Search Results (134)

Epichloë endophytes enhance plant defense and biotic stress resistance through mutualistic interactions in natural hosts. However, whether these protective effects are retained in annual non-native hosts such as barley (Hordeum vulgare L.) remains unclear. An integrated multi-omics approach was used to characterize defense responses in Epichloë bromicola-infected barley (LD1) and endophyte-free barley (CK) following pathogen challenge. The results show that LD1 had 20.0% lower disease incidence and 24.1% lower disease index than CK. LD1 also exhibited 21.1% lower malondialdehyde (MDA) content and higher antioxidant enzyme activities, indicating more effective control of oxidative damage and improved redox homeostasis. Integrated transcriptomic and metabolomic analyses revealed distinct defense responses between LD1 and CK. LD1 showed stronger activation of glutathione metabolism and antioxidant pathways than CK after 6 h of endophyte inoculation. This was consistent with more efficient control of early reactive oxygen species (ROS) dynamics. By 48 h, LD1 preferentially enriched phenylpropanoid biosynthesis and mitogen-activated protein kinase (MAPK) signaling, accompanied by increased accumulation of defense-related metabolites and enhanced structural and chemical defenses. Collectively, the study demonstrates that Epichloë bromicola enhances disease resistance in barley by reprogramming host defense dynamics from early redox regulation to late structural and chemical reinforcement. Full article

Epichloë endophytes can confer diverse benefits to host grasses, but the differences in effects between strains from different populations are poorly understood. In this study, we compared the impacts of two Epichloë bromicola strains isolated from distinct geographic populations of Hordeum bogdanii: GS1 (from Linze County, Gansu Province) and WS1 (from Wensu County, Xinjiang Province). Through controlled inoculation experiments, we established two new symbionts—HE2 (WS1 transferred to endophyte-free GF plants) and HE3 (GS1 transferred to endophyte-free WF plants)—alongside the natural symbionts GI (GS1-harboring) and WI (WS1-harboring) and corresponding endophyte-free controls (GF and WF). Symbiosis was confirmed by microscopic observation of blue-stained hyphae, re-isolation of fungi, and molecular identification using tef and tub gene sequences. Strikingly, the two strains exerted opposite effects on host photosynthesis. GS1-colonized plants (GI and HE3) maintained normal chloroplast ultrastructure, showed increased chlorophyll a, chlorophyll b, and carotenoid contents, and exhibited enhanced net photosynthetic rate, transpiration rate, and stomatal conductance, comparable to or exceeding those of control WF. In contrast, WS1-colonized plants (WI and HE2) had deformed chloroplasts, reduced pigment contents, and depressed gas exchange parameters, similar to control GF. Both newly generated symbionts accumulated more starch grains than their natural counterparts, indicating altered carbon partitioning. Phenotypic patterns were consistent across natural and novel associations, suggesting that fungal genotype drives outcomes. Differing physiological effects caused by strains from the same species and the same host but different populations indicate the importance of strain-level selection in agricultural applications. GS1 shows promise as a growth-promoting bioinoculant to enhance photosynthesis and productivity in forage grasses, particularly under marginal conditions. This study highlights how intraspecific variation and local adaptation shape grass–endophyte interactions and informs targeted use of symbionts in sustainable agriculture. Full article

