Jana Rydlová

Effectiveness of indigenous and non-indigenous isolates of arbuscular mycorrhizal fungi in soils from degraded ecosystems and man-made habitats

Culturing in soils from degraded ecosystems significantly influenced the effectiveness of indigenous arbuscular mycorrhizal fungi (AMF) isolated from disturbed and undisturbed soils. The AMF isolates from degraded or artificially created habitats (acid rain polluted site, power station fly ash deposits, spoil banks, pyrite deposit), were not, in most cases, more effective than those from undisturbed soils, when grown in symbiosis with maize in the disturbed soils. Significant effects of soil or substrate on plant growth were found, while the influence of the AMF inoculant was much less pronounced. The development of AMF isolates was reduced in soils with more adverse chemical properties irrespective of the isolate origin. The length of extraradical mycelium of AMF and NADH-diaphorase activity of the mycelium were good indicators of negative effects of stress factors in the soil.

Surprising spectra of root‐associated fungi in submerged aquatic plants

Similarly to plants from terrestrial ecosystems, aquatic species harbour wide spectra of root-associated fungi (RAF). However, comparably less is known about fungal diversity in submerged roots. We assessed the incidence and diversity of RAF in submerged aquatic plants using microscopy, culture-dependent and culture-independent techniques. We studied RAF of five submerged isoetid species collected in four oligotrophic freshwater lakes in Norway. Levels of dark septate endophytes (DSE) colonization differed among the lakes and were positively related to the organic matter content and negatively related to pH. In total, we identified 41 fungal OTUs using culture-dependent and culture-independent techniques, belonging to Mucoromycotina, Chytridiomycota, Glomeromycota, Ascomycota as well as Basidiomycota. Sequences corresponding to aquatic hyphomycetes (e.g. Nectria lugdunensis, Tetracladium furcatum and Varicosporium elodeae) were obtained. Eight arbuscular mycorrhizal taxa belonging to the orders Archaeosporales, Diversisporales and Glomerales were also detected. However, the vast majority of the fungal species detected (e.g. Ceratobasidium sp., Cryptosporiopsis rhizophila, Leptodontidium orchidicola, and Tuber sp.) have previously been known only from roots of terrestrial plants. The abundance and phylogenetic distribution of mycorrhizal as well as nonmycorrhizal fungi in the roots of submerged plants have reshaped our views on the fungal diversity in aquatic environment.

Differences in AM fungal root colonization between populations of perennial Aster species have genetic reasons

We tested the hypothesis whether differences between plant populations in root colonization by arbuscular mycorrhizal (AM) fungi could be caused by genetic differentiation between populations. In addition, we investigated whether the response to AM fungi differs between plants from different populations and if it is affected by the soil in which the plants are cultivated. We used Aster amellus, which occurs in fragmented dry grasslands, as a model species and we studied six different populations from two regions, which varied in soil nutrient concentration.We found significant differences in the degree of mycorrhizal colonization of plant roots between regions in the field. To test if these differences were due to phenotypic plasticity or had a genetic basis, we performed a greenhouse experiment. The results suggested that Aster amellus is an obligate mycotrophic plant species with a high dependency upon mycorrhiza. Plant biomass was affected only by soil, and not by population or the interaction between the population and the soil. Mycorrhizal colonization was significantly affected by all three factors (soil, population, interaction of soil and population). Plants from the population originating from the soil with lower nutrient availability developed more mycorrhiza even when grown in soil with higher nutrient availability. The correspondence between mycorrhizal colonization of plants in the field and in both soils in the pot experiment suggests that the observed differences in root colonization have a genetic basis.

Cultivation of high-biomass crops on coal mine spoil banks: can microbial inoculation compensate for high doses of organic matter?

Two greenhouse experiments were focused on the application of arbuscular mycorrhizal fungi (AMF) and plant growth promoting rhizobacteria (PGPR) in planting of high-biomass crops on reclaimed spoil banks. In the first experiment, we tested the effects of different organic amendments on growth of alfalfa and on the introduced microorganisms. While growth of plants was supported in substrate with compost amendment, mycorrhizal colonization was suppressed. Lignocellulose papermill waste had no negative effects on AMF, but did not positively affect growth of plants. The mixture of these two amendments was found to be optimal in both respects, plant growth and mycorrhizal development. Decreasing doses of this mixture amendment were used in the second experiment, where the effects of microbial inoculation (assumed to compensate for reduced doses of organic matter) on growth of two high-biomass crops, hemp and reed canarygrass, were studied. Plant growth response to microbial inoculation was either positive or negative, depending on the dose of the applied amendment and plant species.

