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Glyphosate, the active ingredient of glyphosate-based herbicides (GBHs), controls the growth of weeds by inhibiting the shikimate pathway, thereby interrupting amino acid biosynthesis in plants. However, several microbes have the shikimate pathway, and the effect of glyphosate on these non-target organisms is ignored. The action of GBHs is further complicated when used with other agrochemicals, such as phosphate fertilizers, often varying their mode of action depending on soil type or plant species. To address the impact of GBHs and phosphate fertilizers, we simulated agricultural application of GBHs and phosphate fertilizers in a field study, investigating the composition of endophytic microbial communities and correlation of phytohormone concentrations with the microbial diversity of potato (Solanum tuberosum). In leaves, glyphosate residues in soil from GBH treatment alone and in combination with phosphate significantly shifted the bacterial community, whereas phosphate alone and in combination with glyphosate significantly altered the composition of the fungal community. There were no significant changes in microbial communities in roots and tubers. Plants treated with GBHs showed higher ratios of potentially glyphosate-resistant bacteria, with Xanthomonadaceae and Moraxellaceae being more abundant. Additionally, phytohormone concentrations showed various correlations with bacterial and fungal diversity in different treatments. The study highlights the impact of GBH residues in soil, particularly in combination with phosphate fertilizers, on the composition of plant-associated microbial communities. Together with changes in phytohormone concentrations, plant health may be affected. Future studies could provide insights into whether these agrochemicals influence the plant microbiome, leading to changes in phytohormones or vice versa. [Figure: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Abstract Background The herbicide glyphosate is the most widely used active ingredient in pesticides globally. Residues have been found in people, livestock, food and animal feed, and in the environment, but little is known about glyphosate residue in manure-based fertilizer. We describe a feed-feces-fertilizer route of glyphosate contamination with negative impacts for horticultural production. This exposure can harm sensitive plants, such as tomato, and pose a risk to effective waste disposal and nutrient cycling along principles of the circular economy. We review the use and history of glyphosate and present a mixed methods research based on a real-world case from Finland where glyphosate residue in poultry manure fertilizer was suspected of inhibiting commercial organic tomato production. To test the fertilizer, we grew 72 ‘Encore’ variety tomato plants for 14 weeks in a climate-controlled greenhouse according to the practices of the commercial grower. To ascertain awareness and potential contamination mitigation measures, we contacted five fertilizer companies with sales of biogenic fertilizer in Finland, two farming organizations, a feed company, and two government organizations working on nutrient cycling and agricultural circular economy. Results The total harvest of tomatoes grown with fertilizer with the higher content of glyphosate residue was 35% smaller and the yield of first-class tomatoes 37% lower than that of the control, with lower glyphosate concentration. Two of the five fertilizer companies identified poultry manure as a source of glyphosate contamination. Companies with awareness of pesticide residues reported interest in establishing parameters for pesticide residues. Conclusions The extent of glyphosate contamination of recycled fertilizers is unknown, but this study shows that such contamination occurs with negative impacts on crop production. Lack of testing and regulation to ensure that recycled fertilizers are free from harmful levels of glyphosate or other pesticides creates risks for agricultural producers. The issue is particularly acute for certified organic producers dependent on these products, but also for sustainable transitions away from mineral fertilizers in conventional farming. The example from Finland shows that a model of co-production between fertilizer producers and state regulatory agencies to establish safe limits can benefit both fertilizer producers and their customers.

Abstract Introduction Pesticides are increasingly used in intensely managed agro‐environments, with an increasingly acknowledged impact on crop production, root establishment and plant resilience. At the same time management practices are intensified with the goal to maximize productivity. Materials and Methods In a greenhouse, we studied the effects of three mowing regimes (uncut, and cutting 5 or 15 cm) employed three times during the season on root and shoot biomass and chlorophyll content of the cool‐season grass Festuca pratensis (meadow fescue) growing in soil with a history of glyphosate‐based herbicide (GBH) use, the corresponding control soil, and sterilized control soil. Half of the plants hosted a systemic and vertically transmitted fungal endophyte, Epichloë uncinata, which is known to promote host grass growth. Results Endophyte symbiosis did not affect any tested plant parameters. Cutting the plants to 5 cm decreased both root and cumulative shoot biomass. Herbicide soil history, together with intense cutting (5 cm), caused a decrease in shoot biomass and lowered the chlorophyll content. Surprisingly, soil sterilization boosted shoot biomass and chlorophyll concentrations during less intense cutting (15 cm) and noncutting when compared to the control soil. Root biomass reduced in uncut plants when growing in soil with a history of glyphosate use. Conclusion Our results indicate that GBH residues in the soil can diminish shoot biomass when grass is frequently cut. Decreased root biomass caused by soil glyphosate history goes together with a reduction of carbon allocation belowground, which decreases grassland resilience to climate warming, increasingly occurring droughts and extreme weather events.

