Mutualisms as engines for rapid adaptation: Rhizobium evolution facilitates plant drought resistance

Mutualisms play critical roles in organismal stress tolerance; yet environmental stressors may simultaneously alter the evolution of the mutualism itself. Stress may have particularly strong impacts on the evolution of microbial mutualists due to their capacity for rapid genetic change. Here we used a legume-rhizobia mutualism, in which plants exchange carbon for symbiotically fixed nitrogen, to evaluate how mutualisms evolve in response to stressors. We experimentally evolved populations of Rhizobium leguminasarum in a full factorial design, manipulating drought and nitrogen. We quantified genomic changes in Rhizobium populations as well as their quality as partners with their plant host, Trifolium repens. Drought selected for context- dependent stress benefits to the host; drought-adapted Rhizobium strains provided increased benefits to the host under drought, but fewer benefits to the host in well-watered environments. Conversely, nitrogen fertilization selected for decreased Rhizobium partner quality. Comparative genomics indicated that selection on standing structural variants along the symbiotic plasmid may underpin these drought benefits, specifically along genes associated with desiccation tolerance. These results suggest that stress can expand the benefits rhizobia provide to their legume hosts beyond nitrogen fixation, with rapid symbiont evolution an engine in promoting adaptive plant phenotypes. SIGNIFICANCE STATEMENTWhile mutualisms are fundamental to ecosystem functioning, they are often assumed to break down under environmental stress. We showed the opposite can happen. Here, we factorially manipulated drought stress and nitrogen fertilization and experimentally evolved a model legume-rhizobia mutualism, where rhizobial-bacteria trade nitrogen for plant carbon. When droughted, the symbiotic bacteria rapidly evolved new traits that made them better partners under drought, improving plant growth within just a few generations. Evolution occurred mainly on a specialized bacterial plasmid, a mobile piece of DNA distinct from the main genome. By contrast, added nitrogen fertilizer led to decreases in bacterial benefits. These findings suggest that mutualisms are not simply fragile in the face of global change, but can expand mutualistic benefits to buffer partners against stress. CLASSIFICATIONBiological Sciences, Evolution