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BIODIVERSITY OF YEASTS OF LOCAL WINEGRAPES IN TÜRKİYE AND GENOTYPING OF SACCHAROMYCES CEREVISIAE
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28.03.2023 Elnaz Seyedmonir Ph.D Thesis, Adobe.pdf
Date
2023-2-27
Author
Seyedmonir, Elnaz
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Grape must and wine harbors a wide diversity of yeast species responsible for spontaneous alcoholic fermentation and wine aroma. Therefore, the impact of indigenous species and terroir effect on aromatic properties of traditional Turkish wine made from Kalecik Karası, Boğazkere, Öküzgözü, and Dimrit grape varieties collected from three different regions (Ankara, Elazığ and Cappadocia) were investigated, using gas chromatography-mass spectrometry (GC-MS). A total of 56 compounds were identified and quantified in wines from four red Vitis vinifera grape varieties grown in Türkiye. Among the quantitated volatile compounds, different volatiles were found to significantly contribute their flavor notes to the overall aroma of Turkish wines. The native non-Saccharomyces and Saccharomyces yeast diversity in these traditional wine samples made from four red and one white grape varieties at different stages were detected by using sixteen specific oligonucleotide primers in real-time polymerase chain reaction (PCR) analysis. Saccharomyces cerevisiae and three predominant non-Saccharomyces yeasts Hanseniaspora uvarum, Lachancea thermotolerans and Torulaspora delbrueckii were also quantified by quantitative real-time PCR methods. Biodiversity determined with the use of real-time PCR analysis were compared with DNA sequencing result of internal transcribed spacer (ITS) region (ITS1–5.8S rRNA– ITS2) and/or D1/D2 domain of the 26S rRNA gene of the isolates. Moreover, the random amplified polymorphic DNA (RAPD) PCR using one Operon primer (OPA-11) and the mini and microsatellite primed (MSP)-PCR fingerprinting technique using primers as M13, (GTG)5, (ATG)5 were also applied to investigate the intraspecific genetic diversity between the 46 autochthonous S. cerevisiae strains isolated from five different must and wine at different stages. The result indicated that RAPD-PCR and MSP-PCR technique were applicable to the determination of intraspecific genetic diversity between the 46 indigenous S. cerevisiae strains. In addition, karyotyping analyses were carried out for selected S. cerevisiae strains by pulsed-field gel electrophoresis (PFGE) method which distinguished ten different chromosomal patterns between S. cerevisiae strains.
Subject Keywords
Saccharomyces cerevisiae, GC-MS, real-time PCR, RAPD/MSP-PCR, PFGE.
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https://hdl.handle.net/11511/102775
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Graduate School of Natural and Applied Sciences, Thesis
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E. Seyedmonir, “BIODIVERSITY OF YEASTS OF LOCAL WINEGRAPES IN TÜRKİYE AND GENOTYPING OF SACCHAROMYCES CEREVISIAE,” Ph.D. - Doctoral Program, Middle East Technical University, 2023.