Our and literature studies indicated that systemic acquired resistance (SAR) is effectively inducible in greenhouse, and certain artificial light sources cause non-optimal growth of tobacco plants. Therefore, the morphological characteristics, local and systemic resistance response of N. tabacum cv. ‘Xanthi’ nc plants (harbouring NN resistance genes) to tobacco mosaic virus (TMV) infection under three artificial light sources with different spectral distribution were compared with greenhouse conditions. Statistical analysis of data was carried out by R package (R Core Team, 2015). Generally, artificial light sources (especially fluorescent tube and halogen lamp) decreased the local resistance response and caused substantial morphological and developmental differences as compared to greenhouse conditions when plants were kept during their entire life (lifelong experimental regime) under these conditions. On the contrary, no or much less differences were found when plants were transferred from greenhouse to artificial light sources only at six leaf stage (short experimental regime). While induction of SAR frequently decreased TMV lesion size by about 50-60% under greenhouse conditions, two of the three artificial light sources, fluorescent tube and halogen lamp were substantially and significantly less effective under short experimental regime conditions (25-35%). A metal halide light source with similarity to sunshine’s spectral distribution, however, partially mimicked the effect of greenhouse conditions indicating the importance of light spectrum among other factors in SAR induction and prevention of distorted growth of plants. Consequently, the optimization of the effect of artificial light sources is an important factor in experimental design studying signal transduction and biochemistry of SAR.