The virulence evolution of multiple infections of parasites from the same species has been modelled widely in evolution theory, and the trajectories of evolution are relevant to parasite transmission mode, as well as to parasite and host population dynamics. However, experimental studies on this topic remain scarce, particularly regarding multiple infections by different parasite species. In this study, we employed the invasive plant Ageratina adenophora to verify the predictions made by the model. We observed that A. adenophora was a highly susceptible host to phylogenetically diverse foliar pathogens with mixed vertical and horizontal transmission within leaf spots. The pathogen community structure at the leaf spot level was determined by transmission mode. Over time, the pathogen community decreased in diversity; meanwhile, the vertically transmitted pathogens exhibited decreased virulence to the host A. adenophora, but the horizontally transmitted pathogens exhibited increased virulence to the host. Our results demonstrate that the predictions of classical models are still valid in a complex environment. Moreover, we propose that it is very important to determine whether the primary foliar pathogen of a given plant host is relevant to seedborne fungi, as this characteristic is an important factor in understanding pathogen-host interactions.