Phylogeny of the amino acid sequence of all tospoviral RdRps used in this study showed that all the known TSWV isolates including the USA isolate described in this study formed a distinct and a close cluster with that of Impateins necrotic spot virus. Phylogenetic analysis using nucleotide sequences of the complete L RNA showed that TSWV WA-USA isolate clustered with the American and Asian TSWV isolates which formed a distinct clade from Euro-Asiatic Tospoviruses. Two untranslated regions of 241 and 33 nucleotides were present at the 5′ and 3′ termini, respectively that shared conserved tospoviral sequences.
The ORF potentially codes for RNA-dependent RNA polymerase (RdRp) of 330.9 kDa. It consisted of 8914 nucleotides (nt) encoding a single open reading frame of 8640 nts in the viral-complementary sense. The large (L) RNA of a TSWV WA-USA isolate was cloned and sequenced. However the complete sequence of only one TSWV isolate PA01 characterized from pepper in Pennsylvania is available. Tomato spotted wilt virus (TSWV Tospovirus: Bunyaviridae) has been an economically important virus in the USA for over 30 years.
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