Endophytic fungi widely colonize plant tissues without causing severe disease, protect hosts from pathogenic microorganisms, and represent a key potential resource for novel biocontrol agents. To explore the biocontrol potential of endophytic Epichloë fungi and their correlation with alkaloids, 12 Epichloë strains were isolated from six different geographic sites of Festuca sinensis, Achnatherum inebrians, and Hordeum brevisubulatum. The antifungal activity of these strains was evaluated against five phytopathogenic fungi (Alternaria alternata, Bipolaris sorokiniana, Curvularia lunata, Fusarium avenaceum, and Drechslera erythrospila) using dual-culture assays, which measured the inhibition of both colony growth and spore germination. Concurrently, the concentrations of ergonovine and ergine were quantified in the liquid cultures of each Epichloë strain. The results showed that 12 Epichloë strains had varying degrees of inhibitory effects on the colony growth and spore germination of five pathogenic fungi. Among these, strain F2 had the highest inhibition rate on the spore germination of B. sorokiniana (87.73%), while strain H3 had the lowest inhibition rate on F. avenaceum (7.89%). The concentrations of ergonovine and ergine were positively correlated with the inhibition rate of pathogenic fungi, but the degree of these correlations varied among different pathogenic fungi. This study provides further evidence for Epichloë-mediated pathogen inhibition and establishes a basis for their development as biocontrol agents in agricultural systems. Full article

Festulolium hybrids are cool-season forage grasses that form symbiotic relationships with the fungus Epichloë uncinata, which produces loline alkaloids that protect the host from herbivores. This study evaluated the nematotoxicity of shoot and root extracts of Festulolium loliaceum against the stubby root nematode Trichodorus primitivus. Methanolic root and shoot extracts from plants aged 8, 12, 16, and 20 weeks were tested in vitro at five concentrations (312.5–5000 µg mL⁻¹) over 24, 48, and 72 h. Nematode immobility, mortality, and phytochemical profiles, including flavonoids, loline alkaloids, and phenols, were quantified. Extracts from shoots caused significant concentration and time-dependent immobility of T. primitivus (p = 0.001), reaching ≥90% at 5000 µg mL⁻¹ after 72 h in 8–12-week-old plants. Endophyte presence enhanced nematotoxicity, where LD₅₀ values for E+ roots were two-fold lower at 12 weeks and fifty-fold lower at 20 weeks compared with E− root extracts. Shoot extracts of E+ grass had the highest nematicidal activity at 8 weeks, with a significantly lower LD₅₀ value than E− (p < 0.05). Loline alkaloid concentrations increased with plant age, while flavonoids and phenols declined. Nematotoxicity of F. loliaceum extracts was strongly influenced by plant age and endophyte presence. Younger E+ shoots produced the most potent shoot extracts, whereas older plants produced the most potent root extracts. Flavonoid content was negatively correlated with shoot biomass (R = −0.94, p < 0.001). Similarly, phenol content was negatively correlated to both root biomass (R = −0.79, p < 0.001) and shoot biomass (R = −0.67, p < 0.005). Full article

Epichloë sinensis engages in mutualistic symbiosis with Festuca sinensis on the Qinghai–Tibet Plateau. The influence of variation within the Epichloë genus on morphology in this context is poorly understood, as is the influence of environmental factors (e.g., temperature, precipitation, and altitude). Accordingly, a total of 122 fungal endophyte strains were isolated from 270 F. sinensis seeds collected from different locations on the Qinghai–Tibet Plateau, and their morphological characteristics were observed. The colonies were white on the front, dark brown in the center on the back, and light brown or yellow around the PDA medium, exhibiting typical characteristics of E. sinensis. Morphological diversity was categorized into (1) colony features (six types based on texture, shape, and cracks), (2) growth rates (51 strains that produce spores: 0.23–0.78 mm/d; 71 strains that do not produce spores: 0.11–0.93 mm/d), and (3) hyphal width (51 strains that produce spores: 0.60–2.57 μm; 71 strains that do not produce spores: 0.95–2.10 μm). Correlation analyses revealed that temperature and altitude had significant effects on these traits. Phylogenetic relationships showed that 17 strains probably were E. sinensis, and only 4 strains probably were the endophyte E. poae. One strain was haploid and may have originated from E. festucae. All 22 tested strains lacked genes associated with toxic alkaloid biosynthesis (ergot alkaloid) but harbored regulatory genes for the insect-resistant alkaloid peramine, demonstrating potential for use in developing new germplasm in Festuca species. Full article

