Aquatic　products　are　highly　susceptible　to　spoilage　caused　by　adherence　of　spoilage　bacteria　during　storage,　so　it　is　crucial　to　develop　new　microecological　preservatives.　The　probiotic　B.　subtilis　can　achieve　potential　preservation　effects　by　adhering　to　the　fish　gut　to　regulate　the　amount　of　probiotic　colonization　in　the　flora.　Here,　we　demonstrated　the　surface　adhesion　characteristics　of　B.　subtilis　and　the　mechanism　of　adhesion-associated　gene-mediated　adhesion;　the　regulation　of　B.　subtilis　adhesion　ability　by　nitrogen　and　carbon　sources,　and　the　application　of　tilapia　preservation.　Our　results　revealed　10　B.　subtilis　adhesion-related　genes,　among　which　nitrogen　sources　up-regulate　the　adhesion　genes　hag　and　motA,　carbon　sources　up-regulate　sinR　and　fliG.　Based　on　flagellum-associated　high　adhesion　strain　C6　and　high　adhesion　regulation　of　B.　subtilis　C6_H　allowed　the　abundance　of　probiotic　bacteria　Bacillus,　Rhodobacteraceae　to　increase,　attenuate　the　content　of　spoilage　flavor　substances　such　as　1-Octen-3-ol,　15-Crown-5　and　maintain　the　freshness　of　tilapia.　The　results　helped　to　reveal　the　adhesion　mechanism　of　B.　subtilis　and　to　develop　and　apply　microecological　preservatives　with　high　quality　adhesion　ability.　©　2023　The　Authors