Understanding microbial adaptations to the extreme conditions of space is crucial for both astronaut health and the integrity of spacecraft materials. This study comparatively analyses the cosmic radiation resistance and growth responses to simulated microgravity (SMG) of a wild-type strain and an International Space Station (ISS) isolate of Penicillium rubens. Resistance to helium- and iron-ion radiation was determined, alongside growth under SMG using clinorotation. The results revealed that the ISS isolate exhibited higher resistance to both helium- and iron-ion radiation than the wild-type strain, suggesting adaptive mechanisms that enhance survival in space environments. Additionally, while the ISS isolate demonstrated significantly increased growth in SMG compared to normal gravity conditions, the wild-type strain showed no difference between the two conditions. These findings indicate that prolonged exposure to the space environment may select for traits that enhance resistance to cosmic radiation and alter growth dynamics under microgravity. Such adaptations could have implications for microbial monitoring in space habitats, planetary protection policies, and potential biotechnological applications in space. Further investigations into the genetic and metabolic differences between both strains may provide deeper insights into fungal adaptation to space environments.