The host plant Pinus pinaster exerts specific effects on phosphate efflux and polyphosphate metabolism of the ectomycorrhizal fungus Hebeloma cylindrosporum: a radiotracer, cytological staining and 31 P NMR spectroscopy study.

Ectomycorrhizal (ECM) association can improve plant phosphorus (P) nutrition. Polyphosphates (polyP) synthesized in distant fungal cells after P uptake may contribute to P supply from the fungus to the host plant if they are hydrolyzed to phosphate in ECM roots then transferred to the host plant when required. In this study, we addressed this hypothesis for the ECM fungus Hebeloma cylindrosporum grown in vitro and incubated without plant or with host (Pinus pinaster) and non-host (Zea mays) plants, using an experimental system simulating the symbiotic interface. We used 32 P labelling to quantify P accumulation and P efflux and in vivo and in vitro nuclear magnetic resonance (NMR) spectroscopy and cytological staining to follow the fate of fungal polyP. Phosphate supply triggered a massive P accumulation as newly synthesized long-chain polyP in H. cylindrosporum if previously grown under P-deficient conditions. P efflux from H. cylindrosporum towards the roots was stimulated by both host and non-host plants. However, the host plant enhanced 32 P release compared with the non-host plant and specifically increased the proportion of short-chain polyP in the interacting mycelia. These results support the existence of specific host plant effects on fungal P metabolism able to provide P in the apoplast of ectomycorrhizal roots.

Efficiency of acid phosphatases secreted from the ectomycorrhizal fungus Hebeloma cylindrosporum to hydrolyse organic phosphorus in podzols.

Ectomycorrhizal fungi may improve the phosphate nutrition of their host plants by secreting, into the soil solution, acid phosphatases (AcPases) able to release orthophosphate (Pi) from soil organic phosphorus (Po). Using cation-exchange chromatography, we separated four fractions with AcPase activity secreted by the ectomycorrhizal fungus Hebeloma cylindrosporum grown in a pure culture under P-starved conditions. Each AcPase active fraction displayed strong ability in vitro to hydrolyse a wide range of phosphate monoesters, but none of them efficiently hydrolysed phytate. Their efficiency to release Pi from soil NaHCO(3)-extractable Po was studied in a sandy podzol used intact or autoclaved. Soils were collected in a 15-year-old Pinus pinaster stand, receiving regular fertilization or not. Autoclaving increased the NaHCO(3)-extractable Po concentrations by 55% in unfertilized and by 32-43% in fertilized soils. The efficiency of each AcPase fraction was affected significantly by the soil fertilization regime and the soil treatment (intact vs. autoclaved). The proportion of labile Po enzyme ranged from 0% to 11% and 14% to 48% after 1 h of incubation in bicarbonate solutions extracted from intact and autoclaved soils, respectively. This work suggests that AcPases secreted from H. cylindrosporum could be efficient in recycling Po pools from soil microorganisms that may be delivered by soil autoclaving.