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biolistic transformation of chrysanthemum with the nucleocapsid gene of tomato spotted wilt virus.in vitro regeneration and biolistic transformation procedures were developed for several commercial chrysanthemum dendranthema grandiflora tzvelev, syn. chrysanthemum morifolium ramat. cultivars using leaf and stem explants. studies on the effect of several growth regulators and kanamycin on chrysanthemum regeneration were conducted, and a step-wise procedure to optimize kanamycin selection and recovery of transgenic plants was developed. a population of putative transformed chrysanthemum plants ...199524186624
mapping the sw-5 locus for tomato spotted wilt virus resistance in tomatoes using rapd and rflp analyses.the sw-5 locus confers dominant resistance to tomato spotted wilt virus (tswv). to map the location and facilitate the identification of markers linked to sw-5 we developed a pair of near-isogenic lines (nils) and an f2 lycopersicon esculentum x l. pennellii population segregating for resistance to tswv. dna from the nils was analyzed using 748 random 10-mer oligonucleotides to discern linked molecular markers using a random amplified polymorphic dna (rapd) approach. one random primer (gagcacggg ...199524173937
resistance to tospoviruses in nicotiana benthamiana transformed with the n gene of tomato spotted wilt virus: correlation between transgene expression and protection in primary transformants.nicotiana benthamiana was transformed with the nucleoprotein (n) gene of an italian isolate of tomato spotted wilt virus (tswv). forty-five t1 primary transformant lines were analyzed for the expression of n protein and for resistance to tswv and three other tospoviruses: impatiens necrotic spot virus (insv), groundnut bud necrosis virus (gbnv), and groundnut ringspot virus (grsv). thirteen of these lines were further characterized. resistance to all tswv isolates tested was found in two lines. ...19957772806
broad resistance to tospoviruses in transgenic tobacco plants expressing three tospoviral nucleoprotein gene sequences.transgenic tobacco plants have been obtained expressing nucleoprotein (n) gene sequences of three different tospoviruses known to affect vegetable crops: tomato spotted wilt virus (tswv), tomato chlorotic spot virus (tcsv), and groundnut ringspot virus (grsv). the chimeric plant transformation vector used comprised the three viral n gene sequences, each with a copy of the camv 35s promoter and the nos terminator. despite the high levels of homology between the different n gene sequences (74-82%) ...19957772807
completion of the la crosse virus genome sequence and genetic comparisons of the l proteins of the bunyaviridae.la crosse virus is a member of the bunyavirus genus in the family bunyaviridae, viruses with trisegmented rna genomes of mostly negative polarity composed of large (l), medium (m), and small (s) segments. the sequences of the la crosse/original m and s rna segments have been previously characterized. using reverse transcriptase in conjunction with pcr amplification, we have obtained the nucleotide sequence of the l rna segment, which encodes the viral polymerase in a single large open reading fr ...19957831835
an rna-dependent rna polymerase activity associated with virions of tomato spotted wilt virus, a plant- and insect-infecting bunyavirus.an rna-dependent rna polymerase activity was found associated with virions of tomato spotted wilt virus (tswv), a plant- and insect-infecting member of the family bunyaviridae. radiolabeled nucleoside triphosphates were incorporated into trichloroacetic acid-precipitable products by detergent-disrupted, purified tswv virions. incorporation was reduced to near-background levels when rnase was present in the reaction mixture. the predominantly double-stranded rna products were rnase-resistant at h ...19957871744
the nonstructural nsm protein of tomato spotted wilt virus induces tubular structures in plant and insect cells.the expression and subcellular location of the 33.6-kda nonstructural protein nsm of tomato spotted wilt virus (tswv) was analyzed in nicotiana rustica plants and protoplasts as a function of time. immunofluorescent studies in protoplasts isolated from tswv-infected n. rustica leaves showed that this protein could first be detected close to the periphery of the cell, near the plasmamembrane, and later in tubular structures emerging from the cell surface. in situ, these tubules appeared specifica ...19958553550
