Publications

[tomato spotted wilt virus and the characteristics of its biology].		1976	781493
[electron microscope study of tomato spotted wilt virus on tabacco (author's transl)].		1976	1018735
[electron microscopic examination of tomato spotted wilt virus (lycopersicum virus 3)].		1975	1207518
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 ...	1992	1346784
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 ...	1992	1368791
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 ...	1992	1385787
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 ...	1992	1402797
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 ...	1992	1430055
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 ...	1992	1431808
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 ...	1992	1545223
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 ...	1992	1600236
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 ...	1992	1645150
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 ...	1990	1689996
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 ...	1990	1693160
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 ...	1991	1826573
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 ...	1991	1840613
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, ...	1991	1895058
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 ...	1991	1919523
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 ...	1991	1993884
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 ...	1991	2024478
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 ...	1990	2273383
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 ...	1989	2592521
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 ...	1989	2607343
protein composition of tomato spotted wilt virus.		1973	4126840
purification and properties of tomato spotted wilt virus.		1974	4131950
tomato spotted wilt virus.		1968	4872838
an electron microscope study of the distribution of tomato spotted wilt virus in systemically infected datura stramonium leaves.		1970	4923022
antiserum for tomato spotted wilt virus.		1968	4968990
electron microscopy of developmental stages of tomato spotted wilt virus in plant cells.		1971	5543832
a small infective component of tomato spotted wilt virus (strain e).		1967	5621951
serological studies on tomato spotted wilt virus (strains e and r1).		1967	5622015
morphology of tomato spotted wilt virus.		1966	5921647
preparation and properties of tomato spotted wilt virus (strain e).		1966	6005406
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.	1984	6480313
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 ...	1984	6532320
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 ...	1983	6625560
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 ...	1980	7356400
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 ...	1993	7686004
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 ...	1993	7763861
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. ...	1995	7772806
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%) ...	1995	7772807
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 ...	1995	7831835
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 ...	1995	7871744
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 ...	1994	8006111
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 ...	1993	8113346
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 ...	1994	8128638
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 ...	1993	8212587
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 ...	1993	8429298
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 ...	1993	8445364
the nucleotide sequence of the m rna segment of tomato spotted wilt virus, a bunyavirus with two ambisense rna segments.		1993	8468562
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 ...	1995	8553550
sequence analysis of the 5' ends of tomato spotted wilt virus n mrnas.	messenger rnas transcribed from the tomato spotted wilt virus (tswv) rna genome have characteristic extra non-templated heterogeneous sequences at their 5' ends which may be the result of a cap-snatching event involving cellular mrnas. in order to investigate the genetic origin of these extra sequences and to gain more insight in the process of cap-snatching as performed by tswv, nucleocapsid protein (n) mrnas derived from the tswv s rna were cloned and sequenced. twenty clones were obtained whi ...	1996	8629946
engineered rna-mediated resistance to tomato spotted wilt virus is sequence specific.	transgenic plants were produced that expressed a wide range of randomly chosen sequences of the tripartite tomato spotted wilt virus (tswv) rna genome or its complement. testing the progenies of these plants revealed that only transgenic expression of n or ns(m) gene sequences resulted in resistance to tswv.	1996	8672819
