Moleculer Identification of Kalabaki (Gökçeada, Turkey) Wine Grape Yeasts Using ITS1-5.8S-ITS2 rDNA Gene Region


Çakaloğlu Ç., Günay M., Turgut Genç T.

8. International Symposium on Ecology and Environmental Problems (ISEEP)., Çanakkale, Turkey, 4 - 07 October 2017, vol.1, no.1, pp.1

  • Publication Type: Conference Paper / Full Text
  • Volume: 1
  • City: Çanakkale
  • Country: Turkey
  • Page Numbers: pp.1
  • Çanakkale Onsekiz Mart University Affiliated: Yes

Abstract

Yeasts are the most significant organisms to produce fermented products such as wine, beer, etc. They also contribute to beverages’ quality. The grapes are principally source of natural yeasts in wine products. Thus, determining of yeast biota on grapes surface is considerable important for wine-makers. There are a range of moleculer techniques to identify of yeast species. Sequence analysis of ITS1-5.8S-ITS2 rDNA gene region is commonly used for this purpose. In this study, therefore, we have aimed to identify yeast biota using analysis of ITS-5.8S rDNA gene sequence on Kalabaki wine grapes in Gökçeada. In this context, Kalabaki grapes were gathered from vineyard in Gökçeada. After homogenization process, grape extracts were inoculated on the YGCA solid medium and incubated 2-3 days at 30 ºC. Growing yeast strains were inoculated on YPD Agar rich medium and incubated 2-3 days at 30 ºC. Genomic DNA extraction of yeast strains was carried out and ITS1-5.8S-ITS2 rDNA gene regions were amplified using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) primers. According to PCR product lengths and morphological differences, yeast strains were chosen for sequence analysis. Consequently, 34 yeast strains were isolated from Kalabaki grape surface. 3 groups were obtained by PCR products (~450bp, ~600bp and ~750bp) and 11 groups were obtained by morphological features. Totally 14 selected yeast strains were sequenced. Yeast strains were identified as Candida apicola, Metschnikowia pulcherrima, Starmerella bacillaris, Aerobasidium pullulans, Schwanniomyces sp., Sporidiobolus salmonicolor, C. Carpophila and Hanseniaspora uvarum.