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  • 1
    Electronic Resource
    Electronic Resource
    Detection of grapevine leafroll-associated closterovirus III by molecular hybridization (1994)
    Oxford, UK : Blackwell Publishing Ltd
    Plant pathology 43 (1994), S. 0 
    Details
    ISSN: 1365-3059
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Highly purified double-stranded (ds) RNA was obtained from cortical scrapings of mature canes of vines infected by grapevine leafroll-associated closterovirus III (GLRaV III) using phenol-chloroform extraction, chromatography on CF-11 cellulose minicolumns and enzymatic digestion. Complementary (c) DNA fragments of various lengths, obtained by random priming denatured dsRNA templates, were cloned into the plasmid pUC-18 at the Smal site in Escherichia coli strain DH5α. Two 32P-labelled cDNA clones denoted p16ds (c. 1100 bp) and p23ds (c. 1500 bp) were successfully used as probes for detecting GLRaV III sequences in grapevine extracts from leaves and petioles, or cortical tissues. Probe p23ds was virus-specific and did not hybridize with total RNA from healthy controls, or from vines infected by grapevine leafroll-associated closterovirus I (GLRaV I), or with genomic RNA from purified grapevine closterovirus A (GVA) and B (GVB). A riboprobe (pGEM23ds) transcribed from p23ds in transcription vector pGEM3zf specifically recognized GLRaV III sequences, but not GLRaV I or GVA sequences, in extracts from differently infected vines. Moreover, in Northern blots, the same probe hybridized also with smaller dsRNA components, which may be replicative forms of subgenomic RNAs.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1365-3059.1994.tb00557.x
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  • 2
    Electronic Resource
    Electronic Resource
    Nucleotide sequence of the 3′ terminal region of the RNA of two filamentous grapevine viruses (1994)
    Springer
    Archives of virology 137 (1994), S. 249-261 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary The 3′ terminal region of grapevine virus A (GVA) and grapevine virus B (GVB), encompassing 1883 and 2136 nucleotides, respectively, was sequenced by the deoxynucleotide chain termination method. Three putative open reading frames (ORF) were identified in both genomic viral RNAs, denoted 1 to 3 in the 5′ to 3′ direction. ORF 1 encoded a polypeptide with estimated Mr of 31 kDa (GVA) and 36.5 kDa (GVB), possessing the G/D motif of the “30 K superfamily” movement proteins, and showing good alignments with putative movement proteins of trichoviruses and capilloviruses. ORF 2 was identified as the coat protein (CP) cistron, coding for polypeptides with an estimated Mr of 21.5 kDa (GVA) and 21.6 kDa (GVB). These CPs showed substantial sequence homology with one another and with CPs of tricho- and capilloviruses, but not of closteroviruses. ORF 3 potentially coded for two small polypeptides with estimated Mr of 10 kDa (GVA) and 14 kDa (GVB). The ORF 3 product of GVB (14 K), but not that of GVA, shared some homology with the 3′ terminal polypeptides of different plant viruses that exhibit the “zinc finger domain” of proteins with nucleic acid-binding properties. GVA and GVB have many properties in common with trichoviruses but possess an extra open reading frame (ORF 3). Whether this finding may have a bearing on the classification of these viruses is unclear. However, until the taxonomic significance of this difference in genome structure is established, it seems plausible to include GVA and GVB as tentative species in theTrichovirus genus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01309473
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  • 3
    Electronic Resource
    Electronic Resource
    Immunodetection and subcellular localization of the proteins encoded by ORF 3 of grapevine viruses A and B (2000)
    Springer
    Archives of virology 145 (2000), S. 1535-1542 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary.  Polyclonal antisera were raised to Escherichia coli-expressed ORF3 products (putative movement proteins) of Grapevine virus A (GVA) and Grapevine virus B (GVB) (genus Vitivirus), and used for their immunodetection in infected plants. Western blot analysis of subfractionated cellular compartments showed that the distribution of both proteins was comparable to that of plant virus movement proteins, as they were transiently present in a crude membrane fraction and accumulated in a cell wall-enriched fraction. The GVA ORF3-encoded protein, but not the comparable GVB protein, was also present in large amount in a cytoplasmic soluble fraction. Intracellular immunogold labelling localized these proteins in the cell wall and plasmodesmata of infected cells and, especially for GVA, in association with cytoplasmic virus aggregates.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050070074
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  • 4
    Electronic Resource
    Electronic Resource
    Grapevine virus A: nucleotide sequence, genome organization, and relationship in the Trichovirus genus (1997)
