1. EXPLORING BIODIVERSITY POTENTIAL OF WINE ASSOCIATED YEASTSSofia Dashko, 2015, doktorska disertacija Opis: Human exploitation of yeast fermentation dates back to the Neolithic. S. cerevisiae has been the most important yeast used for numerous fermentations of biotechnological interest, including grape fermentation for wine production. Despite its abundant use, the molecular mechanisms controlling alcoholic fermentation are rather unclear and the choice of S. cerevisiae as an inoculum is often the consequence of a mere habit, rather than the result of rational analyses.
In this work we focused on the role of different yeasts in the winemaking process. I was interested in understanding how the specific strain used for inoculum could influence the wine aroma formation. Furthermore, I wished to investigate the yeast genetics and ecology by characterizing their population and strain diversity in one of the wine regions of Slovenia.
To evaluate the effect of the yeast species on the fermentation outcome, we performed successive fermentations with five different species in combination with the industrial strain S. cerevisiae Lalvin T73. The experiment showed that at least two more yeast species, Kazachstania gamospora and Zygosaccharomyces kombuchaensis have good potential to be applied in the winemaking. , The main conclusions of this study are the possibility of: i) expanding the palette of alternative starters to widen the aromatic components ii) co–fermenting using two different yeast species. Mixed yeast culture fermentations are present in the natural context, but this practice has been neglected in biotechnological processes.
Positive results of wine fermentations with non – conventional yeast urged us to explore the diversity of Slovenian wine region natural isolates. The resulting yeast collection counts more than 1200 strains for which phenotype and genotype have been defined. Numerous isolates, including non – Saccharomyces species, showed promising oenological and biotechnological traits because of their capacity of rapid utilization of various carbon sources, growth at low pH and at presence of copper sulfite and potassium metabisulfite. Sampling also revealed sharp discrimination between the ecological niches of S. cerevisiae and S. paradoxus, which is a striking feature of European vineyards. While S. cerevisiae habitats are strongly associated with human activity, S. paradoxus was mainly isolated from the forest sources. Profound analysis of the collected data could give some explanations to the driving forces of S. cerevisiae domestication and S. paradoxus geographic isolation population structure.
Najdeno v: ključnih besedah
Ključne besede: Yeasts, biodiversity, molecular biology, alcoholic fermentation
Objavljeno: 02.11.2015; Ogledov: 7306; Prenosov: 158
 Polno besedilo (41,27 MB) |
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3. Isolation and selection wild yeasts for winemakingNatalia Mikhaylina, 2017, diplomsko delo Opis: The aim of this study was to explore the diversity of fermentative yeast species in a non-vineyard and vineyard environment, and exploit their oenological potential. For this purpose we collected plant and soil material in the vineyard (located in Črniče in Vipava Valley) planted with Vitis vinifera L. and in the forest (forest in Vipava Old Castle), and by using a selective medium with high ethanol and sugar content we managed to obtain 35 isolates. By sequencing the D1/D2 domain of the 26S rDNA we identified nine different yeast species. The oenological potential of identified yeasts was further investigated by employing a series of tests, such as growth at low pH values, tolerance to ethanol and sulfur, and H2S production ability. Based on the results of these tests we selected four different yeast species for the microvinification of Vitis vinifera L. cv. ‘Pinot Noir’ grape juice. A commercial strain Saccharomyces cerevisiae Lalvin D47 was also included as a control in the fermentation experiment. Fermentation monitoring was done by measuring daily weight loss and microbiological analysis in the middle of fermentation. For the final wines, sugars were determined and sensorial evaluation performed. Among four tested yeasts, S. paradoxus IVV 32 and Torulaspora delbrueckii IVV 36 appeared to be the most promising for winemaking, but further studies should be carried out.
Najdeno v: ključnih besedah
Ključne besede: wine, Vitis vinifera L. cv. ‘Pinot Noir’, biodiversity, yeasts, fermentation
Objavljeno: 10.07.2017; Ogledov: 4873; Prenosov: 274
Polno besedilo (2,57 MB) |
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5. Study of yeast biodiversity potential in the development of biological control agents against phytopathogenic fungi in viticultureLorena Butinar, Urban Česnik, Adesida Rowland, Melita Sternad Lemut, 2017, objavljeni povzetek znanstvenega prispevka na konferenci Opis: Despite public’s growing concerns for human health and environmental pollution, synthetic fungicides are still preferred and massively used for the protection of agricultural crop plants against fungal diseases. However, more and more research is focused on finding potential alternatives in the form of effective biological control agents. Although there are reports of yeast’s biocontrol activity, they are up to date poorly commercialized for such purposes.
