Influence of mean stress on fatigue life of the austenitic stainless steel 316 L in air and light water environments (boiling water reactor/hydrogen water chemistry) at 288 °C was determined with a series of tests carried out in load-control mode. Fatigue life was found to increase with application of compressive and tensile mean stress in air and light water reactor environments. Secondary hardening was regarded as the main reason for this behavior. A modified Smith-Watson-Topper (SWT) model was considered to account for mean stress and was shown to predict fatigue life accurately in air and water environments. The reduction of fatigue life in water environment, determined with the SWT curves, was about 2.5. Observations of the end-of-life dislocation arrangements by transmission electron microscopy showed that the dislocation microstructure depends essentially on plastic strain amplitude, which in turn is strongly correlated to stress amplitude and mean stress. The microstructures were found consistent with those usually observed after strain-controlled experiments. At rather low plastic strain amplitudes, corduroy structure consisting of small dislocation loops was observed. Acting as significant obstacle to dislocation motion, corduroy structure affects overall dislocation mobility therefore contributing to notable secondary cyclic hardening.