Updated publication reference for PubMed record(s): 31708949. Plant cultivation on spacecraft or planetary outposts is a promising and actual perspective both for food and bioactive molecules production. To this aim, plants response to ionizing radiations, as an important component of space radiation, must be assessed through on-ground experiments due to the potentially fatal effects on living systems. Hereby we investigated the effects of X-rays and -rays exposure on tomato ‘hairy root’ cultures (HRCs), which represent a solid platform for the production of pharmaceutically relevant molecules, including metabolites and recombinant proteins. In a space application perspective, we used a HRC system previously fortified through the accumulation of anthocyanins, which are known for their anti-oxidant properties. Roots were independently exposed to different photon radiations, namely X-rays (250 kV) and -rays (Co60, 1.25 MeV), both at the intensity of 0.5, 5 and 10 Gy. Molecular changes induced in the proteome of HRCs were investigated by a comparative approach based on Two-Dimensional Difference In-Gel electrophoresis (2D-DIGE) technology and mass spectrometry.