Bacteriocins have been suggested to play an important role in the invasion dynamics of bacteria. Recently, the ‘diversifying selection’ hypothesis has been proposed, which addresses the origin and diversification of one group of bacteriocins, the colicins of . According to this hypothesis, novel colicin gene clusters arise from mutations generating expanded immunity functions. Positive selection, favouring these novel immunities, then rapidly drives strains carrying the evolved colicin gene clusters to fixation in the local population. To test this fixation step driven by selection, invasion experiments were carried out by introducing novel colicinogenic strains into established colicinogenic populations. In all cases, invasion by strains expressing novel immunity functions occurred rapidly, even when initial frequencies of the invader were quite low. These invasions were attributed primarily to colicin killing effect. Other factors, such as growth rate, level of colicin production and stationary-phase survival rate, were shown to play very minor roles in the invasion process. These results provide direct evidence for the hypothesis of diversifying selection acting on colicin gene clusters and shed light on the ecological role of colicins.


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