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isolation of influenza viruses in mdck 33016pf cells and clearance of contaminating respiratory viruses.this paper summarizes results obtained by multiplex pcr screening of human clinical samples for respiratory viruses and corresponding data obtained after passaging of virus-positive samples in mdck 33016pf cells. using the resplexii v2.0 (qiagen) multiplex pcr, 393 positive results were obtained in 468 clinical samples collected during an influenza season in germany. the overall distribution of positive results was influenza a 42.0%, influenza b 38.7%, adenovirus 1.5%, bocavirus 0.5%, coronaviru ...201222119922
potent anti-respiratory syncytial virus activity of a cholestanol-sulfated tetrasaccharide conjugate.a number of different viruses including respiratory syncytial virus (rsv) initiate infection of cells by binding to cell surface glycosaminoglycans and sulfated oligo- and polysaccharide mimetics of these receptors exhibit potent antiviral activity in cultured cells. we investigated whether the introduction of different lipophilic groups to the reducing end of sulfated oligosaccharides would modulate their anti-rsv activity. our results demonstrate that the cholestanol-conjugated tetrasaccharide ...201222101246
seasonal variations of 15 respiratory agents illustrated by the application of a multiplex polymerase chain reaction assay.abstract background: nucleic acid amplification tests are increasingly being used to diagnose viral and bacterial respiratory tract infections. the high sensitivity of these tests affects our understanding of the epidemiology of respiratory tract infections. we have assessed the detection rate of a multiplex real-time polymerase chain reaction (pcr) test, with emphasis on epidemiology and seasonal distribution of the most common respiratory tract infections. methods: seven thousand eight hundr ...201221867470
Human bocavirus and other respiratory viral infections in a 2-year cohort of hospitalized children.Human bocavirus (HBoV) infection is reported worldwide and may cause severe respiratory tract infections. The aim of the present study was to assess the prevalence of HBoV, and other respiratory viral pathogens, in a 2-year retrospective study of children admitted to hospital, and to investigate whether viral loads of HBoV DNA were associated with severity of infection. Between April 2007 and March 2009, 891 respiratory samples from 760 children admitted to hospital with acute respiratory tract ...201222028039
effect of respiratory syncytial virus infection on plasmacytoid dendritic cell regulation of allergic airway inflammation.respiratory syncytial virus (rsv) can infect myeloid dendritic cells (mdcs) and regulate their function in the development of allergy. it has been widely reported that plasmacytoid dcs (pdcs) play a critical role in antiviral innate immunity. in contrast, not much is known about the role of pdcs in the interaction between allergy and viral infection. the purpose of the present study was to investigate the effect of rsv infection on pdc function in the regulation of allergic airway inflammation i ...201221894025
increased replication of respiratory syncytial virus in the presence of cytokeratin 8 and 18.previously, it was reported that productive viral infection, viral protein synthesis, and viral rna replication of respiratory syncytial virus (rsv) operated efficiently in two human epithelial cell lines (hep-2 and a549), but not in a human mast-cell line, hmc-1. based on these observations, it was hypothesized that hmc-1 cells lack the machinery required for rsv replication. to identify the host factors required for rsv replication, cdna subtraction using a549, hep-2, and hmc-1 cells was perfo ...201222170560
microrna-221 modulates rsv replication in human bronchial epithelium by targeting ngf expression.early-life infection by respiratory syncytial virus (rsv) is associated with aberrant expression of the prototypical neurotrophin nerve growth factor (ngf) and its cognate receptors in human bronchial epithelium. however, the chain of events leading to this outcome, and its functional implications for the progression of the viral infection, has not been elucidated. this study sought to test the hypothesis that rsv infection modulates neurotrophic pathways in human airways by silencing the expres ...201222272270
epidemiology of respiratory viral infections in two long-term refugee camps in kenya, 2007-2010.abstract: background: refugees are at risk for poor outcomes from acute respiratory infections (ari) because of overcrowding, suboptimal living conditions, and malnutrition. we implemented surveillance for respiratory viruses in dadaab and kakuma refugee camps in kenya to characterize their role in the epidemiology of ari among refugees. methods: from 1 september 2007 through 31 august 2010, we obtained nasopharyngeal (np) and oropharyngeal (op) specimens from patients with influenza-like illne ...201222251705
[rhinoviruses. frequency in nonhospitalized children with acute respiratory infection].molecular methods for human rhinoviruses (hrv) have increased the sensitivity in their diagnosis. hrv may cause acute respiratory infections (ari) of the upper and lower respiratory tract. hrv infection during childhood is a predictor of asthma development. in this study, the hrv frequency in outpatient children with ari was determined, and their clinical features and previous conditions were evaluated. a total of 186 respiratory samples of children under 6 year old attending the cemic pediatric ...201222257453
regulatory t cells expressing granzyme b play a critical role in controlling lung inflammation during acute viral infection.the inflammatory response to lung infections must be tightly regulated, enabling pathogen elimination while maintaining crucial gas exchange. using recently described "depletion of regulatory t cell" (dereg) mice, we found that selective depletion of regulatory t cells (tregs) during acute respiratory syncytial virus (rsv) infection enhanced viral clearance but increased weight loss, local cytokine and chemokine release, and t-cell activation and cellular influx into the lungs. conversely, infla ...201222236998
molecular evolution of respiratory syncytial virus fusion gene, canada, 2006-2010.to assess molecular evolution of the respiratory syncytial virus (rsv) fusion gene, we analyzed rsv-positive specimens from 123 children in canada who did or did not receive rsv immunoprophylaxis (palivizumab) during 2006-2010. resistance-conferring mutations within the palivizumab binding site occurred in 8.7% of palivizumab recipients and none of the nonrecipients.201222264682
independent structural domains in the paramyxovirus polymerase protein.all enzymatic activities required for genomic replication and transcription of nonsegmented negative strand rna viruses (nnsv or mononegavirales) are believed to be concentrated in the viral polymerase (l) protein. however, our insight into the organization of these different enzymatic activities into a bioactive tertiary structure remains rudimentary. fragments of mononegavirales polymerases analyzed to date cannot restore bioactivity through trans-complementation, unlike the related l proteins ...201222215662
