Chromophoric dissolved organic matter (CDOM), the optically active fraction of dissolved organic matter, is primarily generated by pelagic organisms in the open ocean. In this study, we experimentally determined the quantity and spectral quality of CDOM generated by bacterioplankton using two different substrates (with and without photoproducts) and by Antarctic krill Euphausia superba and evaluated their potential contributions to CDOM dynamics in the peninsular region of the Southern Ocean. CDOM was generated by bacteria in all experiments, and the presence of photoproducts influenced both the quantity and the spectral quality of the resultant CDOM. We confirmed a direct link between bacterial production and CDOM generation, which yielded in situ CDOM duplication times from 31 to 33 d. Antarctic krill as a direct source of CDOM was also confirmed experimentally. We estimated that CDOM generation by krill would lead to CDOM duplication times from 0.48 to 0.80 d within krill swarms. Our findings highlight the potential significance of bacteria and Antarctic krill swarms in the generation of CDOM and underscore the dynamic nature of CDOM in this area.