Isolation of bacteria from ectomycorrhizae of Tuber aestivum Vittad

Fifteen different cultivation media were used to isolate bacteria with the idea to obtain taxa specifically associated with ectomycorrhizae of Tuber aestivum. Ectomycorrhizae were collected at the sampling points previously analyzed for bacterial molecular diversity. We isolated 183 bacterial strains and identified them on the basis of the partial sequence of 16S rDNA. Out of these isolates, only 4 corresponded to operational taxonomic units significantly associated with T. aestivum ectomycorrhizae in previous molecular study. Preliminary study of the effect of 12 selected isolates on growth of T. aestivum mycelium showed no stimulation and one isolate induced the damage of hyphae. Different isolation strategy has to be developed to increase the probability of cultivation of potentially important components of T. aestivum mycorrhizosphere

Utilization of Organic Nitrogen by Arbuscular Mycorrhizal Hyphae in Soil - Zooming into the Hyphosphere Microbiome

Symposium paper Part 1: Function and management of soil microorganisms in agro-ecosystems with special reference to arbuscular mycorrhizal fung

Correlative evidence for co-regulation of phosphorus and carbon exchanges with symbiotic fungus in the arbuscular mycorrhizal Medicago truncatula.

Research efforts directed to elucidation of mechanisms behind trading of resources between the partners in the arbuscular mycorrhizal (AM) symbiosis have seen a considerable progress in the recent years. Yet, despite of the recent developments, some key questions still remain unanswered. For example, it is well established that the strictly biotrophic AM fungus releases phosphorus to- and receives carbon molecules from the plant symbiont, but the particular genes, and their products, responsible for facilitating this exchange, are still not fully described, nor are the principles and pathways of their regulation. Here, we made a de novo quest for genes involved in carbon transfer from the plant to the fungus using genome-wide gene expression array targeting whole root and whole shoot gene expression profiles of mycorrhizal and non-mycorrhizal Medicago truncatula plants grown in a glasshouse. Using physiological intervention of heavy shading (90% incoming light removed) and the correlation of expression levels of MtPT4, the mycorrhiza-inducible phosphate transporter operating at the symbiotic interface between the root cortical cells and the AM fungus, and our candidate genes, we demonstrate that several novel genes may be involved in resource tradings in the AM symbiosis established by M. truncatula. These include glucose-6-phosphate/phosphate translocator, polyol/monosaccharide transporter, DUR3-like, nucleotide-diphospho-sugar transferase or a putative membrane transporter. Besides, we also examined the expression of other M. truncatula phosphate transporters (MtPT1-3, MtPT5-6) to gain further insights in the balance between the "direct" and the "mycorrhizal" phosphate uptake pathways upon colonization of roots by the AM fungus, as affected by short-term carbon/energy deprivation. In addition, the role of the novel candidate genes in plant cell metabolism is discussed based on available literature

Arbuscular mycorrhizal fungi suppress ammonia-oxidizing bacteria but not archaea across agricultural soils

Arbuscular mycorrhizal (AM) fungi are supposedly competing with ammonia-oxidizing microorganisms (AO) for soil nitrogen in form of ammonium. Despite a few studies directly addressing AM fungal and AO interactions, mostly in artificial cultivation substrates, it is not yet clear whether AM fungi can effectively suppress AO in field soils containing complex indigenous microbiomes. To fill this knowledge gap, we conducted compartmentalized pot experiments using four pairs of cropland and grassland soils with varying physicochemical properties. To exclude the interference of roots, a fine nylon mesh was used to separate the rhizosphere and mesh bags, with the latter being filled with unsterile field soils. Inoculation of plants with AM fungus Rhizophagus irregularis LPA9 suppressed AO bacteria (AOB) but not archaea (AOA) in the soils, indicating how soil nitrification could be suppressed by AM fungal presence/activity. In addition, in rhizosphere filled with artificial substrate, AM inoculation did suppress both AOB and AOA, implying more complex interactions between roots, AO, and AM fungi. Besides, we also observed that indigenous AM fungi contained in the field soils eventually did colonize the roots of plants behind the root barrier, and that the extent of such colonization was higher if the soil has previously been taken from cropland than from grassland. Despite this, the effect of experimental AM fungal inoculation on suppression of indigenous AOB in the unsterile field soils did not vanish. It seems that studying processes at a finer temporal scale, using larger buffer zones between rhizosphere and mesh bags, and/or detailed characterization of indigenous AM fungal and AO communities would be needed to uncover further details of the biotic interactions between the AM fungi and indigenous soil AO

Mutabilis in mutabili: Spatiotemporal dynamics of a truffle colony in soil

The functioning of ectomycorrhizal (ECM) symbioses is closely related to the development of the soil mycelial phase the ECM fungi. The properties and spatiotemporal dynamics of such mycelia in ecosystems is, however, poorly understood. Here we show, using a soil colony of summer truffle (Tuber aestivum) as a model, that an ECM mycelium may only grow and colonize newly-opened soil patches when soil temperatures rise above certain threshold, in this case +10 °C, provided other requirements such as sufficient soil moisture are fulfilled. Extension rates of truffle mycelium in the fields was recorded as >0.3 μm min−1, several-fold greater than that predicted from laboratory cultures. Further, we demonstrated that there was a consistent spatial differentiation in mycelium growth patterns within the fungal colony on a decimeter scale, changing from “diffusion” type of growth at the colony margin to “colony-front” pattern further away from the colony margin. This change was clearly accompanied by shifting structure of soil microbial communities with Terrimonas sp. and another unidentified bacterium correlating with the “colony-front” mycelium growth pattern, and Sphingomonas sp. and Lysobacter brunnescens with the “diffusion” type of mycelium growth. Possible implications of the observed truffle colony differentiation are discussed for processes like fruit-body formation and dispersal of this ECM fungus. Our data indicate that the thallus of T. aestivum has to be considered as a principally variable (“mutabilis”) being in space and time, whose behavior correlates with conditions in ever changing soil environment (“in mutabili”)