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  1 / 336 MEDLINE  
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[PMID]:28771638
[Au] Autor:Chou WC; Lin SS; Yeh SD; Li SL; Peng YC; Fan YH; Chen TC
[Ad] Endereço:Department of Biotechnology, Asia University, Wufeng, Taichung, Taiwan.
[Ti] Título:Characterization of the genome of a phylogenetically distinct tospovirus and its interactions with the local lesion-induced host Chenopodium quinoa by whole-transcriptome analyses.
[So] Source:PLoS One;12(8):e0182425, 2017.
[Is] ISSN:1932-6203
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Chenopodium quinoa is a natural local lesion host of numerous plant viruses, including tospoviruses (family Bunyaviridae). Groundnut chlorotic fan-spot tospovirus (GCFSV) has been shown to consistently induce local lesions on the leaves of C. quinoa 4 days post-inoculation (dpi). To reveal the whole genome of GCFSV and its interactions with C. quinoa, RNA-seq was performed to determine the transcriptome profiles of C. quinoa leaves. The high-throughput reads from infected C. quinoa leaves were used to identify the whole genome sequence of GCFSV and its single nucleotide polymorphisms. Our results indicated that GCFSV is a phylogenetically distinct tospovirus. Moreover, 27,170 coding and 29,563 non-coding sequences of C. quinoa were identified through de novo assembly, mixing reads from mock and infected samples. Several key genes involved in the modulation of hypersensitive response (HR) were identified. The expression levels of 4,893 deduced complete genes annotated using the Arabidopsis genome indicated that several HR-related orthologues of pathogenesis-related proteins, transcription factors, mitogen-activated protein kinases, and defense proteins were significantly expressed in leaves that formed local lesions. Here, we also provide new insights into the replication progression of a tospovirus and the molecular regulation of the C. quinoa response to virus infection.
[Mh] Termos MeSH primário: Chenopodium/genética
Chenopodium/virologia
Regulação da Expressão Gênica de Plantas
Genes de Plantas
Interações Hospedeiro-Patógeno
Tospovirus/fisiologia
Transcriptoma
[Mh] Termos MeSH secundário: Genoma Viral
Sequenciamento de Nucleotídeos em Larga Escala
Filogenia
Doenças das Plantas
Folhas de Planta
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171006
[Lr] Data última revisão:
171006
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170804
[St] Status:MEDLINE
[do] DOI:10.1371/journal.pone.0182425


  2 / 336 MEDLINE  
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[PMID]:28768868
[Au] Autor:Komoda K; Narita M; Yamashita K; Tanaka I; Yao M
[Ad] Endereço:Graduate School of Life Sciences, Hokkaido University, Sapporo, Hokkaido, Japan komoda.k@gmail.com yao@castor.sci.hokudai.ac.jp.
[Ti] Título:Asymmetric Trimeric Ring Structure of the Nucleocapsid Protein of Tospovirus.
[So] Source:J Virol;91(20), 2017 Oct 15.
[Is] ISSN:1098-5514
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:(TSWV), belonging to the genus of the family , causes significant economic damage to several vegetables and ornamental plants worldwide. Similar to those of all other negative-strand RNA viruses, the nucleocapsid (N) protein plays very important roles in its viral life cycle. N proteins protect genomic RNAs by encapsidation and form a viral ribonucleoprotein complex (vRNP) with some RNA-dependent RNA polymerases. Here we show the crystal structure of the N protein from TSWV. Protomers of TSWV N proteins consist of three parts: the N arm, C arm, and core domain. Unlike N proteins of other negative-strand RNA viruses, the TSWV N protein forms an asymmetric trimeric ring. To form the trimeric ring, the N and C arms of the N protein interact with the core domains of two adjacent N proteins. By solving the crystal structures of the TSWV N protein with nucleic acids, we showed that an inner cleft of the asymmetric trimeric ring is an RNA-binding site. These characteristics are similar to those of N proteins of other viruses of the family Based on these observations, we discuss possibilities of a TSWV encapsidation model. Tospoviruses cause significant crop losses throughout the world. Particularly, TSWV has an extremely wide host range (>1,000 plant species, including dicots and monocots), and worldwide losses are estimated to be in excess of $1 billion annually. Despite such importance, no proteins of tospoviruses have been elucidated so far. Among TSWV-encoded proteins, the N protein is required for assembling the viral genomic RNA into the viral ribonucleoprotein (vRNP), which is involved in various steps of the life cycle of these viruses, such as RNA replication, virus particle formation, and cell-to-cell movement. This study revealed the structure of the N protein, with or without nucleic acids, of TSWV as the first virus of the genus , so it completed our view of the N proteins of the family .
