The antioxidant balance, photochemical activity of photosystem II (PSII), and photosynthetic pigment content, as well as the expression of genes involved in the light signalling of callus lines of Eutrema salsugineum plants (earlier Thellungiella salsuginea ) under different spectral light compositions were studied. Growth of callus in red light (RL, maximum 660 nm), in contrast to blue light (BL, maximum 450 nm), resulted in a lower H 2 O 2 content and thiobarbituric acid reactive substances (TBARS). The BL increased the activities of key antioxidant enzymes in comparison with the white light (WL) and RL and demonstrated the minimum level of PSII photochemical activity. The activities of catalase (CAT) and peroxidase (POD) had the highest values in BL, which, along with the increased H 2 O 2 and TBARS content, indicate a higher level of oxidative stress in the cells. The expression levels of the main chloroplast protein genes of PSII ( PSBA and PSBD ), the NADPH-dependent oxidase gene of the plasma membrane ( RbohD ), the protochlorophyllide oxidoreductase genes ( POR B, C ) involved in the biosynthesis of chlorophyll, and the key photoreceptor signalling genes ( CIB1, CRY2, PhyB, PhyA , and PIF3 ) were determined. Possible mechanisms of light quality effects on the physiological parameters of callus cells are discussed.