In nature, a significant number of plant species form symbiotic associations with microorganisms, with arbuscular mycorrhizal fungi (AMF) and endophytic fungi being two prevalent groups of these partners. However, the ability to establish such symbioses with AMF and endophytic fungi is limited to a small fraction of native grass species. Nitrogen is a crucial nutrient for plant growth, yet it is often a limiting factor, underscoring the importance of understanding how plants acquire it. AMF enhance plant growth by improving nitrogen uptake efficiency, but the combined effects of endophytic fungi and AMF on plant physiology and ecology remain underexplored. To address this knowledge gap, in the present study, we conducted an indoor randomized block experiment to investigate the influence of endophytic fungi and AMF infection on the physiological and ecological attributes of Festuca rubra under various nitrogen regimes. The findings indicated that AMF inoculation significantly affected the total carbon content of F. rubra and the total sulfur concentration in its underground tissues across different nitrogen conditions. Additionally, dual colonization by AMF and endophytic fungi had a significant impact on the underground total nitrogen content of the plants. Furthermore, the complex interactions among AMF, endophytic fungi, and nitrogen availability emerged as critical determinants influencing underground total carbon content, transpiration rates, intercellular carbon dioxide concentrations, and the activity of soil extracellular enzymes in F. rubra. The activity of soil extracellular enzymes and pH significantly affected the structure and diversity of rhizosphere bacterial, fungal, and archaeal communities. AMF enhanced the richness of rhizosphere bacterial communities under low-nitrogen conditions, whereas endophytic fungi infection increased bacterial diversity. Soil extracellular enzyme activity and pH were closely related to the community structures and diversities of rhizosphere bacteria, fungi, and archaea. This study clarifies the effects of AMF and endophytic fungi infection on the physiological and ecological characteristics of F. rubra, significantly contributing to our understanding of the synergistic mechanisms governing the interactions among AMF, endophytic fungi, and their host plants. Full article

Background: Artificial inoculation of Epichloë endophytes into elite forage germplasm aims to establish beneficial symbioses for developing high-yield, high-quality, and stress-tolerant cultivars, but host specificity of the fungi often causes compatibility issues in non-natural hosts. Methods: The E. bromicola isolated from native wild barley was inoculated into cultivated wild barley (Hordeum brevisubulatum) and cultivated barley (Hordeum valgare), forming Hb+Eb and Hv+Eb. The NHb+Eb (native wild barley naturally infected with E. bromicola) served as a control. We analyzed fungal colonization patterns and symbiotic gene regulation to clarify the compatibility between E. bromicola and non-natural hosts. Results: Compared with NHb+Eb and Hb+Eb, E. bromicola in Hv+Eb showed obvious hyphal vacuolization. E. bromicola colonization altered host trichome morphology and induced stomatal closure. Correspondingly, expression of the siderophore biosynthesis gene sidN and the NADPH oxidase complex genes (NoxA, NoxB, NoxR, RacA) was significantly lower (p < 0.05) in Hv+Eb than in Hb+Eb and NHb+Eb. Conclusions: This study reveals that the incompatibility between cultivated barley and E. bromicola is characterized by altered hyphal morphology, which is linked to the downregulation of sidN and Nox. These findings provide a critical theoretical foundation for developing highly compatible cereal-Epichloë germplasms. Full article

Epichloë is a genus of endophytic fungi that forms systemic, vertically transmitted, and asymptomatic mutualistic associations with grasses in the subfamily Pooideae. These symbioses are non-pathogenic and are of considerable importance in agronomic and livestock systems due to their roles in enhancing host fitness under biotic and abiotic stress. Several studies have reported associations between Epichloë endophytes and species of the genus Bromus, a taxonomically complex group characterized by varying ploidy levels and frequent hybridization. Among its sections, Bromopsis includes the highest number of species naturally colonized by Epichloë fungi, while sections Bromus and Ceratochloa show lower infection rates. In South America, endophytes such as E. pampeana, E. tembladerae, E. typhina, and morphotypes of Neotyphodium spp. have been documented in species including B. auleticus, B. brachyanthera, and B. setifolius, where they appear to contribute to stress resilience. Although most findings originate from Argentina, significant knowledge gaps remain regarding the diversity and distribution of these endophytes in native Bromus species across the continent. This review synthesizes the current understanding of Epichloë–Bromus interactions, emphasizing their ecological and agronomic relevance, particularly in South America. Key factors influencing the establishment of these symbioses are examined, and future research directions are proposed to advance the study of these associations. Full article

