Insect oviposition preference between Epichloë‐symbiotic and Epichloë‐free grasses does not necessarily reflect larval performance

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.     

Altered allocation to roots and shoots in the endophyte‐infected seedlings of Puccinellia distans (Poaceae)

Endophytes play an important role in ecological and evolutionary processes in plants and have marked economic value. Seed‐transmitted fungal endophytes are conventionally regarded as mutualistic symbionts, but their fitness consequences for the offspring of the host are not clear. Puccinellia distans infected with the fungus Epichloë typhina (E+) produces seeds that are several times smaller than normal (E?). This observation suggests that the E+ seedlings face a developmental disadvantage. Our growth chamber experiments compared the germination rates of the small E+ and large E? seeds of P. distans and examined the biomass allocation of seedlings to roots and shoots. The E+ seedlings germinated more slowly and maintained shorter shoots and a smaller root biomass for 30–50 days after sowing. Despite this disadvantage, the E+ plants more quickly increased their total size, attaining a larger shoot and whole‐plant biomass. The shoot:root biomass ratio increased more rapidly through time in the E+ seedlings, attaining a value nine times higher in the E+ than the E? group 50 days after sowing. Such differences between the E+ and E? seedlings were not explained by the growth allometry between shoots and roots. The seedlings of P. distans infected with the Epichloë endophyte were initially handicapped by their postponed emergence, but this disadvantage was quickly overcome by their superior growth capacity. The decrease in the relative allocation to roots may indicate that endophytes increase the performance of roots as resource‐acquiring organs and/or reduce the role of roots in protection against herbivores.     

Plant–fungal symbiosis affects litter decomposition during primary succession

Microbial symbionts of plants can affect decomposition by altering the quality or quantity of host plant tissue (substrate) or the micro‐environment where decomposition occurs (conditioning). In C₃ grasses, foliar fungal endophytes (Clavicipitaceae) can increase plant resistance to drought and/or produce alkaloids that reduce herbivory – effects that may also influence host litter composition and subsequent litter decomposition. We studied the effect of the endophyte Epichloë sp. on litter decomposition in the Great Lakes dunes (USA) using a reciprocal design altering endophyte presence/absence in both American beachgrass Ammophila breviligulata substrate (litter bags) and its conditioning of the decomposition microenvironment. Symbiont treatments were crossed with rain‐out shelters that altered growing season precipitation. The first year of decomposition, senesced leaf substrate from A. breviligulata with Epichloë decomposed 21% faster than endophyte‐free substrate. By the third year, conditioning by live symbiotic plants reduced cumulative decomposition by 33% compared to plots planted with endophyte‐free plants. Of the traits we examined – litter quantity, C:N ratio, mineral composition, fungal colonization, and carbon chemistry – increased litter quantity via greater tiller production was the primary trait shift associated with endophyte symbiosis. Epichloë in A. breviligulata litter also altered litter nitrogen decomposition dynamics, as evidenced by lower nitrogen and protein content in decomposed tissue from plants that hosted the endophyte. Differences in initial litter quality and subsequent colonization by saprotrophic fungi were ruled out as key drivers. Altered precipitation had negligible effects on decomposing processes in the dunes. Grass–Epichloë symbiosis altered nutrient cycling through increasing the rate of litter decomposition when present in the litter and through reducing litter decomposition by conditioning the decomposition microenvironment. Epichloë are widespread symbionts of grasses. Thus, their effects on decomposition could be an important, but often overlooked, driver of nutrient cycling in grass‐dominated ecosystems.     

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。     

Genetic diversity in epichloid endophytes of Hordelymus europaeus suggests repeated host jumps and interspecific hybridizations

Epichloid fungal endophytes (Epichloë and Neotyphodium spp.) are excellent model systems for studying speciation processes because of their variable life history traits that are linked to host grass fitness. Presumed jumps to new hosts and subsequent somatic hybridizations appear to be common among epichloid endophytes resulting in increased genetic variation upon which selection can act and speciation be initiated. In this study, we explored the endophyte diversity of a rare European native woodland grass species, Hordelymus europaeus, along a latitudinal transect covering the entire distribution range of H. europaeus. From 28 populations in six countries, isolates were sampled and molecularly characterized. Based on the sequences of tubB and tefA, six distinct epichloid taxa (interspecific hybrid or cryptic haploid species) were found, of which four were novel and two have been previously reported from this host. Of the novel endophytes, two were presumed to be interspecific hybrids and two of nonhybrid origin. While previously known endophytes of H. europaeus are seed‐born and strictly asexual, one of the novel nonhybrid endophytes found in the glacial refugium of the Apennine peninsula reproduced sexually in cultured plants. This is the first case of a seed‐borne, but sexually reproducing endophyte of this host. We discuss the origin, and possible ancestral species, of the six epichloid taxa using phylogenetic analyses. Repeated host jumps and somatic hybridizations characterize the diversity of the endophytes. To date, no other grass species is known to host a larger diversity of endophytes than H. europaeus.     