Induction of mucosal and systemic immunity against respiratory syncytial virus by inactivated virus supplemented with TLR9 and NOD2 ligands.Respiratory syncytial virus (RSV) infection is the most important viral cause of severe respiratory disease in infants and children worldwide and also forms a serious threat in the elderly. The development of RSV vaccine, however, has been hampered by the disastrous outcome of an earlier trial using an inactivated and parenterally administered RSV vaccine which did not confer protection but rather primed for enhanced disease upon natural infection. Mucosal administration does not seem to prime f ...201222120195
respiratory syncytial virus morbidity, premorbid factors, seasonality, and implications for prophylaxis.objectives: we investigated factors associated with morbidity and pediatric intensive care unit (picu) admission in children with respiratory syncytial virus (rsv) infection and explored seasonality and implication of prophylaxis. methods: a retrospective study between 2006 and 2008 of every child with a laboratory-confirmed rsv infection was included. results: six hundred seventy rsv admissions were identified. ten (1.5%) required picu admissions. children admitted to picu were younger than non ...201222227087
proteins involved in extracellular matrix dynamics are associated with rsv disease severity.severity of respiratory syncytial virus (rsv) infection ranges widely. to what extent the local immune response is involved in rsv disease pathogenesis and which markers of this response are critical in determining disease severity is still matter of debate.the local immune response was studied in nasopharyngeal aspirates (npas) during rsv infection. forty-seven potential markers of disease severity were analysed in a screening cohort of rsv-infected infants with mild disease at home (n=8), hosp ...201222267765
role of neutralizing antibodies in adults with community-acquired pneumonia by respiratory syncytial virus.background. respiratory syncytial virus (rsv) has been implicated in the etiology of adult community-acquired pneumonia (cap). we investigated rsv infection in chilean adults with cap using direct viral detection, real-time reverse-transcription polymerase chain reaction (rtrt-pcr), and serology (microneutralization assay).methods. rsv, other respiratory viruses, and bacteria were studied by conventional and molecular techniques in adults aged ≥18 years presenting with cap to the healthcare faci ...201222238168
a systems approach for identifying resistance factors to rice stripe virus.rice stripe virus (rsv) causes disease that can severely affect the productivity of rice (oryza sativa). several rsv resistant cultivars have been developed. however, host factors conferring rsv resistance in these cultivars are still elusive. here, we present a systems approach for identifying potential rice resistance factors. we developed two near isogenic rsv resistant (nil22) and susceptible (nil37) lines and performed gene expression profiling of the two lines in rsv infected and uninfecte ...201222217248
population diversity of rice stripe virus-derived sirnas in three different hosts and rnai-based antiviral immunity in laodelphgax striatellus.small rna-mediated gene silencing plays evolutionarily conserved roles in gene regulation and defense against invasive nucleic acids. virus-derived small interfering rnas (vsirnas) are one of the key elements involved in rna silencing-based antiviral activities in plant and insect. vsirnas produced after viruses infecting hosts from a single kingdom (i.e., plant or animal) are well described. in contrast, vsirnas derived from viruses capable of infecting both plants and their insect vectors have ...201223029445
role of rice stripe virus nsvc4 in cell-to-cell movement and symptom development in nicotiana benthamiana.our previous work has demonstrated that the nsvc4 protein of rice stripe virus (rsv) functions as a cell-to-cell movement protein. however, the mechanisms whereby rsv traffics through plasmodesmata (pd) are unknown. here we provide evidence that the nsvc4 moves on the actin filament and endoplasmic reticulum network, but not microtubules, to reach cell wall pd. disruption of cytoskeleton using different inhibitors altered nsvc4 localization to pd, thus impeding rsv infection of nicotiana bentham ...201223233857
identification of novel oryza sativa mirnas in deep sequencing-based small rna libraries of rice infected with rice stripe virus.micrornas (mirnas) play essential regulatory roles in the development of eukaryotes. methods based on deep-sequencing have provided a powerful high-throughput strategy for identifying novel mirnas and have previously been used to identify over 100 novel mirnas from rice. most of these reports are related to studies of rice development, tissue differentiation, or abiotic stress, but novel rice mirnas related to viral infection have rarely been identified. in previous work, we constructed and pyro ...201223071571
fine mapping and identification of candidate rice genes associated with qstv11(sg), a major qtl for rice stripe disease resistance.rice stripe disease, caused by rice stripe virus (rsv) is a serious constraint to rice production in subtropical regions of east asia. we performed fine mapping of a rsv resistance qtl on chromosome 11, qstv11 ( sg ), using near-isogenic lines (nils, bc(6)f(4)) derived from a cross between the highly resistant variety, shingwang, and the highly susceptible variety, ilpum, using 11 insertion and deletion (indel) markers. qstv11 ( sg ) was localized to a 150-kb region between indel 11 (17.86 mbp) ...201222751999
processing and intracellular localization of rice stripe virus pc2 protein in insect cells.rice stripe virus (rsv) belongs to the genus tenuivirus and its genome consists of four single-stranded rnas encoding seven proteins. here, we have analyzed the processing and membrane association of pc2 encoded by vcrna2 in insect cells. the enhanced green fluorescent protein (egfp) was fused to the pc2 and used for the detection of pc2 fusion proteins. the results showed that pc2 was cleaved to produce two proteins named pc2-n and pc2-c. when expressed alone, either pc2-n or pc2-c could transp ...201222575054
surface display of rice stripe virus nsvc2 and analysis of its membrane fusion activity.rice stripe virus (rsv) infects rice and is transmitted in a propagative manner by the small brown planthopper. how rsv enters an insect cell to initiate the infection cycle is poorly understood. sequence analysis revealed that the rsv nsvc2 protein was similar to the membrane glycoproteins of several members in the family bunyaviridae and might induce cell membrane fusion. to conveniently study the membrane fusion activity of nsvc2, we constructed cell surface display vectors for expressing nsv ...201222492001
