The aim of this paper is to study the impact of the substrate stiffness on the bacterial adhesion. For this purpose, agarose hydrogels are used as substrates with controlled mechanical properties. Indeed, the elastic modulus of these hydrogels, more precisely the shear storage moduli G', evolves with the agarose concentration (in this study from 0.75% to 3%). Other physico-chemical characteristics of the surface, known to be involved in bacterial adhesion, as hydrophobicity, were confirmed to remain constant. Two marine bacterial strains, a positive Gram Bacillus sp. 4J6 and a negative Gram Pseudoalteromonas sp. D41 were selected. Their retention on the substrates was analysed by confocal laser scanning microscopy and by counting of viable adhered bacteria. It was demonstrated that surface elastic modulus correlated with bacterial retention. Bacteria 041 adhered in higher numbers to rigid surfaces. For 416, bacterial adhesion patterns were changed: clusterings were observed on surfaces with lower elastic modulus. Furthermore, a proteomic study, based on the total soluble proteome of D41 strain, highlights an impact of elastic modulus on proteins synthesis. These data demonstrated an adapted response of adhering bacteria on hydrogels of varying mechanical properties