We studied the relationships between assemblages of soil microfungi and plant communities in the southern boreal mixed-wood forests of Quebec. Sampling took place in 18,100 m2 plots from an existing research site. Plots were separated into three categories based on dominant overstory tree species: (i) trembling aspen, (ii) white birch and (iii) a mixture of white spruce and balsam fir. Within each plot a 1 m2 subplot was established in which the understory herbaceous layer was surveyed and soil cores were collected. Microfungi were isolated from soil cores with the soil-washing technique and isolates were identified morphologically. To support our morphological identifications DNA sequences were obtained for the most abundant microfungi. The most frequently occurring microfungal species were Penicillium thomii, P. spinulosum, P. janthinellum, Penicillium sp., P. melinii, Trichoderma polysporum, T. viride, T. hamatum, Mortierella ramanniana, Geomyces pannorum, Cylindrocarpon didymum, Mortierella sp. and Mucor hiemalis. Multivariate analyses (redundancy analysis followed by variance partitioning) revealed that most of the variation in microfungal communities was explained by understory plant species composition as opposed to soil chemistry or overstory tree species. In this floristically diverse system saprophytic microfungal assemblages were not correlated with the overstory tree species but were significantly correlated with the main understory herbs, thereby reflecting differences at a smaller spatial scale.