analysis of rice stripe virus whole-gene expression in rice and in the small brown planthopper by real-time quantitative pcr.rice stripe virus (rsv) is a typical member of the tenuivirus genus, which is transmitted mainly by the small brown planthopper (sbph) in a persistent, circulative-propagative manner. at present, expression levels of viral genes in rice and in sbph remain unclear. in this study, we investigated rsv gene expression in rsv-infected rice tissues and in viruliferous sbph using a sensitive and reliable real-time quantitative pcr (q-pcr) method. the results revealed that ns3 gene exhibited the highest ...201222404613
the rice stripe virus pc4 functions in movement and foliar necrosis expression in nicotiana benthamiana.the rice stripe virus (rsv) pc4 has been determined as the viral movement protein (mp). in this study, the pc4 gene was cloned into a movement-deficient tobacco mosaic virus (tmv). the resulting hybrid tmv-pc4, in addition to spreading cell to cell in nicotiana tabacum, moved systemically and induced foliar necrosis in nicotiana benthamiana, indicating novel functions of the rsv mp. a systematic alanine-scanning mutagenesis study established the region k(122)-d(258) of the pc4 substantially asso ...201222305130
rnai-directed down-regulation of rsv results in increased resistance in rice (oryza sativa l.).rice stripe disease (rsd), caused by rice stripe virus (rsv), is a serious disease in temperate rice-growing areas. we have created an rnai construct containing coat protein gene (cp) and disease specific protein gene (sp) sequences from rsv. the rnai construct was transformed into two susceptible japonica varieties, suyunuo and guanglingxiangjing, to develop resistance against rsd. the homozygous progeny of rice plants in the t(5) and t(7) generations containing rnai constructs, after self-fert ...201222261864
current insights into research on rice stripe virus.rice stripe virus (rsv) is one of the most destructive viruses of rice, and greatly reduces rice production in china, japan, and korea, where mostly japonica cultivars of rice are grown. rsv is transmitted by the small brown plant-hopper (sbph) in a persistent and circulative-propagative manner. several methods have been developed for detection of rsv, which is composed of four single-stranded rnas that encode seven proteins. genome sequence data and comparative phylogenetic analysis have been u ...201325288949
comparative transcriptome analysis of two rice varieties in response to rice stripe virus and small brown planthoppers during early interaction.rice stripe, a virus disease, transmitted by a small brown planthopper (sbph), has greatly reduced production of japonica rice in east asia, especially in china. although we have made great progress in mapping resistance genes, little is known about the mechanism of resistance. by de novo transcriptome assembling, we gained sufficient transcript data to analyze changes in gene expression of early interaction in response to sbph and rsv infection in rice. respectively 648 and 937 degs were detect ...201324358146
investigation on subcellular localization of rice stripe virus in its vector small brown planthopper by electron microscopy.rice stripe virus (rsv), which is transmitted by small brown planthopper (laodelphax striatellus fallén, sbph), has been reported to be epidemic and cause severe rice stripe disease in rice fields in many east asian countries, including china. investigation on viral localization in the vector is very important for elucidating transmission mechanisms of rsv by sbph. in this study, transmission electron microscopy and immuno-gold labeling technique were used to investigate the subcellular localiza ...201324139455
genetic variability and evolution of rice stripe virus.rice stripe virus (rsv) is the type member of the genus tenuivirus. rsv is known to have four segmented, single-stranded rna molecules and causes rice stripe disease in the rice fields of china, japan, and korea. based on the complete genomic sequences of the determined 6 rsv isolates (from yunnan, jiangsu, zhejiang, and liaoning provinces, china) and 27 other rsv isolates (from yunnan, jiangsu, anhui, henan, and shandong provinces of china, also japan and korea) downloaded from genbank, we prov ...201324101204
production of marker-free and rsv-resistant transgenic rice using a twin t-dna system and rnai.a twin t-dna system is a convenient strategy for creating selectable marker-free transgenic plants. the standard transformation plasmid, pcambia 1300, was modified into a binary vector consisting of two separate t-dnas, one of which contained the hygromycin phosphotransferase (hpt) marker gene. using this binary vector, we constructed two vectors that expressed inverted-repeat (ir) structures targeting the rice stripe virus (rsv) coat protein (cp) gene and the special-disease protein (sp) gene. ...201323938389
transcription of orfs on rna2 and rna4 of rice stripe virus terminate at an auccggau sequence that is conserved in the genus tenuivirus.rice stripe virus, the type member of the genus tenuivirus, has four genomic rnas. rnas 2-4 have an ambisense coding strategy and the noncoding intergenic regions (irs) separating the two orfs are thought to function in termination of transcription. sequencing the 3'-untranslated region of transcripts from rna2 and rna4 in virus-infected oryza sativa (the natural host), nicotiana benthamiana (an experimental host) and laodelphax striatellus (the vector), showed that the sequences of p2 and pc2 t ...201323624227
detection of quantitative trait loci (qtls) for resistances to small brown planthopper and rice stripe virus in rice using recombinant inbred lines.small brown planthopper (sbph) and rice stripe virus (rsv) disease transmitted by sbph cause serious damage to rice (oryza sativa l.) in china. in the present study, we screened 312 rice accessions for resistance to sbph. the indica variety, n22, is highly resistant to sbph. one hundred and eighty two recombinant inbred lines (rils) derived from a cross of n22 and the highly susceptible variety, ussr5, were used for quantitative trait locus (qtl) analysis of resistances to sbph and rsv. in a mod ...201323591851
an insect cell line derived from the small brown planthopper supports replication of rice stripe virus, a tenuivirus.a cell line from the small brown planthopper (sbph; laodelphax striatellus) was established to study replication of rice stripe virus (rsv), a tenuivirus. the sbph cell line, which had been subcultured through 30 passages, formed monolayers of epithelial-like cells. inoculation of cultured vector cells with rsv resulted in a persistent infection. during viral infection in the sbph cell line, the viral non-structural protein ns3 co-localized with the filamentous ribonucleoprotein particles of rsv ...201323468422
