Main Observations from the Plot ✅ Flooding (0W, 150W) decreases microbial diversity • The 0W (orange) and 150W (gray) treatments have the lowest Shannon indices, confirming that prolonged water exposure reduces microbial richness. ✅ Nitrogen addition does not significantly impact diversity • 100D (yellow), 100W (blue), and 150D (red) show similar diversity values, meaning that nitrogen alone does not significantly shift microbial richness. ✅ Highest diversity is in non-flooded control (0D, green) • The 0D treatment (no flooding, no nitrogen) has the highest median Shannon diversity, suggesting that dry conditions support greater microbial diversity.

Dynamic ASV Variability Over Time The top 20 ASVs showed significant fluctuations in abundance across the 11-week period. Some ASVs exhibited persistent presence throughout, while others had sharp increases or decreases at specific time points. Distinct Shifts in ASV Composition Certain ASVs (e.g., ASV107, ASV160, ASV275) displayed peaks in abundance at specific weeks, suggesting a response to environmental changes. Some ASVs were highly active early on (Weeks 1–3), while others dominated in later weeks (e.g., Weeks 9–11). Potential Environmental Influence The shifts in ASV abundance may correlate with external factors such as: Flooding vs. dry conditions Nitrogen application levels Soil chemistry changes over time

EMAI 16srDNA profile beta diversity plot of treatments over 11 sampling weeks

Box Plot - shannon diversity of 16srDNA profile across treatments

PCoA plots visualizing fungal community differences (ITS profiles) between composites, averages and replicate treatment ITS profiles for Week 3

PCoA plots visualizing fungal community differences (ITS profiles) between composites, averages and replicate treatment ITS profiles for Week1

week 3 PERMANOVA compsoites, averages, and replicates

week 1 comparison between averages, composites and replicates

Week1 - Beta diversity measure of community composition similarity between treatment composite DNA, individual reps and their averages - visualized using Principal Coordinate Analysis (PCoA)

Beta diversity measure of community composition similarity between treatment composite DNA, individual reps and their averages - visualized using Principal Coordinate Analysis (PCoA)