allelic relationships between genes for resistance to tomato spotted wilt tospovirus in capsicum chinense.pepper (capsicum chinense jacq.) has been reported to be an important reservoir of resistance genes to tomato spotted wilt virus (tswv). the genes for tswv resistance present in three c. chinense lines ('pi 152225', 'pi 159236' and 'panca') were investigated for allelism. all resistant lines were crossed with each other. parents, f1, backcrosses and f2 populations (including reciprocals) developed from those crosses were mechanically inoculated with a highly virulent tswv isolate. susceptible c. ...199524173797
the detection of tomato spotted wilt virus using the polymerase chain reaction.a polymerase chain reaction (pcr) based assay was used to detected a number of uk tomato spotted wilt virus (tswv) isolates in total rna extractions made from infected plant material. extracts were reverse transcribed and the resultant cdna amplified by pcr, using oligonucleotide primers specific for a 276 base pair fragment of the l rna segment. assay products were electrophoresed on agarose gels and visualised by ethidium bromide staining. the viral origin of the product produced was confirmed ...19948006111
expression and subcellular location of the nsm protein of tomato spotted wilt virus (tswv), a putative viral movement protein.the 33.6-kda nonstructural (nsm) protein gene, located on the ambisense m rna segment of tomato spotted wilt virus (tswv), was cloned and expressed using the escherichia coli pet-11t expression system. the protein thus produced was purified and used for the production of a polyclonal antiserum. western immunoblot analyses of tswv-infected nicotiana rustica plants showed nsm synthesis only during a short period early in systemic infection. although nsm was found associated with cytoplasmic nucleo ...19948128638
detection of the l protein of tomato spotted wilt virus.the 5'-terminal and 3'-terminal parts of the single open reading frame (orf) in the l rna of tomato spotted wilt virus (tswv) were expressed using a prokaryotic expression system. using antibodies raised against the translational products obtained a 330-kda protein could be specifically detected in preparations of purified virions and in nucleocapsid preparations from tswv-infected leaf tissue. the results obtained indicate that the l protein of tswv, though much larger than that of the animal-i ...19938212587
classification of tospoviruses based on phylogeny of nucleoprotein gene sequences.the nucleotide sequences of the nucleoprotein (n) genes of seven tospovirus isolates representing three serogroups were determined and used to establish phylogenetic parameters to delineate species within the tospovirus genus of the bunyaviridae. a high sequence divergence (55.9% identity at the nucleotide level) was observed between isolates of serogroup i (tomato spotted wilt virus) and isolates of serogroup iii (impatiens necrotic spot virus). the serogroup ii isolates take an intermediate po ...19938429298
multiplication of tomato spotted wilt virus in its insect vector, frankliniella occidentalis.the accumulation of two proteins, the nucleocapsid (n) protein and a non-structural (nss) protein both encoded by the s rna of tomato spotted wilt virus (tswv), was followed in larvae during development and in adults of frankliniella occidentalis after ingesting the virus for short periods on infected plants. the amounts of both proteins increased, as shown by elisa and western blot analysis, within 2 days above the levels ingested, indicating multiplication of tswv in these insects. accumulatio ...19938445364
the nucleotide sequence of the m rna segment of tomato spotted wilt virus, a bunyavirus with two ambisense rna segments. 19938468562
a method combining immunocapture and pcr amplification in a microtiter plate for the detection of plant viruses and subviral pathogens.a method for the detection of rna viral and subviral plant pathogens was developed that combines pathogen partial purification by solid-phase adsorbed antibodies, reverse transcriptional-polymerase chain reaction (rt-pcr) and quantitation of the amplified products by fluorescence. the reverse transcription of the rna is performed directly on the retained material without any previous thermal or chemical disruption of the virus particles. the whole procedure can be carried out in a microtiter pla ...19938113346
