In this study, the influence of distinct hydraulic retention times (HRT) and organic loading rates (OLRs) on fungal dynamics during food waste anaerobic digestion in immersed membrane-based bio-reactors (iMBR) were investigated. The organic loading rate 4–8 g VS/L/d (R1) and 6–10 g VS/L/d (R2) were set in two iMBR. T1 (1d), T2 (15d) and T3 (34d) samples collected from each bioreactor were analyzed fungal community by using 18s rDNA. In R2, T2 had the most abundant Ascomycota, Basidiomycota, Chytridiomycota and Mucoromycota. As for R1, T3 also had the richest Cryptomycota except above four kinds of fungi. Subsequently, the Principal Component Analysis (PCA) and Non-Metric Multi-Dimensional Scaling (NMDS) indicated that fungal diversity was varied among the all three phases (T1, T2, and T3) and each treatment (R1 and R2). Finally, the results showed that different OLRs and HRT have significantly influenced the fungal community.