Monitoring species of arbuscular mycorrhizal fungi in planta 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. This revised version was published online in June 2006 with corrections to the Cover Date.