This study reports the biosynthesis of silver nanoparticles (AgNPs) using a Pseudomonas canadensis Ma1 strain isolated from wild-growing mushrooms. Freshly prepared cells of P. canadensis Ma1 incubated at 26-28 °C with a silver nitrate solution changed to a yellowish brown color, indicating the formation of AgNPs, which was confirmed by UV-Vis spectroscopy, scanning electron microscopy (SEM), and X-ray diffraction. SEM analysis showed spherical nanoparticles with a distributed size mainly between 21 and 52 nm, and the XRD pattern revealed the crystalline nature of AgNPs. Also, it provides an evaluation of the antimicrobial activity of the biosynthesized AgNPs against Pseudomonas tolaasii Pt18, the causal agent of mushroom brown blotch disease. AgNPs were found to be bioactive at 7.8 μg/ml showing a minimum inhibitory concentration (MIC) effect against P. tolaasii Pt18 strain. AgNPs at the MIC level significantly reduced virulence traits of P. tolaasii Pt18 such as detoxification of tolaasin, various motility behavior, chemotaxis, and biofilm formation which is important for pathogenicity. Scanning electron microscopy (SEM) revealed that bacterial cells treated with AgNPs showed a significant structural abnormality. Results showed that AgNPs reduced brown blotch symptoms in vivo. This research demonstrates the first helpful use of biosynthesized AgNPs as a bactericidal agent against P. tolaasii.