Trade-off between seed number and weight: Influence of a grass–endophyte symbiosis

Plant fitness is enhanced by resource allocation to seed number (offspring number) or weight (offspring survival). Besides, there is a well known trade-off in resource allocation between both traits. Symbiotic interactions can influence plant resource allocation to reproduction, yet little research has been performed in this direction. We studied the consequences of a grass–fungus symbiosis on the trade-off between seed number and weight, using Lolium multiflorum and the endophyte Neotyphodium occultans as our study system. In ecological terms, we experimentally removed N. occultans from L. multiflorum plants, and compared reproductive allocation to seed number and weight in endophyte-symbiotic vs. non-symbiotic plants at different levels of nutrient availability (small pots vs. large pots). In evolutionary terms, we compared reproductive allocation between symbiotic vs. non-symbiotic plants for different host genotypes. All plants showed a negative association between seed number and weight, once standardized for total reproductive biomass. Under high nutrient availability, endophyte-symbiotic plants showed higher seed weight than non-symbiotic plants for any seed number. However, no differences were observed under low nutrient availability. Endophyte influence also varied according to L. multiflorum genotype; specifically, endophyte-symbiotic plants showed a lower slope in the relationship between seed number and weight than non-symbiotic plants for the ‘Marshall’ genotype but no endophyte influence was found for the “Pampean” genotype. The results implied a higher plasticity in seed weight and lower plasticity in seed number for symbiotic plants. Indeed, endophyte-symbiotic plants showed an overall lower slope in the association between seed number and total reproductive biomass than non-symbiotic plants. Our results suggest that N. occultans induces heavier seeds in L. multiflorum plants under environmental conditions favorable to plant growth or for certain plant genotypes. We propose that symbiotic interactions may influence the evolution of seed number and weight trade-off.     

内生真菌对羽茅生长及光合特性的影响

通过温室栽培实验,以感染两种内生真菌(Neotyphodium sibiricum和Neotyphodium gansuence)和未感染内生真菌的羽茅(Achnatherum sibiricum)为实验材料,分析感染不同种内生真菌对宿主植物的生长及光合特性的影响。结果表明,感染两种内生真菌羽茅的株高和CO₂补偿点显著低于未染菌的羽茅,而染菌羽茅的蒸腾速率和气孔导度显著高于未染菌羽茅,但对于感染不同种内生真菌的羽茅,无论是分蘖数与生物量的积累还是光合生理值之间均无显著差异。     

Production of phenolics and the emission of volatile organic compounds by perennial ryegrass (Lolium perenne L.)/Neotyphodium lolii association as a response to infection by Fusarium poae

Grasses very often form symbiotic associations with Neotyphodium/Epichloë endophytic fungi. These endophytes often allow the host grass to be protected from different pathogens. However, there is little known about the mechanisms of such endophyte influence on the host. Thus, the purpose of this research was to examine the effect of the N. lolii endophyte on the total production of phenolic compounds, VOCs emission and the resistance of three perennial ryegrass genotypes infected by pathogenic Fusarium poae. Analyses of total phenolics content were performed in control (not inoculated) and inoculated plants after 1, 2, 3, 4, 5, and 6 days (DAI) and for VOCs after 0, 3, 6 and 12 DAI. The presence of endophytes significantly reduced the disease index in two of the three genotypes relative to that in E−. Plants infected by N. lolii exhibited higher production of phenolics relative to the E− plants. The highest amounts of phenolics were observed on the second and sixth DAI. Genotype Nl22 showed the strongest effect of the endophyte on the production of phenolics, which increased by over 61%. Both the endophyte infected and non-infected plants emitted most abundantly two GLVs ((Z)-3-hexenal, (Z)-3-hexen-1-yl acetate), three terpenes (linalool, (Z)-ocimene, β-caryophyllene) and three shikimic acid pathway derivatives (benzyl acetate, indole, and methyl salicylate). The endophyte presence and the intervals of VOCs detection were a highly significant source of variation for all emitted volatiles (P < 0.001). The genotype of the perennial ryegrass significantly affected only the emission of methyl salicylate (P < 0.05) and β-caryophyllene (P < 0.05). Most of the VOCs ((Z)-3-hexen-1-yl acetate, (Z)-3-hexenal, linalool and methyl salicylate) reached their highest levels of emission on the sixth DAI, when averaged over genotypes and endophyte status. The results highlight the role of Neotyphodium spp. in the mediation of quadro-trophic interactions among plants, symbiotic endophytes, invertebrate herbivores and plant pathogenic fungi. Our results also confirm the fact that symbiotic plants can activate a defense reaction faster than non-symbiotic plants after a pathogen attack. Thus, N. lolii can be involved in the defense of perennial ryegrass against pathogens and potentially could be central to the host plants’ protection.     

