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Yeasts for the Sustainability in Viticulture and Oenology

Final Report Summary - YESVITE (Yeasts for the Sustainability in Viticulture and Oenology)

The wine world is going through a rapid transformation due to a deep change in consumer preferences, consumption habits and accompanied by an important reduction in economic resources available to the people. Thus, the modern viticulture and oenology has the key role of innovating traditional practices by supporting new choices for a sustainable production of wine. In general, many of the current challenges faced by the world of wine include production quality, making ‘healthier’ wines, and establishing the concept of sustainability in winemaking. Yeasts are among the most important food production microorganisms, especially the yeast Saccharomyces cerevisiae, for the production of wine and several other fermented foods. They exhibit an extremely high biodiversity and they could hide a huge, untapped reservoir for potential innovation in the wine sector helping to establish the meaning of sustainability in the wine World.
The strategic aim of the YeSVitE project is to create a coordinated network that can learn how to manage the topic of sustainability in oenology bringing innovation by exploiting yeasts as the principle resource, a still untapped source in food production.

YeSVitE, starting in January 2014, has carried out research and training activities thanks to the contribution of young scientists (six are females and two males) and experienced researchers (five are females and seven males) which have focused their studies on wine yeasts, with interesting traits for the concept of sustainability, contributing to the general knowledge on yeasts and applications in the wine sector.
YeSVitE consists of seven leading academic laboratories from Italy (Milan and Perugia), Spain, Slovenia, Canada, South Africa and Georgia that provide a unique environment to develop strong academia and new solutions potentially directed to the industry innovation.

YeSVitE general objectives are: i) to assess the evolutionary linkage among wine-related yeasts from ancient and new vine-growing areas; ii) to select promising wild yeasts strains for winemaking; iii) to set up a new international yeast collection for oenological applications; iv) to develop improved strains by the identification of genes associated with aroma pathways or carbon/nitrogen metabolism for the production of quality and/or healthier wines; v) to exploit the metabolic profiles for yeast discrimination at species/strains level (FTIR approach) and to study the impact of fermentative yeast on wine quality (LC-MS approaches); vi) to improve the managing of sulphur dioxide addition during winemaking to reduce spoilage yeasts and decrease the level of sulphite in wines; vii) to reduce ethanol content in wines by the setup of innovative protocols for the inoculation of musts with new starter cultures; viii) to study the role of yeasts in the vineyard ecosystem; ix) to investigate the natural antagonistic potential of various yeasts against moulds to develop a sustainable bio-protection approach; and, x) to select specific yeasts strains for the different cultivar for the optimization of sensory characteristics or to show up nutraceutical features.

During four years of activity, YeSVitE totalized 85.6% of the planned secondments generating data that were used to disseminate/communicate the scientific results of the project. Seventy-one public available research products among which 15 original papers in international peer-reviewed journals, 2 publication in popular press, 51 oral and poster presentations in national and international conferences, 1 Master's and 1 PhD's theses, and the final YeSVitE Conference were released. In general, the YeSVitE consortium has: collected new yeasts for wine fermentation, studied their biodiversity degree (investigating the evolutionary trend by NGS sequencing) and setup of a microbial collection of wine-associated yeasts [2, 11, 15]; setup novel genetic tools for taxonomic analysis of yeasts (Kiloseq method); applied, for the first time, the CRISPR/Cas9 strategy to wine yeasts [6]; exploited yeast metabolome for strain characterisation and wine design (FTIR approach and LC-MS metabolomics); deepened the knowledge on spoilage yeast B. bruxellensis [3, 5, 12, 14] and assessed its sulphite tolerance by RNA-Seq approach; studied new solution to reduce the ethanol content in wines and developed mixed starters for low alcohol wines [1, 10, 13]; proposed attractive yeasts for grape bio-protection [4]; faced the “precision oenology” topic [9, 11]; explored vineyard and fermentation ecosystems to set attractive wine microorganisms [7, 8]; and, explored vineyard and fermentation ecosystems (ARISA and metagenomics studies).

The following management and administrative activities were completed: (i) the kick-off meeting (by web, November 2013); (ii) Consortium Agreement has been signed (iii); (iv) Partnership Agreement has been signed; v) YeSVitE home page, www.yesvite.unimi.it; (vi) 2014 annual meeting in Slovenia (Vipava); (vii) 2015 annual meeting in Italy Perugia; (viii) 2017 final meeting in Italy (Milan); ix) YeSVitE International Conference in Italy (Milan), supported by the University of Milan (Research Support Plan 2015/2017, nmjLinea 2 - Action C, 2017). All the mentioned events represented a wide effort of the members of the consortium to coordinate the network activities and strengthen the scientific collaboration with the final goal of achieving all the promised milestones and deliverables of YeSVitE’s tasks.

