Characterisation of a novel nucleorhabdovirus infecting alfalfa (Medicago sativa).

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  • Additional Information
    • Source:
      Publisher: BioMed Central Country of Publication: England NLM ID: 101231645 Publication Model: Electronic Cited Medium: Internet ISSN: 1743-422X (Electronic) Linking ISSN: 1743422X NLM ISO Abbreviation: Virol J Subsets: MEDLINE
    • Publication Information:
      Original Publication: [London] : BioMed Central, 2004-
    • Subject Terms:
      Genome, Viral*; Cell Nucleus/*virology ; Medicago sativa/*virology ; Rhabdoviridae/*genetics; Austria ; High-Throughput Nucleotide Sequencing ; Microscopy, Electron ; Open Reading Frames ; Plant Diseases/virology ; RNA, Viral/genetics ; Real-Time Polymerase Chain Reaction ; Rhabdoviridae/ultrastructure ; Sequence Analysis, DNA ; Viral Proteins/genetics ; Virion/genetics
    • Abstract:
      Background: Nucleorhabdoviruses possess bacilliform particles which contain a single-stranded negative-sense RNA genome. They replicate and mature in the nucleus of infected cells. Together with viruses of three other genera of the family Rhabdoviridae, they are known to infect plants and can be transmitted by arthropod vectors, during vegetative propagation, or by mechanical means. In 2010, an alfalfa (Medicago sativa) plant showing virus-like symptoms was collected from Stadl-Paura, Austria and sent to Julius Kühn Institute for analysis.
      Methods: Electron microscopy (EM) of leaf extracts from infected plants revealed the presence of rhabdovirus-like particles and was further used for ultrastructural analyses of infected plant tissue. Partially-purified preparations of rhabdovirus nucleocapsids were used for raising an antiserum. To determine the virus genome sequence, high throughput sequencing (HTS) was performed. RT-PCR primers were designed to confirm virus infection and to be used as a diagnostic tool.
      Results: EM revealed bacilliform virions resembling those of plant-infecting rhabdoviruses. HTS of ribosomal RNA-depleted total RNA extracts revealed a consensus sequence consisting of 13,875 nucleotides (nt) and containing seven open reading frames (ORFs). Homology and phylogenetic analyses suggest that this virus isolate represents a new species of the genus Nucleorhabdovirus (family Rhabdoviridae). Since the virus originated from an alfalfa plant in Austria, the name alfalfa-associated nucleorhabdovirus (AaNV) is proposed. Viroplasms (Vp) and budding virions were observed in the nuclei of infected cells by EM, thus confirming its taxonomic assignment based on sequence data.
      Conclusions: In this study, we identified and characterised a new nucleorhabdovirus from alfalfa. It shared only 39.8% nucleotide sequence identity with its closest known relative, black currant-associated rhabdovirus 1. The virus contains an additional open reading frame (accessory gene) with unknown function, located between the matrix protein and the glycoprotein genes. Serological and molecular diagnostic assays were designed for future screening of field samples. Further studies are needed to identify other natural hosts and potential vectors.
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    • Grant Information:
      2015-F-172 International Euphresco; German Egyptian Research Long Term Scholarship International Deutscher Akademischer Austausch Dienst Kairo
    • Contributed Indexing:
      Keywords: alfalfa-associated nucleorhabdovirus; electron microscopy; high throughput sequencing; lucerne; rhabdovirus
    • Accession Number:
      0 (RNA, Viral)
      0 (Viral Proteins)
    • Publication Date:
      Date Created: 20190501 Date Completed: 20190613 Latest Revision: 20200225
    • Publication Date:
      20240105
    • Accession Number:
      PMC6489223
    • Accession Number:
      10.1186/s12985-019-1147-3
    • Accession Number:
      31036009