scholarly journals Complete Genome Sequence of Tomato Spotted Wilt Virus from Paprika in Korea

2013 ◽  
Vol 2 (3) ◽  
pp. 121-136 ◽  
Author(s):  
Jae-Hyun Kim ◽  
Young-Soo Kim ◽  
Soo-Won Jang ◽  
Yong-Ho Jeon

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.

Plant Disease ◽  
2003 ◽  
Vol 87 (1) ◽  
pp. 102-102 ◽  
Author(s):  
S. Adkins ◽  
L. Breman ◽  
C. A. Baker ◽  
S. Wilson

Blackberry lily (Belamcanda chinensis (L.) DC.) is an herbaceous perennial in the Iridaceae characterized by purple-spotted orange flowers followed by persistent clusters of black fruit. In July 2002, virus-like symptoms including chlorotic ringspots and ring patterns were observed on blackberry lily leaves on 2 of 10 plants in a south Florida ornamental demonstration garden. Inclusion body morphology suggested the presence of a Tospovirus. Tomato spotted wilt virus (TSWV) was specifically identified by serological testing using enzyme-linked immunosorbent assay (Agdia, Elkhart, IN). Sequence analysis of a nucleocapsid (N) protein gene fragment amplified by reverse transcription-polymerase chain reaction (RT-PCR) with primers TSWV723 and TSWV722 (1) from total RNA confirmed the diagnosis. Nucleotide and deduced amino acid sequences of a 579 base pair region of the RT-PCR product were 95 to 99% and 95 to 100% identical, respectively, to TSWV N-gene sequences in GenBank. Since these 2-year-old plants were grown on-site from seed, they were likely inoculated by thrips from a nearby source. Together with a previous observation of TSWV in north Florida nursery stock (L. Breman, unpublished), this represents, to our knowledge, the first report of TSWV infection of blackberry lily in North America although TSWV was observed in plants of this species in Japan 25 years ago (2). References: (1) S. Adkins, and E. N. Rosskopf. Plant Dis. 86:1310, 2002. (2) T. Yamamoto and K.-I. Ohata. Bull. Shikoku Agric. Exp. Stn. 30:39, 1977.


1997 ◽  
Vol 87 (11) ◽  
pp. 1168-1173 ◽  
Author(s):  
Alice K. Inoue-Nagata ◽  
Richard Kormelink ◽  
Tatsuya Nagata ◽  
Elliot W. Kitajima ◽  
Rob Goldbach ◽  
...  

The generation of defective interfering (DI) RNA molecules of tomato spotted wilt tospovirus (TSWV) was studied by serially passaging in-ocula from plant to plant under different controlled conditions. DI RNAs were generated at higher rates in plants at 16°C than in plants incubated at higher temperatures. Another factor promoting the TSWV DI RNA generation was the use of high virus concentrations in the inocula. The solanaceous species Capsicum annuum, Datura stramonium, Lycopersicon esculentum, Nicotiana benthamiana, and N. rustica supported the generation of DI RNAs, whereas the virus recovered from the inoculated composite species, Emilia sonchifolia, remained free of any DI RNA under all conditions tested. This study resulted in a strategy to maintain DI RNA-free TSWV isolates, as well as in an efficient way to produce a large population of different DI RNA species. A single DI RNA species usually became dominant in an isolate after a few rounds of serial inoculations. The possible mechanisms involved in TSWV DI RNA generation under different inoculation circumstances are discussed.


Plant Disease ◽  
2001 ◽  
Vol 85 (12) ◽  
pp. 1290-1290 ◽  
Author(s):  
A. R. Golnaraghi ◽  
N. Shahraeen ◽  
R. Pourrahim ◽  
Sh. Ghorbani ◽  
Sh. Farzadfar

During the summers of 1999 and 2000, 3,110 soybean (Glycine max) leaf samples were randomly collected from soybean fields in the Ardebil, Goletan, Khuzestan, Lorestan, and Mazandaran provinces of Iran. Tomato spotted wilt virus (TSWV) was detected in leaf samples by TSWV-specific polyclonal antibody (As-0526 and As-0580, DSMZ, Braunschweig, Germany) in double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Mechanical inoculation of 26 plant species (10 plants per species) and cultivars with extracts of positive leaf samples produced necrotic local lesions in Beta vulgaris, Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Phaseolus vulgaris cv. Talash, Vicia faba, and Vigna unguiculata cv. Mashad; produced systemic chlorosis followed by necrosis in Datura stramonium, D. metel, Nicotiana rustica, N. tabacum cv. Samsun, N. glutinosa, N. bentamiana, and Glycine max cv. Hill; and produced chlorosis, stunting, and bud necrosis in Arachis hypogaea (peanut). Plants developing these symptoms following mechanical inoculation with extracts from original soybean leaves were positive in ELISA for TSWV. ELISA results indicate that the overall incidence of TSWV on soybean in the five provinces was 5.4%. TSWV has been reported in potato (2) and tomato (1) from Iran, but to our knowledge, this is the first report of the occurrence of TSWV on soybean in Iran. References: (1) K. Bananej et al. Iran. J. Plant Pathol. 34:30, 1998. (2) R. Pourrahim et al. Plant Dis. 85:442, 2001.


