scholarly journals Characterization and Establishment of a Recombinase Polymerase Amplification Assay for Rapid Detection of Tomato Spotted Wilt Virus in Pepper

Author(s):  
Kaiqiang Hao ◽  
Ming Gu ◽  
Miaoren Yang ◽  
Xinran Gao ◽  
Zihao Xia ◽  
...  

Abstract Tomato spotted wilt virus (TSWV) is one of the most economically destructive and scientifically challenging plant viruses, which has seriously affected the production of commercial crops. At present, there is no effective strategy to control this virus. Therefore, there is an urgent need for a rapid and simple method to detect TSWV, which is of great significance to prevent its spread. In this study, an isolate of TSWV (TSWV-LNTL) infecting pepper from Liaoning Province of northeast China was obtained. A phylogenetic tree based on neighbor-joining using coat protein (CP) gene was established. A rapid method for detecting TSWV by recombinase polymerase amplification (RPA) was established. The phylogenetic tree based on the nucleotide sequences of coat protein (CP) genes of different TSWV isolates showed that the genetic relationship of TSWV-LNTL was most closely related to that of TSWV-LX-Lettuce-12 (Yunnan) and TSWV-TSHL (Shandong) isolates in China. It can be finished at 39 °C for 20 min and then purified by heating at 65 °C for 10 min. The RPA primers were highly specific and no cross-reactivity was detected with other selected viruses infecting pepper. The results of sensitivity test revealed that the detection limit of RPA is 1.0 × 103 copies/μL, which was tenfold lower than that of PCR method. In addition, the RPA method was successfully applied to detect TSWV in field samples. These results reported the occurrence of TSWV on crop in Liaoning Province of northeast China and demonstrated that the established RPA assay provided an effective molecular diagnostic tool for the accurate and rapid detection of TSWV to prevent its spread.

2020 ◽  
Vol 117 (42) ◽  
pp. 26237-26244
Author(s):  
Yoav Bahat ◽  
Joel Alter ◽  
Moshe Dessau

Tospoviridaeis a family of enveloped RNA plant viruses that infect many field crops, inflicting a heavy global economic burden. These tripartite, single-stranded, negative-sense RNA viruses are transmitted from plant to plant by thrips as the insect vector. The medium (M) segment of the viral genome encodes two envelope glycoproteins, GNand GC, which together form the envelope spikes. GCis considered the virus fusogen, while the accompanying GNprotein serves as an attachment protein that binds to a yet unknown receptor, mediating the virus acquisition by the thrips carrier. Here we present the crystal structure of glycoprotein N (GN) from the tomato spotted wilt virus (TSWV), a representative member of theTospoviridaefamily. The structure suggests that GNis organized as dimers on TSWV’s outer shell. Our structural data also suggest that this dimerization is required for maintaining GNstructural integrity. Although the structure of the TSWV GNis different from other bunyavirus GNproteins, they all share similar domain connectivity that resembles glycoproteins from unrelated animal-infecting viruses, suggesting a common ancestor for these accompanying proteins.


Biljni lekar ◽  
2021 ◽  
Vol 49 (2) ◽  
pp. 148-157
Author(s):  
Marina Ćuk ◽  
Zagorka Savić ◽  
Renata Iličić ◽  
Ferenc Bagi

Tomato spotted wilt virus (TSWV) is the most economically important plant viruses from genus Tospovirus. It has a polyphagous character and infects a wide range of very significant agricultural crops. Vectors of viruses are insects from order Thysanoptera (Thripidae) and till know eight species are known to transmit tospoviruses of which Frankliniella occidentalis is considered to be economically most important vector. TSWV is transmitted by thrips in a persistent and propagative manner. Relationship between vector and TSWV is very specific because vectors acquire the virus in the larval stages, while imago plays a key role in transmission of the virus. TSWV causes wide range of symptoms depending on host plant, external environmental conditions and type of viruses. In addition to affecting the fruit quality of cultivated crops, greatly reduces the yield to agricultural producers. Tomato is the most commonly attacked by TSWV, and after the symptoms manifested on leaves in the form of a bronze color, the virus was name. Protection of agricultural crops is very challenging and difficult due to wide distribution of viruse vectors, their hidden way of life as well as wide range of TSWV hosts.


Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 442-442 ◽  
Author(s):  
R. Pourrahim ◽  
Sh. Farzadfar ◽  
A. A. Moini ◽  
N. Shahraeen ◽  
A. Ahoonmanesh

Severe leaf and stem necrosis before flowering was observed in potato (Solanum tuberosum) fields of Firouzkoh Province, Iran, during the summer of 1998. Infected plants died before the end of the growing season. Necrosis was more severe in cv. Agria than in cvs. Ajaxs and Arinda. A high population of Thrips tabaci was observed in August and September. Tomato spotted wilt virus (TSWV) (1) was detected in affected potatoes by using specific TSWV-IgG (from Bioreba) in double-antibody sandwich enzyme linked immunosorbent assay and by indicator plant reactions. Mechanical inoculation of indicator plants with leaf extracts of symptomatic potatoes produce necrotic local lesions in Chenopodium quinoa, C. amaranticolor, Gomphrena globosa, Vicia faba, Vigna sinensis, Phaseolus aureus var. Gohar, P. vulgaris, and Petunia hybrida. The virus caused systemic necrosis in Capsicum frutescens, Datura stramonium, D. metel, Nicotiana glutinosa, N. rustica, and Trapaeolum majus, preceded by systemic chlorotic spots. TSWV was reported from ornamental crops in Tehran and Absard areas near to Firouzkoh province (2), but this is the first report of TSWV occurrence on potatoes in Iran. References: (1) T. S. Ie. Descriptions of Plant Viruses. No. 39, 1970. (2) A. A. Moeini, et al. Iran. J. Plant Pathol. (In press.)


