Harnessing the Power of eDNA Biodiversity Assessment to Enhance Subsurface Water Flow Pathway Reconstruction

Rainfall runoff contributes to a large proportion of the discharge in streams and therefore, heavily influences stream water quality but also flood generation. Rainfall runoff generation is usually a combination of overland and subsurface flow processes, the latter of which being especially difficult to trace. Here, we explored the viability of environmental DNA (eDNA) for subsurface water flow pathway reconstruction and simultaneous biodiversity assessment. The degree of similarity of community patterns indicates biological and therefore, in principle, also hydrological connectivity. We applied eDNA metabarcoding to characterise 10 drilling cores (0.7-3.2 m depth) on 3 hillslopes (10x50 m) in 4 catchment areas in Germany and Austria. In total, more than 2000 species across taxonomic groups could be identified down to species level. Analysis of alpha and beta diversity in the different catchments showed significant differences in spatial clustering patterns between taxonomic groups, but also between geomorphological and geochemical properties, such as the composition of dissolved organic carbon, of the respective catchment. We could assign indicator species sets in all taxonomic groups to various depth layers and identify habitat-specific communities that can be used as hydrological tracers. Although our results support the potential of eDNA to identify flow pathways and enhance our understanding of subsurface flow processes, we are still at the beginning of understanding the viability of eDNA as a tracer in hydrological research. However, our results show that making use of such naturally occurring tracers can expand our understanding of hydrological phenomena, especially those hidden in the subsurface.