Analysis of effects and factors linked to soil microbial populations and nitrogen cycling under long-term biosolids application.

Information about impacts of long-term biosolids application on soil microbial populations and functional groups and N cycling is important for evaluating soil health and agroecosystem sustainability under long-term biosolids application. Mine spoil plots received annual biosolids application from 1973 to 2010 at low (16.8 Mg ha-1 yr-1), medium (33.6 Mg ha-1 yr-1), and high rates (67.2 Mg ha-1 yr-1). A no-biosolids control received chemical fertilizer at the agronomic rate. Soil samples were collected in three seasons per year spanning 2003-2005 for measuring soil moisture, pH, soil organic C (SOC), total and extractable heavy metals (Cd, Cu, Ni, Zn), NO3-, N mineralization potential (NMP), microbial biomass C (MBC), and populations of three N-cycling bacteria (NCB) groups: ammonia-oxidizing bacteria (AOB), nitrite-oxidizing bacteria (NOB), and denitrifying bacteria (DNB). Soil samples were collected again in 2008 and 2010 for quantifying total and extractable heavy metals, and in 2018 (eight years after biosolids applications ended) for measuring SOC, MBC, NMP, and microbial respiration. During 2003-2005, mean MBC was 315, 554, 794, and 1001 mg kg-1 in the control, low, medium, and high biosolids treatments, respectively. Populations of NCB did not differ among treatments. Biosolids application increased total and extractable metal concentrations but the effect of biosolids rates were much lower on extractable than total concentrations. Soil extractable Cd and Cu concentrations decreased from medium to high applications, likely due to complexing with biosolids organic matter. Partial least squares regression analysis identified a strong positive effect on MBC of SOC and a weak negative effect of Cu, explaining the strong net positive effect of biosolids on MBC. In 2018, the medium and high biosolids treatments maintained higher SOC, MBC, NMP, and microbial respiration than the control. This study provided further evidence that long-term biosolids application has positive effects on soil microbes that persist for years after ending application.

Barcoded NS31/AML2 primers for sequencing of arbuscular mycorrhizal communities in environmental samples.

PREMISE OF THE STUDY: Arbuscular mycorrhizal fungi (AMF) are globally important root symbioses that enhance plant growth and nutrition and influence ecosystem structure and function. To better characterize levels of AMF diversity relevant to ecosystem function, deeper sequencing depth in environmental samples is needed. In this study, Illumina barcoded primers and a bioinformatics pipeline were developed and applied to study AMF diversity and community structure in environmental samples. METHODS: Libraries of small subunit ribosomal RNA fragment amplicons were amplified from environmental DNA using a single-step PCR reaction with barcoded NS31/AML2 primers. Amplicons were sequenced on an Illumina MiSeq sequencer using version 2, 2 × 250-bp paired-end chemistry, and analyzed using QIIME and RDP Classifier. RESULTS: Sequencing captured 196 to 6416 operational taxonomic units (OTUs; depending on clustering parameters) representing nine AMF genera. Regardless of clustering parameters, ∼20 OTUs dominated AMF communities (78-87% reads) with the remaining reads distributed among other OTUs. Analyses also showed significant biogeographic differences in AMF communities and that community composition could be linked to specific edaphic factors. DISCUSSION: Barcoded NS31/AML2 primers and Illumina MiSeq sequencing provide a powerful approach to address AMF diversity and variations in fungal assemblages across host plants, ecosystems, and responses to environmental drivers including global change.