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.


2008 ◽  
Vol 23 (3) ◽  
pp. 153-166 ◽  
Author(s):  
Branka Krstic ◽  
Aleksandra Bulajic ◽  
Ivana Djekic ◽  
Janos Berenji

Tomato spotted wilt virus (TSWV) has one of the largest host ranges among plant viruses and is widespread in all climates. TSWV is responsible for numerous epidemics in many parts of the world in different crops, mainly vegetables, tobacco and ornamentals. Its highly polyphagous nature, effectiveness of virus transmission by the thrips as its vectors, rapidity with which new variants arise, as well as difficulties in controlling the vectors make TSWV one of the most dangerous plant viruses. The ability of this virus to cause such severe losses on a broad range of crops, as well as its intriguing biological and molecular characteristics place TSWV amongst the most extensively studied plant viruses in the world at present. This paper provides a general overview of TSWV, encompassing all the major aspects of its biology and current knowledge on host range, symptomatology, molecular biology, vector relationship, control and diagnosis.


2011 ◽  
Vol 101 (1) ◽  
pp. 147-153 ◽  
Author(s):  
A. C. Kaye ◽  
J. W. Moyer ◽  
E. J. Parks ◽  
I. Carbone ◽  
M. A. Cubeta

Exploring the genetic diversity and evolutionary history of plant viruses is critical to understanding their ecology and epidemiology. In this study, maximum-likelihood and population genetics-based methods were used to investigate the population structure, genetic diversity, and sources of genetic variation in field isolates of Tomato spotted wilt virus (TSWV) from peanut in North Carolina and Virginia. Selected regions of the nucleocapsid, movement, and RNA-dependent RNA polymerase genes were amplified and sequenced to identify haplotypes and infer genetic relationships between isolates of TSWV with heuristic methods. The haplotype structure of each locus consisted of 1 or 2 predominant haplotypes and >100 haplotypes represented by a single isolate. No specific haplotypes were associated with geographic area, peanut cultivar, or year of isolation. The population was panmictic at the regional level and high levels of genetic diversity were observed among isolates. There was evidence for positive selection on single amino acids in each gene on a background of predominant purifying selection acting upon each locus. The results of compatibility analyses and the persistence of specific gene sequences in isolates collected over three field seasons suggest that recombination was occurring in the population. Estimates of the population mutation rate suggest that mutation has had a significant effect on the shaping of this population and, together with purifying selection, these forces have been the predominant evolutionary forces influencing the TSWV population in peanut in North Carolina and Virginia.


Viruses ◽  
2021 ◽  
Vol 13 (3) ◽  
pp. 427
Author(s):  
Jin Zhan ◽  
Huiping Shi ◽  
Weimin Li ◽  
Chao Zhang ◽  
Yongqiang Zhang

Tomato spotted wilt virus (TSWV) is one of the most destructive plant viruses, causing severe losses in many important crops worldwide. The non-structural protein NSm of TSWV is a viral movement protein that induces viral symptoms. However, the molecular mechanisms by which NSm contributes to symptom development are unclear. Here, we present evidence that NSm directly interacts with Nicotiana benthamiana chloroplast thylakoid membrane protein TMP14 (NbTMP14) by yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays. The interaction between NSm and NbTMP14 led to the translocation of the NbTMP14 protein from the chloroplast to the cytoplasm in TSWV-infected plants, and overexpressing NSm decreased NbTMP14 mRNA accumulation. In addition, abnormal chloroplasts and starch accumulation were observed in TSWV-infected plants. Silencing of NbTMP14 by TRV VIGS also showed similar results to those of TSWV-infected plants. Overexpressing NbTMP14 in transgenic N. benthamiana plants impeded TSWV infection, and silencing NbTMP14 in N. benthamiana plants increased disease symptom severity and virus accumulation. To our knowledge, this is the first report showing that the plant chloroplast TMP14 protein is involved in viral infection. Knowledge of the interaction between NSm and NbTMP14 advances our understanding of the molecular mechanisms underlying TSWV symptom development and infection.


2022 ◽  
Author(s):  
Namisha Sharma ◽  
Ashish Prasad ◽  
Manoj Prasad

The Sw5 gene cluster furnishes robust resistance to Tomato spotted wilt virus in tomato, which has led to its widespread applicability in agriculture. Among the five orthologs, Sw5b functions as a resistance gene against a broad-spectrum Tospovirus and is linked with Tospovirus resistance. However, its paralog, Sw5a, has been recently implicated in providing resistance against Tomato leaf curl New Delhi virus , broadening the relevance of the Sw5 gene cluster in promoting defense against plant viruses. We propose that plants have established modifications within the homologs of R genes that permit identification of different effector proteins and provide broad and robust resistance against different pathogens through activation of hypersensitive response and cell death.


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