Consequences of grazing on the vertical transmission of a fungal Neotyphodium symbiont in an annual grass population

Symbiosis between cool‐season grasses and vertically transmitted fungal endophytes are common and significantly impact on ecosystem function. This makes the understanding of the underlying mechanisms to symbiotic individuals frequency in local populations much more interesting. Most studies have been focused on the differential fitness between symbiotic and non‐symbiotic counterparts (relative fitness), barely considering other mechanisms. We performed a microcosms experiment to evaluate whether grazing alters the dynamics of the endophyte Neotyphodium occultans in the annual grass Lolium multiflorum by simultaneously modifying the relative fitness and the endophyte efficiency to be transmitted from host plants to seeds. Grazing was simulated by means of clipping and trampling on symbiotic and non‐symbiotic plants growing separately, in soils obtained from paddocks, differing in their agronomic management history (natural grassland vs. ryegrass promotion). Seed production showed a complex pattern as it depended on the symbiotic status of the plants, the level of grazing and the agro‐ecological context. Grazed plants produced three times fewer seeds than ungrazed plants only in microcosms with endophyte‐symbiotic plants in soils from ryegrass promotion. Endophyte benefits on seed production were exclusively observed in ungrazed plants in the same soil. Symbiotic plants produced symbiotic and non‐symbiotic seeds in all the treatments. While the production of non‐symbiotic seeds by these plants was not affected by grazing and the soil, grazing reduced the production of symbiotic seeds in both contexts. Grazing negative effect on the density of fully infected spikes determined a significant increment in the transmission failures which were not modified by agro‐ecological contexts. Therefore, grazing can modulate symbiosis dynamics through reducing seed production and endophyte transmission efficiency. Transmission has been disregarded, but it is a context‐dependent process that could lead to a gradual reduction in the symbiotic plants frequency in a population if the mutualism effectiveness does not outweigh transmission failures.     

Hybridization in Endophyte Symbionts Alters Host Response to Moisture and Nutrient Treatments

When a host organism is infected by a symbiont, the resulting symbiotum has a phenotype distinct from uninfected hosts. Genotypic interactions between the partners may increase phenotypic variation of the host at the population level. Neotyphodium is an asexual, vertically transmitted endophytic symbiont of grasses often existing in hybrid form. Hybridization in Neotyphodium rapidly increases the symbiotum’s genomic content and is likely to increase the phenotypic variation of the host. This phenotypic variation is predicted to enhance host performance, especially in stressful environments. We tested this hypothesis by comparing the growth, survival, and resource allocation of hybrid and nonhybrid infected host plants exposed to controlled variation in soil moisture and nutrients. Infection by a hybrid endophyte did not fit our predictions of comparatively higher root and total biomass production under low moisture/low nutrient treatments. Regardless of whether the host was infected by a hybrid or nonhybrid endophyte, both produced significantly higher root/total biomass when both nutrient and moisture were high compared to limited nutrient/moisture treatments. However, infection by hybrid Neotyphodium did result in significantly higher total biomass and host survival compared to nonhybrid infected hosts, regardless of treatment. Endophyte hybridization alters host strategies in response to stress by increasing survival in depauperate habitats and thus, potentially increasing the relative long-term host fitness.     

