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Metagenomics for the Identification Of Plant Viruses in the Tallgrass Prairie Preserve

Muthukumar, Vijay
The number of virus species identified by the ICTV is likely to be far less than the actual number of viruses. This study was undertaken to develop a metagenomic technique to discover viruses from plants. Aliquots of plant material were subjected to homogenization and differential centrifugation to isolate virus-like particles. The pellet obtained was treated successively with DNase-I to remove contaminating plant DNA and proteinase-K, SDS to digest the DNase-I and capsid proteins. The viral nucleic acid was amplified by random PCR, cloned and sequenced. The sequences obtained were assembled and characterized by comparison with known viral sequences. Sequences were obtained from 68-plant specimens. Analysis of the sequences suggested the presence of at least 9 different viruses. Sixteen plants contained virus-like sequences and six double infections were found. Plant specimens yielding virus sequences showed no obvious symptoms of infection. Evidence of bacteria and fungi was also found in several samples.