Abstract
We isolated tomato spotted wilt virus (TSWV-KP) from a diseased Capsicum annuum var. grossum with malformed leaves and necrotic spotted fruits. TSWV-KP produced necrosis or necrotic ring spots on inoculated leaves along with mosaic, vein necrosis, or death on the upper leaves on Datura stramonium, Nicotiana clevarandii, N. rustica, and N. tabacum cvs. Ultrastructurally, typical tospovirus particles were observed in the cytoplasm. The virion contained three molecules of genomic RNAs of approximately 9.0, 4.9, and 3.0 kb. The nucleocapsid (N) protein of the purified virion migrated as a single band with ~29 kDa molecular weight in SDS-PAGE. Complete nucleotide sequences of the large (L) genome segments of TSWV-KP were determined. Defective forms of L-RNA containing core polymerase regions were observed. L-RNA (8,917 nucleotides) contained a single open reading frame (ORF) in the viral complementary (vc) strand and encoded a 330-kDa protein. The L-protein had high identity in the “core-polymerase domain” with the corresponding regions of other tospoviruses. The complete nucleotide sequence of TSWV-KP medium-sized (M) RNA comprised 4,768 nucleotides and indicated a typical tospovirus with two genes in ambisense arrangement. The vRNA OFR coded for a potential cell-to-cell movement (NSm) 34.8-kDa protein; and the vcRNA ORF, for the viral glycoprotein (G1/G2) 128.0-kDa precursor. Multiple sequence alignment of the M-RNA showed highest homologies to TSWV-BR01. Amino acid sequences of TSWV-KP NSm and G1/G2 exhibited 48.7–85.3% and 34.9–96.2% identity, respectively. TSWV-KP small (S) RNA comprised 2,991 nucleotides with ambisense coding strategy. The sequence contained two ORFs—one in the viral sense, encoding a protein with predicted 52.4-kDa Mr; and another in the viral complementary sense, encoding the viral nucleocapsid protein of 28.8-kDa Mr. Amino acid sequences of TSWV-KP of S-RNA NSs and N exhibited 35.9–87.9% and 19.9–98.4% identity, respectively.
Highlights
Far, 13 species of the genus Tospovirus have been reported worldwide
Determination of the complete nucleotide sequence of Tomato spotted wilt virus (TSWV)-KP L-RNA demonstrates that TSWV is a negativestrand RNA virus
TSWV-BR-01 strains L-RNA could be folded into a stable panhandle structure, which may be involved in the appearance of circular nucleocapsids in virus particles (Peters et al, 1991), as found for Bunyaviridae (Raju and Kolakofsky 1989)
Summary
COMPLETE GENOME SEQUENCE OF TOMATO SPOTTED WILT VIRUS FROM PAPRIKA IN KOREA aJae-Hyun Kim, bYoung-Soo Kim, aSoo-Won Jang, bYong-Ho Jeon* a Ginseng Resources Research Laboratory, KGC Corp., Daejeon 305-805, Republic of Korea. b Department of Bioresource Sciences, Andong National University, Andong 760-749, Republic of Korea.
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