AbstractSound vibrations (SV) are known to influence molecular and physiological processes that can improve crop performance and yield. In this study, the effects of three audible frequencies (100, 500 and 1000 Hz) at constant amplitude (90 dB) on tomato Micro‐Tom physiological responses were evaluated 1 and 3 days post‐treatment. Moreover, the potential use of SV treatment as priming agent for improved Micro‐Tom resistance to Pseudomonas syringae pv. tomato DC3000 was tested by microarray. Results showed that the SV‐induced physiological changes were frequency‐ and time‐dependent, with the largest changes registered at 1000 Hz at day 3. SV treatments tended to alter the foliar content of photosynthetic pigments, soluble proteins, sugars, phenolic composition, and the enzymatic activity of polyphenol oxidase, peroxidase, superoxide dismutase and catalase. Microarray data revealed that 1000 Hz treatment is effective in eliciting transcriptional reprogramming in tomato plants grown under normal conditions, but particularly after the infection with Pst DC3000. Broadly, in plants challenged with Pst DC3000, the 1000 Hz pretreatment provoked the up‐regulation of unique differentially expressed genes (DEGs) involved in cell wall reinforcement, phenylpropanoid pathway and defensive proteins. In addition, in those plants, DEGs associated with enhancing plant basal immunity, such as proteinase inhibitors, pathogenesis‐related proteins, and carbonic anhydrase 3, were notably up‐regulated in comparison with non‐SV pretreated, infected plants. These findings provide new insights into the modulation of Pst DC3000‐tomato interaction by sound and open up prospects for further development of strategies for plant disease management through the reinforcement of defense mechanisms in Micro‐Tom plants.