Yubao Gao

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.

Increased soil nutrition and decreased light intensity drive species loss after eight years grassland enclosures

Enclosures (fenced, grazing or clipping) within a certain period of years are the most common tools for restoration of degraded grasslands in temperate regions. Short-term enclosures can improve biodiversity and productivity by effectively relieving grazing pressure, while long-term enclosures can reduce species diversity. We therefore carried out a field experiment to investigate the specific causes of the reduced species diversity in Hulunbeier grassland of northern China. After eight years of enclosure, the significantly increased soil available nitrogen (AN) and available phosphorus (AvP) in enclosure community reduced nitrogen (N) limitation but most vegetation was still N limited. Many environmental factors led to decreased species richness, but increased soil AN and decreased light intensity at the community bottom were the most significant ones. Community density decreased independently of soil nutrition but significantly related to decreased species richness. Density of dominant canopy species increased, while dominant understory species decreased during assemblage-level thinning; therefore, the random-loss hypothesis was not supported. The dominant understory species responded to lower light availability by increasing their height, leaf area, and chlorophyll content. Moreover, our results were expected to provide some specific guidance for the restoration mode selection of degraded grasslands in northern China.

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.

Heterogeneous microcommunities and ecosystem multifunctionality in seminatural grasslands under three management modes

Abstract Increasing attention has been paid to the relationship between biodiversity and ecosystem functioning (BEF) because of the rapid increase in species loss. However, over the past 20 years, most BEF studies only focused on the effect of species diversity on one or a few ecosystem functions, and only a few studies focused on ecosystem multifunctionality (i.e., the simultaneous provision of several ecosystem functions). Grassland ecosystems have important economic, environmental, and esthetic value; thus, this study focused on the heterogeneous microcommunities in grasslands under three management modes. The multifunctionality index (M‐index) was assessed at community and microcommunity scales, and the relationship between species diversity and multifunctionality was investigated. The communities were found to be respectively composed of one, three, and six microcommunities in grazing, clipping, and enclosure management, based on a two‐way indicator species analysis (TWINSPAN) and detrended correspondence analysis (DCA) for community structure. Biodiversity and soil indicators showed an apparent degradation of the grazing community, which had the worst M‐index. Clipping and enclosure communities showed no significant difference in biodiversity indices, soil variables, and M‐index; however, these indices were clearly different among microcommunities. Therefore, the microcommunity scale may be suitable to investigate the relationship between vegetation and multifunctionality in seminatural grassland ecosystems. Dominant species richness had more explanatory power for ecosystem multifunctionality than subdominant species richness, rare species richness, and the number of all species. Therefore, it is important to distinguish the role and rank of different species in the species richness–multifunctionality model; otherwise, the model might include redundant and unclear information. Communities with more codominant species whose distribution is also even might have better multifunctionality.

Plant endophytes and arbuscular mycorrhizal fungi alter plant competition

In nature, grasses simultaneously establish multiple symbiotic associations with endophytic fungi and arbuscular mycorrhizal fungi (AMF). The effect of these multiple interactions on competitive interactions between plants remains poorly understood. In this study, we tested whether endophytes and AMF (Glomus mosseae or Glomus etunicatum) alter plant competition between a subordinate plant species that associates with both symbionts (Achnatherum sibiricum) and a dominant plant species, Stipa grandis, that only associates with one symbiont (AMF). And we hypothesized that endophytes can facilitate the coexistence of the subordinate plant species (A.xa0sibiricum) and the dominant plant species (S.xa0grandis). The results demonstrated that endophyte infection significantly enhanced the competitive ability of the subordinate plant species compared to the dominant plant species. The effects of AMF on plant competition were variable and depended on the identity of the AMF species. Glomus etunicatum gave A.xa0sibiricum plants a higher competitive ability, while G.xa0mosseae gave S.xa0grandis a higher competitive ability. Simultaneous infections of both endophytes and AMF in A.xa0sibiricum also altered the competitive relationships with S.xa0grandis. In conclusion, these results suggest that endophytic fungi can facilitate the coexistence of a subordinate plant species with a dominant plant species. Moreover, endophytes could not only affect the competitive ability of the host plant directly but also indirectly by interacting with different AMF to change the growth of competing plant species. A plain language summary is available for this article.

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.