In this study, fatigue limit of ductile cast iron is evaluated based on phenomena of the microscopic observation, such as matrix structure, spheroidal ratio, size of graphite and distribution. Three different ferrite-pearlite matrix structure, GCD 45-), GCD 50, GCD 60 series, all of which contain more than 70% spheroidal ratio of graphite, were used to obtain the correlation between maximum size of graphite and fatigue strength. It was concluded as fellows. (1) In Ductile cast iron of ferrite-pearlite matrix, the fatigue limit of GCD 60 series with 73% pearlite structure was the highest. (2) From observation of the starting point of crack of all specimens, it is noted that the crack initiates, in graphite, goes through ferrite and propagates into pearlite. (3) A good quality of Ductile cast iron used in this experiment can be checked from uniformly distributed graphite. The negligible interaction effect between graphites was verified by microscopic observation and fracture mechanics investigation in surface and interior of the specimen.