Multistrain Infections of the Grass Brachypodium Sylvaticum by Its Fungal Endophyte Epichloë Sylvatica

Endophytes of the genus Epichloë (Clavicipitaceae, Ascomycota) are systemic symbionts of cool-season grasses. Their interactions with grass hosts may vary between mutualistic and pathogenic depending on the mode of endophyte reproduction. Sexual strains prevent flowering and seed set (choke disease) of the host and can be horizontally transmitted by ascospores, while asexual strains remain asymptomatic and are vertically transmitted through seeds. In Switzerland nearly all plants of Brachypodium sylvaticum (Huds.) P.B. are infected by Epichloë sylvatica Leuchtmann & Schardl, but choke symptoms are formed very rarely, and are restricted to particular locations and to a minority of plants at those locations. Earlier research has revealed that E. sylvatica is genetically differentiated into sexual and asexual subpopulations. Given the high level of infection and assuming horizontal transmission of sexual strains, multiple host infections have been predicted. In this study, 25 plants out of 63 examined by isozyme analysis were found to be infected by two or three different endophyte genotypes. In most cases endophyte genotypes appeared to be correlated with the symptom type of a particular tiller, suggesting that the fungal genome controls choke formation and that the sexual and asexual subpopulations are separated at the ramet (tiller) level rather than at the genet (plant) level. These conclusions were further supported by analyses with log-linear models of the population structure of E. sylvatica at four locations where choke symptoms were present. These analyses also revealed a geographic structure in the asexual subpopulation but not in the sexual subpopulation which could be caused by the different dispersal ranges of their propagules. The rare occurrence of sexually reproducing strains and the dominance of a single genotype in asymptomatic plant populations may be explained by the colonization history of B. sylvaticum and its endophyte in Switzerland.