Asexual Neotyphodium endophytes in a native grass reduce competitive abilities

Asexual, vertically transmitted endophytes are well known for increasing competitive abilities of agronomic grasses, but little is known about endophyte–host interactions in native grasses. We tested whether the asexual Neotyphodium endophyte enhances competitive abilities in a native grass, Arizona fescue, in a field experiment pairing naturally infected (E+) and uninfected (E?) plants, and in a greenhouse experiment pairing E+ and E? (experimentally removed) plants, under varying levels of soil water and nutrients. In the field experiment, E? plants had greater vegetative, but not reproductive, growth than E+ plants. In the greenhouse experiment, where plant genotype was strictly controlled, E? plants consistently outperformed their E+ counterparts in terms of root and shoot biomass. Thus, Neotyphodium infection decreases host fitness via reduced competitive properties, at least in the short term. These findings contrast starkly with most endophyte studies involving introduced, agronomic grasses where infection increases competitive abilities, and the interaction is viewed as highly mutualistic.     

Does An Asexual Endophyte Symbiont Alter Life Stage and Long-Term Survival in a Perennial Host Grass?

Asexual, seedborne endophytic fungi in perennial grasses are often viewed as strong mutualists because fitness of the symbiont and host grass are closely coupled. However, at least for some native grasses, the asexual endophyte, Neotyphodium, acts parasitically, yet remains at high frequencies in natural populations. Most previous studies of Neotyphodium effects on host survival have been short term relative to the long life span of the perennial grass host. We therefore tested the hypothesis that Neotyphodium alters the survival in various life stages and long-term survival of adult native Arizona fescue (Festuca arizonica). To test the former, we planted 40 infected (E+) and 40 uninfected (E−, endophyte removed) seeds from four different maternal plants in the field under ambient conditions. We followed survival of seeds, seedlings, and adult plants over a 5-year period. To test the latter, we determined the infection of 1633 adult plants and followed their survival over the next 5–7 years. E+ seeds did not differ from uninfected seeds in terms of overall survival from seed germination to seedling to adult. However, the shape of the survival curve differed, with E+ plants showing higher mortality in early life stages. E+ adult plants did not differ from E− plants in long-term survival. Survival was generally very high during the study, which included a severe and prolonged drought. Infection by asexual Neotyphodium does not increase survival in early life stages or that of adult plants. Because asexual, vertically transmitted symbionts are predicted by evolutionary theory to be strong mutualists, the persistence of high infection frequencies in natural populations without long-term benefits to the host remains enigmatic. One possible explanation is that the long life span of the perennial host and low seedling recruitment may obscure either the costs or benefits of endophyte infection.     

Effects of the Neotyphodium endophyte fungus on dormancy and germination rate of Lolium multiflorum seeds

Abstract Neotyphodium frequently occurs as an endophyte in grasses. Evidence shows enhanced fitness of endophyte infected grasses relative to non‐infected ones. Some studies of seed germination show endophyte enhancement of plant fitness in various environments, but inconsistent results indicate that further studies are needed. So far, experiments have failed to separate the confounded effects of population origin and seed management. For this reason, we evaluated the effects of endophyte infection on seed dormancy and germination in Lolium multiflorum using an experimental design controlling these factors. Depending on the year of seed production, endophyte infection modified seed response to light quality, affecting predominantly seed dormancy levels. Nevertheless, the endophyte did not affect base temperature or thermal time of germination. We concluded that endophytes were not a strong influence on germination behaviour. We speculate from our results that the presence of the endophyte changes germination by an indirect effect, in extending growth of the maternal plant during seed development and ripening. The direct effect of hyphae in the seed on seed behaviour was disregarded, because the difference between infected and non‐infected seed varied within the year of seed production. Future experiments should focus on effects of the endophyte on the canopy of parent plants during seed production and ripening, and, hence, on subsequent dormancy and germination of the seeds.     

Conspicuous epiphytic growth of an interspecific hybrid Neotyphodium sp. endophyte on distorted host inflorescences

Selected Neotyphodium sp. endophytes are now commonly used to enhance pasture persistence and livestock productivity, with seed of perennial ryegrass and tall fescue cultivars with these selected endophytes being commercially available. In a large population of perennial ryegrass plants infected with a Neotyphodium sp. endophyte that was being grown for seed production a small percentage of inflorescences were distorted and covered with a conspicuous white mycelial growth. Within individual plants only a small number of inflorescences were affected and the amount of distortion differed between affected inflorescences. This Neotyphodium sp. is an interspecific hybrid of Epichloë typhina and Neotyphodium. lolii and like nearly all other Neotyphodium spp is symptomless in host grasses. The fungus isolated from distorted inflorescences had colonies that were identical to those isolated from symptomless inflorescences and these were characteristic of this Neotyphodium sp. This is the first report of distorted inflorescences covered with epiphytic hyphal growth on host grasses infected with an interspecific hybrid Neotyphodium sp.     

