We sampled five Achnatherum sibiricum populations and documented the distribution and abundance of endophytes. In total, 438 fungal isolates were obtained. They were classified into 11 morphotypes based on growth rate and morphological characters including colonial morphology, the production of conidiogenous cells, conidial size and dimension on potato dextrose agar medium. Both tub2 and actG sequences identified only a single product in all representative isolates from three dominant morphotypes. All sequences fell in the same main clade, including Neotyphodium gansuense and Neotyphodium gansuense var. inebrians from another Achnatherum species in North China, Achnatherum inebrians, and Neotyphodium guerinii from European grasses Melica ciliate and Melica transsilvanica. Based on both morphological and phylogenetic analyses, isolates in morphotype I were described as representing a novel Neotyphodium species, Neotyphodium sibiricum sp. nov. Isolates in both morphotype II and III were identified as representing N. gansuense. Both tub2 and actG sequences suggested that N. sibiricum and N. guerinii were probably derived from a common Epichloë ancestor.
A survey of grass species associated with Neotyphodium and Epichloë fungal endophytes was made in the permanent grasslands of northern China. A total of 41 grass species, 56 sites and 172 local populations were examined to reveal endophytic infection of native grasses. Twenty-five of the forty-one species of grasses (proportionately 0AE61) were infected by endophytes: twenty-two species had a mean infection rate of 0AE01-0AE50 and only three species had a mean infection >0AE50. Of the 172 local populations examined, however, proportionately 0AE72 were not infected, 0AE181 had an infection rate of 0AE01-0AE50 and 0AE099 had an infection rate of 0AE51-1AE00. Two genera of grasses, Cleistogenes and Koeleria, as well as twenty plant species, were previously unknown endophytic hosts in China. High infection rates (0AE86-1AE00) of plants of Achnatherum sibiricum were detected at all eleven sites studied. Other grasses in which high infection rates were found were Agropyron mongolicum, Roegneria turczaninovii, Agropyron elongate, Poa palustris and Poa angustifolia. It was concluded that endophytes are found widely in natural grass populations and that endophyte-grass interactions may have effects on ecosystems and livestock in the grasslands of northern China.
The growth response of endophyte-infected (EI) and endophyte-free (EF) tall fescue to salt stress was investigated under two growing systems (hydroponic and soil in pots). The hydroponic experiment showed that endophyte infection significantly increased tiller and leaf number, which led to an increase in the total biomass of the host grass. Endophyte infection enhanced Na accumulation in the host grass and improved Na transport from the roots to the shoots. With a 15 g l(-1) NaCl treatment, the phytoextraction efficiency of EI tall fescue was 2.34-fold higher than EF plants. When the plants were grown in saline soils, endophyte infection also significantly increased tiller number, shoot height and the total biomass of the host grass. Although EI tall fescue cannot accumulate Na to a level high enough for it to be termed a halophyte, the increased biomass production and stress tolerance suggested that endophyte/plant associations had the potential to be a model for endophyte-assisted phytoextraction in saline soils.
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.
Random amplified polymorphic DNA was used to assess the level of genetic diversity and genetic structure of Stipa krylovii (Gramineae), an important dominant species in the northern grasslands of China. Genetic diversity was low within S. krylovii populations, and diversity at the population level was associated with precipitation and cumulative temperature variations. There was much genetic differentiation among populations and among habitats as well. A Mantel test indicated no significant correlation between genetic distance and geographic distance of populations. A nonmetric multidimensional scaling analysis revealed some spatial relationships among the 90 individuals in a two-dimensional plot. Habitat fragmentation and degradation throughout the geographic range of S. krylovii could account for the low genetic diversity and high genetic differentiation of the species. Such information will be useful for conservation managers trying to plan an effective strategy to protect this important species.
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