Skip to main content

Nepovirus Isolation and RNA Extraction

  • Protocol
Plant Virology Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 81))

Abstract

Viruses of the nepovirus genus are classified within the family of Comoviridae, along with the comoviruses and the fabaviruses. At the time of writing, there are some 27 confirmed members of the genus, with eight or so viruses that, following further investigation, may be confirmed as members (1). These viruses infect a very wide range of plant species, producing symptoms that include mottles, mosaics, ringspots, and systemic necrosis; often infections are symptomless, particularly in the later stages. All members are transmitted through soil by free-living nematodes, principally of Longidorus or Xiphinema species, feeding on roots. They are also transmitted and disseminated through seed and pollen of both crop species and weeds. It is the combination of a mode of transmission (nematode) and virus morphology (polyhedral) that provides the name of the genus.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Murant, A. F. (1981) Nepoviruses, in Handbook of Plant Virus Infections and Comparative Diagnosis (Kurstak, E., ed.), Elsevier/North Holland, Amsterdam, pp. 197–238.

    Google Scholar 

  2. Roberts, I. M. (1988) The structure of particles of tobacco ringspot nepovirus: evidence from electron microscopy. J. Gen. Virol. 69, 1831–1840.

    Article  Google Scholar 

  3. Murant, A. F., Taylor, M., Duncan, G. H., and Raschke, J. H. (1981) Improved estimates of molecular weight of plant virus RNA by agarose gel electrophoresis and electron microscopy after denaturation with glyoxal. J. Gen. Virol. 53, 321–332.

    Article  CAS  Google Scholar 

  4. Mayo, M. A., Harrison, B. D., Murant, A. F., and Barker, H. (1973) Cross-linking of RNA induced by ultra-violet irradiation of particles of raspberry ringspot virus. J. Gen. Virol. 19, 155–159.

    Article  CAS  Google Scholar 

  5. Greif, C., Hemmer, O., and Fritsch, C. (1988) Nucleotide sequence of tomato black ring virus RNA-1. J. Gen. Virol. 69, 1517–1529.

    Article  CAS  Google Scholar 

  6. Demangeat, G., Greif, C., Hemmer, O., and Fritsch, C. (1990) Analysis of the in vitro cleavage products of the tomato blackring virus RNA-1-encoded 250K polyprotein. J. Gen. Virol. 71, 1649–1654.

    Article  PubMed  CAS  Google Scholar 

  7. Hemmer, O., Greif, C., Dufourcq, P., Reinbolt, J., and Fritsch, C. (1995) Functional characterisation of the proteolytic activity of the tomato black ring nepovirus RNA-1-encoded polyprotein. Virology 206, 362–371.

    Article  PubMed  CAS  Google Scholar 

  8. Demangeat, G., Hemmer, O., Reinbolt, J., Mayo, M. A., and Fritsch, C. (1992) Virus-specific proteins in cells infected with tomato black ring nepovirus: evidence for proteolytic processing in vivo. J. Gen. Virol. 73, 1609–1614.

    Article  PubMed  CAS  Google Scholar 

  9. Meyer, M., Hemmer, O., Mayo, M. A., and Fritsch, C. (1986) The nucleotide sequence of tomato black ring virus RNA-2. J. Gen. Virol. 67, 1257–1271.

    Article  CAS  Google Scholar 

  10. Demangeat, G., Hemmer, O., Fritsch, C., Le Gall, O., and Candresse, T. (1991) In vitro processing of the RNA-2-encoded polyprotein of two nepoviruses: tomato black ring virus and grapevine chrome mosaic virus. J. Gen. Virol. 72, 247–252.

    Article  PubMed  CAS  Google Scholar 

  11. Brault, V., Hibrand, L., Candresse, T., Le Gall, O., and Dunez, J. (1989) Nucleotide sequence and genetic organisation of Hungarian grapevine chrome mosaic nepovirus RNA2. Nucleic Acids Res. 17, 7809–7819.

    Article  PubMed  CAS  Google Scholar 

  12. Serghini, M. A., Fuchs, M., Pinck, M., Reinbolt, J., Walter, B., and Pinck, L. (1990) RNA2 of grapevine fanleaf virus: sequence analysis and coat protein cistron location. J. Virol. 71, 1433–1441.

    Article  CAS  Google Scholar 

  13. Blok, V. C., Wardell, J., Jolly, C. A., Manoukian, A., Robinson, D. J., Edwards, M. L., and Mayo, M. A. (1992) The nucleotide sequence of RNA-2 of raspberry ringspot nepovirus. J. Gen. Virol. 73, 2189–2194.

    Article  PubMed  CAS  Google Scholar 

  14. Rott, M. E., Tremaine, J. H., and Rochon, D. M (1991) Nucleotide sequence of tomato ringspot virus RNA-2. J. Gen. Virol. 72, 1505–1514.

    Article  PubMed  CAS  Google Scholar 

  15. Kreiah, S., Strunk, G., and Cooper, J. I. (1994) Sequence analysis and location of capsid proteins within RNA 2 of strawberry latent ringspot virus. J. Gen. Virol. 75, 2527–2532.

    Article  PubMed  CAS  Google Scholar 

  16. Mayo, M. A., Barker, H., and Harrison, B. D. (1982) Specificity and properties of the genome linked proteins of the nepoviruses. J. Gen. Virol. 59, 149–162.

    Article  CAS  Google Scholar 

  17. Salazar, L. F. and Harrison, B. D. (1978) The relationship of potato black ringspot virus to tobacco ringspot and allied viruses. Ann. Appl. Biol. 90, 387–394.

    Article  Google Scholar 

  18. Rezaian, M. A. and Francki, R. I. B. (1973) Replication of tobacco ringspot virus. I. Detection of a low molecular weight double stranded RNA from infected plants. Virology 56, 238–249.

    Article  PubMed  CAS  Google Scholar 

  19. Harrison, B. D. and Barker, H. (1978) Protease-sensitive structure needed for infectivity of nepovirus RNA. J. Gen. Virol. 40, 711–715.

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 1998 Humana Press Inc.

About this protocol

Cite this protocol

Wood, K.R. (1998). Nepovirus Isolation and RNA Extraction. In: Foster, G.D., Taylor, S.C. (eds) Plant Virology Protocols. Methods in Molecular Biology™, vol 81. Humana Press. https://doi.org/10.1385/0-89603-385-6:197

Download citation

  • DOI: https://doi.org/10.1385/0-89603-385-6:197

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-385-6

  • Online ISBN: 978-1-59259-566-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics