Arbuscular mycorrhizal fungal diversity, external mycelium length, and glomalin-related soil protein content in response to long-term fertilizer management

Arbuscular mycorrhizal (AM) fungi are crucial for ecosystem functioning and can contribute to the formation and maintenance of soil aggregates through the exudation of glomalin by extraradical hyphae. Monitoring fertilization effects on AM fungi may help us to develop sound management strategies. The objectives of this study were to investigate the impacts of long-term fertilization on AM fungal parameters and to find out the key factor that affects the diversity and function of AM fungi. A long-term fertilization experiment established in a sandy loam soil at northern China has received continuous fertilization treatments for 21 years, including control; mineral fertilizers of NK, PK, NP, and NPK; organic manure (OM); and half organic manure N plus half mineral fertilizer N (1/2 OMN). Top soil samples (0-15 cm) from three individual plots per treatment were collected for the analysis of chemical properties and fungal parameters. The population size of soil AM fungi was determined by real-time PCR, and the community composition was analyzed using PCR-denature gradient gel electrophoresis (DGGE), cloning, and sequencing techniques. The external mycelium of AM fungi was assessed using the grid-line intersect method, and the glomalin-related soil protein (GRSP) was extracted with citrate solution using bovine serum albumin as a standard. Long-term fertilization significantly increased (P < 0.05) soil organic C content, AM fungal population, species richness (R), Shannon-Wiener index (H), and GRSP content, except for the P-deficiency (NK) fertilization treatment. OM had a significantly greater (P < 0.05) impact on AM fungal population and GRSP content compared to mineral fertilizers but significantly decreased the length of external mycelium compared to the control (P < 0.05). Fertilization also changed the community composition of AM fungi, and the P-deficiency treatment again had the slightest influence. In addition, most species recovered from the DGGE profiles belonged to three genera, Glomus, Diversispora, and Archaeospora. Redundancy analysis showed that the population size and species richness of AM fungi and the GRSP content all significantly correlated to soil organic C content (P < 0.05). Long-term P-containing fertilization, especially the application of OM, greatly increased the population size, species richness, and species diversity of AM fungi, as well as the contents of GRSP and soil organic C, but tended to decrease the length of external mycelium, while the P-deficiency fertilization had no such effect, suggesting that P was the key factor to maintain soil fertility as well as soil AM fungal diversity in this sandy loam soil.