(A) Simulation of colony development, based on the default model (which incorporates a resource threshold buildup, followed by consumption, switching to a light state, and expansion), compared to a model where the threshold amounts of the resource is removed. Note the final expansion size of the colony. Also see Figure 6—figure supplement 1A–D and Video 5. (B) i) Changes in the availability of the resource as the colony develops, based on the model. Ii) Extracellular amounts of trehalose measured from developing wild-type colonies. Data from three independent colonies. Note: in the model, in ~3–4 days the resource is highest, and reduces sharply after that. In the experimentally obtained data, extracellular trehalose amounts are highest at ~day 4, and then rapidly decreases over day 5. This correlates to when the light cells emerge and expand. (C) A time-course of bright-field images of the developing wild-type colony, illustrating the distribution of dark cells, and the emergence and distribution of light cells. (D) A time-course revealing fluorescence based estimation of the (i) reporter for gluconeogenic activity (dark cells), or (ii) the PPP activity reporter (light cells). Note the delayed, rapid appearance and increase in the PPP activity reporter. (E) Quantification of the increase in the gluconeogenic reporter activity in the colony, and the PPP reporter activity (based on fluorescence intensity) within the colony. The increase in gluconeogenic reporter activity, when plotted, is linear, and saturates. The increase in PPP activity over the first 5 days is highly cooperative (as estimated using a Hill coefficient as a proxy for cooperativity), before saturating (n = 3). Error bars represent standard deviation.