    Springer
    Archives of virology 142 (1997), S. 417-423 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary. The 5′ terminal region of the genomic RNA of grapevine virus A (GVA), a tentative member of the Trichovirus genus, encompassing 5 466 nucleotides, was sequenced. Evidence was obtained that the RNA is capped. Two putative open reading frames (ORF) were identified: ORF 1 that codes for a 194 kDa polypeptide with conserved motifs of replication-related proteins of positive-strand RNA viruses, and ORF 2 that encodes a 19 kDa polypeptide with no significant homology with protein sequences from databases. This polypeptide, however, showed 44% similarity with the product expressed by a comparable ORF present in grapevine virus B (GVB). GVA genome had the same size and structural organization as that of GVB. It also had the same size of the genome of apple chlorotic leaf spot virus (ACLSV), the type species of the Trichovirus genus, but differed substantially in the number (5 versus 3), size, and order of genes. Differences existed also in the degree of sequence homology between polymerases, which did not cluster together in phylogenetic trees. Definitive (ACLSV, PVT) and tentative (GVA, GVB) trichovirus species differ molecularly, biologically and epidemiologically to an extent that warrants the taxonomic revision of the genus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050050088
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  • 5
    Electronic Resource
    Electronic Resource
    Infectious cDNA clones of two grapevine viruses (2000)
    Springer
    Archives of virology 145 (2000), S. 397-405 
    Details
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary.  Full-length cDNA copies of the genomes of Grapevine virus A (GVA) and Grapevine virus B (GVB) under the control of bacteriophage T7 promoter have been synthesized, which were refractory to cloning in Escherichia coli. However, both transcribed cDNAs were infectious when mechanically inoculated to Nicotiana plants. A full-length cDNA copy of GVB was engineered in pCass2, a plasmid containing a partially duplicated copy of the Ca35S promoter, but was rather unstable in Escherichia coli. No infection of Nicotiana plants was obtained following mechanical inoculation but detached Nicotiana leaves, inoculated by particle bombardment, supported the multiplication of a GVB isolate seemingly identical to the wild-type used for cloning. Nicotiana seedling inoculated with sap expressed from these leaves became infected showing typical GVB symptoms. Transient transcription of Ca35S driven cDNA clones was also detected by RT-PCR in leaves of the grapevine hybrid LN33 following inoculation by particle bombardment. The availability of infectious cDNA clones of GVA and GVB constitutes a tool for the study of genome expression and pathogenesis, and for the ultimate establishment of the aetiological role of these viruses.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s007050050031
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  • 6
    Electronic Resource
    Electronic Resource
    Characterization of a pothos (Scindapsus aureus) virus with unusual properties (1995)
    Springer
    European journal of plant pathology 101 (1995), S. 171-182 
    Details
    ISSN: 1573-8469
    Keywords: Scindapsus aureus ; Tombusviridae ; PoLV ; virus purification ; physicochemical properties ; serology ; cytopathology ; dsRNA ; cDNA ; molecular probes
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract A virus for which the name of pothos latent virus (PoLV) is proposed, was isolated by inoculation of sap from symptomless plants ofScindapsus aureus. PoLV had isometric particlesc. 30 nm in diameter, a monopartite genome consisting of a non polyadenylated, single-stranded RNA moleculec. 4,300 nucleotides in length, constitutingc. 17% of the particle weight, and a single type of coat protein subunit with aM r ofc. 40,000 Daltons. The biological properties (host range reactions) of PoLV resembled those ofTombusviridae for it infected most of the artificial hosts locally, inducing symptoms recalling those elicited by several species of the above family. Like tombus- and carmoviruses, PoLV had two subgenomic RNAs which, however, differed in size from those of both genera. The dsRNA pattern was also distinctly different. Cytopathological features recalled those of tombusviruses except for the lack of multivesicular inclusion bodies. PoLV was serologically related to, but distinct from twoCarmovirus (i.e., galinsoga mosaic and Ahlum waterborne viruses) and threeTombusvirus species (i.e. eggplant mottled crinkle, Sikte waterborne and Lato river viruses). Thus, PoLV had properties somewhat intermediate between those ofTombusvirus andCarmovirus genera but bridged the two taxa through the serological relationship with some of their species. The taxonomic position of PoLV is still undetermined. It must await the results of molecular investigations now underway.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01874763
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