the complete nucleotide sequence and genome organization of the m rna segment of peanut bud necrosis tospovirus and comparison with other tospoviruses.	the m rna of peanut bud necrosis virus (pbnv; synonym groundnut bud necrosis virus) is 4801 nucleotides in length. it comprised two orfs in an ambisense organization and terminal inverted repeats. the 3' large orf (3363 nucleotides in the virus-complementary strand) encoded a protein with a predicted size of 127.2 kda which was identified as the glycoprotein precursor (gp) of the g1 and g2 glycoproteins. a comparison of the deduced amino acid sequence of gp revealed 37% identity and 58-59% simil ...	1996	8811036
a polyclonal antiserum against a recombinant viral protein combines specificity with versatility.	a polyclonal rabbit antiserum was obtained to the nucleoprotein of tomato spotted wilt tospovirus expressed as a recombinant fusion protein in e. coli. in indirect plate trapping elisa, the antiserum gave similar titres against purified tswv nucleocapsids in native form, the fusion protein and the carrier protein. the crude antiserum was also tested by western blotting, indirect plate trapping elisa and immunogold electron microscopy of thin sections: purified immunoglobulins were tested by das- ...	1996	8882651
an improved method for the detection of tospoviruses using the polymerase chain reaction.	a reverse transcription-polymerase chain reaction (rt-pcr)-based assay for the detection of tomato spotted wilt virus (tswv) has been improved and extended to enable the detection of additional tospoviruses. in addition to tswv-specific primers, two further pairs of primers have been designed, one pair which specifically detects impatiens necrotic spot virus (insv) and another which detects all tospoviruses tested, including tswv, insv, tomato chlorotic spot virus and groundnut ringspot virus. a ...	1996	8919828
clustering of argentinean tospoviruses with existing species in the genus by sequence analysis of a 450-nucleotide rna region of the n gene.	the genomic diversity of argentine tospoviruses from different geographical areas, and from several distinct crops was analysed here. for each isolate, rt-pcr, cloning and sequencing of a 450 nt fragment of the n gene were performed. comparisons of rna and predicted amino acid sequence identity and similarity were made. a partial sequence of the n gene was able to classify our local isolates within three tospovirus species previously described (tomato spotted wilt virus, tswv; tomato chlorotic s ...	1996	9035370
characterization of rna-mediated resistance to tomato spotted wilt virus in transgenic tobacco plants expressing ns(m) gene sequences.	transgenic nicotiana tabacum plants expressing rna sequences of the tomato spotted wilt virus ns(m) gene, which encodes the putative viral movement protein, were found to be highly resistant to infection with the virus. expression of untranslatable as well as anti-sense rna of the ns(m) gene resulted in resistance levels as high as those in plants expressing translatable rna sequences. for all three types of transgenic plants resistance levels of up to 100% were reached in the s2 progeny. these ...	1997	9037142
serological comparison of tospoviruses with polyclonal antibodies produced against the main structural proteins of tomato spotted wilt virus.	a new purification procedure for the tospoviruses of serogroups ii and iii was developed. sds-polyacrylamide gel electrophoresis of purified tomato spotted wilt virus (tswv; serogroup i), groundnut ringspot virus (grsv: serogroup ii), tomato chlorotic spot virus (tcsv; serogroup ii) and impatiens necrotic spot virus (insv; serogroup iii) showed that the glycoprotein g2 of serogroup ii members differs significantly in size from that of the serogroup i virus. western immunoblot analysis using poly ...	1997	9170504
multiplication of tomato spotted wilt virus in primary cell cultures derived from two thrips species.	primary cell cultures prepared from embryos of the thrips species frankliniella occidentalis and thrips tabaci were tested for their potential to support replication of tomato spotted wilt virus (tswv). using polyclonal antibodies against the viral nucleocapsid protein (n) and indirect immunofluorescent staining, discrete spots with strong signals were observed in the cytoplasm at 48 h post-inoculation in the cell cultures of a f. occidentalis, and a t. tabaci population which failed to transmit ...	1997	9178497
nontarget dna sequences reduce the transgene length necessary for rna-mediated tospovirus resistance in transgenic plants.	rna-mediated virus resistance has recently been shown to be the result of post-transcriptional transgene silencing in transgenic plants. this study was undertaken to characterize the effect of transgene length and nontarget dna sequences on rna-mediated tospovirus resistance in transgenic plants. transgenic nicotiana benthamiana plants were generated to express different regions of the nucleocapsid (n) protein of tomato spotted wilt (tswv) tospovirus. transgenic plants expressing half-gene segme ...	1997	9223349
