25Bacterial-fungal interactions are widespread in nature and there is a growing number of 26 studies reporting distinct fungus-associated bacteria. However, little is known so far about how 27 shifts in the fungus-associated bacteriome will affect the fungal host's lifestyle. In the present 28 study, we describe for the first time the bacterial community associated with the saprotrophic 29 fungus Mucor hiemalis, commonly found in soil and rhizosphere. Two broad-spectrum 30 antibiotics that strongly altered the bacterial community associated with the fungus were 31 applied. Our results revealed that the antibiotic treatment did not significantly reduce the 32 amount of bacteria associated to the fungus but rather changed the community composition 33 by shifting from initially dominating Alpha-Proteobacteria to dominance of Gamma-34
Proteobacteria. A novel approach was applied for the isolation of fungal-associated bacteria 35which also revealed differences between bacterial isolates obtained from the original and the 36 antibiotic-treated M. hiemalis. The shift in the composition of the fungal-associated bacterial 37 community led to significantly reduced fungal growth, changes in fungal morphology, behavior 38 and secondary-metabolites production. Furthermore, our results showed that the antibiotic-39 treated isolate was more attractive and susceptible to mycophagous bacteria as compared to 40 the original isolate. Overall, our study highlights the importance of the fungus-associated 41 bacteriome for the host's lifestyle and interactions and indicate that isolation with antibacterials 42 is not sufficient to eradicate the associated bacteria. 43 44 3