Synthesis of infectious transcripts of blueberry scorch carlavirus in vitro

Lawrence, Diane M.; Hillman, Bradley I.

doi:10.1099/0022-1317-75-9-2509

Cited by 14 publications

(6 citation statements)

References 26 publications

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“…Aphids can also transmit the virus to Chenopodium quinoa and C. amaranticolor [48] as well as N. occidentalis [55]. Mechanical transmission has been successful using infectious transcripts [56]. Blueberries are typically asexually propagated by rooted cuttings.…”

Section: Aphid-bornementioning

confidence: 99%

New and Emerging Viruses of Blueberry and Cranberry

Martín

Polashock

Tzanetakis

2012

Viruses

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Blueberry and cranberry are fruit crops native to North America and they are well known for containing bioactive compounds that can benefit human health. Cultivation is expanding within North America and other parts of the world raising concern regarding distribution of existing viruses as well as the appearance of new viruses. Many of the known viruses of these crops are latent or asymptomatic in at least some cultivars. Diagnosis and detection procedures are often non-existent or unreliable. Whereas new viruses can move into cultivated fields from the wild, there is also the threat that devastating viruses can move into native stands of Vaccinium spp. or other native plants from cultivated fields. The aim of this paper is to highlight the importance of blueberry and cranberry viruses, focusing not only on those that are new but also those that are emerging as serious threats for production in North America and around the world.

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Section: Aphid-bornementioning

confidence: 99%

New and Emerging Viruses of Blueberry and Cranberry

Martín

Polashock

Tzanetakis

2012

Viruses

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“…Among carlaviruses, an infectious RNA transcribed in vitro from the full-length cDNA clone was first reported in BlScV in 1994 [19]. To date, the construction of an infectious full-length cDNA clone has been reported in a limited number of carlaviruses, including poplar mosaic virus [20], PVM [21], chrysanthemum virus B [22], cowpea mild mottle virus [23], and pea streak virus [24].…”

Section: Introductionmentioning

confidence: 99%

Construction and characterization of an infectious cDNA clone of potato virus S developed from selected populations that survived genetic bottlenecks

Hataya

2019

Virol J

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BackgroundInfectious cDNA clones are a powerful tool for studies on RNA viruses using reverse genetics. Potato virus S (PVS) is a carlavirus with a worldwide distribution. Although the complete genome sequences of many PVS isolates have been reported, the construction of an infectious cDNA clone of PVS is yet to be reported. The aim of this study is the development and molecular characterization of an infectious cDNA clone of PVS.MethodsA full-length cDNA clone pPVS-H-FL-AB was constructed by connecting eight cDNA clones of PVS isolate H95. Capped RNA transcripts from pPVS-H-FL-AB and a modified clone pPVS-H-FL-H, containing the consensus genome sequence of PVS-H95, proved to be non-infectious. Therefore, a full-length cDNA clone pPVS-H-FL-β was reconstructed from PVS-H00, isolated from PVS-H95 populations by repeating a single local lesion isolation in Chenopodium quinoa three times; PVS-H00 appeared to be a selected variant that survived genetic bottlenecks. The sequence of cDNA clone pPVS-H-FL-β was determined as the genome sequence of PVS-H00 and compared with the consensus sequence of PVS-H95 genome.ResultsAll Nicotiana occidentalis plants inoculated with ≥0.2 μg capped RNA transcripts from pPVS-H-FL-β developed symptoms on upper leaves, as observed with PVS-H00 inoculation. Similar levels of viral genomic and subgenomic RNAs and coat protein were detected in systemically infected leaves. Sequence comparison of PVS-H95 and PVS-H00 revealed 370 nucleotide polymorphisms (4.4% of the entire genome sequence), causing 91 amino acid substitutions in six open reading frames (ORFs). The infectivity of chimeric RNAs derived from recombinants between the two cDNA clones revealed that the lack of infectivity of pPVS-H-FL-H transcripts was due to ORF1, which encodes replicase and harbors 80 amino acid substitutions compared with pPVS-H-FL-β. Approximately 71.3% amino acid substitutions in replicase were located within the variable region of unknown function between the putative methyltransferase and ovarian tumor-like protease domains.ConclusionsThis is the first report of the development of an infectious cDNA clone of PVS. Our analyses suggest that PVS population within a plant exists as quasispecies and the replicase sequence diversity of PVS obstruct the construction of a full-length infectious cDNA clone.Electronic supplementary materialThe online version of this article (10.1186/s12985-019-1124-x) contains supplementary material, which is available to authorized users.

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“…CVB has slightly flexuous rod shaped particle of 680 9 12 nm and is transmitted by aphid Myzus persiceae in a non-persistent manner and is also sap transmissible. Carlavirus genome comprises of single stranded monopartite, plus sense ssRNA molecule *8.8 kb in size with a poly (A) tail at 3 0 -terminus and a cap structure or a monophosphate at their 5 0 -terminus [4,5,15,31]. The genomic RNA is encapsidated by a single type of coat protein (CP) with molecular weight 34 kDa.…”

Section: Introductionmentioning

confidence: 99%

Intermolecular Interactions of Chrysanthemum virus B Coat Protein: Implications for Capsid Assembly

2011

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Chrysanthemum virus B coat protein constitutes the viral capsid which, besides other functions, encapsulates and protects the viral nucleic acid. We have demonstrated homotypic interaction of the coat protein subunits, essentially important for dimer formation, which is the first step during capsid assembly in vivo and in vitro. Interaction capacity of full length and truncated protein has been investigated and important regions have been identified through protein-protein interaction in yeast and by coimmunoprecipitation assays. Complete coat protein was found to interact strongly with similar subunits. Constructs with 102 amino acids from the N-terminal and 64 amino acids from C-terminal were found to be inconsequential for dimer formation as they did not show any interaction with similar subunits or with full length protein when analysed for b-galactosidase or histidine prototrophy. Results suggest that the region of 98-184 amino acids from the middle plays an important role in the process, probably without the involvement of N-and C-terminals.

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Synthesis of infectious transcripts of blueberry scorch carlavirus in vitro

Cited by 14 publications

References 26 publications

New and Emerging Viruses of Blueberry and Cranberry

New and Emerging Viruses of Blueberry and Cranberry

Construction and characterization of an infectious cDNA clone of potato virus S developed from selected populations that survived genetic bottlenecks

Intermolecular Interactions of Chrysanthemum virus B Coat Protein: Implications for Capsid Assembly

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