Xanthomonas oryzae pv. oryzae (Xoo) is a pervasive bacterial pathogen with global implications, causing bacterial blight in rice and threatening food security. The effective management of Xoo diseases in field conditions, protected farm operations, and international borders relies heavily on the early detection of Xoo. Traditional detection methods, including immunological assays such as direct tissue blot immunoassays (DTBIA) and enzyme-linked immunosorbent assays (ELISA), as well as molecular techniques like loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR), are commonly employed for pathogen identification. However, these methods often require sophisticated instrumentation and trained personnel, resulting in time-consuming and resource-intensive processes that are unsuitable for on-site analysis. To overcome these challenges, we have developed a sensitive electrochemical biosensor for Xoo detection by exploiting the unique properties of a core nano assembly of gold nanoparticles. The biosensor incorporates a specific DNA probe designed based on the PthXo1 gene, a prominent virulence factor of Xoo. This probe is immobilized on the electrode surface, enabling sequence-specific hybridization and subsequent electrochemical transduction for the sensitive and rapid detection of Xoo up to the 1fg/µl level.¹ This innovative approach holds promise for monitoring Xoo, thereby contributing to the development of effective disease management strategies and the preservation of global food security.