dDiurnal temperature cycling is an intrinsic characteristic of many exposed microbial ecosystems. However, its influence on yeast physiology and the yeast transcriptome has not been studied in detail. In this study, 24-h sinusoidal temperature cycles, oscillating between 12°C and 30°C, were imposed on anaerobic, glucose-limited chemostat cultures of Saccharomyces cerevisiae. After three diurnal temperature cycles (DTC), concentrations of glucose and extracellular metabolites as well as CO 2 production rates showed regular, reproducible circadian rhythms. DTC also led to waves of transcriptional activation and repression, which involved one-sixth of the yeast genome. A substantial fraction of these DTC-responsive genes appeared to respond primarily to changes in the glucose concentration. Elimination of known glucose-responsive genes revealed an overrepresentation of previously identified temperature-responsive genes as well as genes involved in the cell cycle and de novo purine biosynthesis. Indepth analysis demonstrated that DTC led to a partial synchronization of the cell cycle of the yeast populations in chemostat cultures, which was lost upon release from DTC. Comparison of DTC results with data from steady-state cultures showed that the 24-h DTC was sufficiently slow to allow S. cerevisiae chemostat cultures to acclimate their transcriptome and physiology at the DTC temperature maximum and to approach acclimation at the DTC temperature minimum. Furthermore, this comparison and literature data on growth rate-dependent cell cycle phase distribution indicated that cell cycle synchronization was most likely an effect of imposed fluctuations of the relative growth rate (/ max ) rather than a direct effect of temperature.T emperature directly influences the kinetics of all biochemical reactions and many cellular processes (1, 2). Within their temperature range for growth, many microorganisms adapt their metabolic networks and biomass composition to optimize their growth rate and viability in response to changing temperatures. Classical examples of temperature adaptation include modifications of membrane fluidity (3, 4) and the expression of heat shock proteins that assist in protein folding at high temperatures (5, 6).Laboratory studies on microbial temperature responses are based largely on two experimental systems. Acclimation, i.e., the result of complete physiological adaptation to a given temperature, has been studied both in batch cultures and in chemostats (7-9). In cold shock and heat shock experiments, the responses of microorganisms to sudden upshifts or downshifts in temperature are studied, typically over a time period ranging from a few minutes to a few hours (3, 10-13).In natural environments, microorganisms are subjected to several types of temperature dynamics. In exposed environments, one of the dominant aspects of temperature dynamics is related to circadian changes, with higher temperatures during the day and lower temperatures during the night. These 24-h temperature cycles, which are superimp...