1. In forest communities, the Janzen–Connell (J–C) model proposes that species diversity is maintained by noncompetitive distance- or density-dependent seedling mortality caused by host-specific natural enemies. Host specificity, however, has not been fully elucidated.
2. We conducted a cross-inoculation experiment to evaluate the host specificity of a pathogenic fungus, Colletotrichum anthrisci. The fungus was isolated from seedlings of four tree species (Prunus grayana, Fraxinus lanuginosa, Cornus controversa and Magnolia obovata), all of which were killed by damping-off disease beneath conspecific adults. Each fungal isolate was then inoculated into seedlings of P. grayana and F. lanuginosa. Molecular identification [internal transcribed spacer (ITS) sequences] also confirmed that all isolates (strains) showed 99–100% similarity with C. anthrisci, irrespective of their origin.
3. In both P. grayana and F. lanuginosa seedlings, all isolates of the pathogen caused damage, irrespective of origin, but the damage was more severe with the isolate from conspecifics rather than from any of the three heterospecifics.
4. In response to infection, callose papillae were deposited on the inner side of the leaf cell wall of seedlings in P. grayana; then, circular abscission layers formed between two layers of leaf cells surrounding the locus of infection. The central area of the infection was completely cut off from the rest of the leaf. In F. lanuginosa, infected leaves of seedlings were shed immediately after inoculation. This defensive behaviour, which may prevent further pathogen invasion, was more frequent in seedlings inoculated with isolates from conspecifics than from heterospecifics in both species.
5. Synthesis. Although the pathogenic fungus C. anthrisci is ubiquitous and attacks a wide range of host species, virulence was much stronger for strains derived from conspecifics rather than from heterospecifics, suggesting local adaptation and development of host specificity. If host specificity is common for several pathogens within a microbial community in a given area occupied by an adult, and the trait is also common for multiple tree species co-occurring within a forest community, the J–C model would be applicable to explain tree species diversity.