One experiment to rule them all? Testing multiple drivers of the temperature-size rule with nonlinear temperature increase

The temperature-size rule (TSR) describes the inverse relationship between organism size and environmental temperature in uni- and multicellular species. Despite the TSR being widespread, the mechanisms for shrinking body size with warming remain elusive. Here, we experimentally test three hypotheses (differential development and growth [DDG], maintain aerobic scope and regulate oxygen supply [MASROS] and the supply-demand hypothesis [SD]) potentially explaining the TSR using the aquatic protist Colpidium striatum in three gradually changing and one constant temperature environment crossed with three different nutrient levels. We find that the constant and slowly warming environments show similar responses in terms of population dynamics, whereas populations with linear and fast warming quickly decline and show a stronger temperature-size response. Our analyses suggest that acclimation may have played a role in observing these differences among treatments. The SD hypothesis is most parsimonious with the data, however, neither the DDG nor the MASROS hypothesis can be firmly dismissed. We conclude that the TSR is driven by multiple ecological and acclimatory responses and hence multicausal. Author statementA.T. designed and led the experiment, and A.T. and A.G. performed the sampling. A.T. and F.P. analyzed and interpreted the data. All authors contributed to the writing of the manuscript.