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Endophytic colonization of entomopathogenic fungi has garnered significant attention for its role in promoting plant growth. Specifically, I. cateniannulatahas shown a positive effect on the germination of F. tataricum (buckwheat) seeds, but the mechanisms underlying this promotion remain unclear. This study aimed to elucidate the process by which I. cateniannulatacolonize F. tataricumseeds, analyze the colonization effect of I. cateniannulata, and explorethe changes in antioxidant enzyme activity associated with this interaction. Additionally, the impactof seedlings grown after colonization on the population of T. urticaewas examined. The results demonstrated for the first time that I. cateniannulatacould successfully colonize germinating F. tataricumseeds during the seed imbibition and germination stages, either by forming dissolution zones with its spores or by germinating and forming mycelia. Complete colonization of all tissues of F. tataricumseeds was achieved within 16 h, with a preferential colonization rate observed as endosperm > embryo > seed coat. Furthermore, the colonization by I. cateniannulataenhanced peroxidase (POD) activity in the embryo and reduced malondialdehyde (MDA) content. Seedlings grown after colonization were also found to effectively reducethe number of eggs laid by T. urticae. These findings provide both theoretical insights and practical foundation for developing a symbiotic system between I. cateniannulataand F. tataricumseeds.