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Competition for root colonization among ectomycorrhizal fungi is well documented, but the mechanisms determining competitive outcomes are not clearly understood. In a previous study, we observed that timing of colonization (i.e., a priority effect) had a significant effect on the outcome of competition between two ectomycorrhizal (EM) fungi in the genus Rhizopogon. In this study, we explicitly tested the role of priority effects in competition among EM fungi by experimentally manipulating the timing of colonization of four Rhizopogon species on Pinus muricata seedlings. In a first experiment, we set up 12 two‐species combinations, in which seedlings were first inoculated from spores with one species, grown for three months, and then inoculated with an equal density of spores of a second species and grown for an additional three months. Root tip occupation in the two‐species treatments was determined by …