Project description:Total RNA versus genomic DNA hybridization on custom arrays designed for all Saccharomyces bayanus genes Total RNA was collected in mid-log phase from Saccharomyces bayanus cells grown in rich medium (abbreviated CM, in house recipe). RNA was then converted to cDNA, Cy3-labeled and hybridized competitively against Cy5 labeled genomic DNA from Saccharomyces bayanus.
Project description:Hybrid progeny can enjoy increased fitness and stress tolerance relative to their ancestral species, a phenomenon known as hybrid vigor. Though this phenomenon has been documented throughout the Eukarya, evolution of hybrid populations has yet to be explored experimentally in the lab. To fill this knowledge gap we created a pool of Saccharomyces cerevisiae and S. bayanus homoploid and aneuploid hybrids, and then investigated how selection in the form of incrementally increased temperature or ethanol impacted hybrid genome structure and adaptation. During 500 generations of continuous ammonia-limited, glucose-sufficient culture, temperature was raised from 25C to 46??C. This selection invariably resulted in nearly-complete loss of the S. bayanus genome, although the dynamics of genome loss differed among independent replicates. Temperature-evolved isolates were significantly more thermal tolerant and exhibited greater phenotypic plasticity than parental species and founding hybrids. By contrast, when the same hybrid pool was subjected to increases in exogenous ethanol from 0% to 14%, selection favored euploid S. cerevisiae x S. bayanus hybrids. Ethanol-evolved isolates exhibited significantly greater ethanol tolerance relative only to S. bayanus and one of the founding hybrids tested. Adaptation to thermal and ethanol stress manifested as heritable changes in cell wall structure demonstrated by resistance to zymolyase or micafungin treatment. This is the first study to show experimentally that the fate of interspecific hybrids critically depends on the type of selection they encounter during the course of evolution.
Project description:This research work investigates the hybridization of the genes of Saccharomyces bayanus var. uvarum CECT 12600 and S. kudriavzevii IFO1802 employing S. cerevisiae microarrays.
Project description:Saccharomyces pastorianus is the yeast used to make lager beer; it is known to be an interspecific hybrid formed by the fusion between S. cerevisiae and S. bayanus genomes. This data set queries 17 S. pastorianus strains, collected at various times over the last 125 years from various breweries located in different geographical locations, which were obtained from CBS and DBVPG culture collections. The data in this set represent array-CGH experiments performed with these strains, using "2-species" custom Agilent arrays (the "2-species" arrays contain probes spaced every ~2 kb across the whole genomes of both S. cerevisiae and S. bayanus; the probes are unique and specific for each genome). The data set also contains 3 self-self hybridizations (S. cerevisiae + S. bayanus DNA mixed together in equimolar amounts, then labeled green or red in separate reactions, then hybridized to the "2-species" arrays) used for normalization in CGH-Miner analysis. A strain or line experiment design type assays differences between multiple strains, cultivars, serovars, isolates, lines from organisms of a single species.