a protoplast system for studying tomato spotted wilt virus infection.	a plant protoplast system for studying tomato spotted wilt tospovirus (tswv) infection was established and tested. using polyethylene glycol-mediated inoculation with highly infectious tswv particles, generally 50% or more of nicotiana rustica protoplasts were infected. in these cells viral rna and viral protein synthesis became detectable at 16 h post-inoculation (p.i.) and continued at least until 90 h p.i. both the structural viral proteins [nucleoprotein (n) and the envelope glycoproteins g1 ...	1997	9225052
cladistic analysis of tospovirus using molecular characters.	the genus tospovirus was thought to be composed only of the tomato spotted wilt virus (tswv), but now at least four tospovirus species have been proposed based on serological and molecular data. a classification of tospoviruses has been proposed taking into account global similarities of the n gene and n protein sequences of 7 isolates of tospovirus. because phylogenetic analyses based on global similarities can lead to classifications which do not mirror the genealogy of the group, we have empl ...	1997	9242593
map-based cloning of the tomato genomic region that spans the sw-5 tospovirus resistance gene in tomato.	two yeast artificial chromosomes (yacs) containing genomic dna from tomato have been isolated using ct220, an rflp marker which is tightly linked to the tomato spotted wilt virus resistance gene, sw-5. high-resolution mapping of the yac ends and internal yac probes demonstrated that one of the yac clones, ty257 (400 kb), spans sw-5. by chromosome walking in a cosmid library, the position of sw-5 has been delimited within the yac to a maximal chromosomal segment of 100 kb, spanned by nine overlap ...	1997	9349703
an antiviral protein from bougainvillea spectabilis roots; purification and characterisation.	an antiviral protein active against mechanical transmission of tomato spotted wilt virus was identified in the root tissues of bougainvillea spectabilis willd. bougainvillea antiviral protein i (bap i) was purified to apparent homogeneity from the roots of bougainvillea by ammonium sulphate precipitation, cm- and deae-sepharose chromatography and reverse phase hplc. bap i is a highly basic protein (pi value > 8.6) with an mr of 28,000. the n-terminal sequence of bap i showed homology with other ...	1998	9612957
early events of tomato spotted wilt transcription and replication in protoplasts.	a protoplast transfection system for tomato spotted wilt tospovirus (tswv) was established by peg-mediated infection of tobacco protoplasts. analysis of viral rna synthesis revealed an asymmetric production of viral (v) and viral-complementary (vc) strands of all three genomic rna segments and a separation in time of subgenomic transcription during the first 72 hr post infection (p.i.). synthesis of vc-rna was detectable 8-10 hrs prior to v-rna production. during the first 72 h of infection the ...	1998	9699207
characterization of the nucleic acid binding properties of tomato spotted wilt virus nucleocapsid protein.	tomato spotted wilt tospovirus (tswv) is the type member of the plant-infecting viruses of the genus tospovirus in the family bunyaviridae. the three tswv rnas are encapsidated with nucleocapsid (n) protein to form ribonucleoprotein (rnp) which serves as the template for viral transcription and replication. regions of the open reading frame coding for the n protein on the small (s) rna were subcloned into pet protein expression vectors and expressed in escherichia coli bl21 (de3) cells. full-len ...	1998	9705250
molecular characterization of tomato spotted wilt virus defective interfering rnas and detection of truncated l proteins.	junction sites of 25 different defective interfering (di) rnas of tomato spotted wilt virus (tswv) were characterized. the di rnas varied in size from 2.0 to 5.2 kilobases (kb) and contained a single internal deletion. the absence of di rnas smaller than 2 kb suggested a size constraint for the survival of tswv di rnas. this hypothesis was reinforced by the finding of a dimeric di rna formed by two 1.6-long monomers linked head to tail. three types of junction sites were found, one type originat ...	1998	9721243
characterization of the large (l) rna of peanut bud necrosis tospovirus.	the nucleocapsids purified from peanut plants systemically infected with peanut bud necrosis virus (pbnv), a member of the genus tospovirus, contained both viral(v) and viral complementary(vc) sense l rnas. defective forms of l rna containing 'core polymerase region' were observed. the full length l rna of pbnv was sequenced using overlapping cdna clones. the 8911 nucleotide l rna contains a single open reading frame (orf) in the vc strand, and encodes a protein of 330 kda. at the 5' and 3' term ...	1998	9930194
