DNA sequence polymorphisms in the genus saccharomyces III. Restriction endonuclease fragment patterns of chromosomal regions in brewing and other yeast strains

Abstract

30 bottom fermenting strains, 13 hybrid lager strains, 11 top fermenting strains. In addition, 24 other yeast species and strains have been analysed in this study. Four DNA regions have been monitored by restriction endonuclease fragment pattern polymorphisms. The regions analysed are: RDN1 (encoding cytosolic ribosomal RNA molecules) located on chromosome XII in S. cerevisiae, HIS4 (histidine 4) and LEU2 (leucine 2) both located on chromosome III and the Ty elements which are distributed at different positions in the genome ofS. cerevisiae. Seven Ty fragment patterns have been detected in the bottom fermenting strains. Patterns I, II, IIa, lib, and llc are present in lager strains while pattern III and IV are found in S. carlsbergensis bottom fermenting strain No. I and in S. monacensis, respectively. Hybridization of the Tyl p14 probe from S. cerevisiae to OFAGE separated chromosomes from bottom fermenting strains show that at least five Ty elements are present in the genomes of the lager strains, at least five in S. carlsbergensis bottom fermenting strain No. I and at least three in S. monacensis. From the restriction fragment patterns and the karyotypes of the bottom fermenting strains it is suggested that these strains are closely related and different from the S. bayanus, S. pastorianus, and S. uvarum group as well as from the S. cerevisiae strains. In spite of their differences in Ty element patterns the lager strains are so homogenous that most likely they all originate from a single strain which is related to S. carlsbergensis and S. monacensis type strains. In contrast to the bottom fermenting strains there is a large variability among the top fermenting strains, especially with regard to the LEU2 gene and Ty elements. It seems that S. bayanus, S. inusitatus, S. pastorianus, S. validus, and S. uvarum are closely related, and none of the studied strains from these taxons contained DNA which hybridized to the Tyl probe from S. cerevisiae. This is a clear difference from the DNA of bottom fermenting strains which consistently hybridized strongly to the Tyl probe.Abbreviations: kb = kilo base pairs; OFAGE = orthogonal field alternation gel electrophoresis; SSC = 0.15 M-NaCI, 15 mM-Na citrate; TBE = 0.089 M-Tris-borate, 0.089 M-boric acid, 0.002 M-EDTA; Tris = tris-(hydroxymethyl)-amino methane Springer-Verlag

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

“…Strains B and C differ by their Ty element patterns. B (Carlsberg lager strain) contains pattern I, while C (Danish lager strain BK2208) contains pattern IIa (27).…”

“…DNA preparations for OFAGE separations were made according to SCHWARTZ and CANTOR (31), with the modifications reported by PEDERSEN (27). The apparatus was constructed after the descriptions of CARLE and OLSON (5, 6).…”

“…The gels were stained with ethidium bromide in a concentration of 1 lag-ml ' in the appropriate electrophoresis buffer. Conditions for OFAGE separations are as in (27). DNA fragments were transferred to nitrocellulose (Millipore HAWP 00010 or to Schleicher and Schuell BA85) by Southern transfer in 20 • SSC (33).…”

“…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.…”

Molecular genetic study of introgression between Saccharomyces bayanus and S. cerevisiae

“…The use of a single-chromosome transfer of chromosomes III, V, X, XII and XIII from the Carlsberg lager production yeast 244 to a haploid S. cerevisiae strain by cytoduction revealed at least two divergent genomes in S. carlsbergensis, one of which is derived from S. cerevisiae, while the origin of the second genome is less clear (Nilsson-Tillgren et al, 1983;Kielland-Brandt et al, 1995;Andersen et al, 1999). Based on the Southern hybridization experiments and sequencing of some non-S. cerevisiae genes, another brewing yeast S. monacensis CBS 1503 was proposed to be a second parent of the lager brewing yeast S. carlsbergensis (Pedersen, 1986;Hansen and Kielland-Brandt, 1994;Børsting et al, 1997). However, recent PCR and genetic analyses of single chromosomes showed that S. monacensis itself is an incomplete hybrid containing a part of the S. cerevisiae genome (Andersen et al, 1999).…”