Morphological and Molecular Tools for Identification of Saccharomyces Boulardii Isolated from Active Dry Yeast
Journal Title: International Journal of Agriculture Innovations and Research - Year 2019, Vol 7, Issue 4
Abstract
Isolation and identification of Saccharomyces boulardii from a product "High Active Dry Yeast" were the aim of this study. Yeast colonies were isolated and purified thrice by repetitive streaking on the YPD agar medium supplemented with 250 μg/ml of Chloramphenicol and 30 μg/ml of Ampicillin. The morphological identification was investigated regarding the shapes, color, and morphology characteristics. S. boulardii clustered in a circular shape, whitish cream color, elliptical morphology, and formed pseudohyphae. Genomic DNA (gDNA) was extracted from S. boulardii by using the TIANamp Yeast DNA Kit and the universal primers were used for species identification. Identification of yeast genetically processed by using the primer pair NL1 and NL4 to amplify and sequence the 26S rDNA gene D1/D2 domain. The internal transcribed spacer was amplified and sequenced by using ITS1 and ITS4 for fungus identification, besides using the primer pair of NS1 and NS6. Discrimination of yeast from bacteria processed by using the primer pair 7F, 1540R, and another primer pair 27F, and 1492R. PCR technique could approve that the S. boulardii was the only frequent strain in the active dry yeast. The photographed pattern by UV transilluminator types the target DNA band to S. boulardii that was achieved by amplifying and sequencing the 26S rDNA region. The amplified band (740 bp) and the resulted sequence length of 26S rDNA gene (585 bp) assigned the isolate to S. boulardii
Authors and Affiliations
Ahmed Hassan Mousa, et al.
Keywords
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