serological comparison of tospovirus isolates from taiwan and india with impatiens necrotic spot virus and different tomato spotted wilt virus isolates.tospovirus isolates from tomato and watermelon in taiwan and from peanut in india reacted neither with antibodies to tomato spotted wilt virus (tswv), nor with antibodies to impatiens necrotic spot virus (insv), in elisa and in electroblot immunoassays. additionally, all three tospovirus isolates had an n protein with a molecular weight of 32 kda as compared to 29 kda for the n protein to tswv and insv. the observed physicochemical and serological differences suggest the generation of a new, thi ...19937686004
different mechanisms protect transgenic tobacco against tomato spotted wilt and impatiens necrotic spot tospoviruses.we generated transgenic tobacco plants expressing the sense or antisense untranslatable n coding sequence of the lettuce isolate of tomato spotted wilt virus (tswv-bl) as well as transgenic plants containing the promoterless n gene of the virus. both sense and antisense untranslatable n gene rnas provided protection against homologous and closely related isolates but not against distantly related tospoviruses. these rna-mediated protections were most effective in plants that synthesized low leve ...19937763861
determination of whether tomato spotted wilt virus replicates in toxorhynchites amboinensis mosquitoes and the relatedness of this virus to phleboviruses (family bunyaviridae).tomato spotted wilt virus (tswv) has been reported to be morphologically, molecularly and structurally similar to viruses in the family bunyaviridae. by various types of enzyme-linked immunosorbent assays (elisa) and western blot hybridizations, we tested tswv with antibodies to 12 viruses in the phlebovirus genus of this family. serological relatedness was not found between tswv and phleboviruses. however, one preparation of antibody to arumowot virus reacted with a 53-kd protein from healthy p ...19921346784
characterization of rna-mediated resistance to tomato spotted wilt virus in transgenic tobacco plants.recently high levels of protection against tomato spotted wilt virus (tswv), a negative-strand rna virus infecting plants, have been obtained by transforming tobacco with viral nucleoprotein (n) gene sequences. here we demonstrate that this protection is primarily due to the presence of n gene transcripts in the cells of transgenic plants, and hence appears to be rna-mediated. further, transgenic tobacco plants are only protected to isolates and strains of tswv and not to other tospoviruses that ...19921368791
the nucleotide sequence of the s rna of impatiens necrotic spot virus, a novel tospovirus.impatiens necrotic spot virus (insv) shares a number of properties with tomato spotted wilt virus (tswv), the type species of the genus tospovirus within the family bunyaviridae. insv, however, differs from tswv in plant host range and serology. in order to define the genomic structure and the taxonomic status of this tswv-like virus, the nucleotide sequence of its genomic s rna segment has been determined. the molecular data obtained demonstrate that, like tswv, insv has an ambisense s rna mole ...19921385787
defective interfering l rna segments of tomato spotted wilt virus retain both virus genome termini and have extensive internal deletions.defective interfering (di) rna molecules derived from the genomic l rna segment of tomato spotted wilt virus (tswv) were generated during sequential passage of the virus at high multiplicity. characterization of di rnas from four distinct isolates by northern blot analysis and sequence determination revealed that both the 5' and 3' genomic termini were retained in these molecules. each di rna contained a single internal deletion of approximately 60% to 80% of the l rna segment. all di rnas studi ...19921402797
immuno-electron microscopical detection of tomato spotted wilt virus and its nucleocapsids in crude plant extracts.tomato spotted wilt virus (tswv) particles were identified in thin sections and in crude extracts of leaves from plants infected with different tswv isolates, using gold labelled protein a and antibodies prepared against purified virus particles or against nucleocapsid preparations. in addition, both in thin sections and in dip preparations aggregates were detected using either gold-labelled antiserum. these aggregates were not detected using antisera against envelope proteins or against the non ...19921430055