As the yeasts represent an important part of the grape microflora, competing with other microorganisms (including pathogens) for nutrients and space, we decided to examine the potential of autochthonous yeasts as "green" alternatives in fighting against phytopathogens such as Botrytis cinerea in viticulture. With this aim we tested biocontrol activity of various strains / species of indigenous wine yeasts that were isolated from the vineyard environment in Slovenia and cross-border Italy. The yeasts were tested for the presence of siderophores, hydrolytic enzymes (chitinase, β-glucosidase and β-glucanase) and sensitivity to fungicides (copper, iprodion, cyprodinil + fludioxonil combination). The tests of phytopathogenic fungi growth inhibition in the presence of antagonistic yeast were also performed. The double Petri dish test was used to verify the inhibition of conidia germination by the formation of antifungal volatile organic compounds (VOC), synthetized by the antagonistic yeasts. The presence of hydrolytic enzymes was confirmed in many wine yeasts, particularly in the strains of Hanseniaspora uvarum and Metschnikowia pulcherrima species and in some strains of the Pichia genus. Using a medium, based on natural diluted grape juice in a double Petri dish test, we found that yeasts of the Debaryomyces hansenii, Lachancea thermotolerans, P. kudriavzevii, Saccharomyces kudriavzevii, S. cerevisiae and Torulaspora delbrueckii species were able to inhibit the germination of fungal conidia at the expense of the VOC formation. However, none of the up to date tested yeasts formed siderophores. On the other hand, the yeasts were generally resistant to the tested fungicide concentrations, thus they could be used as potential biofungicides in the combination with chemical fungicides.
Najdeno v: ključnih besedah
Povzetek najdenega: ...poorly commercialized for such purposes.
As the yeasts represent an important part of the grape...
Ključne besede: yeasts, biocontrol, Botrytis cinerea, viticulture
Objavljeno: 08.11.2017; Ogledov: 3801; Prenosov: 0
Polno besedilo (10,33 MB) |
6. Biodiversity of cider yeasts and their cider-making potentialLorena Butinar, Branka Mozetič Vodopivec, Melita Sternad Lemut, Eivind Vangdal, 2017, objavljeni povzetek znanstvenega prispevka na konferenci Opis: In the area of Hardanger, a part of the fjord region in Western Norway, the production of apple wine (cider) has a long tradition that goes back to the 12th century, when monks introduced apple growing in this area. Nowadays, this is also the main area of fruit production in Norway. Despite the strict regulation of the alcoholic beverage production in Norway, traditional cider is still produced on some farms in this area. By tradition cider is produced by a spontaneous fermentation process of apple juice, performed by naturally occurring indigenous yeasts that originate from the fruit or the surfaces of the processing equipment. Therefore, our aim was primarily to study the ecology and biodiversity of the yeasts associated with the production of traditional cider in the Hardanger area. For two consecutive years, we sampled at 11 different locations in the observed region, where we collected cider samples and surface swabs of processing facilities from the cideries, and also soil and various parts of apple trees in orchards owned by the same producers. Thus, by enriching collected samples with the selective medium with high sugar and ethanol concentration, we managed to isolate about 1,300 yeasts. Based on the multiplex PCR results the yeasts were grouped into the Saccharomyces sensu stricto complex and non-Saccharomyces yeasts. The isolates were determined to the species level by performing the restriction analysis of ITS PCR products, and in some cases identifications were confirmed by sequencing of the D1/D2 domain of the 26S rDNA and/ or ITS region. As expected, non-Saccharomyces yeasts from the genus Metschnikowia and Hanseniaspora mainly populated the orchards, while the Saccharomyces yeasts were isolated in the orchards from the soil and fruits. In contrast, in ciders the species S. uvarum was predominantly found, occasionally also S. cerevisiae, Torulaspora delbrueckii and P. membranifacies. Indigenous cider yeasts were further on characterized in micro-plate format for the most important cider-making technological parameters (tolerance to ethanol, SO2, growth at low pH), for the presence of glucoside hydrolase activity, H2S production ability, and assimilation of malic acid. Based on this screenings the micro-scale fermentations of apple juice were performed with 13 different indigenous cider yeasts as monocultures. The most promising indigenous yeasts, T. delbrueckii and S. uvarum, were also tested as mixed cultures in sequential fermentations. Since the tested strain of T. delbrueckii as monoculture was not able to complete the alcoholic fermentation, better results were obtained in sequential fermentation with the mixed culture in combination with S. uvarum.
Najdeno v: ključnih besedah
Ključne besede: indigenous yeasts, biodiversity, spontaneous fermentation, cider-making
Objavljeno: 08.11.2017; Ogledov: 5159; Prenosov: 0
Polno besedilo (10,33 MB) |
7. Facilitating green care improvements in viticulturePaolo Sivilotti, Tjaša Jug, Urban Česnik, Adesida Rowland, Melita Sternad Lemut, Lorena Butinar, Urška Vrhovšek, 2019, objavljeni znanstveni prispevek na konferenci Opis: The growing demand of consumers for environment-friendly and safe food products already forced many winemakers to restrict the application of synthetic fungicides to earlier season, trying to reduce the risk of residues being detected in wines. Main risk for infection by B. cinerea, however, occurs later in the season. To support an improvement in whole-season B. cinerea green care, the potential of yeasts as biocontrol agent (PBA) was studied. Selected yeasts were tested for their antifungal activities. Furthermore, a field experiment in Pinot noir and Pinot gris vineyards was designed to study different combinations of optimized canopy microclimate manipulation (CMM) techniques and PBA application. In experimental conditions, PBA's ability to maintain appropriate population density for disease prevention was observed. Grape and wine quality parameters were evaluated. Analytical approach of metabolomics was applied to upgrade the understanding of potentially induced changes in plant metabolism.
