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isolation of tobacco necrosis virus from strawberry leaves in the czech republic.leaves of symptomless fragaria ananassa duch cv. cacanská raná were grafted onto fragaria vesca indicator clones. thirty-five of 72 grafted indicator plants developed leaf mottle symptoms. isometric virus-like particles were observed in purified preparations from symptomatic leaves of f. vesca. the latter were mechanically inoculated to herbaceous host plants. a virus was successfully purified from nicotiana occidentalis 37 b symptomatic plants by differential and sucrose density gradient centri ...199810358734
apple chlorotic leaf spot virus 50 kda protein is targeted to plasmodesmata and accumulates in sieve elements in transgenic plant leaves.we investigated the in situ localization of the 50 kda protein encoded by orf2 of apple chlorotic leaf spot virus (aclsv) genome which is thought to be a movement protein. in immunogold electron microscopy of aclsv-infected chenopodium quinoa leaves, the 50 kda protein was localized on plasmodesmata and nearby cytoplasm. observation of transgenic nicotiana occidentalis leaves expressing the 50 kda protein fused to enhanced green fluorescent protein (egfp) by fluorescence and confocal laser scann ...199910664400
intracellular distribution, cell-to-cell trafficking and tubule-inducing activity of the 50 kda movement protein of apple chlorotic leaf spot virus fused to green fluorescent protein.the 50 kda protein (50kp) encoded by orf2 of apple chlorotic leaf spot virus (aclsv) fused to green fluorescent protein (gfp) was expressed transiently in cells of nicotiana occidentalis and chenopodium quinoa leaves. its intracellular distribution, cell-to-cell trafficking in leaf epidermis and tubule formation on the surface of protoplasts were analysed. the 50kp-gfp fluorescence was distributed as small irregular spots or a fibrous network structure on the periphery of epidermal cells and pro ...200010900048
mechanical transmission of poleroviruses.previously, transmission of poleroviruses has relied solely on the use of their aphid vectors. biolistic inoculation allowed for the first time the mechanical transmission of beet western yellows virus (bwyv) and potato leafroll virus (plrv) to several host plants. inoculation with purified preparations and viral rna extracts of plrv resulted in 30-50% systemically infected nicotiana occidentalis p1 plants and 15-30% infected nicotiana clevelandii plants. particle bombardment was also used succe ...200111164501
characterization and complete nucleotide sequence of strawberry mottle virus: a tentative member of a new family of bipartite plant picorna-like viruses.an isolate of strawberry mottle virus (smov) was transferred from fragaria vesca to nicotiana occidentalis and chenopodium quinoa by mechanical inoculation. electron micrographs of infected tissues showed the presence of isometric particles of approximately 28 nm in diameter. smov-associated tubular structures were also conspicuous, particularly in the plasmodesmata of c. quinoa. dsrna extraction of smov-infected n. occidentalis yielded two bands of 6.3 and 7.8 kbp which were cloned and sequence ...200211752720
analysis of the complete genome of peach chlorotic mottle virus: identification of non-aug start codons, in vitro coat protein expression, and elucidation of serological cross-reactions.the entire genome of peach chlorotic mottle virus (pcmv), originally identified as prunus persica cv. agua virus (4n6), was sequenced and analysed. pcmv cross-reacts with antisera to diverse viruses, such as plum pox virus (ppv), genus potyvirus, family potyviridae; and apple stem pitting virus (aspv), genus foveavirus, family flexiviridae. the pcmv genome consists of 9005 nucleotides (nts), excluding a poly(a) tail at the 3' end of the genome. five open reading frames (orfs) were identified wit ...200717891331
complementation analysis of triple gene block of potato virus s (pvs) revealed its capability to support systemic infection and aphid transmissibility of recombinant potato virus x.triple gene block (tgb) sequences derived from isolates of ordinary potato virus s (pvs-o) and chenopodium-systemic (pvs-cs) were analyzed. although the tgb sequences did not reveal any specific difference within the 7k protein, some specific differences within the 25k and 12k orfs were found. in order to investigate a possible functional divergence of pvs-o and pvs-cs tgb variants, these genes were propagated in chimeric potato virus x (pvx). both pvs tgb variants partly complemented pvx tgb in ...200919748533
the 3'-proximal part of the plum pox virus p1 gene determinates the symptom expression in two herbaceous host plants.three major strains of the plum pox virus (ppv) are the most important in europe: ppv-d, ppv-m, and ppv-rec. by combining the genomes of two different strains of ppv (ppv-d with ppv-rec; ppv-d with ppv-m), 20 inter-strain chimeric infectious clones (cicppv) were constructed. biological properties of cicppv were tested by inoculating them on different herbaceous host species susceptible to ppv. four of the seven species tested, exhibited visible symptoms. in nicotiana benthamiana all cicppv induc ...201222367316
