The　outbreak　of　Bacterial　blight　(BB)　caused　by　Xanthomonas　oryzae　(Xoo)　generates　substantial　economic　losses　to　agricultural　production.　Antibiotics　application　is　a　valuable　measure　to　control　this　bacterial　disease.　However,　microbial　antibiotic　resistance　dramatically　reduced　antibiotic　effectiveness.　Identifying　the　resistance　mechanism　of　Xoo　to　antibiotics　and　restoring　antibiotic　susceptibility　is　one　of　the　crucial　ways　to　solve　this　problem.　This　study　employed　a　GC-MS-based　metabolomic　approach　to　reveal　the　differential　metabolomics　between　a　kasugamycin-susceptible　Xoo　strain　(Z173-S)　and　a　kasugamycin-resistant　strain　(Z173-R-KA).　The　metabolic　mechanism　of　kasugamycin　(KA)　resistance　in　Xoo　by　GC-MS　showed　that　the　downregulation　of　the　pyruvate　cycle　(P　cycle)　is　a　crucial　feature　of　Z173-R-KA　resistance　to　KA.　This　conclusion　was　confirmed　by　the　decreased　enzyme　activities　and　the　related　gene　transcriptional　level　in　the　P　cycle.　Furfural　(an　inhibitor　of　pyruvate　dehydrogenase)　can　effectively　inhibit　the　P　cycle　and　increase　the　resistance　of　Z173-R-KA　to　KA.　Moreover,　exogenous　alanine　can　reduce　the　resistance　of　Z173-R-KA　to　KA　by　promoting　the　P　cycle.　Our　work　seems　to　be　the　first　exploration　of　the　mechanism　of　KA　resistance　in　Xoo　by　GC-MS-based　metabonomics　approach.　These　results　provide　a　new　idea　for　developing　metabolic　regulation　to　address　KA　resistance　in　Xoo.