Host genotype overrides endophyte infection effects on growth,physiology, and nutrient content of a native grass,Achnatherum sibiricum

The effect of infection by the fungal endophyte Neotyphodium, host genotype, and their interaction on growth and physiology, as well as photosynthesis, was investigated in the native grass Achnatherum sibiricum. We artificially inoculated the endophyte into mature tillers of endophyte-free A. sibiricum. Plants were clipped to 5 cm height after recording growth traits, and analyzed for total nonstructural carbohydrates (TNC %), the percentage of nitrogen (N %), and carbon (C %) in leaves before and after clipping. In our study, the prominent host genotype–endophyte infection interactions detected in A. sibiricum indicates that, for many growth and storage traits, endophyte infection can impact a little change. However, there is no overriding consistently positive effect of the endophyte on growth or storage in A. sibiricum before or after clipping. Our study showed that the interaction between endophyte and host grasses was highly contingent on plant genotypes. We found host genotype overrode fungal endophyte infection in influencing tiller number and photosynthetic properties of A. sibiricum before clipping. After clipping, host genotype accounted for more of the variation in regrowth and above-ground biomass of A. sibiricum than endophyte infection. Our study showed that host genotype affected the total nonstructural carbohydrates of A. sibiricum before and after clipping, whereas endophyte infection increased the carbon content after clipping. Genotype by infection interactions for plant height, leaf mass, total nonstructural carbohydrates, and photosynthetic characteristics indicated genotype-specific effects of endophytes on A. sibiricum physiology and photosynthetic capacity. The host genotype–endophyte infection interactions detected in A. sibiricum suggest that host genotype overrides fungal endophyte infection on growth, physiology, and nutrient content of this native grass. In contrast, endophyte effects did not appear to positively affect growth, physiology, or photosynthetic capacity before or after clipping.     

Imperfect Vertical Transmission of the Endophyte <Emphasis Type="Italic">Neotyphodium</Emphasis> in Exotic Grasses in Grasslands of the Flooding Pampa

Cool-season grasses establish symbioses with vertically transmitted Neotyphodium endophytes widespread in nature. The frequency of endophyte-infected plants in closed populations (i.e., without migrations) depends on both the differential fitness between infected and non-infected plants, and the endophyte-transmission efficiency. Most studies have been focused on the first mechanism ignoring the second. Infection frequency and endophyte transmission from vegetative tissues to seeds were surveyed in two grasses growing in vegetation units that differ in flood and grazing regimes, and soil salinity. Transmission efficiency and infection frequency for tall fescue did not vary significantly and were 0.98 and 1.00, respectively. For Italian ryegrass, transmission efficiency and infection frequency were 0.88 and 0.57 in humid prairies, and 0.96 and 0.96 in the other vegetation units. Only in humid mesophytic meadows, the observed pattern was irrespective of the presence or absence of grazers. Our results suggest that selection forces for endophyte infection are different for both species. Imperfect transmission was only compensated in tall fescue through an increased fitness of infected plants. Interpreting variations of infection frequency only in terms of differential fitness can be misleading, considering that endophyte transmission can be imperfect and variable in nature. Therefore, this study highlights the importance of measuring transmission efficiency.     

Endophytic fungus fine‐tunes the persistence strategy of its alpine host grass in response to soil resource levels

