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To compare soil viral community composition within and across terrestrial habitats on a regional scale, viromes were generated from 34 near-surface (top 15 cm) soil samples, with a total of 30 viromes included in downstream ecological analyses (see Supplementary Methods). The analyzed viromes were collected from four distinct habitats (wetlands, grasslands, chaparral shrublands, and woodlands, each with 7, 14, 4, and 5 viromes, respectively) across five field sites (Fig. S1 for sampling scheme, Table S1 for soil properties). Following quality filtering, the 30 viromes generated an average of 72,950,833 reads and 416 contigs ≥10 Kbp per virome (Table S2). Wetland viromes yielded more contigs ≥10 Kbp than viromes from other habitats, both in total and on average per virome (Table S2). We used VIBRANT to identify 3490 viral contigs in our assemblies, which were clustered into 3,432 viral operational taxonomic units (vOTUs), defined as ≥10 Kbp viral contigs sharing ≥ 95% average nucleotide identity over 85% contig length [17]. Constrained analysis of principal coordinates (CAP analysis) revealed strong clustering by habitat rather than by site, implying that, where environmental parameters are substantially different, environmental conditions are stronger drivers of viral community composition than geographic distance (Fig. S2).Multiple lines of evidence suggest that wetter soils harbored greater viral diversity than drier soils. We recovered the most vOTUs from wetlands, both in total (56% of all vOTUs were from wetlands) and per virome (on average, 307 vOTUs were recovered per wetland virome, compared to 116 from all habitats) (Fig. 1A). Unsurprisingly, wetlands had significantly greater moisture content than other habitats (Fig. 1B; ANOVA followed by Tukey multiple comparisons of means, p 100 Km distances here. Taken together, we propose that soil viral communities often display high heterogeneity within and among habitats, presumably due to a combination of host adaptations and microdiversity, dispersal limitation, and fluctuating environmental conditions over space and time. More

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