulcherrima/fructicola clade for wine-related applications

Abstract

AIM: Strains belonging to M. pulcherrima/fructicola clade are frequently isolated from flowers, fruits and grape musts, and exhibit a broad spectrum of enzymatic activities and antimicrobial potential (Morata et al., 2019; Sipiczki, 2020; Vicente et al. 2020). By reason of these features, selected strains of this clade have been proposed as non-Saccharomyces starter cultures for winemaking. In this study, with a view to valorise the biotechnological potential of these strains, a new Metschnikowia sp. strain, DBT012, was selected for application in vinification trials of Valpolicella’s fresh and withered typical grapes for reduction of SO2addition and increase of aromatic complexity. Further, in the framework of the investigation on the biodiversity of a collection of strains, distinct pulcherrimin-producing isolates from spontaneous fermentation and grapes were putatively recognized as Metschnikowia spp. and selected for genotypic and phenotypic characterisation.

METHODS: Lyophilised cultures of the reference strain DBT012 were tested in pilot-scale vinifications (mixed-cultures with S. cerevisiae). The isolates were characterised based on enzymatic activities (e.g., sulphite-reductase and beta-glucosidase activities) and growth under different wine-related stress conditions (e.g., ethanol, high sugar content). Genetic fingerprinting techniques (e.g., (GTG)5 and microsatellite) were performed for de-replication of isolates and strain typing. Whole-genome sequencing of the reference strain was carried out to analyse the correlation between genotype and phenotype, and comparative analyses with available type strains were performed to deepen the taxonomic aspects and molecularly identify this strain.

RESULTS: Use of Metschnikowia sp. DBT012 reduced the acetaldehyde content in wines, and positively influenced the wine aroma. About 50 Metschnikowia strains were characterised, which displayed a significant phenotypic diversity in terms of β-glucosidase and esterase activities, H2S production and growth at high sugar concentrations. The relationship of those features with genotypic and genomic distinctiveness of the reference strain was highlighted.

CONCLUSIONS:

The well-characterised thematic collection of strains belonging to M. pulcherrima/fructicola clade presented here constitutes an important reservoir of biodiversity for applications in different wine-related scenarios.

DOI:

Publication date: September 3, 2021

Issue: Macrowine 2021

Type: Article

Authors

Eleonora Troiano,Renato, LEAL BINATI, Ilaria, CHECCHIA, Ilaria, LARINI, Veronica, GATTO, Gianluca, VENERI, Giacomo, ZAPPAROLI, Vittorio, CAPOZZI, Elisa, SALVETTI, Sandra, TORRIANI, Giovanna, E., FELIS

Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy , Department of Biotechnology, University of Verona, Italy, Institute of Sciences of Food Production, National Research Council (CNR), Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy, Department of Biotechnology, University of Verona, Italy

Contact the author

Keywords

metschnikowia, wine microbiology, biodiversity, biotechnological potential

Citation

Related articles…

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.

Combining effect of leaf removal and natural shading on grape ripening under two irrigation strategies in Manto negro (Vitis vinifera L.)

The increasingly frequent heat waves during grape ripening pose challenges for high quality wine grape production. Defoliation is a common practice that can improve the control of diseases in bunches, but also it increases the exposure to sunlight. Grapes exposed to solar radiation reach temperatures over the optimum for berry development and maturation. This makes the development of irrigation and canopy management techniques of great importance to maximize yield and grape quality. A field experiment was carried out during 2021 using Manto negro wine grapes to study the effect of applied irrigation and different light exposure levels on grape quality. Two irrigation treatments were imposed based on the frequency and amount of water doses in a four-block experimental vineyard at Bodega Ribas (Mallorca). Three light exposure treatments were randomly applied in each irrigation plot. The light treatments included exposed clusters from pea size, non-exposed clusters, and shaded clusters after softening. Leaf area index and canopy porosity was estimated every 2 weeks. Midday leaf water potential was measured weekly. Additionally, apparent electrical conductivity was measured between rows to estimate the soil water content variability. Light and temperature sensors were installed at the bunch level to quantify the differences in bunch temperature and light intensity among treatments. The effect of irrigation and cluster light exposure on berry weight, TSS, TA, malic acid, tartaric acid, K+, and pH were analysed at 5 moments along grape ripening. During different heat waves, the natural shading technique decreased the maximum bunch temperature around 10 °C respect to the exposed bunches in both irrigation strategies. The combination of defoliation and shading techniques after softening decreased TSS at harvest and affected most of the quality parameters during the last stages of ripening, showing an interesting technique to delay ripening in warm viticulture areas.