[Mh] Termos MeSH primário: Proteínas do Nucleocapsídeo/química
Tospovirus/química
[Mh] Termos MeSH secundário: Sítios de Ligação
Cristalografia por Raios X
Lycopersicon esculentum/virologia
Modelos Moleculares
Conformação Proteica
Multimerização Proteica
RNA Viral/química
Vírion
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Nucleocapsid Proteins); 0 (RNA, Viral)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171024
[Lr] Data última revisão:
171024
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170804
[St] Status:MEDLINE


  3 / 336 MEDLINE  
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[PMID]:28631603
[Au] Autor:Almási A; Nemes K; Csömör Z; Tóbiás I; Palkovics L; Salánki K
[Ad] Endereço:1​Plant Protection Institute, Centre for Agricultural Research, Hungarian Academy of Sciences, H-1022 Herman Ottó str. 15, Budapest, Hungary.
[Ti] Título:A single point mutation in Tomato spotted wilt virus NSs protein is sufficient to overcome Tsw-gene-mediated resistance in pepper.
[So] Source:J Gen Virol;98(6):1521-1525, 2017 Jun.
[Is] ISSN:1465-2099
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:The nonstructural protein (NSs) of Tomato spotted wilt virus (TSWV) was previously identified as an avirulence determinant for Tsw-based resistance on pepper. The NSs of wild-type (WT) and resistance-breaking (RB) TSWV strains isolated in Hungary had only two amino acid substitutions (104, 461). We have analysed the ability of the NSs and their point mutant variants to trigger Tsw-mediated hypersensitive responses and RNA silencing suppressor (RSS) activity in patch assays. We identified a single amino acid change at position 104 (T-A) that was responsible for the necrosis induction or loss, while a significant difference was not detected in the RSS activity of the two parental strains. We have successfully complemented the infection of the WT strain on resistant pepper cultivar with the infectious S RNA transcript of the RB strain and the WT-T104A point mutant. Our work provides direct evidence that a single amino acid change can induce an RB phenotype.
[Mh] Termos MeSH primário: Capsicum/virologia
Resistência à Doença
Doenças das Plantas/virologia
Mutação Puntual
Tospovirus/patogenicidade
Proteínas não Estruturais Virais/genética
Fatores de Virulência/genética
[Mh] Termos MeSH secundário: Substituição de Aminoácidos
Capsicum/fisiologia
Análise Mutacional de DNA
Teste de Complementação Genética
Hungria
Mutação de Sentido Incorreto
Proteínas não Estruturais Virais/metabolismo
Fatores de Virulência/metabolismo
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Viral Nonstructural Proteins); 0 (Virulence Factors)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170717
[Lr] Data última revisão:
170717
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170621
[St] Status:MEDLINE
[do] DOI:10.1099/jgv.0.000798


  4 / 336 MEDLINE  
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[PMID]:28527340
[Au] Autor:Singh A; Permar V; Jain RK; Goswami S; Kumar RR; Canto T; Palukaitis P; Praveen S
[Ad] Endereço:Division of Biochemistry, Indian Agricultural Research Institute, New Delhi 110012, India.
[Ti] Título:Induction of cell death by tospoviral protein NSs and the motif critical for cell death does not control RNA silencing suppression activity.
[So] Source:Virology;508:108-117, 2017 Aug.