An understanding of hereditary endophytic fungi, and the effects on grass persistence strategies (i.e. relative investment in sexual reproduction and vegetative growth) under natural conditions may help to predict how some alpine ecosystems will respond to environmental change. Grass persistence and endophyte maintenance in host populations are closely related, but could become independent due to endophyte loss mechanisms. We used native grass and endophyte populations to test the hypothesis that fungal endophytes manipulate grass persistence strategies to secure endophyte maintenance in plant populations. Two conditions were required to verify this hypothesis: 1) the fungus caused alterations in host plant strategies; and 2) plant phenotypic changes induced by the fungal endophyte increased endophyte transmission. We compared symbiotic (S) and non‐symbiotic (NS) persistence strategies of Festuca eskia (Poaceae), an alpine grass infected by the asexual form of the fungal endophyte Epichloë festucae. We characterised endophyte transmission efficiency, and described vegetative growth and sexual reproduction in a field population that naturally supports approximately 50% S plants. We built a demographic model to estimate plant vegetative growth rates. A correlation between plant persistence strategy, and fungal maintenance was evaluated by increasing soil resource levels. Under natural conditions, S and NS plants exploited different persistence strategies in the same population; S plants exhibited greater vegetative growth than their NS counterparts, while maintaining the same reproductive output. In response to higher soil resource levels, S plants shifted in persistence strategies and phenology, whereas NS plants maintained the same strategies. Therefore, results suggested the fungal endophyte fine‐tuned host persistence strategies according to soil resource level. Finally, we found no direct relationship between the changes induced by fungal endophyte and endophyte transmission. Consequently, fungal endophytes affected host persistence strategies, but did not directly increase endophyte transmission.     

Genetic Diversity and Structure of Neotyphodium Species and Their Host Achnatherum sibiricum in a Natural Grass–Endophyte System

Achnatherum sibiricum (Poaceae) is a perennial bunchgrass native to the Inner Mongolia Steppe of China. This grass is commonly infected by epichloë endophytes with high-infection frequencies. Previously, we identified two predominant Neotyphodium spp., N. sibiricum and N. gansuense. In the present study, genetic diversity and structure were analyzed for the two predominant Neotyphodium spp. as well as the host grass. We obtained 103 fungal isolates from five populations; 33 were identified as N. sibiricum and 61 as N. gansuense. All populations hosted both endophytic species, but genetic variation was much higher for N. gansuense than for N. sibiricum. The majority of fungal isolates were haploid, and 13% of them were heterozygous at one SSR locus, suggesting hybrid origins of those isolates. Significant linkage disequilibrium of fungal SSR loci suggested that both fungal species primarily propagate by clonal growth through plant seeds, whereas variation in genetic diversity and the presence of hybrids in both endophytic species revealed that although clonal propagation was prevalent, occasional recombination might also occur. By comparing genetic differentiation among populations, we found around 4–7-fold greater differentiation of endophyte populations than host populations, implying more restricted gene flow of endophytes than hosts. We proposed that endophyte infection of A. sibiricum might confer the host some selective advantages under certain conditions, which could help to maintain high-endophyte-infection frequencies in host populations, even when their gene flows do not match each other. Furthermore, we suggested that the same genotype of endophyte as well as host should be confirmed if the objective of the study is to know the influence of endophyte or host genotype on their symbiotic relationship, instead of just considering whether the plant is infected by an endophyte or not, since endophytes from the same host species could exhibit high levels of genetic diversity, which is likely to influence the outcome of their symbiotic relationship.     

Evidence for leaf endophyte regulation of root symbionts: effect of <Emphasis Type="Italic">Neotyphodium</Emphasis> endophytes on the pre-infective state of mycorrhizal fungi

Neotyphodium endophytes and arbuscular mycorrhizal (AM) fungi are common constituents of natural grasslands. The plant–endophyte symbiosis can introduce changes in soil conditions that affect the density and activity of different functional groups of soil organisms. In the present work we performed in vitro assays to evaluate the effect of root and endophyte exudates on the pre-infective state of mycorrhizal fungi (Gigaspora margarita and G. rosea). Plant roots of Bromus setifolius from populations of Patagonia, and four strains of Neotyphodium were used to obtain the exudates. Root exudates of infected plants, at a high concentration, significantly increased AMF hyphal branches and length relative to exudates from naturally endophyte free plants. The effect of Neotyphodium endophyte exudates on AMF mycelial length varied depending on strain and the concentration used, suggesting a differential interaction between endophyte and AMF species. AMF hyphal branches were increased by Neotyphodium fungal exudates in both mycorrhizal species. A few previous studies have suggested that Neotyphodium endophytes can reduce mycorrhizal sporulation and colonization of host roots in commonly-cultivated agronomic hosts. In this study we report the opposite effect in B. setifolius. This study reports the direct and positive effect of root exudates from plants in symbiosis with Neotyphodium, on AMF pre-infective state. Further, identical effects were detected using exudates from Neotyphodium endophytes.     

