Biofumigation has been proposed as an environmentally friendly method of plant protection against soil-borne pathogens, but its effects on microbial communities are still incompletely understood. Using high throughput DNA sequencing, we investigated the effects of oilseed radish residues on the root fungal microbiome of strawberry in the presence of a soil-borne fungal pathogen, Verticillium dahliae. Results of our greenhouse study show that early flowering occurred in response to residue addition, suggesting a plant stress-response and there was a significant decrease in berry yield. The fungal microbiome of roots was significantly restructured by both biofumigation and inoculation with Verticillium. In particular, the abundance of root endophyte- and arbuscular mycorrhizal functional guilds was reduced significantly as a result of biofumigant and V. dahliae addition, whereas the abundance of saprotrophs increased significantly when both treatments were applied together. Alpha diversity analyses of fungi associated with roots indicated a significant increase in species richness following Verticillium inoculation, whereas the biofumigant alone or in the presence of V. dahliae resulted in no significant effect, suggesting that apparently some rare taxa may have been enriched/stimulated in the presence of the pathogen. Further investigations should reveal whether negative effects of biofumigation on potentially beneficial root associated endophytes and arbuscular mycorrhizal fungi are host genotype- or soil-dependent.