Herein, the influence of inter-pass cooling (IC) and conventional two-stage rolling (CTR), on microstructural evolution and plastic deformation behavior of ultra-heavy EH47 plates, is demonstrated. It is reported that the deformation amount and deformation rate, in every deformation pass during rough rolling, at 1/4- and 1/2-thickness of IC steel were higher than the CTR steel. The volume fraction of ferrite and acicular ferrite was 45% and 18%, at 1/4-thickness, and 35% and 50% at 1/2-thickness of IC steel, respectively, whereas the sum of both ferrite phases was smaller than 25% in the CTR steel. The austenite grain boundary area and high-angle grain boundary fraction in the IC steel were higher than the CTR steel. The high density of fine and shapeless pearlite has been observed in IC steel, whereas large-size carbides, with hexagonal structure, have been observed in CTR steel. Compared to the CTR steel, the density of precipitates was apparently lower in IC steel. Two kinds of Nb containing precipitates, such as (Ti, Nb)(C, N) and (Nb, Ti)C, were observed in the tested steels. Total ductility and uniform elongation of the IC steel were higher than the CTR steel. During the tensile process, the crack initiation energy and crack propagation energy of the IC steel were higher than the CTR steel. Moreover, the volume fraction of retained austenite (FCC) was reduced from 7.71% to 0.42% near the tensile fracture in IC steel at 1/4-thickness. In additon, the strain of synergetic plastic deformation of the IC steel was higher than the CTR steel. Meanwhile, compared to the CTR steel, the synergetic plastic deformation of the IC steel occurred at low stress after the yield point, which can be ascribed to the presence of fewer microcracks in the IC steel. Hence, a delayed fracture has been observed in the IC steel plate.