the nucleotide sequence of the m rna segment of tomato spotted wilt virus, a bunyavirus with two ambisense rna segments.the complete sequence of the tomato spotted wilt virus (tswv) m rna segment has been determined. the rna is 4821 nucleotides long and has an ambisense coding strategy similar to that of the s rna segment. the m rna segment contains two open reading frames (orfs), one in the viral sense which encodes a protein with a predicted size of 33.6k, and one in the viral complementary sense which encodes the precursor to the g1 and g2 glycoproteins, with a predicted size of 127.4k. both orfs are expressed ...19921431808
viral rna synthesis in tomato spotted wilt virus-infected nicotiana rustica plants.the synthesis of viral rna species in tomato spotted wilt virus-infected nicotiana rustica plants was followed in terms of time and relative abundance. systemic symptoms were visible after 4 days postinoculation (p.i.), but viral (v) and viral-complementary (vc) strands of all three genomic rna segments [large (l) rna, medium (m) rna and small (s) rna] were detected from 2 days p.i. in addition, two subgenomic mrnas, derived from s rna, were detected. for the l rna segment no subgenomic mrnas we ...19921545223
resistance to tomato spotted wilt virus infection in transgenic tobacco expressing the viral nucleocapsid gene.a recombinant plasmid containing the entire tomato spotted with virus (tswv) nucleocapsid gene, with the exception of nucleotide encoding three n-terminal amino acids, was isolated by screening a complementary dna library, prepared against random primed viral rna, using a specific monoclonal antibody. the insert contained in plasmid ptsw1 was repaired and amplified by polymerase chain reaction, and the complete nucleocapsid protein gene was introduced into nicotiana tabacum 'samsun' by leaf disk ...19921600236
non-viral heterogeneous sequences at the 5' ends of tomato spotted wilt virus mrnas.subgenomic messenger rnas transcribed from the tomato spotted wilt virus (tswv) s rna segment were partially purified from total rna extracts of tswv-infected nicotiana rustica and analysed by primer extension analysis. the data obtained show the presence of non-viral sequences, 12 to 20 nucleotides in length, at the 5' ends of the n and nss mrnas, indicating a cap-snatching mechanism for the initiation of transcription. this is the first report of a plant virus using such a mechanism for transc ...19921645150
the nonstructural protein (nss) encoded by the ambisense s rna segment of tomato spotted wilt virus is associated with fibrous structures in infected plant cells.the open reading frame located in the viral strand of the ambisense s rna of tomato spotted wilt virus (tswv), was cloned into transfer vector pac33dz1 and inserted downstream of the polyhedrin promoter in the autographa californica nuclear polyhedrosis virus genome. recombinant baculoviruses were obtained that showed a high-level expression of a 52.4-kda protein corresponding to the inserted tswv gene. the viral protein thus produced was purified and injected into rabbits to raise antibodies. w ...19911826573
nucleotide sequence of the 3' non-coding region and n gene of the s rna of a serologically distinct tospovirus.a tomato spotted wilt-like virus (tswv-i) is a distinct member of the tospovirus genus of the bunyaviridae and is distinguished from the typical tswv by having a serologically distinct nucleoprotein (n). a cdna clone extending from the 3' terminus of the viral rna through the entire n open reading frame (orf) was sequenced. the tswv-i n orf is capable of encoding a polypeptide of 262 amino acids with a predicted mr of 28.8k. in vitro transcription and translation of the clone produced a protein ...19911840613
tomato spotted wilt virus l rna encodes a putative rna polymerase.the complete nucleotide sequence of the large (l) genome segment of tomato spotted wilt virus (tswv) has been determined. the rna is 8897 nucleotides long and contains complementary 3' and 5' ends, comprising 62 nucleotides at the 5' end and 66 nucleotides at the 3' end. the rna is of negative polarity, with one large open reading frame (orf) located on the viral complementary strand. this orf corresponds to a primary translation product of 2875 amino acids in length, with a predicted mr of 331, ...19911895058