Effects of the Epichloë fungal endophyte symbiosis with Schedonorus pratensis on host grass invasiveness

Initial studies of grass–endophyte mutualisms using Schedonorus arundinaceus cultivar Kentucky‐31 infected with the vertically transmitted endophyte Epichloë coenophiala found strong, positive endophyte effects on host‐grass invasion success. However, more recent work using different cultivars of S. arundinaceus has cast doubt on the ubiquity of this effect, at least as it pertains to S. arundinaceus–E. coenophiala. We investigated the generality of previous work on vertically transmitted Epichloë‐associated grass invasiveness by studying a pair of very closely related species: S. pratensis and E. uncinata. Seven cultivars of S. pratensis and two cultivars of S. arundinaceus that were developed with high‐ or low‐endophyte infection rate were broadcast seeded into 2 × 2‐m plots in a tilled, old‐field grassland community in a completely randomized block design. Schedonorus abundance, endophyte infection rate, and co‐occurring vegetation were sampled 3, 4, 5, and 6 years after establishment, and the aboveground invertebrate community was sampled in S. pratensis plots 3 and 4 years after establishment. Endophyte infection did not enable the host grass to achieve high abundance in the plant community. Contrary to expectations, high‐endophyte S. pratensis increased plant richness relative to low‐endophyte cultivars. However, as expected, high‐endophyte S. pratensis marginally decreased invertebrate taxon richness. Endophyte effects on vegetation and invertebrate community composition were inconsistent among cultivars and were weaker than temporal effects. The effect of the grass–Epichloë symbiosis on diversity is not generalizable, but rather specific to species, cultivar, infection, and potentially site. Examining grass–endophyte systems using multiple cultivars and species replicated among sites will be important to determine the range of conditions in which endophyte associations benefit host grass performance and have subsequent effects on co‐occurring biotic communities.     

内生真菌对氮添加羽茅根际土壤特性和微生物群落的影响

内生真菌不仅能改变与其共生植物的生理和生长指标,还可通过宿主植物间接对土壤的理化性质和微生物群落结构产生影响。以天然禾草——羽茅(Achnatherum sibiricum)为研究材料,探究在不同施氮水平下,内生真菌的种类对不同基因型的宿主植物根际土壤理化性质和微生物群落产生何种影响。结果表明,内生真菌侵染显著提高了羽茅根际土壤的pH值和微生物总量,但降低了土壤中真菌与细菌的比值。同时,土壤的pH值还受到了内生真菌种类的影响,其中感染Epichlo3 sibirica的羽茅根际土壤pH显著高于感染Epichlo3 gansuense-1的羽茅,而感染Epichlo3 gansuense-2的羽茅根际土壤pH与感染E.gansuensis-1、E.sibirica菌的羽茅相比没有显著差异。另外,内生真菌感染与否、内生真菌种类、施氮量以及宿主植物基因型对土壤总碳、总氮、微生物及碳矿化能力均无显著影响。     

Genetic structure of natural populations of the grass endophyte Epichloë festucae in semiarid grasslands

Plants of red fescue (Festuca rubra), a commercially important turf grass, are infected by the fungal endophyte Epichloë festucae in semiarid natural grasslands, known as dehesas, in western Spain. We used amplified fragment length polymorphism (AFLP) markers to analyse the genetic polymorphism existing in two natural populations of Epichloë festucae. Linkage disequilibrium and the presence of clonal lineages indicated that nonrecombinant asexual reproduction predominates in both populations. However, most genetic variation detected was found to occur within populations, with only a moderate amount of genetic differentiation between populations (F_ST: 0.197). Overall, the study suggests that dehesa grasslands are useful reservoirs of Epichloë festucae endophytes, and provides information on population structure which is relevant to design sampling strategies.     