Based on yeasts testing results, biocontrol yeast P. guilliermondii ZIM 624 was selected and after its field application, we were able to detect its suitable density on grapes until harvest. In case of early defoliation for both varieties smaller grape compactness was confirmed together with lower yield. Among grape basic quality parameters, optimized techniques showed positive effect on sugar content, but unexpectedly, in the treatments with biocontrol yeasts the trends toward higher acidity were noticed in Pinot gris. CMM and PBA introduction indicated some effects on grapevine biosynthetic behaviour but the differences in content of these metabolites compared to control were poorly detected in final wines with exception of mainly some flavonols, stilbenes and flavan-3-ols (and consequently total phenolic content).
Najdeno v: ključnih besedah
Povzetek najdenega: ...B. cinerea green care, the potential of yeasts as biocontrol agent (PBA) was studied. Selected...
Ključne besede: sustainable viticulture, Botrytis cinerea, yeasts, biocontrol, canopy microclimate manipulation, metabolomics, grapevine metabolites
Objavljeno: 28.02.2020; Ogledov: 2894; Prenosov: 0
Polno besedilo (7,85 MB) |
8. Synthesis of pyranoanthocyanins from Pinot Noir grape skin extract using fermentation with high pyranoanthocyanin producing yeasts and model wine storage as potential approaches in the production of stable natural food colorantsDorota Korte, Irena Vovk, Alen Albreht, Lorena Butinar, Karin Kovačević Ganić, Natka Ćurko, Jelena Topić, Branka Mozetič Vodopivec, 2020, izvirni znanstveni članek Najdeno v: ključnih besedah
Ključne besede: Pyranoanthocyanins, Anthocyanins, Food colorants, Yeasts, Fermentation
Objavljeno: 03.04.2020; Ogledov: 2464; Prenosov: 0
Polno besedilo (1,03 MB) |
9. Implementation of high performance liquid chromatography coupled to thermal lens spectrometry (HPLC‑TLS) for quantification of pyranoanthocyanins during fermentation of Pinot Noir grapesLorena Butinar, Jelena Topič Božič, Natka Ćurko, Karin Kovačević Ganić, Branka Mozetič Vodopivec, Dorota Korte, Mladen Franko, 2020, izvirni znanstveni članek Opis: In this work high performance liquid chromatography coupled to thermal lens spectrometry (HPLC-TLS) was applied for monitoring of vinylphenolic pyranoanthocyanins formation during the fermentation of Pinot Noir wines. Vinylphenolic pyranoanthocyanins are wine pigments, present in low concentrations, but very important for wine colour stability.
Fermentation process was conducted with four different yeast strains, used as starters, either in sequential fermentation of non-Saccharomyces with S. cerevisiae yeast or as single fermentation with S. cerevisiae yeasts in order to test the applicability of developed method for monitoring of selected compounds in real wine fermentation experiments. The
developed HPLC-TLS method showed higher sensitivity compared to HPLC coupled to diode array detection (DAD) technique for particular wine colour compounds. Obtained limits of detection (LODs), were 6- and 22-times lower in comparison to HPLC–DAD in gradient and isocratic elution mode, respectively, whereas limits of quantification (LOQs)
5 and 18-times lower. Lower LODs enabled earlier observation of vinylphenolic pyranoanthocyanins formation during fermentation (already at day 7) in the case of HPLC-TLS method in gradient mode, while by using HPLC–DAD in gradient elution mode the formation of vinylphenolic pyranoanthocyanins was noticed only after 12 days of fermentation.
Najdeno v: ključnih besedah
Povzetek najdenega: ...or as single fermentation with S. cerevisiae yeasts in order to test the applicability of...
Ključne besede: Thermal lens spectrometry (TLS), High performance liquid chromatography (HPLC), Pyranoanthocyanins, Wine, Yeasts
Objavljeno: 18.06.2020; Ogledov: 2490; Prenosov: 0
Polno besedilo (1,94 MB) |
10. The influence of selected indigenous yeasts on Pinot Noir wine colour propertiesBranka Mozetič Vodopivec, Dorota Korte, Alen Albreht, Mitja Martelanc, Melita Sternad Lemut, Guillaume Antalick, Lorena Butinar, Jelena Topić, 2021, izvirni znanstveni članek Najdeno v: ključnih besedah
Ključne besede: Pinot Noir, wine colour, yeasts, non-Saccharomyces, sensory
Objavljeno: 07.07.2021; Ogledov: 1328; Prenosov: 0
Polno besedilo (783,35 KB) |
