Kusatsu-Shirane volcano has been a particular study field for hydrothermal system and phreatic eruptions with plenty of thermal springs, fumaroles, and a crater lake of Yugama. On 23 January 2018, a phreatic eruption occurred at the Motoshirane cone of Kusatsu-Shirane, where no considerable volcanic activity had been reported in observational and historical records. To understand the eruption process of such a unique event, we examined observed seismic, tilt, and infrasound records. The onset of surface activity accompanied by infrasound signal was preceded by volcanic tremor and inflation of the volcano for 2 minutes. Tremor signals with a frequency of 5–20 Hz remarkably coincide with the rapid inflation. We apply an amplitude source location method to seismic signals in the 5–20 Hz band to estimate tremor source locations. Our analysis locates tremor sources at 1 km north of Motoshirane and at a depth of 0.5–1 km from the surface. Inferred source locations correspond a conductive layer of impermeable cap-rock estimated by magnetotelluric investigations, and an upper portion of the seismogenic region, suggesting hydrothermal activity hosted beneath the cap-rock. Examined seismic signals in the 5–20 Hz band are typically excited by volcano-tectonic events with faulting mechanism. Based on the above characteristics and background, we interpret that excitation of examined volcanic tremor reflects small shear fractures induced by sudden hydrothermal fluid injection to the cap-rock layer. The horizontal distance of 1 km between inferred tremor sources and Motoshirane implies lateral migration of the hydrothermal fluid, although we have not obtained direct evidence. Kusatsu-Shirane has a series of unrest at the Yugama lake since 2014. However, inferred tremor source locations do not overwrap active seismicity beneath Yugama. Therefore, our result suggests that the 2018 eruption was triggered by hydrothermal fluid injection through an independent pathway that has driven unrest activities at Yugama.