tomato spotted wilt virus particle morphogenesis in plant cells.	a model for the maturation of tomato spotted wilt virus (tswv) particles is proposed, mainly based on results with a protoplast infection system, in which the chronology of different maturation events could be determined. by using specific monoclonal and polyclonal antisera in immunofluorescence and electron microscopy, the site of tswv particle morphogenesis was determined to be the golgi system. the viral glycoproteins g1 and g2 accumulate in the golgi prior to a process of wrapping, by which ...	1999	9971812
tissue tropism related to vector competence of frankliniella occidentalis for tomato spotted wilt tospovirus.	the development of tomato spotted wilt tospovirus (tswv) infection in the midgut and salivary glands of transmitting and non-transmitting thrips, frankliniella occidentalis, was studied to elucidate tissue tropism and the virus pathway within the body of this vector. immunohistological techniques used in this study showed that the midgut, foregut and salivary glands were the only organs in which virus accumulated. the first signals of infection, observed as randomly distributed fluorescent granu ...	1999	10073714
functional expression in bacteria and plants of an scfv antibody fragment against tospoviruses.	recombinant antibodies expressed in plants ('plantibodies'), directed against crucial antigens and addressed to the right cell compartment, may be able to protect against viral diseases. moreover, antibody fragments produced in bacteria or plants may provide low cost reagents for immunodiagnosis.	1999	10231089
alfalfa mosaic virus rnas serve as cap donors for tomato spotted wilt virus transcription during coinfection of nicotiana benthamiana.	tomato spotted wilt virus (tswv) was shown to use alfalfa mosaic virus (amv) rnas as cap donors in vivo during a mixed infection in nicotiana benthamiana. by use of nested reverse transcription-pcr, tswv n and nss mrnas provided with capped leader sequences derived from all four amv rnas could be cloned and sequenced. the sequence specificity of the putative tswv endonuclease involved is discussed.	1999	10233983
tomato spotted wilt virus in ornamental plants, vegetables and weeds in the czech republic.	the occurrence of tomato spotted wilt virus (tswv) in horticulture crops and weeds in the czech republic has been studied in 1992-1997. during this period tswv was established in 91 plant species. virus identity was based on the host range, serology and electron microscopy. natural tswv infection was detected in glasshouses where the main vector frankliniella occidentalis was present too. the most frequently tswv-infected plant species were chrysanthemum morifolium and zantedeschia sp. among veg ...	1998	10358737
a minimum length of n gene sequence in transgenic plants is required for rna-mediated tospovirus resistance.	we showed previously that transgenic plants with the green fluorescent protein (gfp) gene fused to segments of the nucleocapsid (n) gene of tomato spotted wilt virus (tswv) displayed post-transcriptional gene silencing of the gfp and n gene segments and resistance to tswv. these results suggested that a chimeric transgene composed of viral gene segments might confer multiple virus resistance in transgenic plants. to test this hypothesis and to determine the minimum length of the n gene that coul ...	2000	10640563
a caps marker to assist selection of tomato spotted wilt virus (tswv) resistance in pepper.	the hypersensitive resistance to tomato spotted wilt virus (tswv) in pepper is determined by a single dominant gene (resistant allele: tsw) in several capsicum chinense genotypes. in order to facilitate the selection for this resistance, four rapd (among 250 10-mer primers tested) were found linked to the tsw locus using the bulked segregant analysis and 153 f2 individuals. a close rapd marker was converted into a codominant cleaved amplified polymorphic sequence (caps) using specific pcr primer ...	2000	10701122
simplified, rapid method for cloning of virus-binding polypeptides (putative receptors) via the far-western screening of a cdna expression library using purified virus particles.	a simplified, alternative method for cloning virus-binding polypeptides (receptor candidates) is described. the method is based on a far-western assay using purified tomato spotted wilt tospovirus (tswv, bunyaviridae) for screening a lambda-phage cdna expression library. the western flower thrips, frankliniella occidentalis pergande, the principal vector of tswv, in which the virus replicates, was used for library construction. using this method several virus-binding polypeptides were identified ...	2000	10785290
