title: Efficient engineering of marker-free synthetic allotetraploids of Saccharomyces.
abstract: Saccharomyces interspecies hybrids are critical biocatalysts in the fermented beverage industry, including in the production of lager beers, Belgian ales, ciders, and cold-fermented wines. Current methods for making synthetic interspecies hybrids are cumbersome and/or require genome modifications. We have developed a simple, robust, and efficient method for generating allotetraploid strains of prototrophic Saccharomyces without sporulation or nuclear genome manipulation. S. cerevisiae×S. eubayanus, S. cerevisiae×S. kudriavzevii, and S. cerevisiae×S. uvarum designer hybrid strains were created as synthetic lager, Belgian, and cider strains, respectively. The ploidy and hybrid nature of the strains were confirmed using flow cytometry and PCR-RFLP analysis, respectively. This method provides an efficient means for producing novel synthetic hybrids for beverage and biofuel production, as well as for constructing tetraploids to be used for basic research in evolutionary genetics and genome stability.
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States; DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53706, United States.
authors.forename: William G
authors.initials: WG
authors.lastname: Alexander
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States; DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53706, United States.
authors.forename: David
authors.initials: D
authors.lastname: Peris
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States.
authors.forename: Brandon T
authors.initials: BT
authors.lastname: Pfannenstiel
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States.
authors.forename: Dana A
authors.initials: DA
authors.lastname: Opulente
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States; Graduate Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, WI 53706, United States.
authors.forename: Meihua
authors.initials: M
authors.lastname: Kuang
authors.affiliation: Laboratory of Genetics, Genome Center of Wisconsin, Wisconsin Energy Institute, J. F. Crow Institute for the Study of Evolution, University of Wisconsin-Madison, Madison, WI 53706, United States; DOE Great Lakes Bioenergy Research Center, University of Wisconsin-Madison, Madison, WI 53706, United States; Graduate Program in Cellular and Molecular Biology, University of Wisconsin-Madison, Madison, WI 53706, United States. Electronic address: cthittinger@wisc.edu.
authors.forename: Chris Todd
authors.initials: CT
authors.lastname: Hittinger
mesh_terms: D001515:Beer; D056804:Biofuels; D019149:Bioreactors; D004271:DNA, Fungal / Q000235:genetics; D005285:Fermentation / Q000235:genetics; D005818:Genetic Engineering / Q000379:methods*; D016681:Genome, Fungal; D006824:Hybridization, Genetic; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D012440:Saccharomyces / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics; D057891:Tetraploidy; D014920:Wine
chemical_list: D056804:Biofuels; D004271:DNA, Fungal
keywords: Biofuels; Brewing; Hybrids; Prototrophic; Saccharomyces; Synthetic zymurgy