Cyanobacteria can grow as biofilms, communities that colonize surfaces and that play a fundamental role in the ecology of many diverse habitats and in the conversion of industrial production to green platforms. Although biofilm growth is known to be significantly affected by several characteristics, the effect of colour surface is an overlooked aspect that has not yet been investigated. In this study, we describe the effect of colour hues (white, red, blue and black) on the growth of cyanobacterial biofilms on air-exposed substrates. We measured growth, architecture, pigment production and levels of ATP and reactive oxygen species in cyanobacterial biofilms formed on different coloured substrates. The study findings demonstrate, for the first time, that the colour of a surface affects biofilm formation at the air-solid interface (with more biomass accumulating on white and red substrates than on blue and black substrates) and also alters the biofilm architecture. In addition, the roles of chromatic adaptation, phototrophic cells and reactive oxygen species as intermediates between colour sensing and biofilm response are discussed. Our results support the importance of colour as a new factor that favours surface colonization by cyanobacteria and its contribution to biofilm formation.