Summary: In a phenotypic and bioinformatic survey of the natural history of Hesperis matronalis across southern Michigan I found that both land cover type and longitude are positively correlated with infection of TuMV at the phenotypic level. However, this finding is likely highly correlated with the land cover types further to the north being categorized as Natural, and land cover types at the southern range of the sampling area Agricultural. Additionally, bioinformatic analysis found that there are differences in the abundance of certain taxa across the diseased and normal phenotypes. The results of an all-by-all BLAST of the longest assemblies of TuMV were visualized and suggestive of strain variation across populations of H. matronalis. These findings are important to our understanding of how biotic and abiotic interactions influence the ecology and evolution of plant viruses in wild populations. To gain a deeper ecological understanding of the frequency of TuMV in wild populations, future studies should focus on the ecology of the aphid vectors.