Dynamics of Neotyphodium endophyte infection in ageing seed pools: incidence of differential viability loss of endophyte,infected seed and non-infected seed

Symbiotic associations between grasses and vertically transmitted endophytic fungi are widespread in nature. Within grass populations, changes in the frequency of infected plants are driven by influence of the endophyte on the fitness of their hosts and by the efficiency of endophyte transmission from parent plants to their offspring. During the seed stage, the endophyte might influence the fitness of its host by affecting the rate of seed viability loss, whereas the efficiency of endophyte transmission is affected by losses of viability of the fungus within viable seeds. We assessed the viability losses of Lolium multiflorum seeds with high and low level of infection of the endophyte Neotyphodium occultans, as well as the loss of viability of the fungus itself, under accelerated seed ageing and under field conditions. Starting with high endophyte-infected accessions of L. multiflorum, we produced their low endophyte-infected counterparts by treating seeds with a fungicide, and subsequently multiplying seeds in adjacent plots allowing pollen exchange. In our accelerated ageing experiments, which included three accessions, high endophyte-infected seeds lost viability significantly faster than their low endophyte-infected counterpart, for only one accession. High endophyte-infected seeds of this particular accession absorbed more water than low endophyte-infected seeds. In contrast, the endophyte lost viability within live seeds of all three accessions, as the proportions of viable seeds producing infected seedlings decreased over time. In our field experiment, which included only one accession, high endophyte-infected seed lost viability significantly but only slightly faster than low endophyte-infected seed. In contrast, the loss of viability of the endophyte was substantial as the proportions of viable seeds producing infected seedlings decreased greatly over time. Moving the seeds from the air to the soil surface (simulating seed dispersion off the spikes) decreased substantially the rate of seed viability loss, but increased the rate of endophyte viability loss. Our experiments suggest that, in ageing seed pools, endophyte viability loss and differential seed mortality determine decreases in the proportions of endophyte-infected seeds in L. multiflorum. Endophyte viability loss within live seeds contributes substantially more to infection frequency changes than differential viability losses of infected and non-infected seeds.     

Are endophyte-mediated effects on herbivores conditional on soil nutrients?

Neotyphodium endophytes are assumed to have mutualistic relationship with their grass hosts, mainly resulting from mycotoxin production increasing plant resistance to herbivores by the fungus that subsists on the plant. To study importance of often ignored environmental effects on these associations, we performed a greenhouse experiment to examine the significance of endophyte infection and nutrient availability for bird-cherry aphid (Rhopalosiphum padi) performance on meadow fescue (Lolium pratense). Naturally endophyte-infected (E+), uninfected (E–), or manipulatively endophyte-free (ME–) half-sib families of meadow fescue were grown on two soil nutrient levels. Endophyte infection reduced aphid performance in general. However, to our knowledge, this is the first study to demonstrate experimentally that herbivore performance decreases on E+ host plants with increasing availability of nutrients in soils. Potential improvement in herbivore performance in high nutrient soils and decreased plant performance in low nutrient soils in ME– plants, compared to E– and E+ plants, suggests that loss of endophyte infection after long coevolutionary relationship may be critical to plant fitness.     

Fungal endophyte infection changes growth attributes in Lolium multiflorum Lam

Abstract Lolium multiflorum is a successful invader of postagricultural succession in the Inland Pampa grasslands in Argentina, becoming a dominant species in the plant community. Individual plants of this annual species are naturally highly infected with fungal endophytes (Neotyphodium sp.) from early successional stages. We assessed the effect of Neotyphodium infection on the biology of L. multiflorum. We evaluated growth attributes between endophyte infected (E+) and uninfected (E–) plants under non‐competitive conditions during the normal growing season. E+ plants produced significantly more vegetative tillers and allocated more biomass to roots and seeds. Although seed germination rates were greater in endophyte free plants, the rate of emergence and the final proportion of emerged seedlings were similar between the biotypes. The greater production of vegetative tillers, and the greater resource allocation to roots and seeds are likely to confer an ecological advantage to E+ plants, thus enabling their dominance over the E– individuals in natural grasslands.     

