The evolution of thermal performance curves in fungi farmed by attine ant mutualists in above-ground or below-ground microclimates.

Fungi are abundant and ecologically important at a global scale, but little is known about whether their thermal adaptations are shaped by biochemical constraints (i.e., the hotter is better model) or evolutionary tradeoffs (i.e., the specialist-generalist model). We tested these hypotheses by generating thermal performance curves of fungal cultivars farmed by six species of Panamanian fungus-farming "attine" ants. These fungi represent evolutionary transitions in farming strategies, as four cultivars are farmed by ants below ground at stable temperatures near 25 °C and two cultivars are farmed above ground at variable temperatures. We generated thermal performance curves using a common garden experiment confining fungal isolates to different temperatures and then used a Bayesian hierarchical modelling approach to compare competing temperature sensitivity models. Some thermal performance traits differed consistently across farming strategies, with above-ground cultivars having: (1) higher tolerance to low temperatures (CTLmin) and (2) higher maximum growth rate at the optimal temperature (rmax). However, two core assumptions shared by the hotter is better model or specialist-generalist model were not supported as above-ground cultivars did not show systematic increases in either their optimal temperature (Topt) or thermal tolerance breadth. These results harness ant farming systems as long-term natural experiments to decouple the effects of environmental thermal variation and innate physiological temperature sensitivity on fungal thermal evolution. The results have clear implications for predicting climate warming-induced breaking points in animal-microbe mutualisms.