This study investigated the response of sweetfish (Plecoglossus altivelis), a species that migrates dynamically throughout a river basin, to two flood events during the summer of 2020 in the Nagara River, located in the central region of Japan. By combining multiple environmental DNA (eDNA) surveys and hydrological modeling, the spatiotemporal distribution of P. altivelis throughout a mountainous river basin was captured and analyzed. The eDNA concentrations at 42 sites in the Nagara River Basin were analyzed five times from August to early October 2020. In addition, Rainfall–Runoff–Inundation model calculations were performed using 1-km resolution precipitation data as input values to analyze the magnitude of the flood disturbance at the eDNA sites. The daily specific discharge Qs (m3/s/km2) was employed as an index of the flood magnitude. The calculation period included Flood Events 1 and 2 with 52 and 38 days of precipitation reaching 1923 and 528 mm, respectively. The results of the eDNA analysis showed that, immediately after Flood Event 1, the spatial distribution of P. altivelis was unevenly distributed in the upper reaches of the Nagara River and some of its tributaries. Subsequently, the distribution expanded to the entire mainstem. The distribution of the maximum daily specific discharge suggested that the river segments with high eDNA concentrations of P. altivelis immediately after the high-magnitude flood event were those with a relatively low intensity of flood disturbance compared with those in the other connected river segments and tributaries. The results of this study indicate that the resilience of riverine communities to extreme floods is supported by the continuity and connectivity between the mainstem and its tributaries in mountainous river basins.