Family issues: fungal endophyte protects host grass from the closely related pathogen Claviceps purpurea

Certain cool season grasses establish systemic and asymptomatic symbioses with clavicipitaceous fungi of the genus Neotyphodium, which affect multiple biotic interactions within host neighborhood. The presumed symbiont-mediated plant resistance to pathogens is mostly based on studies performed under laboratory and greenhouse conditions. Here we investigated, in two outdoor experiments, the relation between two fungi of the same family with opposite effects on Lolium multiflorum plants: the mutualist endophyte Neotyphodium occultans, and the pathogen Claviceps purpurea. Natural infection and its consequences on symbiotic and non-symbiotic plants were studied under varying conditions of stress by herbicide. In both experiments, N. occultans reduced significantly the infection by C. purpurea at population levels (70 % less). The percentage of spikes infected by C. purpurea was almost three times lower in endophyte-symbiotic plants than in non-symbiotic ones. However, the protective effect was not maintained under stress condition. Our results show that constitutive symbionts such as the systemic fungal endophytes mediate the interaction between host grasses and pathogens, although the effect may depend on the level of stress in the environment.     

Neotyphodium endophyte infection frequency in annual grass populations: relative importance of mutualism and transmission efficiency

Persistence and ubiquity of vertically transmitted Neotyphodium endophytes in grass populations is puzzling because infected plants do not consistently exhibit increased fitness. Using an annual grass population model, we show that the problems for matching endophyte infection and mutualism are likely to arise from difficulties in detecting small mutualistic effects, variability in endophyte transmission efficiency and an apparent prevalence of non-equilibrium in the dynamics of infection. Although endophytes would ultimately persist only if the infection confers some fitness increase to the host plants, such an increase can be very small, as long as the transmission efficiency is sufficiently high. In addition, imperfect transmission limits effectively the equilibrium infection level if the infected plants exhibit small or large reproductive advantage. Under frequent natural conditions, the equilibrium infection level is very sensitive to small changes in transmission efficiency and host reproductive advantage, while convergence to such an equilibrium is slow. As a consequence, seed immigration and environmental fluctuation are likely to keep local infection levels away from equilibrium. Transient dynamics analysis suggests that, when driven by environmental fluctuation, infection frequency increases would often be larger than decreases. By contrast, when due to immigration, overrepresentation of infected individuals tends to vanish faster than equivalent overrepresentation of non-infected individuals.     

Co-occurring patterns of endophyte infection and genetic structure in the alpine grass, <Emphasis Type="Italic">Festuca eskia</Emphasis>: implications for seed sourcing in ecological restoration

Choosing the provenance of seed used in ecological restoration could entail its success. An alternative approach to examine local adaptation in seed sourcing is the assessment of genetic structure and diversity based on molecular markers. These types of analyses focus on the genetics of the target plant itself and eliminate the genetic influence of associated organisms, such as Epichloë/Neotyphodium endophytes in grasses. By impacting the fitness of their host, such symbionts may influence population genetic structure and diversity. Therefore, seed sourcing for grasses must consider the influence of their endophytes to increase seed translocation success and minimize the risks associated with this practice. To delineate seed zones for restoration of the alpine fescue Festuca eskia Ramond ex. DC. (Poaceae), we assessed population genetic differentiation and diversity patterns in the species including endophyte occurrence along altitudinal and longitudinal gradients in the Pyrenees Mountains. Twenty-three populations were analysed for endophyte status, and three STS and one SSR marker were used to examine genetic differentiation and diversity patterns. Results showed that F. eskia hosts an asexual form of Epichloë and infection frequency within populations decreased from East to West (100 vs. 8–25%). Molecular markers separated F. eskia into two East and West groups, and endophyte infection and genetic patterns were congruent with molecular data. Little evidence for genetic differentiation or difference in endophyte occurrence associated with altitude was detected. Little variation was found in within population diversity, regardless of provenance altitude and site, and/or endophyte infection frequency. The results of this study suggested the establishment of two distinct management units for F. eskia seed sourcing restoration.