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• This study Previous study Y. lipolytica TEM YL 3 TEM YL 3 Full-fat cheese (9) Y. lipolytica TEM YL 5 TEM YL 5 Semi-fat cheese (9) Y. lipolytica TEM YL 6 TEM YL 6 Semi-fat cheese (9) Y. lipolytica TEM YL 9 TEM YL 9 Full-fat cheese (9) Y. lipolytica TEM YL 10 TEM YL 10 Full-fat cheese (9) Y. lipolytica TEM YL 17 TEM YL 17 Kaşar cheese (9) Y. lipolytica TEM YL 18 TEM YL 18 Kaşar cheese (9) Y. lipolytica TEM YL 19 TEM YL 19 Kaşar cheese (9) Y. lipolytica TEM YL 20 TEM YL 20 Kaşar cheese (9) Y. lipolytica TEM YL 21 TEM YL 21 Low-fat cheese (9) Y. lipolytica TEM ORC 2 Strain Number 2 Waste sludge (15) Y. lipolytica TEM ORC 3 Strain Number 3 Waste water (15) Y. lipolytica TEM ORC 4 Strain Number 4 Soil (15) Y. lipolytica TEM ORC 9 Strain Number 9 Soil (15) Y. lipolytica TEM ORC 10 Strain Number 10 Waste water (15) Y. lipolytica TEM ORC 11 Strain Number 11 Waste water (15) Y. lipolytica TEM ORC 12 Strain Number 12 Waste water (15) Y. lipolytica TEM ORC 13 Strain Number 13 Waste water (15) Y. lipolytica TEM ORC 17 Strain Number 17 Soil (15) Y. lipolytica TEM ORC 20 Strain Number 20 Waste water (15) Y. lipolytica TEM TAN 10 Isolate Number 10 White cheese (16) Y. lipolytica TEM TAN 46 Isolate Number 46 White cheese (17) M T 3 5 6 9 10 17 18 19 20 21 Or2 Or3 Or4 M 1000 bp 900 bp 800 bp 700 bp 600 bp 500 bp 400 bp 300 bp 200 bp 100 bp 1000 bp 900 bp 800 bp 700 bp 600 bp 500 bp 400 bp 300 bp 200 bp 100 bp Figure 1. Amplification of the ITS region of Yarrowia lipolytica strains. Line 1, M, DNA marker (100 bp); Line 2, T, Yarrowia lipolytica CBS6124 type strain; other lines show Yarrowia lipolytica strains, respectively.
• Andrade et al. studied the RFLP analysis of 18S rDNA examined to discriminate yeast species usually found in dry-cured meat products, and they found that the RFLP of the 18S rDNA region allowed discrimination of the species Saccharomyces cerevisiae, Rhodotorula mucilaginosa, and Y. lipolytica (7). However, minimal differences at the strain level were found for all of the species tested. Results found in the literature about suitability of RFLP of the 18S rDNA are contradictory. Thus, this method has been also reported as unsuitable for the differentiation of yeasts while it has been proven as a useful technique for discriminating several yeast species such as Candida stellata, Metschnikowia pulcherrima, Kloeckera apiculata, and Schizosaccharomyces pombe (11). Therefore, this method is not used routinely for the discrimination of yeasts species; however, in our study, it was found to be effective in the discrimination of Y. lipolytica strains. Amplification and sequencing of D1/D2 domain of 26S rDNA region As a result of the amplification of the D1/D2 domain of 26S rDNA region, a single fragment with a molecular size of approximately 550 bp formed for all of the Y. lipolytica strains tested. After sequencing, the sequences obtained were compared with the GenBank database using the BLASTN tool, as presented in Table 3. It was determined that all Y. lipolytica strains tested exhibited a high degree of homology (99% to 100%) with various Y. lipolytica strains present in the National Center for Biotechnology Information, as a result of BLAST analysis of the partial sequence of the 26S rRNA gene. The phylogenetic relationship between the 22 Y. lipolytica strains are shown in the Figure 4. The phylogenetic tree was reconstructed by the neighbor-joining method (24). D1/D2 domain sequencing of 26S rDNA is being used more and more often to identify yeasts with both 1000 bp 900 bp 800 bp 700 bp 600 bp 500 bp 400 bp 300 bp 250 bp 200 bp 150 bp 100 bp 50 bp
• Figure 2. Restriction analysis with the endonucleases RsaI of ITS region. Line 1, M, DNA marker (50 bp); Line 2, T, Yarrowia lipolytica CBS6124 type strain; other lines show
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