P-172 - Computers, Code, and Corn: in silico validation of e-probes targeting 3 phytopathogens of corn (Zea mays) seed

America. In 2023, the US exported USD 13.14 billion of corn worldwide. With increasing phytosanitary requirements placed by national seed health programs and foreign biosecurity at ports of entry, the need for efficient and specific seed pathogen testing is constantly growing. Current detection methods include ELISA, pathogen culturing, and PCR; performing these tests as a conglomerate may be costly, time-consuming, and error-prone. These methods have a limited sample processing capacity and must deal with putative inhibitors from the plant and seed anatomy. The Electronic-probe Diagnostic Nucleic-acid Analysis (EDNA) allows for rapid detection of multiple pathogens using complete or partial metagenomic sequences known as e-probes. This research project aims to develop e-probes for two bacterial pathogens and two fungal pathogens, Clavibacter michiganensis subsp. nebraskensis, Pantoea stewartii subsp. stewartii, Stenocarpella maydis, Stenocarpella macrospora, respectively. A digital database of the targeted pathogens and their taxonomically related pathogens was developed and tested in silico. Further trials include in vitro testing of the target pathogens using PCR to compare and validate results. New primers have been designed and will be tested with a plasmid artificial positive control.