Abstract Increasing agricultural soil carbon sequestration without compromising the productivity of the land is a key challenge in global climate change mitigation. The carbon mitigation potential of grass-based agriculture is particularly high because grasslands represent 70% of the world’s agricultural area. The root systems of grasses transfer large amounts of carbon to below-ground storage, and the carbon allocation to the roots is dependent on the grasses’ photosynthesizing shoot biomass. In a common-garden experiment, Festuca pratensis was used as a model species to study how mowing and weed control practices of perennial cool-season fodder grasses affect total yield and root biomass. Additionally, grass-associated Epichloë endophytes and soil residual glyphosate were tested for their effect on the total yield and root biomass alone or in interaction with mowing. The results demonstrate that elevating the cutting height increases both cumulative yield and root biomass in F. pratensis. Endophyte symbiosis increased the total yield, while glyphosate-based herbicide residues in the soil decreased the root biomass, which indicates a reduction of soil bound carbon sequestration. The findings demonstrate that carbon sequestration and yield quantities on farmed grasslands may significantly be improved by optimizing strategies for the use of plant protection products and adjustment of mowing intensity.

Abstract Variation in plant communities is likely to modulate the feeding and oviposition behavior of herbivorous insects, and plant‐associated microbes are largely ignored in this context. Here, we take into account that insects feeding on grasses commonly encounter systemic and vertically transmitted (via seeds) fungal Epichloë endophytes, which are regarded as defensive grass mutualists. Defensive mutualism is primarily attributable to alkaloids of fungal origin. To study the effects of Epichloë on insect behavior and performance, we selected wild tall fescue (Festuca arundinacea) and red fescue (Festuca rubra) as grass–endophyte models. The plants used either harbored the systemic endophyte (E+) or were endophyte‐free (E−). As a model herbivore, we selected the Coenonympha hero butterfly feeding on grasses as larvae. We examined both oviposition and feeding preferences of the herbivore as well as larval performance in relation to the presence of Epichloë endophytes in the plants. Our findings did not clearly support the female's oviposition preference to reflect the performance of her offspring. First, the preference responses depended greatly on the grass–endophyte symbiotum. In F. arundinacea, C. hero females preferred E+ individuals in oviposition‐choice tests, whereas in F. rubra, the endophytes may decrease exploitation, as both C. hero adults and larvae preferred E− grasses. Second, the endophytes had no effect on larval performance. Overall, F. arundinacea was an inferior host for C. hero larvae. However, the attraction of C. hero females to E+ may not be maladaptive if these plants constitute a favorable oviposition substrate for reasons other than the plants' nutritional quality. For example, rougher surface of E+ plant may physically facilitate the attachment of eggs, or the plants offer greater protection from natural enemies. Our results highlight the importance of considering the preference of herbivorous insects in studies involving the endophyte‐symbiotic grasses as host plants.

Plants host taxonomically and functionally complex communities of microbes. However, ecological studies on plant–microbe interactions rarely address the role of multiple co-occurring plant-associated microbes. Here, we contend that plant-associated microbes interact with each other and can have joint consequences for higher trophic levels. In this study we recorded the occurrence of the plant seed pathogenic fungus Claviceps purpurea and aphids (Sitobion sp.) on an established field experiment with red fescue (Festuca rubra) plants symbiotic to a seed transmitted endophytic fungus Epichloë festucae (E+) or non-symbiotic (E–). Both fungi are known to produce animal-toxic alkaloids. The study was conducted in a semi-natural setting, where E+ and E– plants from different origins (Spain and Northern Finland) were planted in a randomized design in a fenced common garden at Kevo Subarctic Research Station in Northern Finland. The results reveal that 45% of E+ plants were infected with Claviceps compared to 31% of E– plants. Uninfected plants had 4.5 times more aphids than Claviceps infected plants. By contrast, aphid infestation was unaffected by Epichloë symbiosis. Claviceps alkaloid concentrations correlated with a decrease in aphid numbers, which indicates their insect deterring features. These results show that plant mutualistic fungi can increase the infection probability of a pathogenic fungus, which then becomes beneficial to the plant by controlling herbivorous insects. Our study highlights the complexity and context dependency of species–species and multi-trophic interactions, thus challenging the labeling of species as plant mutualists or pathogens.