rice yellow stunt rhabdovirus protein 6 suppresses systemic rna silencing by blocking rdr6-mediated secondary sirna synthesis.the p6 protein of rice yellow stunt rhabdovirus (rysv) is a virion structural protein that can be phosphorylated in vitro. however its exact function remains elusive. we found that p6 enhanced the virulence of potato virus x (pvx) in nicotiana benthamiana and n. tabacum plants, suggesting that it might function as a suppressor of rna silencing. we examined the mechanism of p6-mediated silencing suppression by transiently expressing p6 in both n. benthamiana leaves and rice protoplasts. our resul ...201323634838
construction of single-chain variable fragment antibodies against mcf-7 breast cancer cells.a phage display library of single chain variable fragment (scfv) against mcf-7 breast cancer cells was constructed from c3a8 hybridoma cells. rna from the c3a8 was isolated, cdna was constructed, and variable heavy and light immunoglobulin chain gene region were amplified using pcr. the variable heavy and light chain gene regions were combined with flexible linker, linked to a pcantab 5e phagemid vector and electrophoresed into supe strain of escherichia coli tg1 cells. forty-eight clones demons ...201324301925
migration of rice planthoppers and their vectored re-emerging and novel rice viruses in east asia.this review examines recent studies of the migration of three rice planthoppers, laodelphax striatellus, sogatella furcifera, and nilaparvata lugens, in east asia. laodelphax striatellus has recently broken out in jiangsu province, eastern china. the population density in the province started to increase in the early 2000s and peaked in 2004. in 2005, rice stripe virus (rsv) viruliferous rate of l. striatellus peaked at 31.3%. since then, rice stripe disease spread severely across the whole prov ...201324312081
one-step multiplex reverse transcription-polymerase chain reaction for the simultaneous detection of three rice viruses.rice stripe virus (rsv), rice black-streaked dwarf virus (rbsdv), and rice dwarf virus (rdv) are major rice-infecting viruses in korea that can cause serious crop losses. a one-step multiplex reverse transcription-polymerase chain reaction (mrt-pcr) was developed for the simultaneous detection of these rice viruses. three sets of specific primers targeted to the capsid protein coding genes of rsv, rbsdv, and rdv were used to amplify fragments that were 703 bp, 485 bp, and 252 bp, respectively. t ...201323850700
characterization of rice black-streaked dwarf virus- and rice stripe virus-derived sirnas in singly and doubly infected insect vector laodelphax striatellus.replication of rna viruses in insect cells triggers an antiviral defense that is mediated by rna interference (rnai) which generates viral-derived small interfering rnas (sirnas). however, it is not known whether an antiviral rnai response is also induced in insects by reoviruses, whose double-stranded rna genome replication is thought to occur within core particles. deep sequencing of small rnas showed that when the small brown planthopper (laodelphax striatellus) was infected by rice black-str ...201323776591
strong resistance against rice grassy stunt virus is induced in transgenic rice plants expressing double-stranded rna of the viral genes for nucleocapsid or movement proteins as targets for rna interference.rice grassy stunt virus (rgsv), a member of the genus tenuivirus, causes significant economic losses in rice production in south, southeast, and east asian countries. growing resistant varieties is the most efficient method to control rgsv; however, suitable resistance genes have not yet been found in natural rice resources. one of the most promising methods to confer resistance against rgsv is the use of rna interference (rnai). it is important to target viral genes that play important roles in ...201323190115
interaction between rice stripe virus disease-specific protein and host psbp enhances virus symptoms.rice stripe virus (rsv) causes severe diseases in oryza sativa (rice) in many eastern asian countries. disease-specific protein (sp) of rsv is a non-structural protein and its accumulation level in rice plant was shown to determine the severity of rsv symptoms. here, we present evidence that expression of rsv sp alone in rice or nicotiana benthamiana did not produce visible symptoms. expression of sp in these two plants, however, enhanced rsv- or potato virus x (pvx)-induced symptoms. through ye ...201424214893
[stress effects of imidacloprid on rsv in rice plants].the rice stripe disease is a viral disease transmitted by small brown planthopper, laodelphax striatellus, which outbroke a few years ago in the yangtze river basin, especially jiangsu region, china. to study the effects of imidacloprid stress on rice stripe virus (rsv) in rice plants, the rice seedlings were treated with imidacloprid 1, 2, 3 and 4 times (b1, b2, b3 and b4), respectively, after artificial inoculation by l. striatellus for 48 h, and the expression levels of relative genes includi ...201425876413
[construction of rice stripe virus ns2 and ns3 co-rnai transgenic rice and disease-resistance analysis].ns2 and ns3 are two post-transcriptional gene silencing suppressors that are encoded by rice stripe virus. gene silencing suppressors are always related to the pathogenicity of viruses. in this study, the cdna of ns2 and ns3 were recombined by overlapping pcr assays, ligated to the rnai vector, and inserted into the pxq expression vector using pst i; the expressed vector was transferred into calluses induced from seeds of the japonica rice cultivar, 'nipponbare', using an agrobacterium-mediated ...201425868281
stv11 encodes a sulphotransferase and confers durable resistance to rice stripe virus.rice stripe virus (rsv) causes one of the most serious viral diseases of rice (oryza sativa l.), but the molecular basis of rsv resistance has remained elusive. here we show that the resistant allele of rice stv11 (stv11-r) encodes a sulfotransferase (ossot1) catalysing the conversion of salicylic acid (sa) into sulphonated sa (ssa), whereas the gene product encoded by the susceptible allele stv11-s loses this activity. sequence analyses suggest that the stv11-r and stv11-s alleles were prediffe ...201425203424
heat shock protein 70 is necessary for rice stripe virus infection in plants.heat shock proteins 70 (hsp70s) are a highly conserved family of genes in eukaryotes, and are involved in a remarkable variety of cellular processes. in many plant positive-stranded rna viruses, hsp70 participates in the construction of a viral replication complex and plays various roles during viral infection. here, we found increased expression of hsp70 following infection by rice stripe virus (rsv), a negative-stranded rna virus, in both rice (the natural host) and nicotiana benthamiana (an e ...201424823923
transovarial transmission of a plant virus is mediated by vitellogenin of its insect vector.most plant viruses are transmitted by hemipteroid insects. some viruses can be transmitted from female parent to offspring usually through eggs, but the mechanism of this transovarial transmission remains unclear. rice stripe virus (rsv), a tenuivirus, transmitted mainly by the small brown planthopper (laodelphax striatellus), is also spread to the offspring through the eggs. here, we used the rsv-planthopper system as a model to investigate the mechanism of transovarial transmission and demonst ...201424603905