[Is] ISSN:1096-0341
[Cp] País de publicação:United States
[La] Idioma:eng
[Ab] Resumo:Groundnut bud necrosis virus induces necrotic symptoms in different hosts. Previous studies showed reactive oxygen species-mediated programmed cell death (PCD) resulted in necrotic symptoms. Transgenic expression of viral protein NSs mimics viral symptoms. Here, we showed a role for NSs in influencing oxidative burst in the cell, by analyzing H O accumulation, activities of antioxidant enzymes and expression levels of vacuolar processing enzymes, H O -responsive microRNA 319a.2 plus its possible target metacaspase-8. The role of NSs in PCD, was shown using two NSs mutants: one in the Trp/GH3 motif (a homologue of pro-apototic domain) (NSs ) and the other in a non-Trp/GH3 motif (NSs ). Tobacco rattle virus (TRV) expressing NSs enhanced the PCD response, but not TRV-NSs , while RNA silencing suppression activity was lost in TRV-NSs , but not in TRV-NSs . Therefore, we propose dual roles of NSs in RNA silencing suppression and induction of cell death, controlled by different motifs.
[Mh] Termos MeSH primário: Apoptose
Inativação Gênica
Doenças das Plantas/virologia
Tabaco/citologia
Tabaco/genética
Tospovirus/metabolismo
Proteínas não Estruturais Virais/química
Proteínas não Estruturais Virais/metabolismo
[Mh] Termos MeSH secundário: Motivos de Aminoácidos
Sequência de Aminoácidos
Peróxido de Hidrogênio/metabolismo
Dados de Sequência Molecular
Filogenia
Doenças das Plantas/genética
Explosão Respiratória
Alinhamento de Sequência
Tabaco/metabolismo
Tabaco/virologia
Tospovirus/química
Tospovirus/genética
Proteínas não Estruturais Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (Viral Nonstructural Proteins); BBX060AN9V (Hydrogen Peroxide)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170818
[Lr] Data última revisão:
170818
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170521
[St] Status:MEDLINE


  5 / 336 MEDLINE  
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[PMID]:28260140
[Au] Autor:Huang KS; Tai CH; Cheng YH; Lin SH; Chen TC; Jan FJ
[Ad] Endereço:Department of Plant Pathology, National Chung Hsing University, Taichung, 40227, Taiwan.
[Ti] Título:Complete nucleotide sequences of M and L RNAs from a new pepper-infecting tospovirus, Pepper chlorotic spot virus.
[So] Source:Arch Virol;162(7):2109-2113, 2017 Jul.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:Pepper chlorotic spot virus (PCSV), newly found in Taiwan, was identified as a new tospovirus based on the molecular characterization of its S RNA. In this study, the complete M and L RNA sequences of PCSV were determined. The M RNA has 4795 nucleotides (nts), encoding the NSm protein of 311 aa (34.5 kDa) in the viral (v) strand and the glycoprotein precursor (Gn/Gc) of 1122 aa (127.6 kDa) in the viral complementary (vc) strand. The L RNA has 8859 nts, encoding the RNA-dependent RNA polymerase (RdRp) of 2873 aa (330.8 kDa) in the vc strand. Analyses of the NSm, Gn/Gc and RdRp of PCSV revealed that PCSV is phylogenetically clustered within the watermelon silver mottle virus-related clade. Based on the whole genome sequence, PCSV is closely related to Tomato necrotic ringspot virus and should be classified as a new tospovirus species.
[Mh] Termos MeSH primário: Piper nigrum/virologia
Doenças das Plantas/virologia
RNA Replicase/genética
RNA Viral/genética
Tospovirus/classificação
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Lycopersicon esculentum/virologia
Filogenia
Taiwan
Tospovirus/genética
Tospovirus/isolamento & purificação
Proteínas Virais/genética
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Viral Proteins); EC 2.7.7.48 (RNA Replicase)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170802
[Lr] Data última revisão:
170802
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170306
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3260-1


  6 / 336 MEDLINE  
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[PMID]:28190200
[Au] Autor:Bald-Blume N; Bergervoet JHW; Maiss E
[Ad] Endereço:Section of Phytomedicine, Institute of Horticultural Production Systems, Leibniz Universität Hannover, Hannover, Germany.
[Ti] Título:Development of a molecular assay for the general detection of tospoviruses and the distinction between tospoviral species.