Glyphosate is the most widely used herbicide with a yearly increase in global application. Recent studies report glyphosate residues from diverse habitats globally where the effect on non-target plants are still to be explored. Glyphosate disrupts the shikimate pathway which is the basis for several plant metabolites. The central role of phytohormones in regulating plant growth and responses to abiotic and biotic environment has been ignored in studies examining the effects of glyphosate residues on plant performance and trophic interactions. We studied interactive effects of glyphosate-based herbicide (GBH) residues and phosphate fertilizer in soil on the content of main phytohormones, their precursors and metabolites, as well as on plant performance and herbivore damage, in three plant species, oat (Avena sativa), potato (Solanum tuberosum), and strawberry (Fragaria x ananassa). Plant hormonal responses to GBH residues were highly species-specific. Potato responded to GBH soil treatment with an increase in stress-related phytohormones abscisic acid (ABA), indole-3-acetic acid (IAA), and jasmonic acid (JA) but a decrease in cytokinin (CK) ribosides and cytokinin-O-glycosides. GBH residues in combination with phosphate in soil increased aboveground biomass of potato plants and the concentration of the auxin phenylacetic acid (PAA) but decreased phaseic acid and cytokinin ribosides (CKR) and O-glycosides. Chorismate-derived compounds [IAA, PAA and benzoic acid (BzA)] as well as herbivore damage decreased in oat, when growing in GBH-treated soil but concentrations of the cytokinin dihydrozeatin (DZ) and CKR increased. In strawberry plants, phosphate treatment was associated with an elevation of auxin (IAA) and the CK trans-zeatin (tZ), while decreasing concentrations of the auxin PAA and CK DZ was observed in the case of GBH treatment. Our results demonstrate that ubiquitous herbicide residues have multifaceted consequences by modulating the hormonal equilibrium of plants, which can have cascading effects on trophic interactions.

Plants harbor a large diversity of endophytic microbes. Meadow fescue (Festuca pratensis) is a cool-season grass known for its symbiotic relationship with the systemic and vertically—via seeds—transmitted fungal endophyte Epichloë uncinata, yet its effects on plant hormones and the microbial community is largely unexplored. Here, we sequenced the endophytic bacterial and fungal communities in the leaves and roots, analyzing phytohormone concentrations and plant performance parameters in Epichloë-symbiotic (E+) and Epichloë-free (E-) individuals of two meadow fescue cultivars. The endophytic microbial community differed between leaf and root tissues independent of Epichloë symbiosis, while the fungal community was different in the leaves of Epichloë-symbiotic and Epichloë-free plants in both cultivars. At the same time, Epichloë symbiosis decreased salicylic acid and increased auxin concentrations in leaves. Epichloë-symbiotic plants showed higher biomass and higher seed mass at the end of the season. Our results demonstrate that Epichloë symbiosis alters the leaf fungal microbiota, which coincides with changes in phytohormone concentrations, indicating that Epichloë endophytes affect both plant immune responses and other fungal endophytes. Whether the effect of Epichloë endophytes on other fungal endophytes is connected to changes in phytohormone concentrations remains to be elucidated.