a transmembrane domain determines the localization of rice stripe virus pc4 to plasmodesmata and is essential for its function as a movement protein.the pc4 protein encoded by rice stripe virus (rsv) is a viral movement protein (mp). a transmembrane (tm) domain spanning aas 106-123 of pc4 was identified and shown to be essential for localization of pc4 to plasmodesmata (pd) (but not to chloroplasts) and for its ability to recover the movement of movement-deficient pvx. analysis of alanine-scanning mutants showed that m116a and g120a had a similar localization to wild type pc4, being localized at pd and chloroplasts, but all other mutants wer ...201424560843
transgenic rice expressing rice stripe virus ns3 protein, a suppressor of rna silencing, shows resistance to rice blast disease.the ns3 protein of rice stripe virus (rsv), encoded by the virion strand of rna3, is a viral suppressor of rna silencing (vsr). rice expressing ns3 had a normal phenotype, was initially sensitive to rsv but recovered at the later stages of infection. rsv accumulated slightly more in transgenic than in wild-type plants at the early stage of infection, but accumulation was similar later. transgenic rice expressing ns3 also showed enhanced resistance to the fungus magnaporthe oryzae. meanwhile, exp ...201424557730
nonstructural protein ns4 of rice stripe virus plays a critical role in viral spread in the body of vector insects.rice stripe virus (rsv), a tenuivirus, is transmitted by small brown planthopper (sbph) in a persistent-propagative manner. in this study, sequential infection of rsv in the internal organs of sbph after ingestion of virus indicated that rsv initially infected the midgut epithelium, and then progressed to the visceral muscle tissues, through which rsv spread to the entire alimentary canal. finally, rsv spread into the salivary glands and reproductive system. during viral infection, the nonstruct ...201424523924
interaction study of rice stripe virus proteins reveals a region of the nucleocapsid protein (np) required for np self-interaction and nuclear localization.rice stripe virus (rsv), which belongs to the genus tenuivirus, is an emergent virus problem. the rsv genome is composed of four single-strand rnas (rna1-rna4) and encodes seven proteins. we investigated interactions between six of the rsv proteins by yeast-two hybrid (y2h) assay in vitro and by bimolecular fluorescence complementation (bifc) in planta. y2h identified self-interaction of the nucleocapsid protein (np) and ns3, while bifc revealed self-interaction of np, ns3, and ncp. to identify ...201424463502
intracellular localization of rice stripe virus rna-dependent rna polymerase and its interaction with nucleocapsid protein.the rna-dependent rna polymerase (rdrp) of rice stripe virus (rsv) is critical for both the transcription and replication of the viral genome. despite its importance, little is known about how it functions in cells. in the present study, rsv rdrp was split into three pieces, since expression of the full protein could not be achieved. then, the intracellular localization of these three rdrp fragments and their interactions with nucleocapsid protein (np) were investigated, which is another viral p ...201526560706
functional comparison of rna silencing suppressor between the p5 protein of rice grassy stunt virus and the p3 protein of rice stripe virus.rice grassy stunt virus (rgsv) is a member of the genus tenuivirus, which includes rice stripe virus (rsv), as the type species. a viral suppressor of rna silencing (vsr) of rgsv has not been identified, whereas the p3 protein of rsv (rsvp3) encoded by the viral-sense (v) strand of rna3 has been reported to act as a vsr. in this study, we examined the vsr function of the p5 protein of rgsv (rgsvp5), encoded by vrna5. expecting it to correspond to the vrna3 of rsv, we compared the vsr function of ...201525836276
analysis of genetic variation and diversity of rice stripe virus populations through high-throughput sequencing.plant rna viruses often generate diverse populations in their host plants through error-prone replication and recombination. recent studies on the genetic diversity of plant rna viruses in various host plants have provided valuable information about rna virus evolution and emergence of new diseases caused by rna viruses. we analyzed and compared the genetic diversity of rice stripe virus (rsv) populations in oryza sativa (a natural host of rsv) and compared it with that of the rsv populations ge ...201525852724
rice stripe tenuivirus nonstructural protein 3 hijacks the 26s proteasome of the small brown planthopper via direct interaction with regulatory particle non-atpase subunit 3.the ubiquitin/26s proteasome system plays a vital role in regulating host defenses against pathogens. previous studies have highlighted different roles for the ubiquitin/26s proteasome in defense during virus infection in both mammals and plants, but their role in the vectors that transmit those viruses is still unclear. in this study, we determined that the 26s proteasome is present in the small brown planthopper (sbph) (laodelphax striatellus) and has components similar to those in plants and ...201525653432
alterations in sirna and mirna expression profiles detected by deep sequencing of transgenic rice with sirna-mediated viral resistance.rna-mediated gene silencing has been demonstrated to serve as a defensive mechanism against viral pathogens by plants. it is known that specifically expressed endogenous sirnas and mirnas are involved in the self-defense process during viral infection. however, research has been rarely devoted to the endogenous sirna and mirna expression changes under viral infection if the resistance has already been genetically engineered in plants. aiming to gain a deeper understanding of the rna-mediated gen ...201525559820
interaction of hsp20 with a viral rdrp changes its sub-cellular localization and distribution pattern in plants.small heat shock proteins (shsps) perform a fundamental role in protecting cells against a wide array of stresses but their biological function during viral infection remains unknown. rice stripe virus (rsv) causes a severe disease of rice in eastern asia. oshsp20 and its homologue (nbhsp20) were used as baits in yeast two-hybrid (yth) assays to screen an rsv cdna library and were found to interact with the viral rna-dependent rna polymerase (rdrp) of rsv. interactions were confirmed by pull-dow ...201526359114
time-course rna-seq analysis reveals transcriptional changes in rice plants triggered by rice stripe virus infection.rice stripe virus (rsv) has become a major pathogen of rice. to determine how the rice transcriptome is modified in response to rsv infection, we used rna-seq to perform a genome-wide gene expression analysis of a susceptible rice cultivar. the transcriptomes of rsv-infected samples were compared to those of mock-treated samples at 3, 7, and 15 days post-infection (dpi). from 8 to 11% of the genes were differentially expressed (>2-fold difference in expression) in rsv-infected vs. noninfected ri ...201526305329
