A common contaminant shifts impacts of climate change on a plant-microbe mutualism: effects of temperature, CO 2 and leachate from tire wear particles

Abstract Anthropogenic stressors, such as climate change or chemical pollution, affect individual species and alter species interactions. Moreover, species interactions can modify effects of anthropogenic stressors on interacting species - a process which may vary amongst stressors or stressor combinations. Most ecotoxicological work focuses on single stressors on single species. Here, we test hypotheses about multiple stressors (climate change and tire wear particles) and interacting species, and whether species interactions modify responses. We use duckweed and its microbiome to model responses of plant-microbe interactions. Climate change is occurring globally, and with increasing urbanization, tire wear particles increasingly contaminate road runoff. Their leachate is associated with zinc, PAHs, plastic additives, and other toxic compounds. We crossed perpendicular gradients of temperature and CO 2 in a well plate with factorial manipulation of leachate from tire wear particles and presence of duckweed microbiomes. We measured duckweed and microbial growth, duckweed greenness, and plant-microbe growth correlations. We found that tire leachate and warmer temperatures enhanced duckweed and microbial growth, but microbes diminished positive responses in duck-weed, meaning microbiomes became costly for duckweed. These costs of microbiomes were less-than-additive with warming and leachate, and might be caused by leachate-disrupted endocrine signaling in duckweed. We observed reduced greenness at higher CO 2 without tire leachate, suggesting a relative increase in plant nutrient demand, and possibly underlying positive plant-microbe growth correlations in these conditions, as microbes presumably increase nutrient availability. However, with tire leachate, growth correlations were never positive, and shifted negative at lower CO 2 , further suggesting leachate favors mutualism disruption. In summary, while individual stressors of global change can affect individual species, in ecology we know species interact; and in ecotoxicology, we know stressors interact. Our results demonstrate this complexity: multiple stressors can affect species interactions, and species interactions can alter effects of multiple stressors.