This review focuses on the molecular mechanisms controlling biofilm assembly in *Bacillus subtilis* and briefly summarizes the current understanding of biofilm disassembly. *B. subtilis* has long been a robust model organism for studying biofilm formation, with numerous studies revealing that this process is regulated by integrated pathways. The review highlights the importance of the extracellular matrix, which holds the biofilm together, and discusses the dynamic spatial organization of different cell types within the biofilm. It also covers recent progress in understanding biofilm formation on plant roots, a natural habitat for this soil bacterium. The review delves into the regulatory pathways that control biofilm formation, including the Spo0A pathway, the SlrR-SinR epigenetic switch, and other pathways involving DegS-DegU and YwcC-SlrA. Additionally, it explores triggers of biofilm formation, such as surfactin and cannibalism toxins, and mechanisms of biofilm dispersal, including the release of D-amino acids and norspermidine. The review concludes by discussing the potential of small molecules identified in *B. subtilis* biofilms to control biofilm formation in other species, including pathogenic bacteria, and the implications for therapeutic strategies.This review focuses on the molecular mechanisms controlling biofilm assembly in *Bacillus subtilis* and briefly summarizes the current understanding of biofilm disassembly. *B. subtilis* has long been a robust model organism for studying biofilm formation, with numerous studies revealing that this process is regulated by integrated pathways. The review highlights the importance of the extracellular matrix, which holds the biofilm together, and discusses the dynamic spatial organization of different cell types within the biofilm. It also covers recent progress in understanding biofilm formation on plant roots, a natural habitat for this soil bacterium. The review delves into the regulatory pathways that control biofilm formation, including the Spo0A pathway, the SlrR-SinR epigenetic switch, and other pathways involving DegS-DegU and YwcC-SlrA. Additionally, it explores triggers of biofilm formation, such as surfactin and cannibalism toxins, and mechanisms of biofilm dispersal, including the release of D-amino acids and norspermidine. The review concludes by discussing the potential of small molecules identified in *B. subtilis* biofilms to control biofilm formation in other species, including pathogenic bacteria, and the implications for therapeutic strategies.