Molecular identification of wild non-saccharomyces yeasts from wine by pcr-rflp and strain differentiation by rapd

Allahyari, Nasim
Non-Saccharomyces yeasts are dominant during the early stages of alcoholic fermentation of grape must and play a role in the production of favorable organoleptic traits in the final wine. Identification of indigenous yeasts for using them as starter culture in a sequential inoculation along with Saccharomyces cerevisiae is an effective approach to produce wine with desirable characteristics. In this study, 18 non-Saccharomyces yeasts isolated at different maceration and fermentation times from traditional wine of five different grape types in Turkey were identified by PCR-RFLP (Polymerase Chain reaction- Restriction Fragment Lenghth Polymorphism) at genus and species level and then Lachancea thermotolerans (Lt), Metschnikowia pulcherrima (Mp), Hanseniaspora uvarum (Hu), Hanseniaspora opuntiae (Ho) species were selected for the genotypic differentiation of their strains using RAPD–PCR technique. Amplification of internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) using PCR allowed to identify some yeast isolates at the genus level. However, precise identification at genus and/or species level was possible by PCR-RFLP using appropriate enzymes. In this study, restriction digestion of rDNA ITS region using five endonucleases, HhaІ (CfoI), HaeIII, HinfI, DdeI and DraІ allowed identification of NS yeasts. vi On the other hand, RAPD-PCR using one RAPD primer OPA-03, minisatellite primer M13 and, two microsatellite primers (ATG)5, (GTG)5, were useful for the differentiation of yeast population at the strain level.


The effects of sequential inoculation of non-Saccharomyces and Saccharomyces yeasts from spontaneously fermented grape must varieties on Kalecik Karası wine and molecular characterization of Saccharomyces and Lachancea thermotolerans isolates
Arslan, Gizem Pınar; Gültekin, Güzin Candan; Gözen, Ayşe Gül; Department of Biochemistry (2021-9)
Sequential fermentation is a process based on using non-Saccharomyces yeasts and Saccharomyces cerevisiae to contribute wine quality and improve wine complexity. Identification and characterization of the yeasts are crucial to achieving ideal fermentation. In this study, sequential fermentation of Kalecik Karası must was carried out to indicate potential usage of wild yeasts as starter culture in winemaking. Four selected non-Saccharomyces isolates (Lachancea thermotolerans, Wickerhamomyces anomalus, Metsch...
Seyedmonir, Elnaz; Gültekin, Güzin Candan; Çetinkaya , Asya; Department of Biochemistry (2023-2-27)
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 quantifie...
Effects of a bacteriocin-like substance produced by Leuconostoc mesenteroides subsp. cremoris on spoilage strain Lactobacillus fructivorans and various pathogens
YURDUGÜL, SEYHUN; Bozoglu, Faruk (Wiley, 2008-01-01)
Certain lactic acid bacteria present in the natural flora of the wine fermentation medium perform the malolactic fermentation process and other biochemical activities which lead to the formation of the outstanding aroma and bouquet of wine. However, because of the spoilage phenomena caused also by certain lactic cultures, off-odours formed via metabolic side reactions cause severe economic losses in the wine industry. Several control mechanisms were introduced in order to eliminate these spoilage cultures, ...
Purification and characterization of a bacteriocin from an oenological strain of Leuconostoc mesenteroides subsp cremoris
Dundar, Halil; SALİH, BEKİR; Bozoglu, Faruk (Informa UK Limited, 2016-01-01)
Malolactic fermentation (MLF), which improves organoleptic properties and biologic stability of some wines, may cause wine spoilage if uncontrolled. Bacteriocins were reported as efficient preservatives to control MLF through their bactericidal effect on malolactic bacteria. Leuconostoc mesenteroides subsp. cremoris W3 isolated from wine produces an inhibitory substance that is bactericidal against malolactic bacteria in model wine medium. Treatment of the culture supernatant of strain W3 with proteases eli...
Diversity of Wild Yeasts During Spontaneous Fermentation of Wines from Local Grape Varieties in Turkey
Gültekin, Güzin Candan; Aktuna, Ipek; Seyedmonir, Elnaz (2022-10-01)
This study was conducted to investigate the wild yeast diversity in traditional wine from Turkish grape varieties. Isolated and identified yeast species in wines of grapes from Turkey were compared with those from other countries. The wild yeasts from five different grape varieties were detected by real-time PCR and isolated and/or identified by DNA sequencing techniques. In total, 469 yeast colonies were isolated from selective media at different times from musts and wine samples: 104 from 204 non-Saccharo...
Citation Formats
N. Allahyari, “Molecular identification of wild non-saccharomyces yeasts from wine by pcr-rflp and strain differentiation by rapd,” M.S. - Master of Science, Middle East Technical University, 2020.