Anzhi Ren

Endophyte-mediated effects on the growth and physiology of Achnatherum sibiricum are conditional on both N and P availability.

The interaction of endophyte–grass associations are conditional on nitrogen (N) availability, but the reported responses of these associations to N are inconsistent. We hypothesized that this inconsistency is caused, at least in part, by phosphorus (P) availability. In this experiment, we compared the performance of endophyte-infected (EI) and endophyte-free (EF) Achnatherum sibiricum subjected to four treatments comprising a factorial combination of two levels of N (N+ vs. N−, i.e. N supply vs. N deficiency) and two levels of P (P+ vs. P−, i.e. P supply vs. P deficiency) availability. The results showed that A. sibiricum–Neotyphodium associations were conditional on both N and P availability, but more conditional on N than P. Under N+P− conditions, endophyte infection significantly improved acid phosphatase activity of EI plants, such that the biomass of EI plants was not affected by P deficiency (i.e. similar growth to N+P+ conditions), and resulted in more biomass in EI than EF plants. Under N−P+ conditions, biomass of both EI and EF decreased compared with N+P+; however, EI biomass decreased slowly by decreasing leaf N concentration more rapidly but allocating higher fractions of N to photosynthetic machinery compared with EF plants. This change of N allocation not only improved photosynthetic ability of EI plants but also significantly increased their biomass. Under N−P− conditions, EI plants allocated higher fractions of N to photosynthesis and had greater P concentrations in roots, but there was no significant difference in biomass between EI and EF plants. Our results support the hypothesis that endophyte–grass interactions are dependent on both N and P availability. However, we did not find a clear cost of endophyte infection in A. sibiricum.

Pathogen resistant advantage of endophyte-infected over endophyte-free Leymus chinensis is strengthened by pre-drought treatment

Symbiotic relationships with beneficial microbes often increase the resistance of the host grass to abiotic and biotic stresses. In this study, the resistance of endophyte-infected (E+) and endophyte-free (E-) Leymus chinensis to two species of pathogen fungi, Curvularia lunata and Bipolaris sorokiniana, were compared under normal water and pre-drought treatments. The results showed that endophyte infection significantly reduced leaf lesion area of the host grass on one hand, and on the other hand the maximum quantum efficiency of PSII (Fv/Fm) of E+ plants was higher than E- plants, and these advantages of E+ over E- plants were strengthened by pre-drought stress. Pre-drought treatment led to significantly higher salicylic acid (SA) and lignin accumulation in E+ plants in comparison with E- when exposed to both C. lunata and B. sorokiniana. Additionally, glutamine and phenylalanine concentrations were significantly affected by the interaction between endophyte infection and pathogen inoculation. Overall, endophytes could enhance the pathogen resistance of host by activating a rapid defense reaction of the plant, and this advantage of E+ over E- plants could be strengthened by pre-drought treatment.

Stroma-bearing endophyte and its potential horizontal transmission ability in Achnatherum sibiricum

Stromata occasionally are observed in Achnatherum sibiricum distributed in northern China. However, endophyte species that form stromata on that host have not been studied. Here we identified the first Epichloë sp. endophyte in stroma-bearing A. sibiricum. Isolated colonies of this Epichloë sp. were smoother and more compact than previously described for Epichloë gansuensis and also had longer phialides and faster growth in culture. However, phylogenetic relationships based on intron sequences of genes encoding β-tubulin (tubB) and translation elongation factor 1-α (tefA) grouped all isolates from the stromata in a clade with a close relationship to E. gansuensis. We identified the new isolates as E. gansuensis. The analysis of the stromata revealed no perithecium or ascospores during morphological and paraffin section observation. Furthermore, the ability of conidia formed on stromata to germinate and initiate infection of new seedlings was tested. After 3 mo 20% endophyte-free seedlings became infected by E. gansuensis, whereas the control group showed no endophyte infection. The results indicated the potential of cultures from conidia to mediate horizontal transmission.

Endophyte species influence the biomass production of the native grass Achnatherum sibiricum (L.) Keng under high nitrogen availability

