The effect of photosynthetically available radiation (PAR; 400–700 nm) and ultraviolet radiation (UVR; 280–400 nm) on marine bacterial heterotrophic activity was assessed monthly throughout a seasonal cycle in Blanes Bay (northwestern Mediterranean Sea). Seawater samples amended with 3 H-leucine were exposed to solar radiation under three radiation treatments: PAR + UVR (280–700 nm), PAR + UVA (320–700 nm), and PAR only. Parallel reference incubations in the dark and under a fixed artificial light source (PAR only) were also performed. Exposure to high UVR doses caused strong inhibition of 3 H-leucine incorporation rates (LIR), whereas natural PAR doses did not cause overall significant effects. Within UVR, UVA radiation accounted for most of the reduction in LIR, and this effect was modulated by the proportionality of the experimental light to the previous light exposure history of the samples. Constant (artificial) PAR-only exposure led to a general but seasonally variable increase in bacterial heterotrophic production compared to the dark controls, with large increases in spring and lower changes during summer. This pattern was likely caused by the stimulation of the bacterial group Gammaproteobacteria , which showed higher numbers of cells active in 3 H-leucine uptake after light exposure. Again, the previous light history of the samples seemed to partly explain the measured effects. Overall, our results show variable responses of bacterial activities to light manipulations, depending on seasonally changing light conditions and communities, and stress the importance of realistic simulation of light exposure conditions for ecosystem-relevant photobiological studies with microbial plankton.