Effects of long-term mineral fertilization and manuring on the biomass of arbuscular mycorrhizal fungi (AMF) were studied in a field experiment. Mineral fertilization reduced the growth of AMF, as estimated using both measurements of hyphal length and the signature fatty acid 16:1omega5, whereas manuring alone increased the growth of AMF. The results of AMF root colonization followed the same pattern as AMF hyphal length in soil samples, but not AMF spore densities, which increased with increasing mineral and organic fertilization. AMF spore counts and concentration of 16:1omega5 in soil did not correlate positively, suggesting that a significant portion of spores found in soil samples was dead. AMF hyphal length was not correlated with whole cell fatty acid (WCFA) 18:2omega6,9 levels, a biomarker of saprotrophic fungi, indicating that visual measurements of the AMF mycelium were not distorted by erroneous involvement of hyphae of saprotrophs. Our observations indicate that the measurement of WCFAs in soil is a useful research tool for providing information in the characterization of soil microflora.

Tuber aestivum is the most common European truffle with significant commercial exploitation. Its production originates from natural habitats and from artificially inoculated host tree plantations. Formation of Tuber ectomycorrhizae in host seedling roots is often inefficient. One possible reason is the lack of indigenous associative microbes. Here we aimed at metagenetic characterization and cultivation of indigenous prokaryotes associated with T. aestivum in a field transect cutting through the fungus colony margin. Several operational taxonomic units (OTUs) showed close association with the T. aestivum in the ectomycorrhizae and in the soil, but there was no overlap between the associative prokaryotes in the two different habitats. Among those positively associated with the ectomycorrhizae, we identified several bacterial genera belonging to Pseudonocardineae. Extensive isolation efforts yielded many cultures of ectomycorrhizae-associative bacteria belonging to Rhizobiales and Streptomycineae, but none belonging to the Pseudonocardineae. The specific unculturable Tuber-associated prokaryotes are likely to play important roles in the biology of these ectomycorrhizal fungi, including modulation of competition with other symbiotic and saprotrophic microbes, facilitation of root penetration and/or accessing mineral nutrients in the soil. However, the ultimate proof of this hypothesis will require isolation of the microbes for metabolic studies, using novel cultivation approaches.

Arbuscular mycorrhizal (AM) fungi lack efficient exoenzymes to access organic nutrients directly. Nevertheless, the fungi often obtain and further channel to their host plants a significant share of nitrogen (N) and/or phosphorus from such resources, presumably via cooperation with other soil microorganisms. Because it is challenging to disentangle individual microbial players and processes in complex soil, we took a synthetic approach here to study 15N-labelled chitin (an organic N source) recycling via microbial loop in AM fungal hyphosphere. To this end, we employed a compartmented in vitro cultivation system and monoxenic culture of Rhizophagus irregularis associated with Cichorium intybus roots, various soil bacteria, and the protist Polysphondylium pallidum. We showed that upon presence of Paenibacillus sp. in its hyphosphere, the AM fungus (and associated plant roots) obtained several-fold larger quantities of N from the chitin than it did with any other bacteria, whether chitinolytic or not. Moreover, we demonstrated that adding P. pallidum to the hyphosphere with Paenibacillus sp. further increased by at least 65% the gain of N from the chitin by the AM fungus compared to the hyphosphere without protists. We thus directly demonstrate microbial interplay possibly involved in efficient organic N utilisation by AM fungal hyphae.

Arbuscular mycorrhizal (AM) symbiosis is heavily and positively implicated in phosphorus (P) acquisition from soil to plants, including many important agricultural crops. Its role in plant nitrogen (N) nutrition is generally not as prominent or beneficial, with exception of some situations when N is available predominantly in organic forms. Yet the AM fungi (AMF) are, due to their poor exo-enzymatic repertoire, unlikely to degrade organic compounds on their own, therefore they possibly depend on other microorganisms to liberate nutrients contained in those materials. Here, we review current knowledge on the roles played by the AMF in plant N nutrition in general and uptake of N from organic compounds in particular, with a specific reference to microbes and processes involved in liberation and AM fungal utilization of N from organic compounds. Future research needs and directions are outlined, as is the agronomic and societal context of such research.