osrdr6 plays role in host defense against double-stranded rna virus, rice dwarf phytoreovirus.rnai is a major antiviral defense response in plant and animal model systems. rna-dependent rna polymerase 6 (rdr6) is an essential component of rnai, which plays an important role in the resistance against viruses in the model plants. we found previously that rice rdr6 (osrdr6) functioned in the defense against rice stripe virus (rsv), and rice dwarf phytoreovirus (rdv) infection resulted in down-regulation of expression of rdr6. here we report our new findings on the function of osrdr6 against ...201526165755
itraq-based quantitative proteomics analysis of rice leaves infected by rice stripe virus reveals several proteins involved in symptom formation.rice plants infected by rice stripe virus (rsv) usually leads to chlorosis and death of newly emerged leaves. however, the mechanism of rsv-induced these symptoms was not clear.201526113023
assessment of reference gene stability in rice stripe virus and rice black streaked dwarf virus infection rice by quantitative real-time pcr.stably expressed reference gene(s) normalization is important for the understanding of gene expression patterns by quantitative real-time pcr (rt-qpcr), particularly for rice stripe virus (rsv) and rice black streaked dwarf virus (rbsdv) that caused seriously damage on rice plants in china and southeast asia.201526497487
rice stripe tenuivirus p2 may recruit or manipulate nucleolar functions through an interaction with fibrillarin to promote virus systemic movement.rice stripe virus (rsv) is the type species of the genus tenuivirus and represents a major viral pathogen affecting rice production in east asia. in this study, rsv p2 was fused to yellow fluorescent protein (p2-yfp) and expressed in epidermal cells of nicotiana benthamiana. p2-yfp fluorescence was found to move to the nucleolus initially, but to leave the nucleolus for the cytoplasm forming numerous distinct bright spots there at later time points. a bimolecular fluorescence complementation (bi ...201525431002
the rna-binding properties and domain of rice stripe virus nucleocapsid protein.the nucleocapsid protein (np) of rice stripe virus (rsv) encapsidates viral genomic rnas to form virion. the binding of np with rna is essential for the formation of virus particle. in this study, the binding specificity of rsv np to rna and the domains within the np that mediate this interaction were investigated by gel electrophoretic mobility shift assays and northwestern blot analysis. the results demonstrated that rsv np was able to bind to all synthetic rnas and dnas without sequence speci ...201526250446
first discovery of acetone extract from cottonseed oil sludge as a novel antiviral agent against plant viruses.a novel acetone extract from cottonseed oil sludge was firstly discovered against plant viruses including tobacco mosaic virus (tmv), rice stripe virus (rsv) and southern rice black streaked dwarf virus (srbsdv). gossypol and β-sitosterol separated from the acetone extract were tested for their effects on anti-tmv and analysed by nuclear magnetic resonance (nmr) assay. in vivo and field trials in different geographic distributions and different host varieties declared that this extract mixture w ...201525705894
a simplified method for simultaneous detection of rice stripe virus and rice black-streaked dwarf virus in insect vector.rice stripe virus (rsv) and rice black-streaked dwarf virus (rbsdv) are transmitted by their common vector small brown planthopper (sbph) that cause serious crop losses in china. a simple reverse transcription-pcr method was developed for the simultaneous detection of rsv and rbsdv in single sbph. three primers targeted to rsv-rna4 and rbsdv-s2 segments were designed to amplify respectively 1114-bp and 414-bp fragments in a reaction. the method is reliable, rapid and inexpensive for detecting th ...201525455902
rice stripe virus counters reduced fecundity in its insect vector by modifying insect physiology, primary endosymbionts and feeding behavior.virus-vector relationships can be complex and diverse as a result of long-term coevolution. understanding these interactions is crucial for disease and vector management. rice stripe virus (rsv) is known to be transovarially transmitted within its vector, laodelphax striatellus, and causes serious rice stripe disease. in rsv-infected l. striatellus, we found contrasting changes in vector fecundity, physiology, primary endosymbionts (i.e. yeast-like symbionts, yls) and feeding behavior that can i ...201526211618
proteomic analysis of interaction between a plant virus and its vector insect reveals new functions of hemipteran cuticular protein.numerous viruses can be transmitted by their corresponding vector insects; however, the molecular mechanisms enabling virus transmission by vector insects have been poorly understood, especially the identity of vector components interacting with the virus. here, we used the yeast two-hybrid system to study proteomic interactions of a plant virus (rice stripe virus, rsv, genus tenuivirus) with its vector insect, small brown planthopper (laodelphax striatellus). sixty-six proteins of l. striatellu ...201526091699
rice stripe virus affects the viability of its vector offspring by changing developmental gene expression in embryos.plant viruses may affect the viability and development process of their herbivore vectors. small brown planthopper (sbph) is main vector of rice stripe virus (rsv), which causes serious rice stripe disease. here, we reported the effects of rsv on sbph offspring by crossing experiments between viruliferous and non-viruliferous strains. the life parameters of offspring from different cross combinations were compared. the hatchability of f1 progeny from viruliferous parents decreased significantly, ...201525601039
facilitation of rice stripe virus accumulation in the insect vector by himetobi p virus vp1.the small brown planthopper (sbph) is the main vector for rice stripe virus (rsv), which causes serious rice stripe disease in east asia. to characterize the virus-vector interactions, the sbph cdna library was screened with rsv ribonucleoprotein (rnp) as bait using a gal4-based yeast two-hybrid system. the interaction between rsv-rnp and the himetobi p virus (hipv, an insect picorna-like virus) vp1 protein was identified. the relationships between hipv and rsv in sbph were further investigated, ...201525807055
roles of the laodelphax striatellus down syndrome cell adhesion molecule in rice stripe virus infection of its insect vector.the arthropod down syndrome cell adhesion molecule (dscam) mediates pathogen-specific recognition via an extensive protein isoform repertoire produced by alternative splicing. to date, most studies have focused on the subsequent pathogen-specific immune response, and few have investigated the entry into cells of viruses or endosymbionts. in the present study, we cloned and characterized the cdna of laodelphax striatellus dscam (lsdscam) and investigated the function of lsdscam in rice stripe vir ...201626991800