Claviceps purpurea is a specialized phytopathogenic fungus that infects grasses and produces pharmacologically active compounds, attracting considerable interest in genomic research. In this study, we assembled and annotated the complete mitogenomes of 15 C. purpurea strains isolated from different host plants, including seven newly sequenced isolates from China. Analysis of the C. purpurea pan-mitogenome demonstrated that the accessory regions, with an average proportion of 48.23%, are the main contributor to mitogenome variation. Analysis of the 14 protein-coding genes revealed Ka/Ks ratios below 1, indicating strong purifying selection. Notably, the atp9 gene was absent in all strains, suggesting a potential adaptive gene loss. Structural variations were predominantly located in the intergenic region between rns and rnl. Phylogenetic analysis based on concatenated mitochondrial genes placed Claviceps as most closely related to the genus Epichloë. The 15 C. purpurea strains grouped into five well-supported subclades, with Chinese and non-Chinese isolates forming distinct lineages. Among these, the Chinese strains ACCC 37001 and KUNCC 11030 represented the earliest diverging lineages. This study elucidates the intraspecific variation and evolutionary patterns of the mitogenome in C. purpurea and highlights the value of mitogenome in resolving phylogenetic relationships. Full article

Epichloë endophytes are mutualistic associates with grasses, conferring host plants with enhanced competitiveness, improved stress tolerance, and increased ecological dominance. Epichloë can produce any of several classes of bioactive alkaloids, of which indole-diterpenes and ergot alkaloids are toxic to invertebrate and mammalian herbivores; peramine acts as an insect-feeding deterrent; and loline alkaloids possess potent insecticidal activity. Here, it was characterized as Epichloë endophytes inhibiting the Elymus species, El. cylindricus, and El. tangutorum from the Qinghai–Tibet Plateau, China. Based on the results of four types of alkaloid synthesis genes, the 30 isolates were divided into five types; they exhibited distinct alkaloid synthesis capabilities, highlighting intraspecific diversity within E. bromicola regarding its alkaloid-producing potential. Considering the toxicity of these isolates to the safety of herbivorous livestock, the above five types of isolates can be divided into two categories. Category I includes five animal-safe strains of type V, which do not produce alkaloids. Category II includes the remaining 25 strains, which could produce indole-diterpene (paspaline) and/or ergot alkaloids (chanoclavine I, D-lysergic acid, ergovaline) that are toxic to herbivorous livestock. Morphology and phylogenetic analysis confirmed all 30 isolates were Epichloë bromicola; mating type gene detection shows that all belonged to mating type A. Overall, this study has laid a solid foundation for the scientific and rational utilization of Epichloë endophyte resources. Furthermore, the presence of ergovaline in El. cylindricus and El. tangutorum poses a potential concern for livestock managers who conduct grazing. Full article

Research has evaluated the efficacy of various methods for eliminating endophytes from grass seeds, as well as changes in endophyte viability during seed storage under different conditions, indicating significant variation in different procedures and cultivars. Chemical seed treatment (tebuconazole and thiram) completely eliminated viable fungal mycelia, leaving no trace in any tested cultivar. Non-chemical methods, such as drying and microwave treatment, only partially reduced mycelial viability by 30.3% and 33.1%, respectively, with no statistically significant difference between them. A significant positive correlation was observed between the initial mycelial viability and its reduction. Lolium perenne cv. Vigor showed no impact from non-chemical methods, while Festuca rubra cv. Anielka exhibited the greatest reduction (79% after microwave treatment). Seed storage also impacted endophyte survival. Storage at +7 °C, +23 °C, and −20 °C reduced viability by 27.4%, 31.7%, and 37.3%, respectively. Positive correlations existed between initial viability and post-storage reductions. Similarly to elimination methods, cv. Vigor showed resistance to storage conditions. However, −20 °C storage proved least favorable for endophyte survival, particularly for Festuca pratensis cv. Artema, cv. Anielka, and Festuca ovina cv. Jolka. To maintain the viability of beneficial endophytes during seed storage, we must carefully control storage conditions, especially ambient temperature. Full article

