Two Subgroups within the Saccharomyces bayanus Species Evidenced by PCR Amplification and Restriction Polymorphism of the Non-Transcribed Spacer 2 in the Ribosomal DNA Unit

• Published in: Systematic and applied microbiology Vol. 20; no. 2; pp. 286 - 294
• Main Authors: Nguyen, Huu-Vang, Gaillardin, C.
• Format: Journal Article
• Language: English
• Published: Jena Elsevier GmbH 1997; Elsevier
• Subjects: ADN; ANALISIS MICROBIOLOGICO; ANALYSE MICROBIOLOGIQUE; Biological and medical sciences; CARIOTIPOS; CARYOTYPE; DNA; Fundamental and applied biological sciences. Psychology; IDENTIFICACION; IDENTIFICATION; KARYOTYPES; karyotyping; Life Sciences; MICROBIOLOGICAL ANALYSIS; Microbiology; Microbiology and Parasitology; Mycological methods and techniques used in mycology; Mycology; NUCLEOTIDE SEQUENCE; POLIMERIZACION; POLIMORFISMO; Polymerase Chain Reaction; POLYMERISATION; POLYMERIZATION; POLYMORPHISM; POLYMORPHISME; Ribosomal-DNA; RIBOSOMAS; RIBOSOME; RIBOSOMES; rRNA 18S gene; rRNA 5S gene; SACCHAROMYCES; Saccharomyces bayanus; Saccharomyces cerevisiae; Saccharomyces paradoxus; Saccharomyces pastorianus; Saccharomyces sensu stricto; Saccharomyces uvarum; SECUENCIA NUCLEOTIDICA; SEQUENCE NUCLEOTIDIQUE; SOUS ESPECE; SUBESPECIES; SUBSPECIES; TAXONOMIA; TAXONOMIE; TAXONOMY; Identification; Saccharomyces; Ribosomal-DNA; karyotyping; Taxonomy; Polymerase Chain Reaction; Fungi; Polymerase chain reaction; Ribosomal DNA; Spacer DNA; Ascomycetes; Species specificity; Method; Saccharomyces bayanus; Thallophyta
• ISSN: 0723-2020; 1618-0984
• DOI: 10.1016/S0723-2020(97)80075-6

A PCR-RFLP method has been improved for the rapid identification of the four species of Saccharomyces sensu stricto. We used the NTS2-ETS sequence of the rDNA as target and the amplification reaction was realized by chosing a pair of primers that could anneal to the end and the beginning of the 5S and 18S conserved sequences, respectively. PCR products obtained from S. cerevisiae, S. bayanus, S. paradoxus and S. pastorianus (synonym S. carlsbergensis) type strains displayed a clear polymorphism when digested with several restriction enzymes. Using only three enzymes: Ban I, Alu I and Taq I we were able to differentiate S. cerevisiae, S. paradoxus and the S. bayanus/S. pastorianus complex which shared the same pattern. The S. bayanus taxon now regroups the former species S. abuliensis, S. globosus, S. heterogenicus, S. intermedius, S. inusitatus, S. uvarum. PCR-RFLP of these species showed that, except for S. abuliensis and S. uvarum, they shared the same patterns as S. bayanus type strain. That lead us to conclude that there are two subgroups in S. bayanus and that S. abuliensis and S. uvarum belong to a distinct subgroup. Remarkably, wine strains that were previously identified as S. bayanus by genetic hybridization techniques, as well as strains isolated from cider fermentation, belong to the S. uvarum subgroup.