Direct and indirect effects of the fungal endophyte <Emphasis Type="Italic">Epichloë uncinatum</Emphasis> on litter decomposition of the host grass, <Emphasis Type="Italic">Schedonorus pratensis</Emphasis>

Microbial plant symbionts have been suggested to mediate plant-soil feedback and affect ecosystem functions. Systemic Epichloë fungal endophytes of grasses are found to mediate litter decomposition. These effects are often linked to alkaloids produced by Epichloë species, which are hypothesized to negatively affect decomposers. Although endophytes have been found to affect plant community and soil biota, direct (through litter quality) and indirect (through the environment) effects of fungal endophytes on litter decomposition have been scarcely scrutinized. We placed litterbags with endophyte-symbiotic (E+) and non-symbiotic (E?) Schedonorus pratensis plant litter in plots dominated by E+ or E? plants of the same species, and followed the dynamics of mass losses over time. We predicted the endophyte would hinder decomposition through changes in litter quality and that both types of litter would decompose faster in home environments. E+ litter decomposed faster in both environments. The mean difference between decomposition rate of E+ and E? litter tended to be higher in E? plots. Nitrogen and phosphorus, two elements usually associated with high decomposition rates, were significantly lower in E+ litter. We also detected a higher proportion of C in the cellulose form in E+ litter. Contrary to the general assumption, we found that symbiosis with Epichloë fungal endophytes can be associated with higher decomposition of plant litter. Since direct effects of Epichloë fungi were still stronger than indirect effects, it is suggested that besides the alkaloids, other changes in plant biomass would explain in a context-dependent manner, the endophyte effects on the litter decomposition.     

感染内生真菌的羽茅对大针茅的化感作用

以羽茅(Achnatherum sibiricum)为材料,采用室内生物测定法研究感染不同内生真菌的羽茅浸提液对大针茅(Stipa grandis)种子萌发和幼苗生长的化感作用,并采用气相色谱-质谱(GC-MS)联用技术对羽茅浸提液中的化学成分进行分析。结果表明:(1)内生真菌感染可缓解宿主羽茅对群落优势种大针茅的化感作用;(2)内生真菌感染对宿主羽茅化感作用的影响与内生真菌种类有关,具体表现为相比于未染菌羽茅,感染Epichloё gansuensis可显著缓解宿主羽茅对大针茅的抑制作用,而感染Neotyphodium sibiricum、Neotyphodium gansuensis有缓解羽茅对大针茅化感作用的趋势,但未达到显著影响;(3)羽茅浸提液中含有2,4-二叔丁基苯酚、邻苯二甲酸、邻苯二甲酸二丁酯和硬脂酸甲酯等潜在化感物质,这4种物质相对含量的差异可能是不同内生真菌感染状态的羽茅产生不同化感效应的主要原因;(4)对羽茅浸提液中4种潜在化感物质的化感作用进行验证实验,结果表明,2,4-二叔丁基苯酚对大针茅的化感作用基本表现为低浓度无影响,高浓度抑制,邻苯二甲酸二丁酯的化感作用表现为"低促高抑"的效应趋势,而邻苯二甲酸和硬脂酸甲酯对大针茅均表现出"剂量效应"的化感作用趋势,即4种潜在化感物质在高浓度时均可显著抑制大针茅的种子萌发和幼苗生长。     

Contribution of ergot alkaloids to suppression of a grass-feeding caterpillar assessed with gene knockout endophytes in perennial ryegrass

Neotyphodium and Epichloë species (Ascomycota: Clavicipitaceae) are fungal symbionts (endophytes) of grasses. Many of these endophytes produce alkaloids that enhance their hosts’ resistance to insects or are toxic to grazing mammals. The goals of eliminating from forage grasses factors such as ergot alkaloids that are responsible for livestock disorders, while retaining pasture sustainability, and of developing resistant turf grasses, require better understanding of how particular alkaloids affect insect herbivores. We used perennial ryegrass Lolium perenne L. (Poaceae) symbiotic with Neotyphodium lolii × Epichloë typhina isolate Lp1 (a natural interspecific hybrid), as well as with genetically modified strains of Lp1 with altered ergot alkaloid profiles, to test effects of ergot alkaloids on feeding, growth, and survival of the black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), a generalist grass‐feeding caterpillar. Neonates or late instars were provided clippings from glasshouse‐grown plants in choice and rearing trials. Wild‐type endophytic grass showed strong antixenosis and antibiosis, especially to neonates. Plant‐endophyte symbiota from which complex ergot alkaloids (ergovaline and lysergic acid amides such as ergine) or all ergot alkaloids were eliminated by endophyte gene knockout retained significant resistance against neonates. However, this activity was reduced compared to that of wild‐type Lp1, providing the first direct genetic evidence that ergot alkaloids contribute to insect resistance of endophytic grasses. Similarity of larval response to the two mutants suggested that ergovaline and/or ergine account for the somewhat greater potency of wild‐type Lp1 compared to the knockouts, whereas simpler ergot alkaloids contribute little to that added resistance. All of the endophyte strains also produced peramine, which was probably their primary resistance component. This study suggests that ergot alkaloids can be eliminated from an endophyte of perennial ryegrass while retaining significant insect resistance.     