A study was conducted to examine the effects of the spray application of nine antifungal products, including microbial-derived fungicides, plant-derived fungicides, and chemical fungicides, on the grass Achnatherum inebrians that was either host to Epichloë gansuensis (E+) or E. gansuensis-free (E−) and that was exposed to Blumeria graminis, the fungal pathogen causing powdery mildew. The Epichloë endophyte is a seed-borne mutualistic biotrophic fungus whose growth is fully synchronized with the host grass. Bl. graminis is a biotrophic pathogen that continually infects leaves and stems via conidia, the formation of appressoria, leading to the presence of haustoria in epidermal cells. Prior to fungicide application, the presence of endophytes significantly increased the resistance of A. inebrians to powdery mildew and was able to increase the chlorophyll content. However, the positive effects of the Epichloë endophyte on the plant were suppressed with the use of some fungicides and the increase in the number of sprays, but the reciprocal relationship between the Epichloë endophyte and the plant was not significantly disrupted. Full article

Perennial ryegrass (Lolium perenne), an important forage and turfgrass species, can establish a mutualistic symbiosis with the fungal endophyte Epichloë festucae var. lolii. Although the physiological and ecological impacts of endophyte infection on ryegrass have been extensively investigated, the response of the soil microbial community and nitrogen-cycling gene to this relationship has received much less attention. The present study emphasized abundance and diversity variation in the AOB-amoA, nirK and nosZ functional genes in the rhizosphere soil of the endophyte–ryegrass symbiosis following litter addition. We sampled four times: at T₀ (prior to first litter addition), T₁ (post 120 d of 1st litter addition), T₂ (post 120 d of 2nd litter addition) and T₃ (post 120 d of 3rd litter addition) times. Real-time fluorescence quantitative PCR (qPCR) and PCR amplification and sequencing were used to characterize the abundance and diversity of the AOB-amoA, nirK and nosZ genes in rhizosphere soils of endophyte-infected (E+) plants and endophyte-free (E−) plants. A significant enhancement of total Phosphorus (P), Soil Organic Carbon (SOC), Ammonium ion (NH₄⁺) and Nitrate ion (NO₃⁻) contents in the rhizosphere soil was recorded in endophyte-infected plants at different sampling times compared to endophyte-free plants (p ≤ 0.05). The absolute abundance of the AOB-amoA gene at T₀ and T₁ times was higher, as was the absolute abundance of the nosZ gene at T₀, T₁ and T₃ times in the E+ plant rhizophere soils relative to E− plant rhizosphere soils. A significant change in relative abundance of the AOB-amoA and nosZ genes in the host rhizophere soils of endophyte-infected plants at T₁ and T₃ times was observed. The experiment failed to show any significant alteration in abundance and diversity of the nirK gene, and diversity of the AOB-amoA and nosZ genes. Analysis of the abundance and diversity of the nirK gene indicated that changes in soil properties accounted for approximately 70.38% of the variation along the first axis and 16.69% along the second axis, and soil NH₄⁺ (p = 0.002, 50.4%) and soil C/P ratio (p = 0.012, 15.8%) had a strong effect. The changes in community abundance and diversity of the AOB-amoA and nosZ genes were mainly related to soil pH, N/P ratio and NH₄⁺ content. The results demonstrate that the existence of tripartite interactions among the foliar endophyte E. festucae var. Lolii, L. perenne and soil nitrogen-cycling gene has important implications for reducing soil losses on N. Full article

