Abstract
A general method to convert homothallic strains of the yeast Saccharomyces cerevisiae to heterothallism is described which is applicable to genetically well-behaved diploids, as well as to strains that sporulate poorly or produce few viable and mating-competent spores. The heterothallic (ho) allele was introduced into three widely used wine strains through spore × cell hybridization. The resultant hybrids were sporulated, and heterothallic segregants were isolated for use in successive backcrosses. Heterothallic progeny of opposite mating type and monosomic for chromosome III produced by sixth-backcross hybrids or their progeny were mated together to reconstruct heterothallic derivatives of the wine strain parents. A helpful prerequisite to the introduction of ho was genetic purification of the parental strains based on repeated cycles of sporulation, ascus dissection, and clonal selection. A positive selection to isolate laboratory-wine strain hybrids requiring no prior genetic alteration of the industrial strains, coupled with a partial selection to reduce the number of spore progeny needed to be screened to isolate heterothallic segregants of the proper genotype made the procedure valuable for genetically intractable strains. Trial grape juice fermentations indicated that introduction of ho had no deleterious effect on fermentation behavior.
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- Campbell D., Doctor J. S., Feuersanger J. H., Doolittle M. M. Differential mitotic stability of yeast disomes derived from triploid meiosis. Genetics. 1981 Jun;98(2):239–255. doi: 10.1093/genetics/98.2.239. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Dawes I. W., Hardie I. D. Selective killing of vegetative cells in sporulated yeast cultures by exposure to diethyl ether. Mol Gen Genet. 1974;131(4):281–289. doi: 10.1007/BF00264859. [DOI] [PubMed] [Google Scholar]
- Harashima S., Nogi Y., Oshima Y. The genetic system controlling homothallism in Saccharomyces yeasts. Genetics. 1974 Aug;77(4):639–650. doi: 10.1093/genetics/77.4.639. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hicks J. B., Herskowitz I. Interconversion of Yeast Mating Types I. Direct Observations of the Action of the Homothallism (HO) Gene. Genetics. 1976 Jun;83(2):245–258. doi: 10.1093/genetics/83.2.245. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hisatomi T., Yanagishima N., Ban-no I. Induction of heterothallic strains and their genetic and physiological characterization in a homothallic strain of the yeast Saccharomyces exiguus. Curr Genet. 1986;10(12):887–892. doi: 10.1007/BF00398285. [DOI] [PubMed] [Google Scholar]
- JOHNSTON J. R., MORTIMER R. K. Use of snail digestive juice in isolation of yeast spore tetrads. J Bacteriol. 1959 Aug;78:292–292. doi: 10.1128/jb.78.2.292-292.1959. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kostriken R., Heffron F. The product of the HO gene is a nuclease: purification and characterization of the enzyme. Cold Spring Harb Symp Quant Biol. 1984;49:89–96. doi: 10.1101/sqb.1984.049.01.012. [DOI] [PubMed] [Google Scholar]
- Mortimer R. K., Schild D. Genetic map of Saccharomyces cerevisiae, edition 9. Microbiol Rev. 1985 Sep;49(3):181–213. doi: 10.1128/mr.49.3.181-213.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Oshima T., Takano I. Mutants Showing Heterothallism from a Homothallic Strain of SACCHAROMYCES CEREVISIAE. Genetics. 1980 Apr;94(4):841–857. doi: 10.1093/genetics/94.4.841. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Parry E. M., Cox B. S. The tolerance of aneuploidy in yeast. Genet Res. 1970 Dec;16(3):333–340. doi: 10.1017/s0016672300002597. [DOI] [PubMed] [Google Scholar]
- Romano P., Soli M. G., Suzzi G., Grazia L., Zambonelli C. Improvement of a Wine Saccharomyces cerevisiae Strain by a Breeding Program. Appl Environ Microbiol. 1985 Oct;50(4):1064–1067. doi: 10.1128/aem.50.4.1064-1067.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shaffer B., Brearley I., Littlewood R., Fink G. R. A stable aneuploid of Saccharomyces cerevisiae. Genetics. 1971 Apr;67(4):483–495. doi: 10.1093/genetics/67.4.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Spencer J. F., Spencer D. M. Genetic improvement of industrial yeasts. Annu Rev Microbiol. 1983;37:121–142. doi: 10.1146/annurev.mi.37.100183.001005. [DOI] [PubMed] [Google Scholar]
- Spencer J. F., Spencer D. M. The use of mitochondrial mutants in the isolation of hybrids involving industrial yeast strains. Mol Gen Genet. 1980 Jan;177(2):355–358. doi: 10.1007/BF00267451. [DOI] [PubMed] [Google Scholar]