The diversity of viruses associated with non-cultivated plants was assessed from plant samples collected in the Tallgrass Prairie Preserve of northeastern Oklahoma, USA. The samples were processed to determine the sequences of nucleic acids extracted from the virus-like particle fraction of plant homogenates. Sequences from 95 specimens of 52 plant species included those of probable origin from the genomes of plants (including retroelements), bacteria, fungi, other organisms, and viruses. Virus-like sequences were identified in sequences from 25% of the specimens, coming from 19% of the plant species. Evidence of a member of the genus Tymovirus was found in 16 specimens of 6 plant species, making it the most predominant virus associated with the sampled plants. There was evidence of the presence of more than one virus in each of six specimens.