Author(s):  
Casey L. Ruark-Seward ◽  
Brian Bonville ◽  
George Kennedy ◽  
David A. Rasmussen

AbstractTomato spotted wilt virus (TSWV) is a generalist pathogen with one of the broadest known host ranges among RNA viruses. To understand how TSWV adapts to different hosts, we experimentally passaged viral populations between two alternate hosts, Emilia sochifolia and Datura stramonium, and an obligate vector in which it also replicates, western flower thrips (Frankliniella occidentalis). Deep sequencing viral populations at multiple time points allowed us to track the evolutionary dynamics of viral populations within and between hosts. High levels of viral genetic diversity were maintained in both plants and thrips between transmission events. Rapid fluctuations in the frequency of amino acid variants indicated strong host-specific selection pressures on proteins involved in viral movement (NSm) and replication (RdRp). While several genetic variants showed opposing fitness effects in different hosts, fitness effects were generally positively correlated between hosts indicating that positive rather than antagonistic pleiotropy is pervasive. These results suggest that high levels of genetic diversity together with the positive pleiotropic effects of mutations have allowed TSWV to rapidly adapt to new hosts and expand its host range.


2020 ◽  
Vol 11 ◽  
Author(s):  
Punya Nachappa ◽  
Jean Challacombe ◽  
David C. Margolies ◽  
James R. Nechols ◽  
Anna E. Whitfield ◽  
...  

Several plant viruses modulate vector fitness and behavior in ways that may enhance virus transmission. Previous studies have documented indirect, plant-mediated effects of tomato spotted wilt virus (TSWV) infection on the fecundity, growth and survival of its principal thrips vector, Frankliniella occidentalis, the western flower thrips. We conducted thrips performance and preference experiments combined with plant gene expression, phytohormone and total free amino acid analyses to determine if systemically-infected tomato plants modulate primary metabolic and defense-related pathways to culminate into a more favorable environment for the vector. In a greenhouse setting, we documented a significant increase in the number of offspring produced by F. occidentalis on TSWV-infected tomato plants compared to mock-inoculated plants, and in choice test assays, females exhibited enhanced settling on TSWV-infected leaves. Microarray analysis combined with phytohormone signaling pathway analysis revealed reciprocal modulation of key phytohormone pathways under dual attack, possibly indicating a coordinated and dampening defense against the vector on infected plants. TSWV infection, alone or in combination with thrips, suppressed genes associated with photosynthesis and chloroplast function thereby significantly impacting primary metabolism of the host plant, and hierarchical cluster and network analyses revealed that many of these genes were co-regulated with phytohormone defense signaling genes. TSWV infection increased expression of genes related to protein synthesis and degradation which was reflected in the increased total free amino acid content in virus-infected plants that harbored higher thrips populations. These results suggest coordinated gene networks that regulate plant primary metabolism and defense responses rendering virus-infected plants more conducive for vector colonization, an outcome that is potentially beneficial to the vector and the virus when considered within the context of the complex transmission biology of TSWV. To our knowledge this is the first study to identify global transcriptional networks that underlie the TSWV-thrips interaction as compared to a single mechanistic approach. Findings of this study increase our fundamental knowledge of host plant-virus-vector interactions and identifies underlying mechanisms of induced host susceptibility to the insect vector.


2002 ◽  
Vol 92 (6) ◽  
pp. 603-609 ◽  
Author(s):  
Elisavet K. Chatzivassiliou ◽  
Dick Peters ◽  
Nikolaos I. Katis

Arrhenotokous and thelytokous populations of Thrips tabaci from tobacco or leek plants were evaluated for their ability to transmit Tomato spotted wilt virus (TSWV) and for their host preference. Transmission efficiencies were comparatively studied using leaf disks of Petunia hybrida, Datura stramonium, and Nicotiana tabacum cv. Basmas. Adults of arrhenotokous populations collected on infected tobacco plants in the field were efficient transmitters (up to 48.5% transmission) and remained so when maintained on tobacco for several generations. Arrhenotokous T. tabacipopulations from leek plants were poor transmitters (up to 3.1% transmission), whereas no transmission was obtained with thelytokous populations from leek. All populations could infest leek, however none of the arrhenotokous and thelytokous populations from leek plants was able to infest tobacco. TSWV could be acquired by both first and second larval instars of a T. tabacipopulation from tobacco. However, the transmission by adults decreased with the age at which the virus was acquired by larvae. The highest efficiencies (61% of males and 51% of females transmitted) were obtained when newborn (0- to 24-h old) larvae acquired the virus. The majority of thrips started to transmit after becoming adult and rates were positively correlated with the temperature at which the thrips were kept. The median latent period values found for adults decreased with increasing temperature. The median acquisition access period (AAP50) of the population was 41 min, whereas the AAP50 was 65 min for males and 35 min for females. The median inoculation access period of males was 246 and 365 min on tobacco and petunia, respectively, and 96 and 345 min for females. The results show that T. tabaci forms a complex in terms of host preference, reproductive strategy, and ability to transmit TSWV. The transmission parameters show that the thrips of arrhenotokous populations infesting tobacco are highly efficient vectors.