Epichloë festucae is a common symbiont of the perennial and widely distributed cool season grass, Festuca rubra. The symbiosis is highly integrated involving systemic growth of the fungus throughout above-ground host parts and vertical transmission from plant to its offspring via host seeds. However, the nature of symbiosis is labile ranging from antagonistic to mutualistic depending on prevailing selection pressures. Both the loss of fungus in the maternal host lineage and horizontal transmission through sexual spores within the host population may partly explain the detected variation in symbiosis in wild grass populations. Epichloë species are commonly considered as pathogens when they produce sexual spores and partly castrate their host plant. This is the pathogenic end of the continuum from antagonistic to mutualistic interactions. Here we examined the population genetic structure of E. festucae to reveal the gene flow, importance of reproduction modes, and alkaloid potential of the symbiotic fungus in Europe. Epichloë-species are highly dependent on the host in survival and reproduction whilst benefits to the host are largely linked to defensive mutualism attributable to fungal-origin bioactive alkaloids that negatively affect vertebrate and/or invertebrate herbivores. We detected decreased genetic diversity in previously glaciated areas compared to non-glaciated regions during the last glacial maximum period and found three major genetic clusters in E. festucae populations: southern, northeastern and northwestern Europe. Sexual reproduction may have a higher role than expected in Spanish E. festucae populations due to the predominance of unique genotypes and presence of both mating types in the region. In contrast, asexual reproduction via host seeds predominates in the Faroe Island and Finland in northern Europe due to the presence of biased mating-type ratios and large dominant genotypes in the E. festucae populations within the region. A substantially larger variation of alkaloid genotypes was observed in the fungal populations than expected, although the variability of the alkaloid genotypes within populations is considerably lower in northern than Spanish populations in southern Europe. E. festucae populations consist of different combinations of alkaloid classes from the gene clusters of ergot alkaloid and indole-terpenes, and from pyrrolopyrazine alkaloid gene. We suggest that the postglacial distribution history of the host grass, prevailing reproduction strategies of E. festucae, and local selection pressures likely explain a large part of the genetic variation observed in fungal populations among geographic regions. The identified alkaloid genotypes can be used by turfgrass breeders to improve resistance against herbivores in red fescue varieties and to develop new sustainable cultivars in Europe.

Vertically transmitted endophytic fungi can mitigate the negative effects of salinity encountered by their host grass and alter the competitive interactions between plant individuals. To experimentally study the interactive effects of the fungal endophyte Epichloë coenophiala on salt tolerance and intraspecific competition of its host plant, tall fescue Festuca arundinacea, we subjected 15 maternal lines of each Epichloë associated (E+) and Epichloë free (E−) tall fescue to salt treatment and competition in the greenhouse and common garden. Then, to explore variation in endophyte incidence in natural populations of tall fescue, we surveyed 23 natural populations occurring on or near the Baltic Sea coast in Aland islands in southwestern Finland for endophyte incidence, distance to shore, and competitive environment. Under salinity in the greenhouse, E− plants grew larger than E+ plants, but there was no size difference in the control treatment. E− plants grew taller and were more likely to flower than E+ plants when grown in benign conditions in the common garden but not with salinity or competition. The frequency of Epichloë incidence was high (90%) in natural populations, and it decreased towards the shore and risk of salt exposure. These results demonstrate a negative effect of Epichloë endophyte on the salt tolerance of its host. The high incidence of Epichloë in natural populations of tall fescue in the northern part of the species distribution range is likely due to factors other than salinity.

Abstract We conducted a large‐scale population genetic survey of genetic diversity of the host grass Festuca rubra s.l., which fitness can be highly dependent on its symbiotic fungus Epichloë festucae, to evaluate genetic variation and population structure across the European range. The 27 studied populations have previously been found to differ in frequencies of occurrence of the symbiotic fungus E. festucae and ploidy levels. As predicted, we found decreased genetic diversity in previously glaciated areas in comparison with nonglaciated regions and discovered three major maternal genetic groups: southern, northeastern, and northwestern Europe. Interestingly, host populations from Greenland were genetically similar to those from the Faroe Islands and Iceland, suggesting gene flow also between those areas. The level of variation among populations within regions is evidently highly dependent on the postglacial colonization history, in particular on the number of independent long‐distance seed colonization events. Yet, also anthropogenic effects may have affected the population structure in F. rubra. We did not observe higher fungal infection rates in grass populations with lower levels of genetic variability. In fact, the fungal infection rates of E. festucae in relation to genetic variability of the host populations varied widely among geographical areas, which indicate differences in population histories due to colonization events and possible costs of systemic fungi in harsh environmental conditions. We found that the plants of different ploidy levels are genetically closely related within geographic areas indicating independent formation of polyploids in different maternal lineages.

Glyphosate is the world’s most widely used agrochemical. Its use in agriculture and gardening has been proclaimed safe because humans and other animals do not have the target enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS). However, increasing numbers of studies have demonstrated risks to humans and animals because the shikimate metabolic pathway is present in many microbes. Here, we assess the potential effect of glyphosate on healthy human microbiota. Our results demonstrate that more than one-half of human microbiome are intrinsically sensitive to glyphosate. However, further empirical studies are needed to determine the effect of glyphosate on healthy human microbiota.