generation of envelope and defective interfering rna mutants of tomato spotted wilt virus by mechanical passage.during a series of mechanical transfers of tomato spotted wilt virus, two distinct types of mutants were generated. firstly, a morphologically defective isolate was obtained which had lost the ability to produce the membrane glycoproteins and, as a consequence, was not able to form enveloped particles. analysis of the genomic rnas of this isolate suggested that this defect was caused by either point mutations or very small deletions in the medium genomic rna segment. secondly, isolates were obta ...19911919523
cloning and sequencing of the s rna from a bulgarian isolate of tomato spotted wilt virus.libraries of cloned cdna were prepared from complete genomic rna and isolated s rna of the bulgarian l3 isolate of tomato spotted wilt virus (tswv-l3). northern blotting of tswv genomic rna detected clones specific for the l, m and s rnas in the library from complete rna. s rna-specific clones selected from both libraries covered approximately 2.8 kb (about 95%) of the s rna. sequencing of these clones showed tswv-l3 s rna to be ambisense. it contains two open reading frames (orfs); one of 1401 ...19911993884
nucleotide sequence and rna hybridization analyses reveal an ambisense coding strategy for maize stripe virus rna3.the 2357-nt sequence of maize stripe virus (mstv) rna3 was determined. two nonoverlapping open reading frames (orfs) of opposite polarities are contained in rna3. a 591-nt orf is located near the 5' end of mstv rna3, while a second orf of 948 nt is located near the 3' end in the viral complementary rna (vcrna). in vitro translation of transcripts derived from cdna clones representing regions of each orf was used to identify the respective proteins. a ca. 22,000 mr protein was translated from the ...19912024478
serological differentiation of 20 isolates of tomato spotted wilt virus.twenty tomato spotted wilt virus (tswv) isolates were serologically compared in elisa employing five different procedures using a rabbit polyclonal antiserum against nucleocapsid proteins (nuabr) and mouse monoclonal antibodies (mabs), two directed to nucleocapsid proteins (n1 and n2) and four directed to glycoproteins g1 to g4. all the antisera were raised against tswv-cnph1. the 20 isolates were differentiated into two distinct serogroups. serogroup i consisting of 16 isolates strongly reacted ...19902273383
detection of tomato spotted wilt virus using monoclonal antibodies and riboprobes.the immunoreactivity of a panel of monoclonal antibodies raised to tomato spotted wilt virus (tswv) was examined in enzyme-linked immunosorbent assays (elisa) and dot immunobinding assays (diba) procedures. mabs 6.12.15 and 2.9 were specific for the nucleocapsid protein of tswv. the sensitivity of the two immunoassays was compared with that of a dot-blot hybridization technique using riboprobes (rna transcripts) to tswv m rna. using deproteinized plant extracts or purified virus preparations, as ...19901689996
the s rna segment of tomato spotted wilt virus has an ambisense character.the complete nucleotide sequence of the s rna of tomato spotted wilt virus (tswv) was determined. the rna is 2916 nucleotides long and has an ambisense coding strategy. the sequence contains two open reading frames (orfs), one in the viral sense which encodes a protein with a predicted mr of 52.4k and one in the viral complementary sense which encodes the viral nucleocapsid protein of mr 28.8k. both proteins are expressed by translation of two subgenomic rna species that possibly terminate at a ...19901693160
penicillinase-based enzyme-linked immunosorbent assay for the detection of plant viruses.a penicillinase (pnc)-based, enzyme-linked immunosorbent assay (elisa) was standardized to detect maize mosaic virus (mmv) in sorghum leaf extracts, peanut mottle virus (pmv) in pea leaf extracts, and tomato spotted wilt virus (tswv) in peanut leaf extracts. rabbit fc-specific antibodies were conjugated with pnc by a single step glutaraldehyde bridge. among several indicators tested, bromothymol blue (btb) was found suitable for measuring pnc activity under simulated conditions. two reagents, st ...19892592521
molecular cloning and terminal sequence determination of the s and m rnas of tomato spotted wilt virus.complementary dna to the genomic rna of tomato spotted wilt virus (tswv) was synthesized and cloned in either puc19 or lambda gt10. restriction endonuclease maps were constructed from cdna clones specific for the s and the m rna segments, extending 3.0 and 5.4 kbp respectively. the nucleotide sequences of the 3' and 5' termini of both rna molecules were determined. the s and m rnas contain inverted complementary repeats at their termini, probably involved in rna replication and in the formation ...19892607343