Extraradical mycelium of arbuscular mycorrhizal fungi radiating from large plants depresses the growth of nearby seedlings in a nutrient deficient substrate

The effect of arbuscular mycorrhiza (AM) on the interaction of large plants and seedlings in an early succession situation was investigated in a greenhouse experiment using compartmented rhizoboxes. Tripleurospermum inodorum, a highly mycorrhiza-responsive early coloniser of spoil banks, was cultivated either non-mycorrhizal or inoculated with AM fungi in the central compartment of the rhizoboxes. After two months, seedlings of T. inodorum or Sisymbrium loeselii, a non-host species colonising spoil banks simultaneously with T. inodorum, were planted in lateral compartments, which were colonised by the extraradical mycelium (ERM) of the pre-cultivated T. inodorum in the inoculated treatments. The experiment comprised the comparison of two AM fungal isolates and two substrates: spoil bank soil and a mixture of this soil with sand. As expected based on the low nutrient levels in the substrates, the pre-cultivated T. inodorum plants responded positively to mycorrhiza, the response being more pronounced in phosphorus uptake than in nitrogen uptake and growth. In contrast, the growth of the seedlings, both the host and the non-host species, was inhibited in the mycorrhizal treatments. Based on the phosphorus and nitrogen concentrations in the biomass of the experimental plants, this growth inhibition was attributed to nitrogen depletion in the lateral compartments by the ERM radiating from the central compartment. The results point to an important aspect of mycorrhizal effects on the coexistence of large plants and seedlings in nutrient deficient substrates.

The response of Aster amellus (Asteraceae) to mycorrhiza depends on the origins of both the soil and the fungi.

PREMISE OF STUDY Previous studies testing pairwise interactions between plants, arbuscular mycorrhizal fungi (AMF) and soil have shown that the effectiveness of such interactions depends on the origins of the plants, soil, and AMF. Surprisingly, no study has yet looked at the three-way interaction between plants, soil, and AMF originating from the same and from different sites. Such knowledge could elucidate the determinants of local adaptations of plants and thus might help in various revegetation attempts. METHODS Six populations of the obligately mycorrhizal plant species Aster amellus from two isolated regions were combined with the soil and the AMF ecotype from their sites and plant growth were monitored over 16 months. KEY RESULTS For each combination of soil and native AMF, plants grown with their native AMF in their native soil had higher aboveground biomass, invested more to aboveground biomass and had higher numbers of fl ower heads than the other plants. The specifi city of the relationship among plant populations, AMF, and soil was also observed for percentage of root colonization. CONCLUSIONS The study extends our knowledge of the specifi c interaction between plants and AMF by demonstrating that the positive effect of native AMF occurs only when the plants are also grown in their native soil. Thus, when attempting to facilitate plant growth, we need to consider the origin of the plants, the soil, and the AMF, because all three components are adapted to each other.

The Effect of AMF Suppression on Plant Species Composition in a Nutrient-Poor Dry Grassland

Arbuscular mycorrhizal fungi (AMF) are expected to be one of the key drivers determining the diversity of natural plant communities, especially in nutrient-poor and dry habitats. Several previous studies have explored the importance of AMF for the composition of plant communities in various types of habitats. Surprisingly, studies of the role of AMF in nutrient-poor dry grassland communities dominated by less mycotrophic plant species are still relatively rare. We present the results of a 3-year study in which a plant community in a species-rich dry grassland was subjected to the fungicide carbendazim to suppress AMF colonization. We tested the effect of the fungicide on the following parameters: the plant species composition; the number of plant species; the cover of the rare, highly mycorrhiza-dependent species Aster amellus; the cover of the dominant, less mycorrhiza-dependent species Brachypodium pinnatum; and the cover of graminoids and perennial forbs. In addition, we examined the mycorrhizal inoculation potential of the soil. We found that the suppression of AMF with fungicide resulted in substantial changes in plant species composition and significant decrease in species richness, the cover of A. amellus and the cover of perennial forbs. In contrast the species increasing their cover after fungicide application were graminoids—the C3 grasses B. pinnatum and Bromus erectus and the sedge Carex flacca. These species appear to be less mycorrhiza dependent. Moreover, due to their clonal growth and efficient nutrient usage, they are, most likely, better competitors than perennial forbs under fungicide application. Our results thus suggest that AMF are an essential part of the soil communities supporting a high diversity of plant species in species-rich dry grasslands in nutrient-poor habitats. The AMF are especially important for the maintenance of the populations of perennial forbs, many of which are rare and endangered in the area.

