User Ratings

Easy to Follow Achieved Expected Results Overall Rating Add your comments

Aphid Transmission of Plant Viruses

Stewart M. Gray¹

¹USDA, 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

GO TO THE FULL TEXT:

PDF or HTML at Wiley Interscience

Are you the author of this protocol? Login or register and return to this page.

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

GO TO THE FULL PROTOCOL:

PDF or HTML at Wiley Interscience

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

GO TO THE FULL PROTOCOL:

PDF or HTML at Wiley Interscience

Materials

Basic Protocol: Transmission from Infected Plant Material

Materials

Aphids (Support Protocol) 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. 16B.1.1)
Fine white sand (sandbox sand, available in garden supply stores), optional

Alternate Protocol: In Vitro–Transmission Assay

Additional Materials (also see Basic Protocol)

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. 16B.1.2) 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: 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. 16B.1.1)

Looking for materials? Suppliers Guide

GO TO THE FULL PROTOCOL:

PDF or HTML at Wiley Interscience

Figures

Figure 16B.1.1

Examples of insect cages appropriate for small insects. Cages are used to retain aphids on healthy plants during inoculation. They may enclose an entire plant or group of plants, or they may confine aphids to a leaf or area of a single plant.

View Image
Figure 16B.1.2

Device for feeding aphids on solutions through a Parafilm membrane.

View Image

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.

GO TO THE FULL PROTOCOL:

PDF or HTML at Wiley Interscience

Looking for Answers?

Do you have tips, tricks, or improvements to share?

Join the Conversation

saif (not verified)

Posted on June 23, 2009 - 3:08am

Studies must be conducted to find out weather the whlole body of aphids contains virus or only the mouth parts cos Their parasitoids are not reported to have the virus

User Ratings