Microbes have driven eco-evolutionary adaptations organizing biodiversity from the origin of life. They are ubiquitous and abundant, facilitating the biochemical processes that make Earth habitable and shape ecosystem structures, functions, and services. Recent studies reveal that commensalistic and beneficial microbes associated with wild and domesticated plants may aid in establishing sustainable agriculture for a changing climate. However, developing microbe-based biotechnologies and ecosystem services requires a thorough understanding of the diversity and complexity of microbial interactions with each other and with higher organisms. We discuss the hot and blind spots in contemporary research on plant microbiomes, and how the latest molecular biological techniques and empirical eco-evolutionary approaches could elevate our perception of microbe–plant interactions through multidisciplinary studies.

Many grasses (Poaceae) have symbiotic fungal endophytes, which affect livestock by producing unpalatable or harmful secondary compounds. Less is known about the repelling effects of fungal endophytes on avian grazers despite potential wildlife management implications. Herbivorous goose (Branta spp.) species may become a nuisance in recreational use areas via fecal littering. Planting these areas with grasses that avian grazers avoid may help mitigate this damage. In 2016, we studied the foraging preference of the barnacle geese (B. leucopsis) with endophytic (E+) or endophyte-free (E-) red fescue (Festuca rubra) and/or tall fescue (Schedonorus phoenix) in 2 sites in Finland that had a history of nuisance geese damage. In the high grazing pressure site, we planted both grass species, while in the low grazing pressure site only tall fescue was used. Geese preference was measured as the percentage of the area grazed, the height of the residual grass grazed, and the number of fecal droppings in the grass plots. Geese foraging did not differ between E- and E+ grasses, but red fescues were preferred over tall fescues. This supports previous findings that tall fescues or other coarse species could reduce the attractiveness of recreational areas to geese.

Recent work on microbiomes is revealing the wealth and importance of plant-microbe interactions. Microbial symbionts are proposed to have profound effects on fitness of their host plants and vice versa, especially when their fitness is tightly linked. Here we studied local adaptation of host plants and possible fitness contribution of such symbiosis in the context of abiotic environmental factors. We conducted a four-way multi-year reciprocal transplant experiment with natural populations of the perennial grass Festuca rubra s.l. from northern and southern Finland, Faroe Islands and Spain. We included F. rubra with and without transmitted symbiotic fungus Epichloë that is vertically transmitted via host seed. We found local adaptation across the European range, as evidenced by higher host fitness of the local geographic origin compared with nonlocals at three of the four studied sites, suggesting that selection pressures are driving evolution in different directions. Abiotic factors did not result in strong fitness effects related to Epichloë symbiosis, indicating that other factors such as herbivory are more likely to contribute to fitness differences between plants naturally occurring with or without Epichloë. Nevertheless, in the case of asymmetric symbiosis that is obligatory for the symbiont, abiotic conditions that affect performance of the host, may also cause selective pressure for the symbiont.

Climate change may facilitate shifts in the ranges and the spread of insect pests, but a warming climate may also affect herbivorous insects adversely if it disrupts the locally adapted synchrony between the phenology of insects and that of their host plant. The ability of a pest species to colonize new areas depends on its ability to adjust the timing of phenological events in its life cycle, particularly at high latitudes where there is marked seasonality in temperature and day length. Here we incubated eggs of three species of geometrid moth, Epirrita autumnata, Operophtera brumata and Erannis defoliaria from different geographical populations (E. autumnata and O. brumata from Northern Finland, E. autumnata and E. defoliaria from Southern Finland and all three species from Germany) in a climate chamber at a constant temperature to determine the relative importance of geographic origin in the timing of egg hatch measured in terms of cumulative temperature sums (degree days above 5°C, DD5); i.e. the relative importance of local adaptation versus phenotypic plasticity in the timing of egg hatch. In all three species, eggs from northern populations required a significantly lower temperature sum for hatching than eggs from southern populations, but the differences between them in temperature sum requirements varied considerably among species, with the differences being largest for the earliest hatching and northernmost species, E. autumnata, and smallest for the southern, late-hatching E. defoliaria. In addition, the difference in hatch timing between the E. autumnata eggs from Southern Finland and Germany was many times greater than the difference between the two Finnish populations of E. autumnata, despite the fact that the geographical distances between these populations is similar. We discuss how these differences in hatching time may be explained by the differences in hatch-budburst synchrony and its importance for different moth species and populations. We also briefly reflect on the significance of photoperiod, which is not affected by climate change. It is a controller that works parallel or in addition to temperature sum both for egg hatch in moths and bud burst of their host plants.

