Novel breeding method, matα2-PBT, to construct isogenic series of polyploid strains of Saccharomyces cerevisiae

How ploidy is determined in organisms is an important issue in bioscience. Polyploidy is believed to be relevant to useful traits of domesticated plants and microorganisms. As such, polyploidy is central to many applications in biotechnology. However, studies of polyploidy are poorly advanced because no methodologies to construct desired polyploid have been developed for any organism. Herein we describe the development of a novel breeding technology, matα2-PBT, to generate polyploid strains of Saccharomyces cerevisiae. S. cerevisiae has two mating types, a and α, determined by MATa and MATα gene each of which consists of a1 and a2 and α1 and α2 cistrons. This novel technology exploits an interesting feature of a specific mutation, matα2-102, in the MATα2 gene. Unlike the MATα wild-type strain, which gives a non-mating phenotype when mated with MATa cells, the matα2-102 strain confers an α mating-type to a-type strains when mated with a-type strains. We constructed plasmid with the cloned matα2-102 mutant gene. An a-type cells harboring this plasmid displayed an α mating-type and mated with a-type cells. Because the resultant hybrid displays an α mating-type, it can mate again with a-type cells. By repeating this procedure, we have constructed an isogenic series of haploid to tetraploid of S. cerevisiae. Although whether even higher polyploid than tetraploid can be constructed by using this technology remains to be determined in the future, we believe that it became possible for the first time with matα2-PBT method to investigate whether higher polyploid than tetraploid can be constructed.