Exponential cells of the Saccharomyces cerevisiae tps1 mutant underwent a rapid loss of viability upon a non-lethal heat exposure (from 28 to 42 degrees C). However, a further more severe heat stress (52.5 degrees C 5 min) induced an increase in the fraction of viable cells. This mutant can not synthesize trehalose either at 28 degrees C or at 42 degrees C due to the lack of a functional trehalose-6P synthase complex. In control experiments carried out with the wild-type W303-1B, heat-stressed exponential phase cultures grown on YPgal at 28 degrees C acquired thermotolerance to a higher extent than identical cultures grown on YPD, although in both cultures the level of stored trehalose was negligible. These data suggest that the bulk of trehalose accumulated in yeast upon mild heat treatments is not sufficient to account for the acquisition of thermotolerance.