Şaraptan izole edilmiş doğal non-saccharomyces mayalarin PCR-RFLP ile moleküler tanımlanması ve RAPD ile suş farklilaştirmasi

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. 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 strains belonging to same species (starins belonging to Lt, Hu, Ho, Mp species) .