[So] Source:Arch Virol;162(6):1519-1528, 2017 Jun.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:A Luminex xTAG-based assay for plant-infecting tospoviruses was developed. The test enables the detection of tospoviruses in general and the differentiation of the four important member species of this genus: Tomato spotted wilt virus, Impatiens necrotic spot virus, the proposed 'Capsicum chlorosis virus' and Watermelon silver mottle virus. The generic tospovirus primers used in this method are also applicable for detection of tospoviruses by basic RT-PCR. We also describe an economic alternative method for the distinction of the four tospoviruses mentioned and of additional member viruses, based on a restriction fragment length polymorphism (RFLP). The sophisticated Luminex xTAG technology allows the simultaneous detection of various targets. This study is part of a project that aims to develop a method for the simultaneous detection of various plant pathogens (viral, bacterial and fungal) in plant material.
[Mh] Termos MeSH primário: Imunoensaio/métodos
Reação em Cadeia da Polimerase Multiplex/métodos
Tospovirus/genética
Tospovirus/isolamento & purificação
[Mh] Termos MeSH secundário: Primers do DNA
Doenças das Plantas/virologia
Plantas/virologia
Polimorfismo de Fragmento de Restrição
RNA Viral/análise
Mapeamento por Restrição/métodos
Tospovirus/classificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (DNA Primers); 0 (RNA, Viral)
[Em] Mês de entrada:1707
[Cu] Atualização por classe:170720
[Lr] Data última revisão:
170720
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170213
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3256-x


  7 / 336 MEDLINE  
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[PMID]:28155192
[Au] Autor:Ciuffo M; Nerva L; Turina M
[Ad] Endereço:Istituto per la Protezione Sostenibile delle Piante, CNR, Strada delle Cacce 73, 10135, Turin, Italy.
[Ti] Título:Full-length genome sequence of the tospovirus melon severe mosaic virus.
[So] Source:Arch Virol;162(5):1419-1422, 2017 May.
[Is] ISSN:1432-8798
[Cp] País de publicação:Austria
[La] Idioma:eng
[Ab] Resumo:The complete genome sequence of melon severe mosaic virus (MSMV), genus Tospovirus, family Bunyaviridae, was determined. The small segment is 3283 nucleotide (nt) long and contains two open reading frames in an ambisense organization. The medium segment is 4873 nt long and also encodes two proteins in an ambisense organization. The large segment is 9811 nt long and contains a single, negative-sense ORF. Phylogenetic analysis of each of the five encoded proteins compared to those of tospoviruses present in the databases reveals the same topology for each tree, suggesting that the MSMV genome did not result from recombination or reassortment. Sequence variants present in the RNA population of an infected leaf are described.
[Mh] Termos MeSH primário: Cucumis melo/virologia
Genoma Viral/genética
Vírus do Mosaico/genética
Doenças das Plantas/virologia
RNA Viral/genética
Tospovirus/classificação
Tospovirus/genética
Proteínas Virais/genética
[Mh] Termos MeSH secundário: Sequência de Aminoácidos
Sequência de Bases
Variação Genética
Fases de Leitura Aberta/genética
Filogenia
Folhas de Planta/virologia
Análise de Sequência de RNA
Tospovirus/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral); 0 (Viral Proteins)
[Em] Mês de entrada:1704
[Cu] Atualização por classe:170428
[Lr] Data última revisão:
170428
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170204
[St] Status:MEDLINE
[do] DOI:10.1007/s00705-017-3237-0


  8 / 336 MEDLINE  
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[PMID]:28129432
[Au] Autor:Asano S; Hirayama Y; Matsushita Y
[Ad] Endereço:Nara Prefecture Agricultural Research and Development Center, Sakurai, Japan.
[Ti] Título:Distribution of Tomato spotted wilt virus in dahlia plants.
[So] Source:Lett Appl Microbiol;64(4):297-303, 2017 Apr.
