Intrinsically robust and scalable biofilm segmentation under diverse physical growth conditions

Developmental patterning is a shared feature across biological systems ranging from vertebrates to bacterial biofilms. While vertebrate patterning benefits from well-controlled homeostatic environments, bacterial biofilms can grow in diverse physical contexts. What mechanisms provide developmental robustness under diverse environments remains an open question. We show that a native clock-and-wavefront mechanism robustly segments biofilms in both solid-air and solid-liquid interfaces. Biofilms grown under these distinct physical conditions differ 4-fold in size yet exhibit robust segmentation. The segmentation pattern scaled with biofilm growth rate in a mathematically predictable manner independent of habitat conditions. We show that scaling arises from the coupling between wavefront speed and biofilm growth rate. In contrast to the complexity of scaling mechanisms in vertebrates, our data suggests that the minimal bacterial clock-and-wavefront mechanism is intrinsically robust and scales in real time. Consequently, bacterial biofilms robustly segment under diverse conditions without requiring cell-to-cell signaling to track system size.