should tomato spotted wilt virus be considered as a possible member of the family bunyaviridae?from a comparison of published data on the properties of tomato spotted wilt virus and viruses in the family bunyaviridae, it is concluded that tomato spotted wilt virus should be considered as a possible member of the family.19846480313
effect of tiazofurin on tomato plants infected with tomato spotted wilt virus.tiazofurin (2-beta-d-ribofuranosylthiazole-4-carboxamide) was examined for its activity against tomato-spotted wilt virus (tswv) in tomato plants. solutions containing 50, 100, 200, 400 and 800 mg/l of the drug were sprayed onto the leaves. the results showed that 100 and 200 mg/l were the most efficient concentrations to suppress tswv infection, thereby delaying the appearance of systemic symptoms. the drug was more effective in controlling tswv infection when applied after than before virus in ...19846532320
inhibitory effect of distamycin-a and a pyrazino-pyrazine derivative on tomato spotted wilt virus.distamycin-a hydrochloride, a synthetic antibiotic, and 2,3-dihydroxy-6-bromo-pyrazino (2,3-beta) pyrazine derivative, were used against tomato spotted wilt virus (tswv) in tobacco plants. the drugs were applied to the leaves at concentrations of 200 and 400 mg/l. the results showed that both drugs delayed virus spread within the plant, retarding the appearance of systemic symptoms. a virus recovery test, carried out on primary leaves of phaseolus vulgaris cv. manteiga, showed that tswv replicat ...19836625560
effect of virazole (ribavirin) on tomato spotted wilt virus in two systemic hosts, tomato and tobacco.virazole (1, beta-d-ribofuranosyl-1,2,4-triazole-3-carboxamide = ribavirin), a synthetic nucleoside analogue, has been successfully used against tomato spotted wilt virus (tswv) in tomato and tobacco plants. in tomato, the most efficient concentration to control tsww was 500 mg/l while in tobacco, concentrations of 100 mg/l were sufficient to control systemic infection. when the plants did not show a systemic reaction virus could not be recovered from inoculated and treated tomato and tobacco pl ...19807356400
[tomato spotted wilt virus and the characteristics of its biology]. 1976781493
[electron microscope study of tomato spotted wilt virus on tabacco (author's transl)]. 19761018735
[electron microscopic examination of tomato spotted wilt virus (lycopersicum virus 3)]. 19751207518
purification and properties of tomato spotted wilt virus. 19744131950
protein composition of tomato spotted wilt virus. 19734126840
electron microscopy of developmental stages of tomato spotted wilt virus in plant cells. 19715543832
an electron microscope study of the distribution of tomato spotted wilt virus in systemically infected datura stramonium leaves. 19704923022
antiserum for tomato spotted wilt virus. 19684968990
tomato spotted wilt virus. 19684872838
a small infective component of tomato spotted wilt virus (strain e). 19675621951
serological studies on tomato spotted wilt virus (strains e and r1). 19675622015
morphology of tomato spotted wilt virus. 19665921647
preparation and properties of tomato spotted wilt virus (strain e). 19666005406
electron microscopy of vira-cabeca virus (brazilian tomato spotted wilt virus) within the host cell. 196514293317
purification and electron microscope studies of tomato spotted wilt virus (tswv) from tomato roots. 196414166124
electron microscopy of strain e of tomato spotted wilt virus and comments on its probable biosynthesis. 196414204698
partial amino acid composition of tomato spotted wilt virus, (strain e). 196414240390
purification and electron microscopy of tomato spotted-wilt virus. 196313971368
recombination experiments with strains a and e of tomato spotted wilt virus. 196113868589
further evidence for the transfer of character-determinants (recombination) between strains of tomato spotted wilt virus. 195513317855
physiological studies on thrips in relation to transmission of tomato spotted wilt virus. 195413219035
cross protection by strains of tomato spotted wilt virus and a new theory to explain it. 195413229839
thermal inactivation of tomato spotted wilt virus. 194620985881
inactivation of tomato spotted wilt virus by salicylate. 194620985882
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