genetic mapping of the tsw locus for resistance to the tospovirus tomato spotted wilt virus in capsicum spp. and its relationship to the sw-5 gene for resistance to the same pathogen in tomato.	the tsw gene conferring dominant resistance to the tospovirus tomato spotted wilt virus (tswv) in capsicum spp. has been tagged with a random amplified polymorphic dna marker and mapped to the distal portion of chromosome 10. no mapped homologues of sw-5, a phenotypically similar dominant tswv resistance gene in tomato, map to this region in c. annuum, although a number of sw-5 homologues are found at corresponding positions in pepper and tomato. the relationship between tsw and sw-5 was also ex ...	2000	10830267
comparative genetics of disease resistance within the solanaceae.	genomic positions of phenotypically defined disease resistance genes (r genes) and r gene homologues were analyzed in three solanaceous crop genera, lycopersicon (tomato), solanum (potato), and capsicum (pepper). r genes occurred at corresponding positions in two or more genomes more frequently than expected by chance; however, in only two cases, both involving phytophthora spp., did genes at corresponding positions have specificity for closely related pathogen taxa. in contrast, resistances to ...	2000	10835406
immunoprecipitation of a 50-kda protein: a candidate receptor component for tomato spotted wilt tospovirus (bunyaviridae) in its main vector, frankliniella occidentalis.	a 50-kda protein that binds to viral particles in solid-phase assays and that is recognized by anti-idiotypic antibodies made against anti-viral glycoproteins g1/g2 (anti-ids) has been proposed as a receptor candidate for tomato spotted wilt tospovirus (tswv) in its main thrips vector, frankliniella occidentalis pergande (bandla et al., 1998. phytopathology 88, 98-104). here we show the immunoprecipitation of the 50-kda protein by anti-ids and by an anti-g1/g2-tswv conjugate - a new immunoprecip ...	2000	10867190
a single chimeric transgene derived from two distinct viruses confers multi-virus resistance in transgenic plants through homology-dependent gene silencing.	we showed previously that 218 and 110 bp n gene segments of tomato spotted wilt virus (tswv) that were fused to the non-target green fluorescent protein (gfp) gene were able to confer resistance to tswv via post-transcriptional gene silencing (ptgs). n gene segments expressed alone did not confer resistance. apparently, the gfp dna induced ptgs that targetted n gene segments and the incoming homologous tswv for degradation, resulting in a resistant phenotype. these observations suggested that mu ...	2000	10900050
real-time rt-pcr fluorescent detection of tomato spotted wilt virus.	a real-time reverse transcription-polymerase chain reaction assay based on taqman chemistry was developed for the detection and quantification of tomato spotted wilt virus (tswv). this method enabled sensitive, reproducible and specific detection of tswv in 'leaf soak' and total rna extracts from infected plants. taqman reliably detected tswv in as little as 500 fg total rna. the assay was 10-fold more sensitive than visualisation of ethidium bromide-stained bands following agarose gel electroph ...	2000	10921836
tomato spotted wilt virus glycoproteins exhibit trafficking and localization signals that are functional in mammalian cells.	the glycoprotein precursor (g1/g2) gene of tomato spotted wilt virus (tswv) was expressed in bhk cells using the semliki forest virus expression system. the results reveal that in this cell system, the precursor is efficiently cleaved and the resulting g1 and g2 glycoproteins are transported from the endoplasmic reticulum (er) to the golgi complex, where they are retained, a process that could be blocked by tunicamycin. expression of g2 alone resulted in transport to and retention in the golgi c ...	2001	11134314
gst fusion proteins cause false positives during selection of viral movement protein specific single chain antibodies.	glutathione s-transferase (gst) fusion proteins are used frequently for investigating protein-protein and protein-dna interactions. the present study demonstrates that the use of gst fusion proteins caused false positives during selection of phage-displayed single-chain antibody fragments (scfvs) specific for three domains of the movement protein (ns(m)) of tomato spotted wilt virus (tswv). to identify and exclude the false positives when using gst as a fusion partner linked to the antigen of in ...	2001	11164495
