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The aim of this study was to analyze the diversity and genetic relationship of ethanol tolerant yeasts isolated from rice wine starters (Loog-pang) in Chiang Mai province, Thailand. In total, 35 isolates from 9 samples of Loog-pang were identified by polymerase chain reaction (PCR) technique. The internal transcribed spacer 1 (ITS1) and ITS2 regions and the 5.8S ribosomal DNA (rDNA) region of the yeasts were amplified by using universal primers ITS1 and ITS4. According to the PCR product sequence, 21, 11, and 3 isolates were identified as Saccharomyces cerevisiae, Pichia kudriavzevii, and Candida glabrata, respectively. A phylogenetic analysis using neighbour-joining method was conducted in MEGA 7.0.18 using 2,000 replication (bootstrap). The phylogram of S. cerevisiae and P. kudriavzevii strains were divided into 3 and 2 groups, respectively, but the C. glabrata phylogram with bootstrap could not be carried out. Even though the results demonstrated that the yeasts from each sample of Loog-pang were genetically closely related but three isolates of C. glabrata from same district (San-sai) showed a high genetic diversity.
Keywords: ethanol tolerant yeast; rice wine starter; genetic relationship; ITS; DNA marker
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