[Is] ISSN:1472-765X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:Tomato spotted wilt virus (TSWV) causes significant losses in the production of the ornamental plant Dahlia variabilis in Japan. The purpose of this study was to examine the distribution of TSWV in dahlia plants and identify plant parts that can be used in the selection of TSWV-free plants. The distribution of TSWV was investigated using reverse transcriptional polymerase chain reaction (RT-PCR) and tissue blot immunoassay. The detection rate of TSWV in latent infected compound leaves was the highest in the petiole, and it decreased from the veins and rachis to the lamina. The tissue blot immunoassays of the leaflets showed an uneven distribution of TSWV, especially along the edge of the leaf blade. In stems, the detection rate of TSWV was high partway up the stem compared to that in the upper and the lower parts of the stem during the vegetative growth stage. A highly uneven distribution was observed in the bulb. Our results indicated that middle parts of the stem as well as the petioles, rachis, and veins of compound leaves are suitable for detection of TSWV in dahlias. This study is the first to report uneven distribution of TSWV in dahlia plants. SIGNIFICANCE AND IMPACT OF THE STUDY: In this study, the distribution of Tomato spotted wilt virus (TSWV) in various parts of dahlia plants was investigated for the first time. The distribution of TSWV was uneven in compound leaves, leaflets, stems, and bulbs. The middle parts of the stem or the petiole and leaf veins should be sampled to detect TSWV when selecting healthy plants.
[Mh] Termos MeSH primário: Dahlia/virologia
Doenças das Plantas/virologia
Tospovirus/isolamento & purificação
[Mh] Termos MeSH secundário: Japão
Folhas de Planta/virologia
Raízes de Plantas/virologia
Caules de Planta/virologia
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Tospovirus/genética
Tospovirus/imunologia
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Em] Mês de entrada:1706
[Cu] Atualização por classe:170614
[Lr] Data última revisão:
170614
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170128
[St] Status:MEDLINE
[do] DOI:10.1111/lam.12720


  9 / 336 MEDLINE  
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[PMID]:28082165
[Au] Autor:Marshall SH; Adegbola RO; Adkins S; Naidu RA
[Ad] Endereço:Washington State University, Department of Plant Pathology, Irrigated Agricultural Research and Extension Center, Prosser, WA 99350, United States.
[Ti] Título:An efficient and high fidelity method for amplification, cloning and sequencing of complete tospovirus genomic RNA segments.
[So] Source:J Virol Methods;242:22-26, 2017 Apr.
[Is] ISSN:1879-0984
[Cp] País de publicação:Netherlands
[La] Idioma:eng
[Ab] Resumo:Tospoviruses (genus Tospovirus, family Bunyaviridae) are responsible for major losses in an extensive range of crops worldwide. New species of these single-stranded, ambisense RNA viruses regularly emerge and have been shown to maintain heterogeneous populations with individual isolates having quite variable biological and virulence characteristics. Most tospovirus phylogenetic studies have focused on analysis of a single gene, most often the nucleocapsid protein gene. Complete genomic RNA segment amplification as a single fragment would facilitate more detailed analyses of genome-wide sequence variability, but obtaining such sequences for a large number of tospovirus isolates using traditional methods of amplification and cloning of small overlapping fragments is tedious, time consuming and expensive. In this study, protocols were optimized to amplify, clone and sequence full-length M- and S-RNA genome segments of Tomato spotted wilt virus and Impatiens necrotic spot virus. The strategy presented here is straightforward, scalable and offers several advantages over the previously commonplace and overlapping amplicon-based approach. Use of whole genome segments, instead of individual gene sequences or defined portions of genome segments, will facilitate a better understanding of the underlying molecular diversity of tospoviruses in mixed infections.
[Mh] Termos MeSH primário: Clonagem Molecular/métodos
Genoma Viral
Técnicas de Amplificação de Ácido Nucleico
RNA Viral/genética
Tospovirus/genética
[Mh] Termos MeSH secundário: Bunyaviridae/genética
Nucleocapsídeo/genética
Filogenia
RNA Viral/isolamento & purificação
Análise de Sequência de DNA/métodos
Tospovirus/isolamento & purificação
[Pt] Tipo de publicação:JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral)
[Em] Mês de entrada:1710
[Cu] Atualização por classe:171016
[Lr] Data última revisão:
171016
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170114
[St] Status:MEDLINE


  10 / 336 MEDLINE  
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[PMID]:28081705
[Au] Autor:Liu LY; Ye HY; Chen TH; Chen TC
[Ad] Endereço:Department of Plant Industry, National Pingtung University of Science and Technology, Pingtung, 91201, Taiwan.