Human assisted plant invasions from Europe to North America have been more common than the reverse. We tested endophyte-mediated performance of tall fescue in parallel three year experiments in Europe and the USA using endophyte infected and uninfected wild and cultivated plants. Experimental plants were subjected to nutrient and water treatments. Whereas endophyte infection increased tall fescue performance in general, the effects of endophytes on plant growth and reproduction varied among plant origins under different environmental conditions. Naturally endophyte-free Finnish cultivar 'Retu' performed equally well as 'Kentucky-31' in both geographic locations. All Eurasian origin plants performed well in the US. In Finland, plants established well and both cultivars survived over the first winter. However, winter mortality of 'Kentucky-31' plants was higher, particularly in fertilized soils in the subsequent winters. Our results suggest that tall fescue ecotype 'Kentucky-31' that flourishes in North America is poorly adapted to Northern European conditions.

Polyploidy and symbiotic Epichloë fungal endophytes are common and heritable characteristics that can facilitate environmental range expansion in grasses. Here we examined geographic patterns of polyploidy and the frequency of fungal endophyte colonized plants in 29 Festuca rubra L. populations from eight geographic sites across latitudes from Spain to northernmost Finland and Greenland. Ploidy seemed to be positively and negatively correlated with latitude and productivity, respectively. However, the correlations were nonlinear; 84% of the plants were hexaploids (2n = 6x = 42), and the positive correlation between ploidy level and latitude is the result of only four populations skewing the data. In the southernmost end of the gradient 86% of the plants were tetraploids (2n = 4x = 28), whereas in the northernmost end of the gradient one population had only octoploid plants (2n = 8x = 56). Endophytes were detected in 22 out of the 29 populations. Endophyte frequencies varied among geographic sites, and populations and habitats within geographic sites irrespective of ploidy, latitude or productivity. The highest overall endophyte frequencies were found in the southernmost end of the gradient, Spain, where 69% of plants harbored endophytes. In northern Finland, endophytes were detected in 30% of grasses but endophyte frequencies varied among populations from 0% to 75%, being higher in meadows compared to riverbanks. The endophytes were detected in 36%, 30% and 27% of the plants in Faroe Islands, Iceland and Switzerland, respectively. Practically all examined plants collected from southern Finland and Greenland were endophyte-free, whereas in other geographic sites endophyte frequencies were highly variable among populations. Common to all populations with high endophyte frequencies is heavy vertebrate grazing. We propose that the detected endophyte frequencies and ploidy levels mirror past distribution history of F. rubra after the last glaciation period, and local adaptations to past or prevailing selection forces such as vertebrate grazing.

Epichloë endophytes are common symbionts living asymptomatically in pooid grasses and may provide chemical defences against herbivorous insects. While the mechanisms underlying these fungal defences have been well studied, it remains unknown whether endophyte presence affects the host's own defences. We addressed this issue by examining variation in the impact of Epichloë on constitutive and herbivore-induced emissions of volatile organic compounds (VOC), a well-known indirect plant defence, between two grass species, Schedonorus phoenix (ex. Festuca arundinacea; tall fescue) and Festuca pratensis (meadow fescue). We found that feeding by a generalist aphid species, Rhopalosiphum padi, induced VOC emissions by uninfected plants of both grass species but to varying extents, while mechanical wounding failed to do so in both species after one day of damage. Interestingly, regardless of damage treatment, Epichloë uncinata-infected F. pratensis emitted significantly lower quantities of VOCs than their uninfected counterparts. In contrast, Epichloë coenophiala-infected S. phoenix did not differ from their uninfected counterparts in constitutive VOC emissions but tended to increase VOC emissions under intense aphid feeding. A multivariate analysis showed that endophyte status imposed stronger differences in VOC profiles of F. pratensis than damage treatment, while the reverse was true for S. phoenix. Additionally, both endophytes inhibited R. padi population growth as measured by aphid dry biomass, with the inhibition appearing greater in E. uncinata-infected F. pratensis. Our results suggest, not only that Epichloë endophytes may play important roles in mediating host VOC responses to herbivory, but also that the magnitude and direction of such responses may vary with the identity of the Epichloë-grass symbiosis. Whether Epichloë-mediated host VOC responses will eventually translate into effects on higher trophic levels merits future investigation.

