DNA sequence polymorphisms in the genus Saccharomyces. I. Comparison of theHIS4 and ribosomal RNA genes in lager strains, ale strains and various species

Abstract

The region of chromosome XII containing the RDNI gene which encodes the cytosolic ribosomal RNA molecules and the region of chromosome Ill containing the H1S4 (histidine 4) gene were analysed in 30 lager yeast strains, 11 ale strains and 20 strains from a number of different species in the genus Saccharomyces.With the aid of restriction endonuclease fragment patterns and cloned probes to the RDN1 gene ofS. cerevisiae three forms of this gene were identified, two of them corresponding to the previously known forms I and II and a third one characterized by an additional HindlIl site located in the 3' spacer region. A more distantly related form of the RDNlgene containing a single HindlIl restriction site was found in Saccharomyces fermentati and one form without any Hindlll site in a wild yeast contaminant.With the help of the restriction endonuclease fragments derived from the HIS4 region seven genotypes can be recognized. They result from various combinations of three restriction endonuclease fragment patterns designated I, II and III, each pattern represents a chromosome.All lager strains are homozygous for form II of the ribosomal RNA gene and heterozygous for patterns I and II of the HIS4 gene. An exception is one German brewing strain which is homozygous for pattern II.With one exception the ale strains were homozygous for form II of the RDN1 gene and for pattern I of the HIS4 gene. One British strain contains form I of the RDN1 gene. Bakers yeast, S. diastaticus and S. italicus are homozygous for form I of the RDN1 gene and for pattern I of the HIS4 gene. In S. bayanus and S. pastorianus homozygosity for form III of the RDN1 gene was combined with heterozygosity for patterns II and III of the HIS4 gene. S. uvarum is homozygous for both form III of the RDN1 gene and pattern III of the HIS4 gene. Form III of the RDN1 gene and patterns I and ti of the HIS4 gene were combined in a Chinese brewing strain and a strain designated as a type strain ofS. carlsbergensis.The nucleotide sequence polymorphisms are useful markers for strain characterization in addition to the generally used fermentation properties.Abbreviations: bp = basepairs; kb = kilobase; SSC = 0.15 M-NaCI, 15 mM-Na citrate; Tris -tris-(hydroxymethyl)-amino methane.

DNA sequence polymorphisms in the genus Saccharomyces IV. Homoeologous chromosomes III of Saccharomyces bayanus, S. carlsbergensis, and S. uvarum

“…The similarity of the karyotypes of S. bayanus, S. inusitatus, and S. uvarum is attested by the restriction endonuclease fragment patterns of RDN1, HIS4, LEU2, and DNA-DNA reassociation kinetics (4,23,24,25,27,30). Since S. bayanus NCYC374 shows composite restriction endonuclease fragment patterns at the HIS4 locus (24,25,27), and independent segregation of these patterns in spore clones more than one chromosome III are present in the genome ofS. bayanus.…”

“…S. pastorianus and S. bayanus are considered according to VAN DER WALT (35) to be strains of the same species and restriction endonuclease fragment length polymorphisms at RDN1, H1S4, and LEU2 support this view (24,27). The four strains included in the S. bayanus group (Table I) In contrast to the species just discussed the three S. carlsbergensis strains have more than one chromosome band containing a HIS4 gene ( Figure Figure 4).…”

“…Base composition, DNA-RNA homology, DNA-DNA reassociation kinetics and restriction endonuclease fragment length polymorphisms have proven useful in the taxonomic evaluations of genera and species as well as in the diagnosis of closely related yeast strains (3,4,15,21,23,24,25,26,27,28,30,40). For taxonomic as well as breeding purposes the establishment of genetic linkage groups and their assignment to Abbreviations: OFAGE = Orthogonal field alternation gel electrophoresis; TBE = 0.089 M-Tris-borate, 0.089 M-boric acid, 0.002 M-EDTA; SSC = 0.15 M-NaC1, 15 mM-Na citrate.…”

Cloning and analysis of two alleles of theILV3 gene from Saccharomyces carlsbergensis

“…The results presented here with ILF3 are also consistent with reports on gene polymorphism between S. cerevisiae and S. carlsbergensis in the CYC7, HIS4, ILV1. LEU2, MAT, and URA3 loci (15,27,28,29,30).…”

“…Accordingly, the Saccharomyces carlsbergensis alleles display restriction endonuclease site polymorphisms in molecular hybridization studies using radioactively labelled Saccharomyces cerevisiae gene probes. Examples include HIS4 (15,27,29), LEU2 (15,30), and regions adjacent to MAT (15) on chromosome III and ILV1, URA3, and CYC7 on chromosome V (28). In the case of the RDNI gene on chromosome XII, no gene polymorphism has yet been uncovered in Saccharomyces carlsbergensis, which suggests that chromosomal organization and nucleotide sequences of this gene is very similar to that in Saccharomyces cerevisiae.…”