This chapter aims to provide the reader with a one-stop reference to the basic procedures needed to access, grow, store, mate, and sporulate yeast cells. It starts with an introduction to the Web-based yeast resources, which are becoming increasingly important in the investigation of S. cerevisiae in the post-genomic era. It then goes on to describe recipes for the different types of media and to explain how cells are grown to the appropriate cell numbers at the correct stage in the growth cycle. It also provides a detailed explanation on both short- and long-term storage of yeast cells. It ends by explaining how to set up genetic crosses with special advice on the demanding technique of diploid cell sporulation and spore isolation.
Mycophenolic acid inhibited the growth of Candida albicans. Cultures exposed to a concentration of 8.4 micrograms ml-1 mycophenolic acid were found to exhibit cell cycle arrest with two or more buds. Nuclear staining revealed that these were nucleate implying a possible defect in cytokinesis. The results are discussed in relation to the possible mode of action of mycophenolic acid.
~~ ~Saccharomyces cerevisiae was grown in a glucose-limited chemostat at dilution rates equivalent to mass doubling times of 2 to 16 h. Phase contrast microscopy revealed that cultures with mass doubling times greater than 4.5 h contained two sub-populations of cells, phase-dark and phaselight. The latter were shown to be predominantly unbudded daughter cells, which were not proliferating, and had properties similar to those of stationary phase cells in batch culture. Phase-dark cells were equivalent to cells growing exponentially in batch culture, The results suggest that the formation of phase-light cells represents cell cycle specific differentiation into stationary phase. The consequences of this phenomenon for the interpretation of results obtained with S. cerevisiae in the chemostat are discussed.
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