[Ti] Título:Development of a microarray for simultaneous detection and differentiation of different tospoviruses that are serologically related to Tomato spotted wilt virus.
[So] Source:Virol J;14(1):1, 2017 Jan 10.
[Is] ISSN:1743-422X
[Cp] País de publicação:England
[La] Idioma:eng
[Ab] Resumo:BACKGROUND: Tospoviruses, the plant-infecting genus in the family Bunyaviridae, are thrips borne and cause severe agricultural losses worldwide. Based on the serological relationships of the structural nucleocapsid protein (NP), the current tospoviruses are divided into six serogroups. The use of NP-antisera is convenient for virus detection, but it is insufficient to identify virus species grouped in a serogroup due to the serological cross-reaction. Alternatively, virus species can be identified by the N gene amplification using specific primers. Tomato spotted wilt virus (TSWV) is the type species of the genus Tospovirus and one of the most destructive plant viruses. Eight known tospoviruses, Alstroemeria necrotic streak virus (ANSV), Chrysanthemum stem necrosis virus (CSNV), Groundnut ringspot virus (GRSV), Impatiens necrotic spot virus (INSV), Melon severe mosaic virus (MeSMV), Pepper necrotic spot virus (PNSV), Tomato chlorotic spot virus (TCSV) and Zucchini lethal chlorosis virus (ZLCV), sharing serological relatedness with TSWV in NP, are grouped in the TSWV serogroup. Most of the TSWV-serogroup viruses prevail in Europe and America. An efficient diagnostic method is necessary for inspecting these tospoviruses in Asia, including Taiwan. METHODS: A microarray platform was developed for simultaneous detection and identification of TSWV-serogroup tospoviruses. Total RNAs extracted from Chenopodium quinoa leaves separately inoculated with ANSV, CSNV, GRSV, INSV, TCSV and TSWV were used for testing purposes. The 5'-biotinylated degenerate forward and reverse primers were designed from the consensus sequences of N genes of TSWV-serogroup tospoviruses for reverse transcription-polymerase chain reaction (RT-PCR) amplification. Virus-specific oligonucleotide probes were spotted on the surface of polyvinyl chloride (PVC) chips to hybridize with PCR products. The hybridization signals were visualized by hydrolysis of NBT/BCIP with streptavidine-conjugated alkaline phosphatase. The microarray was further applied to diagnose virus infection in field crop samples. RESULTS: Amplicons of approximately 0.46 kb were amplified from all tested TSWV-serogroup tospoviruses by RT-PCR using the degenerate primer pair Pr-dTS-f/Pr-dTS-r. Virus species were identified on chips by hybridization of PCR products with respective virus-specific probes. The microarray was successfully used to diagnose TSWV infection in field pepper samples. CONCLUSIONS: In this study, a rapid, sensitive and precise microarray method has been developed to simultaneously detect and identify six TSWV-serogroup tospoviruses. The microarray platform provides a great potential to explore tospoviruses that can help researchers and quarantine staff to prevent invasions of tospoviruses.
[Mh] Termos MeSH primário: Análise em Microsséries/métodos
Técnicas de Diagnóstico Molecular/métodos
Vírus de Plantas/classificação
Vírus de Plantas/genética
Tospovirus/classificação
Tospovirus/genética
Virologia/métodos
[Mh] Termos MeSH secundário: Hibridização de Ácido Nucleico
Análise de Sequência com Séries de Oligonucleotídeos
Doenças das Plantas/virologia
RNA Viral/genética
RNA Viral/isolamento & purificação
Reação em Cadeia da Polimerase Via Transcriptase Reversa
Sensibilidade e Especificidade
Fatores de Tempo
[Pt] Tipo de publicação:EVALUATION STUDIES; JOURNAL ARTICLE
[Nm] Nome de substância:
0 (RNA, Viral)
[Em] Mês de entrada:1708
[Cu] Atualização por classe:170828
[Lr] Data última revisão:
170828
[Sb] Subgrupo de revista:IM
[Da] Data de entrada para processamento:170114
[St] Status:MEDLINE
[do] DOI:10.1186/s12985-016-0669-1



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