Anthocyanin profile is differentially affected by high temperature, elevated CO2 and water deficit in Tempranillo (Vitis vinifera L.) clones

Anthocyanin potential of grape berries is an important quality factor in wine production. Anthocyanin concentration and profile differ among varieties but it also depends on the environmental conditions, which are expected to be greatly modified by climate change in the future. These modifications may significantly modify the biochemical composition of berries at harvest, and thus wine typicity. Among the diverse approaches proposed to reduce the potential negative effects that climate change may have on grape quality, genetic diversity among clones can represent a source of potential candidates to select better adapted plant material for future climatic conditions. The effects of individual and combined factors associated to climate change (increase of temperature, rise of air CO2 concentration and water deficit) on the anthocyanin profile of different clones of Tempranillo that differ in the length of their reproductive cycle were studied. The aim was to highlight those clones more adapted to maintain specific Tempranillo typicity in the future. Fruit-bearing cuttings were grown in controlled conditions under two temperatures (ambient temperature versus ambient temperature + 4ºC), two CO2 levels (400 ppm versus 700 ppm) and two water regimes (well-watered versus water deficit), both in combination or independently, in order to simulate future climate change scenarios. Elevated temperature increased anthocyanin acylation, whereas elevated CO2 and water deficit favoured the accumulation of malvidin derivatives, as well as the acylation and tri-hydroxylation level of anthocyanins. Although the changes in anthocyanin profile observed followed a common pattern among clones, such impact of environmental conditions was especially noticeable in one of the most widely distributed Tempranillo clones, the accession RJ43.

Measurement of redox potential as a new analytical winegrowing tool

Excell laboratory has initiated the development of an analytical method based on electrochemistry to evaluate the ability of wines to undergo or resist to oxidative phenomena. Electrochemistry is a powerful tool to probe reactions involving electron transfers and offers possibility of real-time measurements. In that context, the laboratory has implemented electrochemical analysis to assess oxidation state of different wine matrices but also in order to evaluate oxidative or reduced character of leaf and soil. Initially, our laboratory focused on dosage of compounds involved in responses of plant stresses and we were also interested in microbiological activity of soils. These analyses were compared with the measurement of redox potential (Eh) and pH which are two fundamental variables involved in the modulation of plant metabolism. Indeed, the variation of redox states of the plant reflects its biological activity but also its capacity to absorb nutriments. The Eh-pH conditions mainly determine metabolic processes involved in soil and leaf and our goal is to determine if this combined analytical approach will be sufficiently precise to detect biological evolutions (plant health, parasitic attack…).

Elevational range shifts of mountain vineyards: Recent dynamics in response to a warming climate

Increasing temperatures worldwide are expected to cause a change in spatial distribution of plant species along elevational gradients and there are already observable shifts to higher elevations as a consequence of climate change for many species. Not only naturally growing plants, but also agricultural cultivations are subject to the effects of climate change, as the type of cultivation and the economic viability depends largely on the prevailing climatic conditions. A shift to higher elevations therefore represents a viable adaptation strategy to climate change, as higher elevations are characterized by lower temperatures. This is especially important in the case of viticulture because a certain wine-style can only be achieved under very specific climatic conditions. Although there are several studies investigating climatic suitability within winegrowing regions or longitudinal shifts of winegrowing areas, little is known about how fast vineyards move to higher elevations, which may represent a viable strategy for winegrowers to maintain growing conditions and thus wine-style, despite the effects of climate change. We therefore investigated the change in the spatial distribution of vineyards along an elevational gradient over the past 20 years in the mountainous wine-growing region of Alto Adige (Italy). A dataset containing information about location and planting year of more than 26000 vineyard parcels and 30 varieties was used to perform this analysis. Preliminary results suggest that there has been a shift to higher elevations for vineyards in general (from formerly 700m to currently 850 m a.s.l., with extreme sites reaching 1200 m a.s.l.), but also that this development has not been uniform across different varieties and products (i.e. vitis vinifera vs hybrid varieties and still vssparkling wines). This is important for climate change adaptation as well as for rural development. Mountain areas, especially at mid to high elevations, are often characterized by severe land abandonment which can be avoided to some degree if economically viable and sustainable land management strategies are available.