Monitoring species of arbuscular mycorrhizal fungi <i>in</i> <i>planta</i> and in soil by nested PCR: application to the study of the impact of sewage sludge

Nested PCR is a highly sensitive procedure for monitoring species of arbuscular mycorrhizal (AM) fungi and for determining their abundance in planta and in soil. DNA sequence variability in the D1 and D2 domains of the large ribosomal subunit is sufficient to design primers which discriminate between AM fungi at the species level. The usefulness of this molecular approach is illustrated in the present study on the differential impact of sewage sludges on a community of three AM fungi ( Glomus mosseae , Glomus intraradices , Gigaspora rosea). Nested PCR was applied to trypan blue-stained mycorrhizal root fragments and soil mycelium from pot cultures of Medicago truncatula inoculated with the three fungi separately or together, and grown in sand containing sewage sludge that had been enriched or not with metallic or organic pollutants. G. intraradices and Gig. rosea varied in behaviour depending on whether they were inoculated alone or as a mixed community. G. mosseae showed a similar sensitivity towards each sewage sludge whether in community or alone, making it a potential candidate for ecotoxicological tests using M. truncatula to evaluate the quality or potential toxicity of sewage sludges which are widely used as fertilizers in agricultural lands</p

Evaluation moléculaire in planta de la diversité des champignons mycorhizogènes dans des champs amendés avec des boues d'épuration

National audienc

Evaluation moléculaire in planta de la diversité des champignons mycorhizogènes dans des champs amendés avec des boues d'épuration

National audienc

25S rDNA-based molecular monitoring of glomalean fungi in sewage sludge-treated field plots

International audienc

Heavy metals in sewage sludges contribute to their adverse effects on the arbuscular mycorrhizal fungus Glomus mosseae

International audienc

25S rDNA-based molecular monitoring of glomalean fungi in sewage sludge-treated field plots

Recycling of sewage wastes in agriculture is likely to affect the biological activity of soils through contamination of ecosystems by pathogens and metallic or organic micropollutants. The impact of sewage sludge spreading under field conditions on arbuscular mycorrhiza (AM) formation by a community of glomalean fungi was evaluated using a nested polymerase chain reaction (PCR) and discriminating primers based on 25S rDNA polymorphisms to detect different fungal species within root systems. Medicago truncatula was grown in soil of field plots amended or not with a composted sewage sludge, spiked or not with organic or metallic micropollutants. Overall AM development in roots decreased with sewage sludge application, and the relative abundance of five AM fungal morphotypes in root fragments was modified by the input of composted sludges. Sewage sludge spiked or not with organic pollutants had a generally positive effect on the relative diversity of AM fungal populations in planta, whereas after spreading of the sludge spiked with metallic pollutants, no variation was observed in the abundance of different species

Four Swedish long-term field experiments with sewage sludge reveal a limited effect on soil microbes and on metal uptake by crops

Purpose: This study aims to study the effect of sewage sludge amendment on crop yield and on microbial biomass and community structure in Swedish agricultural soils. Materials and methods Topsoil samples (0-0.20 m depth) from four sites where sewage sludge had been repeatedly applied during 14-53 years were analysed for total C, total N, pH and phospholipid fatty acids (PLFAs). Heavy metals were analysed in both soil and plant samples, and crop yields were recorded. Results and discussion At all four sites, sewage sludge application increased crop yield and soil organic carbon. Sludge addition also resulted in elevated concentrations of some heavy metals (mainly Cu and Zn) in soils, but high concentrations of metals (Ni and Zn) in plant materials were almost exclusively found in the oldest experiment, started in 1956. PLFA analysis showed that themicrobial community structure was strongly affected by changes in soil pH. At those sites where sewage sludge had caused low pH, Gram-positive bacteria were more abundant. However, differences in community structure were larger between sites than between the treatments. Conclusions: At all four sites, long-term sewage sludge application increased the soil organic carbon and nitrogen content, microbial biomass and crop yield. Long-term sewage sludge application led to a decrease in soil pH. Concentrations of some metals had increased significantly with sewage sludge application at all sites, but the amounts of metals added to soil with sewage sludge were found not to be toxic for microbes at any site