Arbuscular mycorrhizal (AM) fungi enhance plant uptake of available phosphorus (P) from soil through their extraradical hyphae. The mechanism underlying this P uptake enhanced by AM fungi is the increase in the surface area for absorption of available P. Little is known about utilization of unavailable P by AM fungi. We investigated whether extraradical hyphae of AM fungi release acid phosphatase (ACP). Sterilized Andosol was packed in pots that were separated into the mycorrhizal and hyphal compartments with a nylon net of 30-μm pore size. Seeds of Allium fistulosum L. were inoculated or uninoculated with the AM fungus Rhizophagus clarus (Nicolson & Schenck) Walker & Schüßler. Mullite ceramic tubes were buried in the soil of each compartment, and soil solution was collected. A. fistulosum L. and Linum usitatissimum L. inoculated with R. clarus were grown in sand culture and in vitro monoxenic culture, respectively. Uninoculated A. fistulosum L was grown in hydroponic culture to collect root exudate. The soil solution, hyphal extracts, root extract and root exudates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) analysis. Shoot P concentration, shoot P content and shoot dry weight were higher in the inoculated treatment than in the uninoculated treatment. Activity staining of the gel revealed that ACP activity at 187 kDa was observed in the soil solution in the inoculation treatment, and in the hyphal extract collected from sand culture and in vitro monoxenic culture, but neither in the root exudate of non-mycorrhizal plant grown in the hydroponic culture nor in the root extracts irrespective of mycorrhizal status. Those results provide strong evidence that the corresponding activity in the soil solutions in soil culture is of R. clarus CK001 origin. These findings suggest that the fungus releases ACP from extraradical hyphae into the hyphosphere.

Root exudates are derived from plant metabolites and their composition is affected by the plant nutritional status. The purpose of this study was to examine soybean root exudates under phosphorus (P) deficiency using a metabolite profiling technique. Glycine max L. (cv. Suzuyutaka) was grown in a culture solution at P concentrations of 0 (P0) and 8 (P8) mg P L⁻¹ for 1, 5, 10 and 15 days after transplanting. Shoot extract, root extract and root exudates were collected and their metabolites were determined by capillary electrophoresis/time-of-flight mass spectrometry (CE-TOF MS). The shoot P concentration and dry weight of soybean plants grown at P0 were lower than those at P8. One hundred and eight, 116 and 79 metabolites were identified in the shoot extract, root extract and root exudates, respectively. The concentrations of several metabolites including amino acids and organic acids in root exudates were higher at P0 than at P8, irrespective of the P concentration in the shoot or root extract. These findings suggest that soybean roots actively release metabolites in response to P deficiency.

Root exudates improve the nutrient acquisition of plants and affect rhizosphere microbial communities. The plant nutrient status affects the composition of root exudates. The purpose of this study was to examine common bean (Phaseolus vulgaris L.) root exudates under phosphorus (P) deficiency using a metabolite profiling technique. Common bean plants were grown in a culture solution at P concentrations of 0 (P0), 1 (P1) and 8 (P8) mg P L⁻¹ for 1, 10 and 20 days after transplanting (DAT). Root exudates were collected, and their metabolites were determined by capillary electrophoresis time-of-flight mass spectrometry (CE-TOF MS). The shoot P concentration and dry weight of common bean plants grown at P0 were lower than those grown at P8. One hundred and fifty-nine, 203 and 212 metabolites were identified in the root exudates, and 16% (26/159), 13% (26/203) and 9% (20/212) of metabolites showed a P0/P8 ratio higher than 2.0 at 1, 10 and 20 DAT, respectively. The relative peak areas of several metabolites, including organic acids and amino acids, in root exudates were higher at P0 than at P8. These results suggest that more than 10% of primary and secondary metabolites are induced to exude from roots of common bean by P deficiency.