inherent properties not conserved in other tenuiviruses increase priming and realignment cycles during transcription of rice stripe virus.two tenuiviruses rice stripe virus (rsv) and rice grassy stunt virus (rgsv) were found to co-infect rice with the same reovirus rice ragged stunt virus (rrsv). during the co-infection, both tenuiviruses recruited 10-21 nucleotides sized capped-rna leaders from the rrsv. a total of 245 and 102 rrsv-rgsv and rrsv-rsv chimeric mrna clones, respectively, were sequenced. an analysis of the sequences suggested a scenario consistent with previously reported data on related viruses, in which capped lead ...201627393974
rice responses and resistance to planthopper-borne viruses at transcriptomic and proteomic levels.rice (oryza sativa) is one of the most important cereal crops in the world, especially in asian areas. rice virus diseases are considered as the most serious threat to rice yields. most rice viruses are transmitted by hemipteran insects such as planthoppers and leafhoppers. in asia five rice viruses are transmitted mainly by three planthopper species in a persistent manner: rice stripe virus, rice black-streaked dwarf virus, rice ragged stunt virus, rice grassy stunt virus, and southern rice bla ...201626363817
time-course small rna profiling reveals rice mirnas and their target genes in response to rice stripe virus infection.it has been known that many micrornas (mirnas) are involved in the regulation for the plant development and defense mechanism by regulating the expression of the target gene. several previous studies has demonstrated functional roles of mirnas in antiviral defense mechanisms. in this study, we employed high-throughput sequencing technology to identify rice mirnas upon rice stripe virus (rsv) infection at three different time points. six libraries from mock and rsv-infected samples were subjected ...201627626631
artificial feeding rice stripe virus enables efficient virus infection of laodelphax striatellus.rice stripe virus (rsv), the causative agent of rice stripe disease, is transmitted by laodelphax striatellus in a persistent-propagative manner. efficient virus acquisition is primary for studies of virus transmission and virus-insect vector interactions. however, under greenhouse conditions, less than 30% of the l. striatellus population, on average, become viruliferous during feeding on rsv-infected plants. here, we explored a method for efficient rsv acquisition by feeding the insects with a ...201627283882
differential proteomics profiling of the ova between healthy and rice stripe virus-infected female insects of laodelphax striatellus.rice stripe virus-infected females of the small brown planthopper (sbph, laodelphax striatellus) usually lay fewer eggs with a longer hatch period, low hatchability, malformation and retarded or defective development compared with healthy females. to explore the molecular mechanism of those phenomena, we analyzed the differential proteomics profiling of the ova between viruliferous and healthy female insects using an isobaric tag for relative and absolute quantitation (itraq) approach. we obtain ...201627277140
seasonal changes in the percentage of rice stripe virus viruliferous laodelphax striatellus (hemiptera: delphacidae) in paddy fields in japan.rice stripe disease, which is caused by rice stripe virus (rsv), is one of the most serious viral diseases of rice. rsv is transmitted in a persistent manner bylaodelphax striatellus(fallén). the incidence of the disease can be estimated from the density of viruliferous vectors. understanding seasonal changes of the percentage of viruliferousl. striatelluscan facilitate forecasting and controlling the disease. in paddies, the percentage of viruliferous insects fluctuated in phase with the rate o ...201627099363
mapping of the regions involved in self-interaction of rice stripe virus p3 protein.rice stripe virus (rsv) protein p3 is a suppressor of rna silencing in plants. p3 has been shown by biomolecular fluorescence complementation assay to self-interact in planta but the regions responsible for homotypic interaction have not been determined. here we analyzed the domains for the self-interaction of p3 by using yeast two-hybrid, co-immunoprecipitation and fluorescence experiments. the results showed that p3 was also able to interact with itself in yeast and insect cells. the domain re ...201626982473
viruliferous rate of small brown planthopper is a good indicator of rice stripe disease epidemics.rice stripe virus (rsv), its vector insect (small brown planthopper, sbph) and climatic conditions in jiangsu, china were monitored between 2002 and 2012 to determine key biotic and abiotic factors driving epidemics of the disease. average disease severity, disease incidence and viruliferous rate of sbph peaked in 2004 and then gradually decreased. disease severity of rsv was positively correlated with viruliferous rate of the vector but not with the population density of the insect, suggesting ...201626898155
a signaling cascade from mir444 to rdr1 in rice antiviral rna silencing pathway.plant rna-dependent rna polymerase1 (rdr1) is a key component of the antiviral rna-silencing pathway, contributing to the biogenesis of virus-derived small interfering rnas. this enzyme also is responsible for producing virus-activated endogenous small interfering rnas to stimulate the broad-spectrum antiviral activity through silencing host genes. the expression of rdr1 orthologs in various plants is usually induced by virus infection. however, the molecular mechanisms of activation of rdr1 exp ...201626858364
integrative analysis of the micrornaome and transcriptome illuminates the response of susceptible rice plants to rice stripe virus.rice stripe virus (rsv) is one of the most serious rice viruses in east asia. to investigate how rice responds to rsv infection, we integrated mirna expression with parallel mrna transcription profiling by deep sequencing. a total of 570 mirnas were identified of which 69 mirnas (56 up-regulated and 13 down-regulated) were significantly modified by rsv infection. digital gene expression (dge) analysis showed that 1274 mrnas (431 up-regulated and 843 down-regulated genes) were differentially expr ...201626799317
organ-specific transcriptome response of the small brown planthopper toward rice stripe virus.rice stripe virus (rsv) causes rice stripe disease and is transmitted by the small brown planthopper (laodelphax striatellus, sbph) in a persistent, circulative, and propagative manner. the alimentary canal and salivary gland of sbph play important roles in viral replication and transmission. however, little is known about the underlying molecular functions of these two organs in the interaction between rsv and sbph. in this study, organ-specific transcriptomes of the alimentary canal and saliva ...201626678499
