Endophytes of the genus Epichloë (Clavicipitaceae, Ascomycota) frequently occur within cool-season grasses and form interactions with their hosts that range from mutualistic to antagonistic. Many Epichloë species have arisen via interspecific hybridization, resulting in species with two or three subgenomes that retain all or nearly all of their original parental genomes, a process termed allopolyploidization. Here, we characterize Epichloë hybrida, sp. nov., a mutualistic species that has increasingly become a model system for investigating allopolyploidy in fungi. The Epichloë species so far identified as the closest known relatives of the two progenitors of E. hybrida are E. festucae var. lolii and E. typhina. We confirm that the nuclear genome of E. hybrida contains two homeologs of most protein-coding genes from E. festucae and E. typhina, with genome-wide gene expression analysis indicating a slight bias in overall gene expression from the E. typhina subgenome. Mitochondrial DNA is detectable only from E. festucae, whereas ribosomal DNA is detectable only from E. typhina. Inheriting ribosomal DNA from just one parent might be expected to preferentially favor interactions with ribosomal proteins from the same parent, but we find that ribosomal protein genes from both parental subgenomes are nearly all expressed equally in E. hybrida. Finally, we provide a comprehensive set of resources for this model system that are intended to facilitate further study of fungal hybridization by other researchers.

In identifying endophytes for use in pastures in New Zealand there have been two strategies used, either exclusion of ergopeptine and lolitrem alkaloids or, retaining some ergopeptine expression to enhance the stability of natural defences against invertebrate pests. Both have their strengths and weaknesses. It is a matter of balancing these to ensure the end‐user has access to a product that satisfies their needs – a persistent pasture with low to nil animal toxicity in terms of animal production and welfare. The range and intensity of ergot alkaloids in grazed pasture on both pasture and ruminants is reviewed, with emphasis on New Zealand. Ergot alkaloids associated with pasture are produced by associations between certain Epichloë endophyte strains and temperate grasses. Ergot alkaloids have been shown to improve persistence of pasture through providing resistance/deterrence to insect pests as well as deterring grazing animals. However, ergovaline is toxic to grazing animals. Some commercially available ryegrass‐endophyte associations can produce ergovaline concentrations close to those found in associations between ryegrass and the standard endophyte. It is feasible to eliminate ergot alkaloids from pasture grasses in New Zealand as endophyte strains are commercially available that do not express ergovaline and yet still provide excellent pest resistance.

The aphid Aploneura lentisci is widespread in Australia and New Zealand, living all year round on roots of its secondary grass hosts. The fungal endophyte (Epichloë festucae var. lolii), strain AR37 in Lolium perenne is known to greatly reduce populations and was a likely reason for the superior growth and persistence of this association previously observed in the field. Aphid populations were quantified in a field trial near Ballarat, comparing yields of perennial ryegrass infected with eight different endophyte strains and an endophyte-free (Nil) control in a common ryegrass background (Grasslands Samson (G. Samson)). AR37 and another endophyte strain, AR5, had significantly fewer aphids than all other endophytes. These differences were significantly related to yield increases taken before and after sampling that persisted until the end of the trial. In a pot trial comparing commercially available ryegrass-endophyte combinations with equivalent Nil controls, aphid numbers were lower on G. Samson AR37 and Banquet II with AR5 (Endo®5) than on all other cultivar-endophyte combinations. Compared with Nil controls, the common toxic strain in G. Samson, and two strains in Trojan also reduced aphid numbers. The AR5 endophyte produces the alkaloid ergovaline but high concentrations of this in roots of potted plants could not account for differences in root aphid numbers. Root concentrations of epoxyjanthitrems, the only known alkaloids produced by AR37, were low and unlikely to be the cause of resistance to A. lentisci.

Plants deploy diverse anti‐herbivore defences which reduce feeding and performance of herbivores. Temperate grasses use silicon (Si) accumulation and Epichloë‐endophytes for physical and chemical (i.e. endophytic‐alkaloids) defence against insect herbivores. Recent studies suggest that Epichloë‐endophytes increase Si accumulation in their host grass. It is unknown, however, how this affects Si‐deposition on the leaf surface, their impacts on insect herbivore feeding efficiency and their immunity to potential infection/parasitism. To address this knowledge gap, we grew tall fescue (Festuca arundinacea) hydroponically with and without Si, in the absence or presence of the novel AR584 Epichloë‐strain. We exposed plants to Helicoverpa armigera (Lepidoptera: Noctuidae) in both in situ (intact leaves) and ex situ (excised leaves) feeding trials and determined the effects of Si and endophyte defences on herbivore feeding efficiency, growth rates and immunity against potential infection/parasitism. Endophytic plants supplied with Si showed 110% and 143% increases in leaf silica density and leaf Si concentrations, respectively, when exposed to herbivory, compared to non‐endophytic plants that were herbivore‐free. Despite the endophyte‐mediated increases in Si concentrations, H. armigera was only affected by Si supply; growth rates decreased by 87% and most feeding efficiency indices decreased by at least 30%. Si supply also increased mandibular wear by 16%, which was negatively correlated with H. armigera growth rates. Cellular and humoral immunity of H. armigera were negatively affected by both Si and endophytes. Endophytic‐loline alkaloid concentrations were unaffected by Si supply or herbivory, whereas herbivory increased peramine concentrations by 290%. To our knowledge, this is the first report of Si defences and Epichloë‐endophyte derived alkaloids compromising insect immunity via reduced melanisation response. Using tall fescue and H. armigera, our study suggests that deploying both physical (i.e. Si accumulation) and chemical (i.e. endophytic‐alkaloids) defences acting against multiple insect herbivore traits, including feeding efficiency, growth and immunity, may be a successful defence strategy in temperate grasses. This multi‐faceted defence may be particularly difficult for insect herbivores to overcome. Read the free Plain Language Summary for this article on the Journal blog.