Xanthomonas　oryzae　pv.　oryzicola　(Xoo)　is　a　plant　pathogenic　bacterium　that　can　cause　rice　bacterial　blight　disease,　which　results　in　a　severe　reduction　in　rice　production.　Antimicrobial-dependent　microbial　controlling　is　a　useful　way　to　control　the　spread　and　outbreak　of　plant　pathogenic　bacteria.　However,　the　abuse　and　long-term　use　of　antimicrobials　also　cause　microbial　antimicrobial　resistance.　As　far　as　known,　the　mechanism　of　antimicrobial　resistance　in　agricultural　plant　pathogenic　bacteria　still　lacks　prospecting.　In　this　study,　we　explore　the　mechanism　of　Zhongshengmycin　(ZSM)-resistance　in　Xoo　by　GC-MS-based　metabolomic　analysis.　The　results　showed　that　the　down-regulation　of　the　TCA　cycle　was　characteristic　of　antimicrobial　resistance　in　Xoo,　which　was　further　demonstrated　by　the　reduction　of　activity　and　gene　expression　levels　of　key　enzymes　in　the　TCA　cycle.　Furthermore,　alanine　was　proven　to　reverse　the　ZSM　resistance　in　Xoo　by　accelerating　the　TCA　cycle　in　vivo.　Our　results　are　essential　for　understanding　the　mechanisms　of　ZSM　resistance　in　Xoo　and　may　provide　new　strategies　for　controlling　this　agricultural　plant　pathogen　at　the　metabolic　level.　©　2023　by　the　authors.