Publications
1. Tronchoni J, Curiel JA, Saenz-Navajas MP, Morales P, de-la-Fuente-Blanco A, Fernandez-Zurbano P, Ferreira V, Gonzalez R (2018) Aroma profiling of an aerated fermentation of natural grape must with selected yeast strains at pilot scale. Food Microbiol 70: 214-223. doi.org/10.1016/j.fm.2017.10.008
2. Vigentini I, Barrera Cardenas S, Valdetara F, Faccincani M, Panont C, Picozzi C, Foschino R (2017) Use of Native Yeast Strains for In-Bottle Fermentation to Face the Uniformity in Sparkling Wine Production. Front Microbiol, 8: 1225. doi: 10.3389/fmicb.2017.01225
3. Valdetara F, Fracassetti D, Campanello A, Costa C, Foschino R, Compagno C, Vigentini I (2017) A Response Surface Methodology Approach to Investigate the Effect of Sulfur Dioxide, pH, and Ethanol on DbCD and DbVPR Gene Expression and on the Volatile Phenol Production in Dekkera/Brettanomyces bruxellensis CBS2499. Front Microbiol, 8: 1727. doi: 10.3389/fmicb.2017.01727
4. Cordero-Bueso G, Mangieri N, Maghradze D, Foschino R, Valdetara F, Cantoral JM, Vigentini I (2017) Wild Grape-Associated Yeasts as Promising Biocontrol Agents against Vitis vinifera Fungal Pathogens. Front Microbiol. 2017; 8: 2025, doi: 10.3389/fmicb.2017.02025
5. Romano D, Valdetara F, Zambelli P, Galafassi S, De Vitis V, Molinari F, Compagno C, Foschino R, Vigentini I (2017) Cloning the putative gene of vinyl phenol reductase of Dekkera bruxellensis in Saccharomyces cerevisiae. Food Microbiol, 63, 92-100, doi: 10.1016/j.fm.2016.11.003
6. Vigentini I, Gebbia M, Belotti A, Foschino R, Roth FP (2017) CRISPR/Cas9 system as a valuable genome editing tool applicable to wine yeasts: the decrease of urea production as a case of study. Front Microbiol, 8: 2194. doi: 10.3389/fmicb.2017.02194
7. Curiel JA, Morales P, Gonzalez R, Tronchoni J (2017) Different Non-Saccharomyces Yeast Species Stimulate Nutrient Consumption in S. cerevisiae Mixed Cultures. Front. Microbiol, 8:2121. doi: 10.3389/fmicb.2017.02121
8. Vigentini I, Praz A, Domeneghetti D, Zenato S, Picozzi C, Barmaz A, Foschino R (2016) Characterization of malolactic bacteria isolated from Aosta Valley wines and evidence of psychrotrophy in some strains. J Appl Microbiol, 120, 934-945, doi: 10.1111/jam.13080
9. Vigentini I, Maghradze D, Petrozziello M, Bonello F, Mezzapelle V, Valdetara F, Failla O, Foschino R (2016) Indigenous Georgian wine-associated yeasts and grape cultivars to edit the wine quality in a precision oenology perspective. Front Microbiol, 7: 352, doi: 10.3389/fmicb.2016.00352
10. Rodrigues AJ, Raimbourg T, Gonzalez R, Morales P (2016) Environmental factors influencing the efficacy of different yeast strains for alcohol level reduction in wine by respiration. LWT - Food Science and Technology, 65 1038-1043. http://dx.doi.org/10.1016/j.lwt.2015.09.046
11. Vigentini I, Gardana C, Fracassetti D, Gabrielli M, Foschino R, Simonetti P, Tirelli A, Iriti M (2015) Yeast contribution in melatonin, melatonin isomers and tryptophan-ethylester during alcoholic fermentation of grape musts. J Pineal Res, 58, 388-396. doi: 10.1111/jpi.12223
12. V. Fabrizio, Vigentini I., Parisi N., Picozzi C., Compagno C., Foschino R. (2015). Heat inactivation of wine spoilage yeast Dekkera bruxellensis by hot water treatment. Letters in Applied Microbiology, 61, p.186-191 ISSN: 0266-8254, doi: 10.1111/lam.12444
13. Morales P, Rojas V, Quirós M, Gonzalez R (2015) The impact of oxygen on the final alcohol content of wine fermented by a mixed starter culture. Appl Microbiol Biotechnol 99:3993–4003. DOI 10.1007/s00253-014-6321-3
14. Fracassetti D. and Vigentini I. (2017) Occurrence and analysis of sulfur compounds in wine. In: "Grapes and Wines - Advances in Production, Processing, Analysis and Valorization" Ed: Jordão A.M. and Cosme F. In: TechOpen. ISBN: 978-953-51-5583-6
15. Cordero-Bueso G, Vigentini I, Foschino R, Maghradze D, Manuel Cantoral J (2017) Genetic diversity of yeasts isolated from Eurasian populations of Vitis vinifera ssp. sylvestris (Gmelin) Hegi. 40th World Congress of Vine and Wine. BIO Web of Conferences 9, 02019. DOI: 10.1051/bioconf/20170902019.