It is well recognized, that outcomes of mutualistic plant-microorganism interactions are often context dependent and can range from mutualistic to antagonistic depending on conditions. Instead, seemingly pathogenic associations are generally considered only harmful to plants. The ergot fungus (Claviceps purpurea) is a common seed pathogen of grasses and cereals. Ergot sclerotia contain alkaloids which can cause severe toxicity in mammals when ingested, and thus the fungal infection might provide protection for the host plant against mammalian herbivores. Theoretically, the net effect of ergot infection would positively affect host seed set if the cost is not too high and the defensive effect is strong enough. According to our empirical data, this situation is plausible. First, we found no statistically significant seed loss in wild red fescue (Festuca rubra) inflorescences due to ergot infection, but the seed succession decreased along increasing number of sclerotia. Second, in a food choice experiment, sheep showed avoidance against forage containing ergot. Third, the frequency of ergot-infected inflorescences was higher in sheep pastures than surrounding ungrazed areas, indicating a protective effect against mammalian grazing. We conclude that, although ergot can primarily be categorized as a plant pathogen, ergot infection may sometimes represent indirect beneficial effects for the host plant. Ergot may thus serve as a conditional defensive mutualist for its host grass, and the pathogenic interaction may range from antagonistic to mutualistic depending on the situation.

Plant-associated micro-organisms such as mycotoxin-producing endophytes commonly have direct negative effects on herbivores. These effects may be carried over to natural enemies of the herbivores, but this has been rarely explored. We examined how feeding on Neotyphodium endophyte infected (E+) and endophyte free (E-) meadow ryegrass (Scherodonus pratensis) affects body mass, population size and mobility of sibling voles (Microtus levis), and whether the diet mediates the vulnerability of voles to least weasel (Mustela nivalis nivalis) predation. Because least weasels are known to be olfactory hunters, we also examined whether they are able to distinguish olfactory cues of voles fed on E+ and E- diets. Neither body mass of voles nor population size differed between diets. However, contrary to our prediction, least weasels preyed more often on voles fed with E- grass than on voles fed with E+ grass. The mobility of voles fed on E+ grass was reduced compared to voles fed on E- grass, but this effect was unrelated to risk of predation. Least weasels appeared unable to distinguish between excrement odours of voles between the two treatments. Our results suggest that consumption of endophytic grass is not directly deleterious to sibling voles. What's more, consumption of endophytes appears to be advantageous to voles by reducing risk of mammalian predation. Our study is thus the first to demonstrate an effect of plant-associated microbial symbionts on herbivore-predator interactions in vertebrate communities.

Even highly mutually beneficial microbial-plant interactions, such as mycorrhizal- and rhizobial-plant exchanges, involve selfishness, cheating and power-struggles between the partners, which depending on prevailing selective pressures, lead to a continuum of interactions from antagonistic to mutualistic. Using manipulated grass-endophyte combinations in a five year common garden experiment, we show that grass genotypes and genetic mismatches constrain genetic combinations between the vertically (via host seeds) transmitted endophytes and the out-crossing host, thereby reducing infections in established grass populations. Infections were lost in both grass tillers and seedlings in F(1) and F(2) generations, respectively. Experimental plants were collected as seeds from two different environments, i.e., meadows and nearby riverbanks. Endophyte-related benefits to the host included an increased number of inflorescences, but only in meadow plants and not until the last growing season of the experiment. Our results illustrate the importance of genetic host specificity and trans-generational maternal effects on the genetic structure of a host population, which act as destabilizing forces in endophyte-grass symbioses. We propose that (1) genetic mismatches may act as a buffering mechanism against highly competitive endophyte-grass genotype combinations threatening the biodiversity of grassland communities and (2) these mismatches should be acknowledged, particularly in breeding programmes aimed at harnessing systemic and heritable endophytes to improve the agriculturally valuable characteristics of cultivars.