The　abuse　of　antibiotics　on　animals　could　induce　the　development　of　antibiotic　resistant　genes　(ARGs)　and　antibiotic　resistant　bacteria　(ARB),　and　acesulfame　potassium　(ACE)　is　the　widely　used　artificial　sweetener　in　animal　feed.　Generally　speaking,　ACE　and　ARB　often　coexist　in　livestock　wastewater,　however,　the　impact　of　the　co-occurrence　of　ACE　and　ARB　on　the　transmission　of　ARGs　is　still　unknown.　In　this　study,　the　effects　of　ACE　on　vertical　gene　transfer　(VGT)　and　horizontal　gene　transfer　(HGT)　were　both　evaluated.　For　VGT,　ACE　may　hinder　the　spread　of　sul　gene　in　Pseudomonas　HLS-6　by　blocking　ARB　growth.　As　for　HGT　(from　Escherichia　coli　DH5　alpha　to　Pseudomonas　HLS-6),　environmentally　relevant　ACE　concentration　could　facilitate　the　conjugative　transfer.　The　underlying　mechanisms　of　HGT　were　characterized　by　enhanced　cell　membrane　permeability,　reactive　oxygen　species　overproduction,　SOS　response,　energy　supply,　which　were　all　further　verified　by　the　changes　in　transcription　levels　of　related　genes.　Interestingly,　intracellular　Mg2+　in　donor　strain　was　found　for　the　first　time　as　an　indicator　for　the　conjugation　occurrence　in　ACE　treated　mating　system.　This　study　may　provide　new　insights　into　the　role　of　ACE　on　ARGs　proliferation　and　highlight　its　potential　environmental　impacts.