Hidden viral players: Diversity and ecological roles of viruses in groundwater microbiomes

Groundwater ecosystems harbor diverse microbial communities adapted to energy-limited, light-deprived conditions, yet the role of viruses in these environments remains poorly understood. Here, we analyzed 1.26 terabases of metagenomic and metatranscriptomic data from seven wells in the Hainich Critical Zone Exploratory (CZE) to characterize groundwater viromes. We identified 257,252 viral operational taxonomic units (vOTUs) ([≥]5 kb), with 99% classified as novel, highlighting extensive uncharacterized viral diversity. Viruses exhibited a distinct host range, primarily targeting Proteobacteria, Candidate Phyla Radiation (CPR) bacteria, and DPANN archaea. Notably, CPR lineages displayed low virus-host ratios and viral CRISPR targeting multiple hosts, suggesting a "virus decoy" mechanism where they may absorb viral pressure, protecting bacteria hosts. Additionally, 3,378 vOTUs encoded auxiliary metabolic genes (AMGs) linked to carbon, nitrogen, and sulfur cycling, with viruses targeting 31.5% of host metabolic modules. These findings demonstrate viruses influence on microbial metabolic reprogramming and nutrient cycling in groundwater, shaping subsurface biogeochemistry.