host-specific accumulation and temperature effects on the generation of dimeric viral rna species derived from the s-rna of members of the tospovirus genus.polygonum ringspot virus (polrsv) is a recently characterized tospovirus reported in italy. northern blot analyses of polrsv infections in nicotiana benthamiana and tomato plants showed that a viral rna species with nearly twice the length of the small genomic rna (s-rna) accumulated abundantly in the former host, but was not detected in the latter. additional assays confirmed that biogenesis of this novel rna species was common to all polrsv isolates tested and also to an isolate of tomato spot ...201627600541
lucerne transient streak virus; a recently detected virus infecting alfafa (medicago sativa) in central saudi arabia.a survey was conducted to determine the status of lucerne transient streak virus (ltsv) in three high-yielding alfalfa regions in central saudi arabia (riyadh, qassim, and hail) during 2014. three hundred and eight symptomatic alfalfa, and seven sonchus oleraceus samples were collected. das-elisa indicated that 59 of these samples were positive to ltsv. two isolates of ltsv from each region were selected for molecular studies. rt-pcr confirmed the presence of ltsv in the selected samples using a ...201728167887
suppression of aggressive strains of 'candidatus phytoplasma mali' by mild strains in catharanthus roseus and nicotiana occidentalis and indication of similar action in apple trees.to study antagonistic interactions of 'candidatus phytoplasma mali' strains, graft inoculation of catharanthus roseus and nicotiana occidentalis was performed with mild strains 1/93vin and 1/93tab as suppressors and three aggressive strains as challengers. inoculation of the suppressors was carried out in either the cross-protection modus prior to grafting of the challengers or by co-inoculating suppressors and challengers. monitoring using multiplex real-time polymerase chain reaction assays re ...201424724815
strawberry crinkle virus, a cytorhabdovirus needing more attention from virologists.summary taxonomic relationship: a member of nonsegmented, negative-sense, single-stranded rna viruses of the genus cytorhabdovirus (type member: lettuce necrotic yellows virus), family rhabdoviridae, order mononegavirales. members of the family rhabdoviridae can infect vertebrates, invertebrates and plants. physical properties: virions are bacilliform, 74-88 nm in diameter and 163-383 nm in length with surface projections probably composed of trimers of the glycoprotein g, occurring in the cytop ...200020572980
interference of long-distance movement of grapevine berry inner necrosis virus in transgenic plants expressing a defective movement protein of apple chlorotic leaf spot virus.abstract transgenic nicotiana occidentalis plants expressing a movement protein (p50) and partially functional deletion mutants (deltaa and deltac) of the apple chlorotic leaf spot virus (aclsv) showed resistance to grapevine berry inner necrosis virus (ginv). the resistance is highly effective and ginv was below the level of detection in both inoculated and uninoculated upper leaves. in contrast, ginv accumulated in inoculated and uninoculated leaves of nontransgenic (nt) plants and transgenic ...200618943419
detection and characterisation of two novel vitiviruses infecting actinidia.two co-infecting novel vitiviruses from actinidia chinensis were identified from mechanically inoculated nicotiana occidentalis. both virus genomes were sequenced and share 64% nucleotide identity. their overall structure is typical of vitiviruses, with five open reading frames (orfs) and a polyadenylated 3' end. open reading frame 4 (orf4) encodes the coat protein, the most conserved gene of the vitiviruses, in which they share 75% amino acid identity, 61-68% with grapevine virus b, 55-59% with ...201222274622
the fifty distal amino acids of the 2a(hp) homing protein of grapevine fanleaf virus elicit a hypersensitive reaction on nicotiana occidentalis.avirulence factors are critical for the arm's race between a virus and its host in determining incompatible reactions. the response of plants to viruses from the genus nepovirus in the family secoviridae, including grapevine fanleaf virus (gflv), is well characterized although the nature and characteristics of a viral avirulence factor remains elusive. by using infectious clones of gflv strains f13 and ghu in a reverse genetics approach with wild-type, assortant and chimeric viruses, the determi ...201728387986
