Proposal of INTRODUCTIONZygosaccharomyces bailii is a widely distributed yeast species that is often associated with food spoilage, particularly of acidified, preserved foods containing high concentrations of fermentable sugars (Thomas & Davenport, 1985;Cole & Keenan, 1987;Makdesi & Beuchat, 1996;James & Stratford, 2011). The yeast has been proposed as a new host for several biotechnological processes (e.g. Branduardi et al., 2004) due to its ability to tolerate such environments at relatively high temperatures, which could improve the efficiency of these processes under restrictive conditions. Moreover, the high growth rate of Z. bailii and its high biomass yield make this yeast particularly attractive for heterologous protein and metabolite production (e.g. Sousa et al., 1996Sousa et al., , 1998.Despite their well-known role in food/beverage spoilage, accurate identification of Z. bailii and related yeasts to the species level using conventional taxonomic tests remains problematic. An inability to ferment and assimilate many of the carbon compounds typically used in yeast identification, as well as ambiguous tests results due to strain variability, often hampers identification (James & Stratford, 2011). Furthermore, significant intraspecific variation in internal transcribed spacer (ITS) sequences was also reported among some strains of the species (James et al., 1996), which may cause difficulties for the use of this barcode region for identifying the species (Schoch et al., 2012). We hypothesized that polyphasic analyses of the yeasts encompassed by Zygosaccharomyces bailii sensu lato may lead to a more accurate understanding of their phylogenetic relationship and taxonomic status. Here we report the molecular, physiological and morphological characterization of these yeasts, and propose two novel species near Z. bailii. METHODSYeast strains and characterization. Strains of Z. bailii sensu lato and related taxa were selected from the ATCC Mycology Collection or were provided by bioMérieux, Inc. (Table 1). Morphological observations and metabolic tests comprising the yeast standard Abbreviations: ITS, internal transcribed spacer; LSU, large subunit; SSU, small subunit.
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