Biochemical and molecular characterization of microbial communities from a metal contaminated and reclaimed region.

Abstract

Metal contamination in the Greater Sudbury Region (GSR) resulted in severe environmental degradation. Soil liming and tree planting have been the main approaches to restoring the damaged ecosystem. The specific objective of the present study was to assess the effects of soil metal contamination and liming on 1) microbial biomass and abundance, 2) bacterial and fungal diversity, and 3) enzymatic activities and soil respiration. Phospholipid fatty acid (PLFA) analysis and 454 pyrosequencing were used to address these research objectives. Total biomasses for bacteria, arbuscular fungi (AM fungi), other fungi and eukaryotes were significantly lower in metal contaminated compared to uncontaminated reference areas. Analysis of bacterial communities revealed Chao1 index values of 232 and 273 for metal contaminated and reference soils, respectively. For fungi, the Chao index values were 23 for metal contaminated and 45 for reference sites. There was a significant increase of total microbial biomass in limed sites (342.15 ng/g) compared to unlimed areas (149.89 ng/g). Chao1 estimates followed the same trend. But the total number of OTUs (Operational Taxonomic Units) in limed (463 OTUs) and unlimed (473 OTUs) soil samples for bacteria were similar. For fungi, OTUs were 96 and 81 for limed and unlimed soil samples, respectively. Bacterial and fungal groups that were specific to either limed or unlimed sites were identified. Bradyrhizobiaceae family with 12 genera including the nitrogen fixing Bradirhizobium genus was more abundant in limed sites compared to unlimed areas. For fungi, Ascomycota was the most predominant phylum in unlimed soils (46.00%) while Basidiomycota phylum represented 85.74% of all fungi in the limed areas. Detailed analysis of the data showed that although soil liming increases significantly the amount of microbial biomass, the level of species diversity remained statistically unchanged. Soil respiration rates were higher in limed soils (65 ppm) compared to unlimed soils (35 ppm). They were significantly lower in metal contaminated sites (55 ppm) compared to reference sites (90 ppm). β-glucosidase (BG), cellobiohydrolase (CBH), β-N-acetylglucosaminidase (NAGase), aryl sulfatase (AS), acid phosphatase (AP), alkaline phosphatase (AlP), glycine aminopeptidase (GAP), and leucine aminopeptidase (LAP) activites were significantly higher in limed compared to unlimed sites. Metal contamination significantly reduced the activities of these enzymes with the exception of LAP. An opposite trend was observed for peroxidase (PER) enzyme activity that was higher in unlimed and metal contaminated sites compared to limed and reference areas.