Dynamics of the peptidoglycan biosynthetic machinery in the stalked budding bacterium Hyphomonas neptunium

Summary Most commonly studied bacteria grow symmetrically and divide by binary fission, generating two siblings of equal morphology. An exception to this rule are budding bacteria, in which new offspring emerges de novo from a morphologically invariant mother cell. Although this mode of proliferation is widespread in diverse bacterial lineages, the underlying mechanisms are still incompletely understood. Here, we report the first molecular‐level analysis of growth and morphogenesis in the stalked budding alphaproteobacterium Hyphomonas neptunium. Peptidoglycan labeling shows that, in this species, buds originate from a stalk‐like extension of the mother cell whose terminal segment is gradually remodeled into a new cell compartment. As a first step toward identifying the machinery mediating the budding process, we performed comprehensive mutational and localization studies of predicted peptidoglycan biosynthetic proteins in H. neptunium. These analyses identify factors that localize to distinct zones of dispersed and zonal growth, and they suggest a critical role of the MreB‐controlled elongasome in cell morphogenesis. Collectively, our work shows that the mechanism of growth in H. neptunium is distinct from that in related, polarly growing members of the order Rhizobiales, setting the stage for in‐depth analyses of the molecular principles regulating the fascinating developmental cycle of this species. The stalked budding bacterium Hyphomonas neptunium grows by generating buds at the tip of a stalk‐like cellular extension. This study reports the first molecular‐level analysis of cell wall biosynthesis in this species. We show that its mode of growth differs significantly from that in related polarly growing species and identify factors critical to proper morphogenesis. These findings set the stage for in‐depth mechanistic studies of a fascinating but poorly understood mode of bacterial proliferation.