Restriction polymorphisms in the internal transcribed spacers and 5.8S rDNA ofSaccharomyces

Abstract

Polymorphisms in enzymatically amplified ribosomal DNA (rDNA) were examined in 18 strains of Saccharomyces. Restriction patterns generated from the region spanning the internal transcribed spacers (ITS) and the 5.8S rDNA produced two clusters corresponding to S. bayanus and S. cerevisiae. The type culture of S. carlsbergensis (ATCC 76529), which could not be separated from the S. cerevisiae group by small subunit (SSU) rDNA patterns, showed a ScrfI profile that was distinct from all the other strains. The type culture of S. intermedius var. turicensis (ATCC 76519) is assigned to S. bayanus on the basis of the combined results of SSU and ITS restriction analyses. S. kluyveri occurred at a separate branch of the distance tree and is unrelated to any of the strains. Results were in general agreement with reported DNA homologies and are discussed in relation to other molecular and genetic data.

Characterization of Technologically Utilized Saccharomyces Yeast

“…A fragment with the size of 855 bp was formed in all strains (Table 3). Molina et al (1992) found that the size of the PCR product with primers complementary to the ITS region in the species S. cerevisiae, S. bayanus and S. pastoriunus is approximately 850 bp, while the S. kluyveri (currently Lachancea kluyveri) provides a fragment of about 700 bp. Restriction analysis of the ITS region using enzymes has been performed to distinguish between species (Valente et al, 1996;Pham et al, 2011).…”

Rapid Characterization of Four Species of the Saccharomyces Sensu Stricto Complex According to Mitochondrial DNA Patterns

“…Furthermore, the 900-base sequences from S. pastonanus and S. bayanus were identical, and those from S. cerevisiae and S. paradoxus differed in only five nucleotides. rDNA restriction analysis from PCR-amplified fragments (13,(23)(24)(25) has also been used recently to differentiate between strains belonging to S. cerevisiae, S. bayanus, and S. pastorianus.…”

“…These include DNA fingerprinting (5,(31)(32)(33)(34)42), chromosomal DNA profiles (7,9,14,15,21,27), mitochondrial DNA (mtDNA) restriction analysis (22,48), and rRNA and ribosomal DNA (rDNA) sequencing (18) as well as restriction analysis (13,(23)(24)(25). Until now, only electrophoretic karyotyping, rRNA sequencing and rDNA restriction analysis have proved useful for differentiating Saccharomyces species.…”

Restriction analysis of the ITS region for characterization of the Debaryomyces species.

“…The generic term ITS represents the rDNA sequences ITS1, 5.8S, and ITS2, amplified together in the polymerase chain reaction (PCR) experiments (White et al, 1990 and Orilio Leoncini* within a species and could be used as an exclusionary characteristic for detecting misidentified isolates (Valente et al, 1996(Valente et al, , 1997, but it alone cannot identify the isolate at the species level because more than one species can have the same ITS product length. A restriction analysis of this product has already been used for black yeast and Saccharomyces identification (Molina et al, 1992;Uijthof and de Hoog, 1995;Yurlova et al, 1996) and seems to be a useful tool for differentiating phenotypically similar species that have ITS products of about the same size.…”