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.     

Effects of the endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue in different saline-alkali soils

Soil salinization is detrimental to plant growth and yield in agroecosystems worldwide. Epichloë endophytes, a class of clavicipitaceous fungi, enhance the resistance of host plants to saline-alkali stress. This study explored the effects of the systemic fungal endophyte Epichloë coenophiala on the root microbial community and growth performance of tall fescue (Lolium arundinaceum) growing under different saline-alkali stress conditions. Structural equation modeling (SEM) was conducted to analyze the direct and indirect effects (mediated by root microbial community diversity and soil properties) of the endophyte on the growth of tall fescue under saline-alkali stress. The endophyte-infected plants produced higher shoot and root biomass compared to endophyte-free plants under saline-alkali stress (200 and 400 mM). Endophyte infection increased the fungal community diversity and altered its composition in the roots, decreasing the relative abundance of Ascomycota and increasing that of Glomeromycota. Furthermore, endophyte infection decreased the bacterial community diversity and the relative abundance of dominant Proteobacteria. SEM showed that endophyte infection increased the shoot and root biomass under saline-alkali stress (200 and 400 mM) by increasing the arbuscular mycorrhizal fungal diversity in the roots, and soil total nitrogen and phosphorus concentrations. Therefore, it is important to examine aboveground microbes as factors influencing plant growth in saline-alkali stress by affecting belowground microbes and soil chemical properties.     

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.     

Can seed-borne endophytes promote grass invasion by reducing host dependence on mycorrhizas?

Symbiotic interactions between plants and microorganisms have recently become the focus of research on biological invasions. However, the interaction between different symbionts and their consequences in host-plant invasion have been seldom explored. Here, we propose that vertically transmitted fungal endophytes could reduce the dependency of invasive grasses on mycorrhizal fungi allowing host establishment in those environments where the specific mutualist may be not present. Through analyzing published studies on nine grass species, we evaluated the effect of seed-borne Epichloë endophytes on the relationship of invasive and non-invasive grasses with arbuscular mycorrhizal fungi (AMF), a symbiosis known to be fundamental for plant fitness and invasion success. The endophyte effect on AMF colonization differed between invasive and non-invasive grasses, reducing mycorrhization only on invasive species but with no impact on their biomass. These results allowed us to propose that Epichloë endophytes could reduce the dependency of host plants on the mutualism with AMF, promoting host grass establishment and subsequent invasion. Simultaneous interactions with different types of mutualists may have profound effects on the host-plant fitness facilitating its range expansion. Our findings suggest that some specific mutualistic fungi such as epichloid endophytes facilitate host invasion by reducing the requirements of the benefits derived from other mutualisms.     

Soil microbial and chemical responses to foliar Epichloë fungal infection in Lolium perenne,Hordeum brevisubulatum and Achnatherum inebrians

We tested whether the host species identity in grass-Epichloë symbioses affected soil chemical and microbial properties. We grew endophyte infected (E+) and endophyte free (E−) Lolium perenne, Hordeum brevisubulatum and Achnatherum inebrians for 18 months in field plots. In E+ soil of all three grasses, available phosphorus was lower whereas total soil nitrogen was higher. Endophyte effects on soil pH, microbial biomass nitrogen, total carbon and organic carbon as well as bacteria and fungi abundance were host species dependent. Ammonia oxidizing bacteria abundance was higher in E+ soils for all species. Bacterial community composition of E+ and E− soils were different only for Lolium perenne with soil pH being the key factor. Fungal community composition of E+ and E− soils was not different for the three grasses. This study confirmed that the effects of foliar Epichloë infection on belowground properties depended on host species identity.     

Consequences of simultaneous interactions of fungal endophytes and arbuscular mycorrhizal fungi with a shared host grass

Plants simultaneously associate with multiple microbial symbionts throughout their lifetimes. To address the question of whether the effects of simultaneous symbionts are contingent on the specific identities, we conducted a greenhouse experiment manipulating the presence and identities of arbuscular mycorrhizal fungi (AMF) and fungal endophytes on the shared host grass Elymus hystrix. Each plant host was inoculated with one of two AMF species having varying effects on host growth, or a sterile soil control. Further, we used naturally occurring endophyte‐infected (E+) and uninfected (E–) individuals from two populations of the endophyte Epichloë elymi that varied in their interaction with E. hystrix. We then measured responses of plants, AMF, and fungal endophytes. Overall, we found that the combined effects of AMF and fungal endophytes on plant growth were additive, reflecting the mutualistic quality of each symbiont independently interacting with host plants. However, fungal endophyte infection differentially altered hyphal colonization of the two AMF species and the identity of the coinfecting AMF species affected fungal endophyte fitness traits. The results of this study demonstrate that the outcome of interspecific symbiotic interactions varies with partner identity such that the effects of simultaneous symbioses can not be generalized.     

