Epichloë typhina (fungus) - Botanophila lobata (fly) interaction : an invasive "pollinator" system in its introduced range in western Oregon

Abstract

Epichloë typhina (Ascomycetes: Clavicipitaceae) is an endophytic fungus that infects perennial Pooid grasses and is the causal agent of choke disease. It is endemic to Europe and was inadvertently introduced into orchardgrass seed production fields in western Oregon. Choke disease, which was first recorded in Oregon in 1996, currently infects ~90% of cultivated orchardgrass seed fields in the region, resulting in yield losses >65%. Infective propagules (i.e. ascospores) are produced sexually by the bipolar heterothallic fungus and gamete outcross has been shown to be facilitated in the wild by Botanophila spp. (Diptera: Anthomyiidae), including Botanophila lobata. The fly - fungus interaction is generally considered to be one of obligatory mutualism based on studies conducted in areas endemism. However, recent evidence suggests that the fungus is able to sexually outcross in cultivated Oregon orchardgrass fields without the aid of fly "pollinators." Additionally, ascosporic fertilization has recently been implicated as an alternative mechanism for gamete transfer and might have important impacts on fungal reproduction in Oregon. The objectives of this study were to: 1) explore how two tightly linked species, which appear to have an obligate mutualistic relationship in areas of endemism, interact in a non-native context; 2) quantitatively examine the seasonal and diurnal presence of E. typhina ascospores in a cultivated Oregon orchardgrass field; and 3) test alternative transfer mechanisms of fungal spermatia for E. typhina. To address objective 1), the spatial variability and reproductive success of E. typhina and B. lobata were estimated during surveys of ten cultivated orchardgrass fields in 2008 and four fields in 2009. Fungal distributions were spatially aggregated at five of the study sites in 2008 and three in 2009. Fly distributions were spatially aggregated at three sites in 2008 and one in 2009. Botanophila lobata density exhibited a positive linear relationship with E. typhina density, suggestive of positive density dependence of fly oviposition with fungal density. However, fungal reproductive success was not affected by fly density or fungal density within the range of distributions observed in this study. To address objective 2), airborne ascospores were monitored in a single cultivated orchardgrass field during 2008 and 2009 using a Burkard volumetric spore trap. Ascospore production began in early to middle May and continued into late July during both years of the study. Daily ascospore production exhibited a circadian rhythm, with production peaking on average at 1:08 am and 12:36 am, in 2008 and 2009, respectively. The prolonged duration and high intensity of ascospore production during the growing season suggest a large window within which new plants are at risk to infection, and within which preventative management strategies must be adopted. To address objective 3), splash fertilization, contact fertilization and B. lobata fertilization were tested in the greenhouse. Although only two replicates were completed, both contact and splash fertilization appeared to be viable mechanisms of sexual outcross for E. typhina. The results of this study strongly indicate that E. typhina can successfully reproduce without the presence of B. lobata. Splash and contact fertilization, as well as ascosporic fertilization, provide opportunities for reproduction of the fungus in absence of fly pollinators. It appears that the fly - fungus interaction has shifted from an obligatory mutualism to facultative mutualism or simple fungivory within the introduced range in western Oregon.