Ohranitev genetskega potenciala kvasovk Primorskega vinorodnega okoliša in njihova genetska tipizacija

Jerovšek, Ines (avtor), Čadež, Neža (mentor), Butinar, Lorena (komentor)

Najpomembnejša kvasovka v biotehnologiji je Saccharomyces cerevisiae, ki si primarno ekološko nišo deli z ozko sorodno vrsto Saccharomyces paradoxus. V magistrskem delu smo želeli ohraniti genetski potencial 386 sevov kvasovk Vipavskega vinorodnega okoliša in določiti njihovo genetsko raznolikost. Šest sevov kvasovk smo shranili pri –80 °C in ob tem določili njihovo stopnjo preživetja po enem in štirih mesecih. Stopnja preživetja kvasovk je s časom upadala. Najvišjo stopnjo preživetja so imele kvasovke Dekkera bruxellensis, S. cerevisiae, Metschnikowia pulcherrima in Rhodotorula glutinis. Na podlagi rezultatov PCR-RFLP regije ITS smo razlikovali S. cerevisiae in S. paradoxus od drugih vrst kvasovk. Izmed 204 Saccharomyces sevov je bilo 5 % izolatov S. cerevisiae in 9 % S. paradoxus iz naravnega okolja, ostali so bili iz vinograda. Ostale vrste kvasovk rodov Candida, Debaryomyces, Hanseniaspora, Kluyveromyces, Metschnikowia, Meyerozyma, Lachancea, Pichia, Schwanniomyces, Torulaspora in Zygosaccharomyce smo identificirali s sekvenciranjem regije ITS, ter jim določili genetsko raznolikost na podlagi vzorca pomnožkov z (GTG)5x. S povezovanjem genetske raznolikosti v mikrosatelitskih hipervariabilnih regijah vrst Saccharomyces in njihovim fenotipom smo pokazali obstoj dveh skupin vinogradniških sevov S. cerevisiae, prve odporne proti visokim koncentracijam etanola in glukoze, druga pa protu kalijevemu metabisulfitu.

• Jezik: Slovenski jezik
• Ključne besede: kvasovke, Saccharomyces cerevisiae, Primorski vinorodni okoliš, krioprezervacija, stopnja preživetja, PCR-RFLP, ITS, (GTG)5x, mikrosateliti
• Vrsta gradiva: Magistrsko delo/naloga
• Tipologija: 2.09 - Magistrsko delo
• Organizacija: BF - Biotehniška fakulteta
• Založnik: [I. Jerovšek]
• Leto izida: 2019
• PID: 20.500.12556/RUL-107733
• UDK: 579.25:582.282.23:577.2.08
• COBISS.SI-ID: 5050744
• Datum objave v RUL: 18.05.2019

JEROVŠEK, Ines, 2019, Ohranitev genetskega potenciala kvasovk Primorskega vinorodnega okoliša in njihova genetska tipizacija [na spletu]. Magistrsko delo. I. Jerovšek. [Dostopano 10 april 2025]. Pridobljeno s: https://repozitorij.uni-lj.si/IzpisGradiva.php?lang=slv&id=107733

Sekundarni jezik

• Jezik: Angleški jezik
• Naslov: Preservation of yeasts genetic potential from Primorska wine region and their genetic typing

Izvleček

The most important yeast in biotechnology is Saccharomyces cerevisiae, which shares its primary ecological niche with a closely related species Saccharomyces paradoxus. The purpose of our work was to preserve genetic potential of 386 yeast strains isolated in Vipava valley and to determine their genetic diversity. Six yeast strains were preserved at –80 °C and their survival rate was determined after one and four months. Yeasts survival rate declined over time. The highest survival rates were found in species Dekkera bruxellensis, S. cerevisiae, Metschnikowia pulcherrima and Rhodotorula glutinis. Based on PCR-RFLP results of the ITS region we could differentiate S. cerevisiae and S. paradoxus from other yeast species. Five and nine percent of S. paradoxus and S. cerevisiae out of 204 isolates, respectively, were of non-vineyard origin. Other yeast species of genera Candida, Debaryomyces, Hanseniaspora, Kluyveromyces, Metschnikowia, Meyerozyma, Lachancea, Pichia, Schwanniomyces, Torulaspora and Zygosaccharomyce were identified by sequencing of the ITS region and their genetic diversity was determined by (GTG)5x fingerprinting. By combining genetic diversity in microsatellite hypervariable regions of the Saccharomyces species and their phenotypes, we determined two distinct groups of S. cerevisiae vineyard strains, one resistant to high concentration of ethanol and glucose, and the second to potassium metabisulphite.

• Ključne besede: yeasts, Saccharomyces cerevisiae, Primorska wine region, kriopreservation, survival rate, PCR-RFLP, ITS, (GTG)5x, microsatellites