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Epichloë endophytes have been shown to increase tolerance to biotic and abiotic stresses in many cool-season grasses. We investigated the impact of endophyte infection of Festuca sinensis, on root metabolic activity, photosynthetic pigments, leaf relative water content (RWC) and soluble carbohydrates in a field experiment carried out during chilling and irrigation with Ca(NO3)2. A highly significant (P < 0.001) correlation for Epichloë endophytes was observed for root metabolic activity. Ca(NO3)2affectedverysignificantly root metabolic activity and total chlorophyll (P < 0.001). Low temperature led to highly significant (P < 0.001) reductions in root metabolic activity, RWC, total chlorophyll, chlorophyll a/b ratio, and carotenoid contents. In addition, the fructose concentrations of shoots were greater on the 14th day than on the 28th day and before treatment, whilst the glucose concentration of roots was much higher on the 28th day than before and after 14 days treatment. Moreover, our results indicated that the addition of calcium nitrate contributed to higher levels of total chlorophylls, soluble sugars, sucrose, fructose or glucose in the shoots and roots in both E+ and E- plants during long periods of chilling. These results suggest that Epichloë endophyte infection and/or exogenous calcium nitrate can confer better tolerance to cold stress.

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Selenium (Se) is an essential micronutrient with many beneficial effects for humans and other living organisms. Numerous microorganisms in culture systems enrich and convert inorganic selenium to organic selenium. In this study, Epichloë sp. from Festuca sinensis was exposed to increasing Na2SeO3 concentrations (0, 0.1, 0.2, 0.3, and 0.4 mmol/L) in Petri dishes with potato dextrose agar (PDA) for 8 weeks. Epichloë sp. mycelia were immediately collected after mycelial diameters were measured at 4, 5, 6, 7, and 8 weeks of cultivation, respectively. Gas chromatography-mass spectrometer (GC-MS) analysis was performed on different groups of Epichloë sp. mycelia. Different changes were observed as Epichloë sp. was exposed to different selenite conditions and cultivation time. The colony diameter of Epichloë sp. decreased in response to increased selenite concentrations, whereas the inhibitory effects diminished over time. Seventy-two of the 203 identified metabolites did not differ significantly across selenite treatments within the same time point, while 82 compounds did not differ significantly between multiple time points of the same Se concentration. However, the relative levels of 122 metabolites increased the most under selenite conditions. Specifically, between the 4th and 8th weeks, there were increases in 2-keto-isovaleric acid, uridine, and maltose in selenite treatments compared to controls. Selenium increased glutathione levels and exhibited antioxidant properties in weeks 4, 5, and 7. Additionally, we observed that different doses of selenite could promote the production of carbohydrates such as isomaltose, cellobiose, and sucrose; fatty acids such as palmitoleic acid, palmitic acid, and stearic acid; and amino acids such as lysine and tyrosine in Epichloë sp. mycelia. Therefore, Epichloë sp. exposed to selenite stress may benefit from increased levels of some metabolite compounds.

Selenium (Se) is not only an essential trace element critical for the proper functioning of an organism, but it is also an abiotic stressor that affects an organism’s growth and metabolite profile. In this study, Epichloë sp. from Festuca sinensis was exposed to increasing concentrations of Na2SeO3 (0, 0.1, and 0.2 mmol/L) in a liquid media for eight weeks. The mycelia and fermentation broth of Epichloë sp. were collected from four to eight weeks of cultivation. The mycelial biomass decreased in response to increased Se concentrations, and biomass accumulation peaked at week five. Using gas chromatography-mass spectrometry (GC-MS), approximately 157 and 197 metabolites were determined in the fermentation broth and mycelia, respectively. Diverse changes in extracellular and intracellular metabolites were observed in Epichloë sp. throughout the cultivation period in Se conditions. Some metabolites accumulated in the fermentation broth, while others decreased after different times of Se exposure compared to the control media. However, some metabolites were present at lower concentrations in the mycelia when cultivated with Se. The changes in metabolites under Se conditions were dynamic over the experimental period and were involved in amino acids, carbohydrates, organic acids, fatty acids, and nucleotides. Based on these results, we conclude that selenite concentrations and culture time influence the growth, extracellular and intracellular metabolite profiles of Epichloë sp. from F. sinensis.