rapid and sensitive detection of apple stem pitting virus in apple trees through rna amplification and probing with fluorescent molecular beacons.abstract currently, detection of apple stem pitting virus (aspv; genus foveavirus) in apple trees for certification purposes occurs by woody indexing. this method requires a minimum of 12 to 24 weeks in greenhouse testing to up to 2 years in field testing. in this paper, the development of a single tube amplidet rna system for the rapid gel-free detection of aspv in apple tree tissues is described. the system relies on the specific amplification of the viral rna by nucleic acid sequence-based am ...200118943445
transgenic nicotiana occidentalis plants expressing the 50-kda protein of apple chlorotic leaf spot virus display increased susceptibility to homologous virus, but strong resistance to grapevine berry inner necrosis virus.abstract the 50-kda protein (p50) encoded by the open reading frame 2 of apple chlorotic leaf spot virus (aclsv), a putative movement protein, was expressed in transgenic nicotiana occidentalis plants. p50 in transgenic plants was mainly detected in a modified form in the cell wall fraction, similar to that in infected leaves. the p50-expressing plants (p50 plants) complemented the systemic spread of the p50-defective mutants of an infectious cdna clone of aclsv (pclsf), indicating that p50 in t ...200018944625
the 50-kda protein of apple chlorotic leaf spot virus interferes with intracellular and intercellular targeting and tubule-inducing activity of the 39-kda protein of grapevine berry inner necrosis virus.to understand why transgenic nicotiana occidentalis plants expressing a functional movement protein (mp) of apple chlorotic leaf spot virus (aclsv) show specific resistance to grapevine berry inner necrosis virus (ginv), the mps of aclsv (50kp) and ginv (39kp) were fused to green, yellow, or cyan fluorescent proteins (gfp, yfp, or cfp). these fusion proteins were transiently expressed in leaf cells of both transgenic (50kp) and nontransgenic (nt) plants, and the intracellular and intercellular t ...200312650450
cymbidium mosaic virus coat protein gene in antisense confers resistance to transgenic nicotiana occidentalis.the nucleotide sequence of the 3'-terminal region of the korean isolate of cymbidium mosaic virus (cymv-ca) from a naturally infected cattleya was determined. the sequence contains an open reading frame (orf) coding for the viral coat protein (cp) at the 3'-end and three other orfs (triple gene block or movement protein) of cymv. the cp gene encodes a polypeptide chain of 220 amino acids with a molecular mass of 23,760 da. the deduced cp sequence showed a strong homology with those of two cymvs ...199910672926
a simple and reliable protocol for the detection of apple stem grooving virus by rt-pcr and in a multiplex pcr assay.primers were identified which amplify specifically a 499 bp fragment in the coat protein coding region of apple stem grooving virus (asgv) genome. these primers were used in various rt-polymerase chain reaction (pcr) analyses for the detection of asgv in chenopodium quinoa, nicotiana occidentalis, and in species of malus and pyrus. isolates of asgv in malus and pyrus from locations in canada, china, israel, japan, nepal, pakistan, south africa, and the u.s.a. were reliably detected in leaf and b ...199910598077
properties of a filamentous virus isolated from grapevines affected by corky bark.a virus with highly flexuous filamentous particles c. 800 nm long, showing distinct transverse striations was isolated with high frequency (60%) by inoculation of nicotiana occidentalis with sap from grapevine accessions indexing positive for corky bark. the virus, for which the name grapevine virus b (gvb) is proposed, has an ssrna genome with mol. wt. of c. 2.5 x 10(6) da (c. 7600 nt) and coat protein subunits with mol. wt. of c 23,000 da. gvb has a very restricted herbaceous host range and wa ...19938503778
development of a full-length infectious clone of sunflower chlorotic mottle virus (sucmov).a full-length cdna clone (p35sucmov) of the sunflower chlorotic mottle virus common strain (sucmov-c) genomic rna was constructed. three cdna fragments covering the whole genome of sucmov-c were cloned between a cauliflower mosaic virus 35s promoter and a nopaline synthase terminator. mechanical inoculation of sunflower and nicotiana occidentalis seedlings with p35sucmov dna led to systemic infection. symptoms induced by p35sucmov were similar to those caused by the wild-type sucmov-c but appear ...201323081677
development of a full-genome cdna clone of citrus leaf blotch virus and infection of citrus plants.citrus leaf blotch virus (clbv), a member of the family flexiviridae, has a ~9-kb single-stranded, positive-sense genomic rna encapsidated by a 41-kda coat protein. clbv isolates are associated with symptom production in citrus including leaf blotching of dweet tangor and stem pitting in etrog citron (dweet mottle disease), and some isolates are associated with bud union crease on trifoliate rootstocks, but koch's postulates for this virus were not fulfilled. a full-genome cdna of clbv isolate s ...200819019007
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