Location: Soil Management and Sugarbeet Research
Project Number: 3012-12210-001-023-I
Project Type: Interagency Reimbursable Agreement
Start Date: Jan 1, 2024
End Date: Jan 1, 2025
Objective:
The goal of this research is to utilize DNA metabarcoding to determine fungal and bacterial community composition in forest soils in the USA. This research will support the overall goal of this IA to assess soil microbial communities in relation to biochar amendment in forest soils. Specific outcomes will include the determination of community richness, diversity, phyla relative abundance, and composition. Further, absolute abundance of specific microbial populations of interest can be measured using quantitative PCR (qPCR) if it serves the overall purpose of this research.
Approach:
• Study sites: ca. 9 – 10 field sites will be selected (e.g., 3 sites in northwestern USA, 3 sites in midwestern USA, and 3 sites in southeastern USA).
• Soil collection in the field/site: ca. 30 soil samples will be collected per site (10 samples from control, 10 samples from biochar-amended soils, and 10 samples from adjacent forests)
• Soil DNA extraction: Qiagen DNeasy PowerSoil kits will be used to extract DNA from the soil samples.
• Metabarcoding sequencing and qPCR: Large subunit (LSU, ~1,700bp) for fungi and 16S (V4-V5 region, ~470bp) rRNA for bacteria will be sequenced using the Oxford Nanopore device or similar technology.
• Bioinformatic analyses: Community richness, diversity, phyla relative abundance, and composition, etc. will be determined using various genetic software.