Effects of cross host species inoculation of nitrogen‐fixing endophytes on growth and leaf physiology of maize

Plants that grow and thrive under abiotic stress often do so with the help of endophytic microorganisms. Although nitrogen‐fixing (diazotrophic) endophytes colonize many wild plants, these natural relationships may be disrupted in cultivated crop species where breeding and genotype selection often occur under conditions of intensive fertilization and irrigation. Many energy crops including corn may still benefit from diazotrophic endophyte inoculations allowing for more efficient biomass production with less input of petroleum‐derived fertilizer. A selection of diazotrophic endophytes isolated from willow (Salix sitchensis, Sitka willow) and poplar (Populus trichocarpa, black cottonwood) growing in nutrient‐poor river sides were used as inoculum in three experiments testing the effect on plant growth and leaf level physiology of a sweet corn variety under various levels of applied nitrogen fertilizer. We report substantial growth promotion with improved leaf physiology of corn plants in response to diazotrophic endophyte inoculations. Significant gains of early biomass with a greater root : shoot ratio were found for plants receiving endophytic inocula over the uninoculated control groups regardless of the nitrogen level. Furthermore, inoculated plants exhibited consistently higher rates of net CO₂ assimilation than did those without endophytic inoculation. These results have beneficial implications for enhanced plant growth in a low‐input system on nutrient‐poor sites. The immediate increase of root mass observed in endophyte inoculated plants has the potential to provide better establishment and early growth in resource‐limited environments. The initial results of this study also indicate that the beneficial effect from endophytes isolated from poplar and willow species is not restricted to the species from which they were initially isolated.     

Role of foliar fungal endophytes in litter decomposition among species and population origins

Litter decomposition is a key ecosystem process which returns nutrients from dead plant material to mineral forms in the soil. We examined whether systemic fungal endophytes modulate recycling of nutrients directly by altering litter decomposition. We studied litter decomposition mediated by Epichloë endophytes in litter-bag experiments. We examined direct endophyte effects on litter decomposition in wild populations and cultivars of Schedonorus phoenix and Schedonorus pratensis. In the first experiment, endophyte presence tended to increase litter decomposition rate in cultivars of the two grass species (S. phoenix and S. pratensis). However, in the second experiment plant origin had a stronger influence than endophyte symbiosis in S. phoenix. Interestingly, the initial level of alkaloids was associated positively with decomposition in S. phoenix populations. Characteristics associated with litter quality were not clearly related to either endophytes or decomposition rate. Our results suggest that endophytes can enhance litter breakdown but their role in nutrient cycling is far more complex depending on plant population origin.     

An ecological framework for understanding the roles of Epichloë endophytes on plant defenses against fungal diseases

Plants harbor a wide diversity of microorganisms in their tissues. Some of them have a long co-evolutionary history with their hosts, likely playing a pivotal role in regulating the plant interaction with other microbes such as pathogens. Some cool-season grasses are symbiotic with Epichloë fungal endophytes that grow symptomless and systemically in aboveground tissues. Among the many benefits that have been ascribed to endophytes, their role in mediating plant interactions with pathogens has been scarcely developed. Here, we explored the effects of Epichloë fungal endophytes on the interaction of host grasses with fungal pathogens. We made a meta-analysis that covered a total of 18 host grass species, 11 fungal endophyte species, and 22 fungal pathogen species. We observed endophyte-mediated negative effects on pathogens in vitro and in planta. Endophyte negative effects on pathogens were apparent not only in laboratory but also in greenhouse and field experiments. Epichloë fungal endophytes had negative effects on pathogen growth and spores' germination. On living plants, endophytes reduced both severity and incidence of the disease as well as colonization and subsequent infection of seeds. Symbiosis with endophytes showed an inhibitory effect on debilitator and killer pathogens, but not on castrators, and this effect did not differ among biotrophic or necrotrophic lifestyles. We found that this protection can be direct through the production of fungistatic compounds, the competition for a common resource, or the induction of plant defenses, and indirect associated with endophyte-generated changes in the abiotic or the biotic environment. Several mechanisms operate simultaneously and contribute differentially to the reduction of disease within grass populations.