Bacterial differentiation within Moraxella bovis colonies growing at the interface of the agar medium with the Petri dish.

Moraxella bovis was found to colonize the interface between agar and the polystyrene Petri dish, producing circular colonies when the inoculum was stabbed at a single point. The bacteria occurred in a thin layer of nearly uniform thickness, and colonial expansion occurred in at least two temporal phases. In the first phase, the radial colonial expansion was slow and non-linear. In the second phase, the radial expansion was linear. The interfacial colonies possessed three characteristic concentric growth zones. At the periphery was a narrow ring zone that enclosed another wider ring zone, which, in turn, surrounded a central circular zone. Different bacterial phase variants were recovered from these zones. The two outer ring zones yielded bacteria that formed agar surface colonies of spreading-corroding morphology, while cells from the innermost zone always yielded colonies with a different morphology. The uniform thickness of the colonies implied that replication was restricted to the outermost ring, and that the bacteria within the inner ring and inner circle had entered a quiescent state. The inner ring appeared to represent the lag in time needed for the replicative form to differentiate into the quiescent form. A different kind of variant was associated with wedge-shaped sectors within the colonies. The greatest number of these clonal variants appeared shortly after inoculation and their frequency decreased after the onset of linear growth. The period of slowest colonization coincided with highest frequency of clonal variant expression. It is proposed that the proliferative rate of the parental bacterial population exerted selective pressure on the expression of new clonal variants.