overcoming host- and pathogen-mediated resistance in tomato and tobacco maps to the m rna of tomato spotted wilt virus.	a viral genetic system was used to map the determinants of the ability of tomato spotted wilt virus (tswv) to overcome the r gene (sw-5) in tomato and the resistance conferred by the nucleocapsid gene of tswv (n gene) in tobacco. a complete set of reassortant genotypes was generated from tswv isolates a and d. tswv-a was able to overcome the sw-5 gene in tomato and the tswv n gene in tobacco, whereas tswv-d was repressed by both forms of resistance. the ability to overcome both forms of resistan ...	2001	11204788
constitutive expression of interferon-induced human mxa protein in transgenic tobacco plants does not confer resistance to a variety of rna viruses.	mxa is a key component in the interferon-induced antiviral defense in humans. after viral infections, mxa is rapidly induced and accumulates in the cytoplasm. the multiplication of many rna viruses, including all bunyaviruses tested so far, is inhibited by mxa. these findings prompted us to express mxa in plants in an attempt to create resistance to tospoviruses. here, we report the generation of transgenic tobacco plants that constitutively express mxa under the control of the 35 s cauliflower ...	2000	11206971
variability of the n-protein and the intergenic region of the s rna of tomato spotted wilt tospovirus (tswv).	the genetic heterogeneity of the n protein gene and the intergenic region (igr) of the s rna from tomato spotted wilt virus (tswv) isolates, collected in bulgaria, were compared with isolates from other parts of the world. the results substantiated the highly conserved nature of the n protein. twenty six independent sequences revealed only seven variable amino acid positions, common to all isolates. the type of amino acids present in these positions seems to be independent of the geographical or ...	2001	11346302
in vivo analysis of the tswv cap-snatching mechanism: single base complementarity and primer length requirements.	requirements for capped leader sequences for use during transcription initiation by tomato spotted wilt virus (tswv) were tested using mutant alfalfa mosaic virus (amv) rnas as specific cap donors in transgenic nicotiana tabacum plants expressing the amv replicase proteins. using a series of amv rna3 mutants modified in either the 5'-non-translated region or in the subgenomic rna4 leader, sequence analysis revealed that cleaved leader lengths could vary between 13 and 18 nucleotides. cleavage oc ...	2001	11350944
rt-pcr for detecting five distinct tospovirus species using degenerate primers and dsrna template.	rt-pcr procedures for detection of multiple species of tospovirus from plant tissues were investigated. downstream primers were designated from the 3' untranslated sequences of the s rna. an upstream primer was designated from the degenerated sequences of the nucleocapsid protein. approximately 450 bp dna fragments were detected when tomato spotted wilt virus (tswv)- or impatiens necrotic spot virus (insv)- infected tissues were examined. approximately 350 bp dna fragments were detected when wat ...	2001	11445145
visual representation by atomic force microscopy (afm) of tomato spotted wilt virus ribonucleoproteins.	atomic force microscopy (afm) allows the observation of biological material without fixation procedures. here we present afm images of ribonucleoproteins (nucleocapsids) derived from a plant infecting rna virus (tomato spotted wilt virus, tswv), which have been recorded in contact mode. the nucleocapsids, prepared from systemically infected leaves of tobacco, were spreaded on a glass surface and dried in air, and appeared as regularly formed rings, resembling the proposed pseudocircular and panh ...	2001	11767945
factors determining vector competence and specificity for transmission of tomato spotted wilt virus.	the competence of a frankliniella occidentalis and a thrips tabaci population to transmit tomato spotted wilt virus (tswv) was analysed. adults of the f. occidentalis population transmitted this virus efficiently, whereas those of the thelytokous t. tabaci population failed to transmit. tswv replicated in the midgut of the larvae of both populations after ingestion of virus; however, lower amounts accumulated in t. tabaci larvae than in f. occidentalis larvae. the virus was almost undetectable i ...	2002	11842261
the detection of tomato spotted wilt virus (tswv) in individual thrips using real time fluorescent rt-pcr (taqman).	tomato spotted wilt virus (tswv) is an important virus, economically in the uk, causing damaging disease in ornamental and vegetable crops. the virus is vectored by several species of thrips, most importantly the western flower thrips (frankliniella occidentalis pergande [thysanoptera: thripidae]). the vector thrips themselves constitute a damaging pest and are difficult to control completely. monitoring thrips numbers is an important part of the control of virus, but does not give information o ...	2002	11849682