Effect ofGlomus intraradices isolated from Pb-contaminated soil on Pb uptake byAgrostis capillaris is changed by its cultivation in a metal-free substrate

Development and heavy metal tolerance of two cultivation lineages of the indigenous isolate of arbuscular mycorrhizal fungus (AMF)Glomus intraradices PH5 were compared in a pot experiment in soil from lead (Pb) smelter waste deposits. One lineage was sub-cultured in original Pb-contaminated soil; the second one was maintained for 13 months in an inert substrate (river sand) without Pb stress. The contribution of these cultivation lineages to the Pb uptake and accumulation by the host plantAgrostis capillaris was investigated. The experiment was conducted in a compartmented system where the lateral compartments withAgrostis seedlings were separated from the central pot containing 4-week olderAgrostis plants by a nylon mesh for allowing out-growing of extraradical mycelium (ERM) from the pot.No differences in mycorrhizal colonization, ERM length and viability were observed between the two lineages ofG. intraradices PH5 in the soil of the isolate origin. However, the ability to support plant growth and Pb uptake differed between the lineages and also between the plants in the central pots and the lateral compartments. The growth of the plants in the central pots was positively affected by AMF inoculation. The plants inoculated with the lineage maintained in original soil showed larger shoot biomass and higher shoot P content as compared to the other inoculation treatments. The shoot Pb concentration of these plants was lower when compared to the plants inoculated with the lineage sub-cultured in the inert substrate. However the concentration did not differ from non-mycorrhizal control or from the reference isolateG. intraradices BEG75 from non-contaminated soil. Also shoot Pb contents were similar for all inoculation treatments. The development ofG. intraradices BEG75 in the contaminated soil was very poor; this isolate was not able to initiate colonization of seedlings in lateral compartments. In lateral compartments, growth of seedlings in contaminated soil was inhibited by theG. intraradices PH5 lineage maintained in the inert substrate. Pb translocation from the seedling roots to shoots was increased for plants inoculated with either lineage as compared to the non-mycorrhizal control; however, the increase for the lineage cultivated in the inert substrate was significantly higher in comparison with that maintained in the original soil. After 13 months of cultivation in a metal free substrate, theG. intraradices isolate from Pb contaminated soil did not lose its tolerance to Pb as regards colonization of plant roots and growth of ERM in the soil of its origin. However, its ability to support plant growth and to prevent Pb translocation from the roots to the shoots was decreased.

Associations of dominant plant species with arbuscular mycorrhizal fungi during vegetation development on coal mine spoil banks

Among plants colonizing mine spoil banks in Northern Bohemia the first colonizers, mainly ruderal annuals fromChenopodiaceae andBrassicaceae were found not to be associated with arbuscular mycorrhizal fungi (AMF). These species cultivated in pots with soil from four sites in different succession stages of the spoil bank did not respond to the presence of native or non-native AMF. All grass species studied (Elytrigia repens, Calamagrostis epigejos andArrhenatherum elatius) were found moderately colonized in the field and they seemed to be facultative mycotrophs, because their response to mycorrhiza in the experiment was negligible.Carduus acanthoides was found to be highly colonized in the field, however, it did not show growth response to AMF in the pot experiment. The AMF native in four sites on the spoil banks showed high infectivity but low effectiveness in association with colonizing plants compared to the non-native isolateG. fistulosum BEG23. In general, dependence on AMF in the cultivation experiment was rather low, regardless of the fact that plants were found to be associated with AMF either in the field or in pots. Occurrence and effectiveness of mycorrhizal associations might relate primarily to the mycotrophic status of each plant species rather than to the age of the spoil bank sites studied.

Ploidy-specific interactions of three host plants with arbuscular mycorrhizal fungi: Does genome copy number matter?

UNLABELLED PREMISE OF THE STUDY Polyploidy has been shown to affect different plant traits and modulate interactions between plants and other organisms, such as pollinators and herbivores. However, no information is available on whether it can also shape the functioning of mycorrhizal symbiosis. • METHODS The mycorrhizal growth response was assessed for three angiosperms with intraspecific ploidy variation. Different cytotypes of Aster amellus, Campanula gentilis, and Pimpinella saxifraga were either left uninoculated or were inoculated with arbuscular mycorrhizal (AM) fungi in a pot experiment. After 3 mo of cultivation in a greenhouse, plant growth, phosphorus concentration in the shoot biomass, and development of the AM symbiosis were evaluated. • KEY RESULTS No significant ploidy-specific differences in AM development were recorded. The inoculation led to consistently greater phosphorus uptake; however, the effect on plant growth differed considerably among plant species, populations, ploidy levels, and AM species. A salient ploidy-specific response was observed in A. amellus. Whereas diploid plants benefited from AM inoculation, the hexaploids consistently showed negative or no-growth responses (depending on the AM species). In contrast to A. amellus, no interactions between inoculation and ploidy were observed in C. gentilis and P. saxifraga. • CONCLUSIONS The first evidence is provided of a ploidy-specific response of a mycotrophic plant to AM fungi. Our results demonstrate the complexity of interaction between plants and associated AM fungi, with the ploidy level of the host plant being one component that may modulate the functioning of the symbiosis.