Open AccessMolecular Characterization ofYarrowia lipolyticaandCandida zeylanoidesIsolated from Poultry
Author(s) -
T. Deák,
J. Chen,
Larry R. Beuchat
Publication year - 2000
Publication title -
applied and environmental microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.552
H-Index - 324
eISSN - 1070-6291
pISSN - 0099-2240
DOI - 10.1128/aem.66.10.4340-4344.2000
Subject(s) - yarrowia , biology , spacer dna , intergenic region , rapd , genetics , trichosporon , dna , yeast , microbiology and biotechnology , ribosomal rna , internal transcribed spacer , genome , gene , genetic diversity , population , demography , sociology
Yeast isolates from raw and processed poultry products were characterized using PCR amplification of the internally transcribed spacer (ITS) 5.8S ribosomal DNA region (ITS-PCR), restriction analysis of amplified products, randomly amplified polymorphic DNA (RAPD) analysis, and pulsed-field gel electrophoresis (PFGE). ITS-PCR resulted in single fragments of 350 and 650 bp, respectively, from eight strains ofYarrowia lipolytica and seven strains ofCandida zeylanoides . Digestion of amplicons withHin fI andHae III produced two fragments of 200 and 150 bp fromY. lipolytica and three fragments of 350, 150, and 100 bp fromC. zeylanoides , respectively. Although these fragments showed species-specific patterns and confirmed species identification, characterization did not enable intraspecies typing. Contour-clamped heterogeneous electric field PFGE separated chromosomal DNA ofY. lipolytica into three to five bands, most larger than 2 Mbp, whereas six to eight bands in the range of 750 to 2,200 bp were obtained fromC. zeylanoides . Karyotypes of both yeasts showed different polymorphic patterns among strains. RAPD analysis, using enterobacterial repetitive intergenic sequences as primers, discriminated between strains within the same species. Cluster analysis of patterns formed groups that correlated with the source of isolation. For ITS-PCR, extraction of DNA by boiling yeast cells was successfully used.