Abstract Research on the interaction of endophytes and native grasses normally takes infection status into account, but less often considers the species of endophyte involved in the interaction. Here, we examined the effect of endophyte infection, endophyte species, nitrogen availability, and plant maternal genotype on the performance of a wild grass, Achnatherum sibiricum. Six different Epichloë‐infected maternal lines of A. sibiricum were used in the study; three lines harbored Epichloë gansuensis (Eg), while three lines harbored Epichloë sibirica (Es). These endophytes are vertically transmitted, while Eg also occasionally produces stromata on host tillers. We experimentally removed the endophyte from some ramets of the six lines, with the infected (E+) and uninfected (E−) plants grown under varying levels of nitrogen availability. Eg hosts produced more aboveground biomass than Es hosts only under high nitrogen supply. Endophyte species did not show any influence on the maximum net photosynthetic rate (P max), photosynthetic nitrogen use efficiency, or total phenolics of A. sibiricum under all nitrogen conditions. However, the plant maternal genotype did influence the P max and shoot biomass of A. sibiricum. Our results show that endophyte species influenced the shoot biomass of A. sibiricum, and this effect was dependent on nitrogen supply. As with most coevolutionary interactions, A. sibiricum that harbored Eg and Es may show pronounced geographic variation in natural habitats with increased nitrogen deposition. In addition, stroma‐bearing endophyte (Eg) provides positive effects (e.g., higher biomass production) to A. sibiricum plants during the vegetative growth stage.

The advantages of endophyte-infected over uninfected tall fescue in the growth and pathogen resistance are counteracted by elevated CO 2

Atmospheric CO2 concentrations are predicted to double within the next century. Despite this trend, the extent and mechanisms through which elevated CO2 affects grass-endophyte symbionts remain uncertain. In the present study, the growth, chemical composition and pathogen resistance of endophyte-infected (E+) and uninfected (E−) tall fescue were compared under elevated CO2 conditions. The results showed that the effect of endophyte infection on the growth of tall fescue was significantly affected by elevated CO2. Significant advantage of E+ over E− tall fescue in tiller number, maximum net photosynthetic rate and shoot biomass occurred only under ambient CO2. With CO2 concentration elevated, the beneficial effect of endophyte infection on the growth disappeared. Similarly, endophyte infection reduced lesion number and spore concentration of Curvularia lunata only under ambient CO2. These results suggest that the beneficial effect of endophyte infection on the growth and pathogen resistance of tall fescue could be counteracted by elevated CO2. An explanation for the counteraction may be found in a change in photosynthesis and nutritive quality of leaf tissue.

Arbuscular mycorrhizal fungus inoculation reduces the drought-resistance advantage of endophyte-infected versus endophyte-free Leymus chinensis

Grasses can be infected simultaneously by endophytic fungi and arbuscular mycorrhizal (AM) fungi. In this study, we tested the hypothesis that endophyte-associated drought resistance of a native grass was affected by an AM fungus. In a greenhouse experiment, we compared the performance of endophyte-infected (EI) and endophyte-free (EF) Leymus chinensis, a dominant species native to the Inner Mongolia steppe, under altered water and AM fungus availability. The results showed that endophyte infection significantly increased drought resistance of the host grass, but the beneficial effects were reduced by AM fungus inoculation. In the mycorrhizal-non-inoculated (MF) treatment, EI plants accumulated significantly more biomass, had greater proline and total phenolic concentration, and lower malondialdehyde concentration than EF plants. In the mycorrhizal-inoculation (MI) treatment, however, no significant difference occurred in either growth or physiological characters measured between EI and EF plants. AM fungus inoculation enhanced drought resistance of EF plants but had no significant effect on drought resistance of EI plants, thus AM fungus inoculation reduced the difference between EI and EF plants. Our findings highlight the importance of interactions among multiple microorganisms for plant performance under drought stress.

Effect of Endophyte Infection and Clipping Treatment on Resistance and Tolerance of Achnatherum sibiricum

It is well-documented that endophytes can enhance the resistance of agronomical grasses, such as tall fescue and perennial ryegrass to herbivory. For native grasses, however, the related reports are limited, and the conclusions are variable. Achnatherum sibiricum is a grass native to the Inner Mongolian steppe. This grass is highly infected by endophytes but does not produce detectable endophyte-related alkaloids known under normal conditions. In this study, the contributions of endophytes to the resistance of A. sibiricum to Locusta migratoria were studied. We found that locusts preferred EF (endophyte-free) plants to EI (endophyte-infected) plants, and the weight of locusts fed on EI plants was significantly lower than those fed on EF plants. Hence, endophyte infection significantly enhanced the resistance of the host to L. migratoria. Endophyte infection significantly decreased the concentration of soluble sugar and amino acids while significantly increased the concentration of total phenolic content, and these metabolites may contribute to herbivore resistance of the host. The clipping treatment further strengthened the locust resistance advantage of EI over EF plants. After clipping, the weight of the locusts fed on EI plants significantly decreased compared with those fed on unclipped plants, whereas the weight of the locusts fed on EF plants increased significantly. The results suggested that endophyte infection could increase herbivore resistance while decreasing the tolerance of the host grass by mechanisms apart from endophyte-conferred alkaloid defense.