Identification of two yeast strains from oil-polluted environment by RFLP on ITS-5.8S rDNA and RAPD analyses

Two yeast strains DP3 and DP4, isolated from oil-polluted environment and previously studied using morpho-physiological tests, presented 91.40% similarity with Issatchenkia orientalis CBS5147 (DP3) and, respectively, 83.80% similarity with Rhodotorula glutinis CBS20T (DP4). For a more accurate characterization, we performed restriction of the amplified region of ITS1-5,8S-ITS2, and compared the results with those from laboratory reference yeast strains, MYCOBANK and similar experiments described in literature. Strain DP3 ITS-PCR and ARDRA profiles were almost identical with I. orientalis CMGB224 and I. orientalis CBS5147, while an obvious similarity was observed between the DP4 patterns and those from R. glutinis ICCF and R. glutinis CBS20T. For the RAPD studies, we used eight primers: OPA01, OPA03, OPA10, SOY, RP4-2, M13 for DP3, and M13, OPH15, OPH19 for DP4. Further dendograms were constructed using UPGAMA method from Quantity One (Bio-Rad) program and OPA03 profiles for DP3, DP4 and reference yeast strains. The dendrograms revealed a high degree of relatedness between DP3 and Issatchenkia orientalis CMGB224, DP4 and Rhodotorula glutinis ICCF.