The effect of the intercritical temperature on the microstructure and mechanical properties of a newly developed quenching and partitioning steel using martensitic microstructure prior to the heat treatment process was studied. Such a quenching and partitioning process possessed a unique microstructure evolution, especially during intercritical annealing after prequenching. Excellent mechanical properties were obtained due to this unique multiphase microstructure. Significant amount of interlath-retained austenite was acquired and the relationship between the microstructure and work-hardening behaviors was proposed. The martensite/austenite islands increased at elevated annealing temperature, which deteriorated the total elongation and increased the tensile strength as hard constituents when it was excessive. The result indicated that the present full martensitic microstructure before the intercritical annealing is probably more suitable to an industrial application and is a better way to produce high strength steels with suitable ductility.