Aphid Transmission of Plant Viruses

Stewart M. Gray1

1 USDA, ARS, Department of Plant Pathology, Cornell University, Ithaca, New York
Publication Name:  Current Protocols in Microbiology
Unit Number:  Unit 16B.1
DOI:  10.1002/9780471729259.mc16b01s10
Online Posting Date:  August, 2008
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Abstract

A majority of plant viruses are transmitted between hosts by insect vectors, and it is often important to use insect transmission in the laboratory to maintain virus isolates or to study virus‐vector‐plant interactions. Although many of these viruses can also be mechanically transmitted in the laboratory using infected sap, maintenance by mechanical transmission can often lead to changes in the virus, either minor changes in gene sequences or, in some cases, major deletions of genome sequences. These can affect both virus‐vector and virus‐host interactions. This unit describes some simple and practical methods for conducting virus transmission experiments using sap‐sucking insects. Curr. Protoc. Microbiol. 10:16B.1.1‐16B.1.10. © 2008 by John Wiley & Sons, Inc.

Keywords: plant virus; transmission

     
 
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Table of Contents

  • Introduction
  • Strategic Planning
  • Basic Protocol 1: Transmission from Infected Plant Material
  • Alternate Protocol 1: In Vitro–Transmission Assay
  • Support Protocol 1: Rearing Aphids
  • Commentary
  • Literature Cited
  • Figures
     
 
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Materials

Basic Protocol 1: Transmission from Infected Plant Material

  Materials
  • Aphids ( protocol 3) or other insects
  • Infected plant material
  • 1.5‐ml tubes, optional
  • Parafilm, optional
  • Healthy plants
  • Appropriate insecticide: insecticidal soaps, contact insecticides (e.g., pyrethroids), or systemic insecticides (e.g., organophosphates and carbamates)
  • Fine‐tip paint brush (e.g., round camel hair #2)
  • Shallow plastic or glass dishes with tight‐fitting lids, ∼10 cm in diameter and 2 cm deep
  • Cages to retain aphids on healthy plants during inoculation (e.g., see Fig. )
  • Fine white sand (sandbox sand, available in garden supply stores), optional

Alternate Protocol 1: In Vitro–Transmission Assay

  • Test solution of virus or test proteins: prepare in an appropriate buffer or water (vehicle determined by virus or protein used and tested alone to determine any effect on insect feeding or survival) containing 15% (w/v) sucrose and sterilize by passing through a 0.2‐µm filter
  • Feeding chamber (see Fig. ) consisting of:
    • Parafilm: sterilized with UV light (from several hours to overnight)
    • 1‐ to 2‐cm diameter plastic or glass tubes ∼2 to 3 cm in length
    • Rigid foam block
    • Screening material that prevents the smallest life stage of aphid from escaping but allows air movement; muslin may be used
    • Contact cement or other adhesive
  • Yellow light, optional

Support Protocol 1: Rearing Aphids

  Materials
  • Aphids, either collected from the wild or obtained from other researchers' colonies
  • Leaves from healthy plants (or moist filter papers)
  • Healthy plants
  • Fine‐tip paint brush (e.g., round camel hair #2)
  • Small containers with tight‐fitting lids
  • Plant cages (see Fig. )
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Figures

Literature Cited

Literature Cited
   Ammar, E.D. and Nault, L.R. 2002. Virus transmission by leafhoppers, planthoppers, and treehoppers (Auchenorrhyncha, Homoptera). Adv. Bot. Res. 36:141‐167.
   Blackman, R.L. and Eastop, V.F. 2000. Aphids on the World's Crops: An Identification and Information Guide. John Wiley & Sons, Chichester, United Kingdom.
   Blackman, R.L. and Eastop, V.F. 2006. Aphids on the World's Herbaceous Plants and Shrubs. John Wiley & Sons, Chichester, United Kingdom.
   Brown, J.K. and Czosnek, H. 2002. Whitefly transmission of plant viruses. Adv. Bot. Res. 36:65‐100.
   Dixon, A.F.G. 2005. Insect Herbivore‐Host Dynamics: Tree‐Dwelling Aphids. Cambridge University Press, Cambridge.
   Gray, S. and Gildow, F.E. 2003. Luteovirus‐aphid interactions. Annu. Rev. Phytopathol. 41:539‐566.
   Hogenhout, S.A., Redinbaugh, M.G., and Ammar, E.D. 2003. Plant and animal rhabdovirus host range: A bug's view. Trends Microbiol. 11:264‐271.
   Katsar, C. and Gray, S.M. 1999. Rearing aphids to use in virus‐vector studies. In Maintenance of Human Animal and Plant Pathogen Vectors (K. Maramorosch and F. Mahmood, eds.) pp. 183‐195. Science Publishers, Enfield, N.H.
   Kunkel, H. 1977. Membrane feeding systems in aphid research. In Aphids as Virus Vectors (K.F. Harris and K. Maramorosch, eds.) pp. 311‐338. Academic Press, New York.
   Ng, J.C.K. and Perry, K.L. 2004. Transmission of plant viruses by aphid vectors. Mol. Plant Pathol. 5:505‐511.
   Ullman, D.E., Whitfield, A.E., and German, T.L. 2005. Thrips and tospoviruses come of age: Mapping determinants of insect transmission. Proc. Natl. Acad. Sci. U.S.A. 102:4931‐4932.
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