RDJLΦ1 is a marine siphophage infecting Roseobacter denitrificans OCh114. In this study, host responses of R. denitrificans OCh114 to phage infection were investigated through in situ real-time atomic force microscopy (AFM) and proteomics approaches. As seen from the AFM observations, during phage infection processes, depression areas appeared on the host cell surface in a few minutes after infection and expanded in both diameter and depth over time and finally led to the collapse of host cells within 30 min. The two-dimensional polyacrylamide gel electrophoresis revealed significant changes in the proteomic composition of the host cells during infection. The expression of 91 proteins, including some involved in DNA transcription regulation and substrate transportation, was changed with at least twofold up- or downregulation as compared to the control without phage infection. This observed rapid lysis of host cells and the great changes in protein expression caused by phage infection added more perspectives to the documented important roles of viruses in mediating carbon cycling in the ocean.