Plant Disease ◽  
2006 ◽  
Vol 90 (3) ◽  
pp. 376-376 ◽  
Author(s):  
Y. Abou-Jawdah ◽  
C. El Mohtar ◽  
H. Sobh ◽  
M. K. Nakhla

During the spring and summer of 2004, an epidemic of Tomato spotted wilt virus (TSWV) (genus Tospovirus, family Bunyaviridae) was observed in an isolated tomato field at an elevation of 1,000 m in Lebanon. Symptoms were characteristic of TSWV (2). Seedlings came from a nursery in the coastal area of Byblos. In the spring of 2005, TSWV-like symptoms (2) appeared on tomato in the same mountainous area, as well as on tomato, pepper, and lettuce crops in the Byblos coastal area. Initial diagnosis using TSWV ImmunoStrip Tests (Agdia, IN) gave positive results on tomato and lettuce samples. When these samples were analyzed using reverse transcription-polymerase chain reaction, a specific band (619 nt) was observed in symptomatic samples but not in healthy controls (1). Amplicons were cloned into the pGEM-T easy vector (Promega, Madison, WI) and three clones were sequenced in both directions (GenBank Accession No. DQ131804). Sequence analysis revealed more than 99% nucleotide identity (GenBank Accession Nos. AY744476, AJ297611, and AJ418781) and 99% amino acid identity and 100% amino acid similarity (GenBank Accession Nos. AAU95409, CAA85356, and CAD11452) to the nucleocapsid protein of several TSWV isolates. To our knowledge, this is the first report of TSWV in Lebanon. To prevent rapid spread, farmers were informed about the disease, its vector, and appropriate preventive control measures. References: (1) S. Adkins and E. N. Rosskopf. Plant Dis. 86:1310, 2002. (2) G. Marchoux et al. Plant Pathol. 40:347, 1991.


2008 ◽  
Vol 89 (8) ◽  
pp. 1811-1818 ◽  
Author(s):  
Daniela Ribeiro ◽  
Ombretta Foresti ◽  
Jurgen Denecke ◽  
Joan Wellink ◽  
Rob Goldbach ◽  
...  

Tomato spotted wilt virus (TSWV) particles are spherical and enveloped, an uncommon feature among plant infecting viruses. Previous studies have shown that virus particle formation involves the enwrapment of ribonucleoproteins with viral glycoprotein containing Golgi stacks. In this study, the localization and behaviour of the viral glycoproteins Gn and Gc were analysed, upon transient expression in plant protoplasts. When separately expressed, Gc was solely observed in the endoplasmic reticulum (ER), whereas Gn was found both within the ER and Golgi membranes. Upon co-expression, both glycoproteins were found at ER-export sites and ultimately at the Golgi complex, confirming the ability of Gn to rescue Gc from the ER, possibly due to heterodimerization. Interestingly, both Gc and Gn were shown to induce the deformation of ER and Golgi membranes, respectively, also observed upon co-expression of the two glycoproteins. The behaviour of both glycoproteins within the plant cell and the phenomenon of membrane deformation are discussed in light of the natural process of viral infection.


Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 647 ◽  
Author(s):  
Xin Qian ◽  
Qing Xiang ◽  
Tongqing Yang ◽  
Hongyu Ma ◽  
Xin Ding ◽  
...  

Tospovirus is a tripartite negative stranded RNA virus and is considered as one of the most devastating plant viruses. Successful virus infection in plant requires many host factors. To date, very few host factors have been identified as important in Tospovirus infection in plants. We reported earlier that NSm protein encoded by Tomato spotted wilt virus (TSWV), a type species of the genus Orthotospovirus, plays critical roles in viral cell-to-cell and long-distance movement. In this study, we determined that molecular co-chaperone NbSGT1 interacted with TSWV NSm in Nicotiana benthamiana. TSWV infection significantly upregulated the expression of NbSGT1 gene and transient overexpression of NbSGT1 in N. benthamiana leaves accelerated TSWV infection. In contrast, silencing the NbSGT1 gene expression using a virus-induced gene silencing (VIGS) approach strongly inhibited TSWV NSm cell-to-cell movement, as well as TSWV local and systemic infection in N. benthamiana plants. Furthermore, NbSGT1 was found to regulate the infection of both American and Euro/Asia type tospoviruses in N. benthamiana plant. Collectively, our findings presented in this paper and the results published previously indicated that molecular co-chaperone NbSGT1 plays important roles in modulating both positive stranded and tripartite negative stranded RNA virus infection in plants.


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