Records from 2012 to 2019 for two herds were analyzed to determine how tall fescue (Schedonorus arundinaceus (Schreb.) Dumont) endophyte (Epichloë coenophialum) status affected the productivity of spring-calving cows and calves. Pastures either contained tall fescue with wildtype endophyte (high levels of ergot alkaloids) or novel- or endophyte-free tall fescue (largely ergot alkaloid free). The experimental design was a randomized complete block with year as the replication unit. Forage samples from the farm with toxic endophyte-infected tall fescue contained 1136 ± 413 ppb total ergot alkaloids, while forage from the non-toxic pastures on the second farm contained 118 ± 83 ppb total ergot alkaloids. Artificial insemination pregnancy rates and calving rates were greater (p < 0.05) for cows that grazed non-toxic tall fescue (51.2 ± 2.8% and 93.5 ± 1.4%, respectively) than for cows on toxic endophyte-infected tall fescue (43.3 ± 2.8% and 88.8 ± 1.4%, respectively). Birth weights and weaning weights were greater (p < 0.05) for calves from the non-toxic tall fescue system (37 ± 1 kg and 278 ± 8 kg, respectively) than for calves from the toxic endophyte-infected tall fescue system (33 ± 1 kg and 254 ± 8 kg, respectively). Raising cattle on tall fescue without the toxic endophyte improved cow and calf productivity. Full article

Despite chemical exchange often serving as the first step in plant–microbe interactions, the specialized chemical metabolites produced by grass–Epichloë endophyte symbiosis as mediators of host growth, nutrient acquisition, and modulators of the rhizosphere community under low-nitrogen conditions are areas lacking in knowledge. In this study, we investigated the plant growth-promoting effects of the Epichloë endophyte strain and identified the growth of the Epichloë strain under different types of nitrogen source treatments. In addition to the in vitro test, we evaluated growth performance for Epichloë endophyte–infected plants (E+) and Epichloë endophyte–free plants (E−) in a pot trial under 0.01 mol/L urea treatment. Seedlings from E+ and E− groups were collected to analyze the plant bacterial microbiome and root metabolites. The E. gansuensis endophyte strain was found not to produce indoleacetic acid (IAA), pectinase, or contain ferritin. The nitrogenase gene, essential for nitrogen fixation, was also absent. These results suggest that E. gansuensis endophyte strains themselves do not contain attributes to promote plant growth. Concerning N fertilization, it was observed an increase in the colony diameter of E. gansuensis strain was observed only in the NO₃⁻-N (NN) treatment, while inhibition was observed in the urea-N (UN) treatment. E. gansuensis endophyte symbiosis significantly increased tiller number and plant dry weight. Overall, our results suggest that the E+ plants had more root forks and greater average root diameter compared to E− plants under the UN treatment. In a pot experiment using UN, data from 16S rRNA amplicon sequencing revealed that E. gansuensis endophyte infection significantly altered the bacterial community composition in shoot and root, and significantly increased Shannon (p < 0.001) and Chao 1 (p < 0.01) indexes. The relative abundance of Acidobacteriota, Actinomycetota, Cyanobacteriota, Fibrobacterota, Myxococcota, and Patescibacteria in the shoot, and Cyanobacteriota, Pseudomonadota, and Verrucomicrobiota in the root were significantly increased by E. gansuensis endophyte infection. Similarly, E. gansuensis endophyte symbiosis shifted the metabolite composition of the host plants, with the E+ plants showing a higher number of metabolites than the E− plants. In addition, co-metabolism network analysis revealed that the positive relevance between exudates and microorganisms in the root of the E+ plants is higher than that of the E− plants. These findings provide valuable insights into the knowledge of the effects of the symbiotic relationship between host plants and Epichloë endophyte on interspecific interactions of plant microbiome, beneficial for harnessing endophytic symbiosis, promoting plant growth. Full article