expression of the movement protein of tomato spotted wilt virus in its insect vector frankliniella occidentalis.	tomato spotted wilt virus (tswv) is able to infect both its botanical hosts and its insect vector (thrips). in plant tissue the ns(m) protein of tswv functions as viral movement protein (mp), aggregating into plasmodesma-penetrating tubules to establish cell-to-cell movement. as upon heterologous expression ns(m) was able to form similar tubules on the surface of insect (spodoptera frugiperda) cells, we have now investigated the expression and cellular manifestation of this protein in infected t ...	2002	12038691
induction of phenylpropanoid metabolism by pseudomonas fluorescens against tomato spotted wilt virus in tomato.	pseudomonas fluorescens (two native strains, one collection strain and their strain mixtures in all possible combinations) when applied through seed, seedling dip, soil and on leaf significantly reduced the tomato spotted wilt virus (tswv) disease. in p. fluorescens-treated plants, the peroxidase and phenylalanine ammonia-lyase activity increased. accumulation of phenolic compounds and lignin were shown to be increased in the p. fluorescens-treated plants. isoperoxidase native page indicated tha ...	2002	12058389
evaluation of metabolic detoxifying enzyme activities and insecticide resistance in frankliniella occidentalis.	the western flower thrips frankliniella occidentalis (pergande) is a very significant pest of a number of different agricultural crops in the south-east of spain. the importance of thrips as a pest is not due mainly to the direct damage inflicted on the plant, but to the loss in commercial value which occurs as a consequence of the development of dark spots caused by the tomato spotted wilt virus (tswv) which they transmit. the economic threshold is therefore almost zero, which enhances the prob ...	2002	12233183
the route of tomato spotted wilt virus inside the thrips body in relation to transmission efficiency.	the route of tomato spotted wilt virus (tswv) in the body of its vectors, frankliniella occidentalis and thrips tabaci (thysanoptera: thripidae) was studied during their development. first instar larvae were allowed, immediately upon hatching, to acquire virus from mechanically infected datura stramonium plants for 24 h. the rate of transmission by adults was determined in inoculation access feeding test on emilia sonchifolia leaf disks. thrips tissues were analysed for infection at 24 h interva ...	2002	12417949
identification of a novel rna silencing suppressor, nss protein of tomato spotted wilt virus.	rna silencing or post-transcriptional gene silencing (ptgs) in plants is known as a defense system against virus infection. several plant viruses have been shown to encode an rna silencing suppressor. using a green fluorescent protein-based transient suppression assay, we show that nss protein of tomato spotted wilt virus (tswv) has rna silencing suppressor activity. tswv nss protein suppressed sense transgene-induced ptgs but did not suppress inverted repeat transgene-induced ptgs. tswv nss pro ...	2002	12459466
purified tomato spotted wilt virus particles support both genome replication and transcription in vitro.	purified tomato spotted wilt virus particles were shown to support either genome replication or transcription in vitro, depending on the conditions chosen. transcriptional activity was observed only upon addition of rabbit reticulocyte lysate, indicating a dependence on translation. under these conditions rna molecules of subgenomic length were synthesized that hybridized to strand-specific probes for the n and nss genes. cloning of these transcripts demonstrated the presence of nonviral leader ...	2002	12490389
negative-strand tospoviruses and tenuiviruses carry a gene for a suppressor of gene silencing at analogous genomic positions.	posttranscriptional silencing of a green fluorescent protein (gfp) transgene in nicotiana benthamiana plants was suppressed when these plants were infected with tomato spotted wilt virus (tswv), a plant-infecting member of the bunyaviridae: infection with tswv resulted in complete reactivation of gfp expression, similar to the case for potato virus y, but distinct from that for cucumber mosaic virus, two viruses known to carry genes encoding silencing suppressor proteins. agrobacterium-based lea ...	2003	12502849
an unusual large intergenic region in the s-rna of a bulgarian tomato spotted wilt virus isolate.	the complete s-rna sequences of four bulgarian and one german isolate of tomato spotted wilt tospovirus were determined. all isolates show a high conservation in their n proteins, while the nss proteins and the intergenic regions (igr) are more variable. the bulgarian isolate 10hk96 has the largest s-rna (3364 nucleotides) among tomato spotted wilt viruses reported so far. the enlargement is based on an insertion of 365 nts in the igr that may have resulted from stuttering of the viral polymeras ...	2003	12536305