different pathogenicities of rice stripe virus from the insect vector and from viruliferous plants.persistent plant viruses usually depend on insects for their transmission; they cannot be transmitted between plants or through mechanical inoculation. however, the mechanism by which persistent viruses become pathogenic in insect vectors remains unknown. in this study, we used rice stripe virus (rsv), its insect vector laodelphax striatellus and host plant (oryza sativa) to explore how persistent viruses acquire pathogenicity from insect vectors. rsv acquired phytopathogenicity in both the alim ...201626585422
identification and regulation of host genes related to rice stripe virus symptom production.viral infections cause plant chlorosis, stunting, necrosis or other symptoms. the down-regulation of chloroplast-related genes (chrgs) is assumed to be responsible for chlorosis. we identified the differentially expressed genes (degs) in rice stripe virus (rsv)-infected nicotiana benthamiana, and examined the contribution of 75 down-regulated degs to rsv symptoms by silencing them one by one using tobacco rattle virus (trv)-induced gene silencing. silencing of 11 of the 75 down-regulated degs ca ...201626487490
rna-seq-based digital gene expression analysis reveals modification of host defense responses by rice stripe virus during disease symptom development in arabidopsis.virus infection induces and suppresses host gene expression on a global level. rice stripe virus (rsv) is the type species of the genus tenuivirus and infects rice and arabidopsis plants. microarray-based and next generation sequencing-based transcriptomic approaches have been used to study rice-rsv interactions. however, our knowledge of the response of arabidopsis plants to rsv infection is limited, and it requires further investigation to determine the similarities (or differences) in virus-h ...201627912765
overexpression of bsr1 confers broad-spectrum resistance against two bacterial diseases and two major fungal diseases in rice.broad-spectrum disease resistance against two or more types of pathogen species is desirable for crop improvement. in rice, xanthomonas oryzae pv. oryzae (xoo), the causal bacteria of rice leaf blight, and magnaporthe oryzae, the fungal pathogen causing rice blast, are two of the most devastating pathogens. we identified the rice broad-spectrum resistance 1 (bsr1) gene for a bik1-like receptor-like cytoplasmic kinase using the fox hunting system, and demonstrated that bsr1-overexpressing (ox) ri ...201627436950
development of a simplified rt-pcr without rna isolation for rapid detection of rna viruses in a single small brown planthopper (laodelphax striatellus fallén).the small brown planthopper (sbph) is an important pest of cereal crops and acts as a transmission vector for multiple rna viruses. rapid diagnosis of virus in the vector is crucial for efficient forecast and control of viral disease. reverse transcription polymerase chain reaction (rt-pcr) is a rapid, sensitive and reliable method for virus detection. the traditional rt-pcr contains a rna isolation step and is widely used for virus detection in insect. however, using the traditional rt-pcr for ...201728468626
bacterial microbiota in small brown planthopper populations with different rice viruses.the small brown planthopper (sbph) is an important virus vector, transmitting rice stripe virus (rsv), and rice black-streaked dwarf virus (rbsdv). insect symbionts play an essential role in the insect fitness, however, it is still unclear about their contributions to viral transmission by sbph. here, we investigated endosymbiont communities in non-viruliferous, rsv-infected, and rbsdv-infected sbph populations using illumina 16s rrna gene miseq sequencing. in total, 281,803 effective sequences ...201728471518
the c-jun n-terminal kinase pathway of a vector insect is activated by virus capsid protein and promotes viral replication.no evidence has shown whether insect-borne viruses manipulate the c-jun n-terminal kinase (jnk) signaling pathway of vector insects. using a system comprising the plant virus rice stripe virus (rsv) and its vector insect, the small brown planthopper, we have studied the response of the vector insect's jnk pathway to plant virus infection. we found that rsv increased the level of tumor necrosis factor-α and decreased the level of g protein pathway suppressor 2 (gps2) in the insect vector. the vir ...201728716183
quantitative analysis of rice stripe virus in a transovarial transmission cycle during the development and reproduction of its vector, laodelphax striatellus.the amount of rice stripe virus (rsv) maintained through transovarial transmission was analyzed during the development and reproduction of its vector, laodelphax striatellus. reverse transcription quantitative pcr analysis was used to quantify rna expressed from the rsv coat protein (cp) gene as an estimate of rsv content in nymphs and adults of l. striatellus at various developmental stages. the 18s ribosome rna gene of l. striatellus was chosen as the reference for calculating rsv cp expressio ...201728589385
identification of residues or motif(s) of the rice stripe virus ns3 protein required for self-interaction and for silencing suppressor activity.rice stripe virus (rsv) is an important pathogen of rice. the rsv genome consists of four single-stranded rna segments that encode seven viral proteins. a previous report found that ns3 is a viral suppressor of rna silencing and self interacts. using a model that predicts protein structure, we identified amino acid residues or motifs, including four α-helix motifs, required for ns3 self-interaction. we then used yeast two-hybrid (y2h) and bimolecular fluorescence complementation (bifc) assays to ...201728392445
evolution of rice stripe virus.rice stripe virus (rsv) is an insect-borne tenuivirus of economical significance. it is endemic to the rice-growing regions of east asia and exhibits more genetic diversity in yunnan province of china. to gain more insights into the molecular epidemiology and evolution of rsv, recombination analyses were conducted and potential events were detected in each of the four rna segments of rsv. bayesian coalescent method was then applied to the time-stamped coding sequences of the cp gene. the nucleot ...201728189616
rice stripe virus infection alters mrna levels of sphingolipid-metabolizing enzymes and sphingolipids content in laodelphax striatellus.sphingolipids and their metabolites have been implicated in viral infection and replication in mammal cells but how their metabolizing enzymes in the host are regulated by viruses remains largely unknown. here we report the identification of 12 sphingolipid genes and their regulation by rice stripe virus in the small brown planthopper (laodelphax striatellus fallén), a serious pest of rice throughout eastern asia. according to protein sequence similarity, we identified 12 sphingolipid enzyme gen ...201728130458
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