Multi-step substrate consumption pathways can promote microbial biodiversity via cross-feeding. If one cell-type preferentially consumes a primary substrate rather than the subsequently formed intermediates, then other cell-types can specialize at consuming the intermediates. While this mechanism for promoting biodiversity is established, predicting the long-term persistence of such cross-feeding interactions remains challenging. Under what conditions will the interaction (and thus biodiversity) persist or disappear? To address this question, we propagated co-cultures of two isogenic strains of the bacterium Pseudomonas stutzeri. One completely reduces nitrate to nitrogen gas but preferentially reduces nitrate rather than nitrite (referred to as the generalist) while the other only reduces nitrite to nitrogen gas (referred to as the specialist). We found that the two strains coexist via nitrite cross-feeding when grown together, but the initial ratio of specialist-to-generalist (rS/G) determines the long-term dynamics of the co-culture. Co-cultures with large initial rS/Gs converge to the same rS/G and persist thereafter. Co-cultures with small initial rS/Gs also converge to the same rS/G but then become increasingly dominated by the generalist. The likely cause of these different dynamics is that the initial rS/G determines the initial environment, which in turn determines the initial selection pressures and phenotypes acquired by the generalist. Our results demonstrate that initial community composition controls the long-term dynamics and persistence of a cross-feeding interaction, and is therefore an important factor for community development and for engineering communities to achieve desired outcomes.
