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The Journal of Visualized Experiments (JoVE) is a peer reviewed, PubMed-indexed video journal. Our mission is to increase the productivity of scientific research.

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Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

1, 1, 2

1Plant Pathology, Cornell University, 2Boyce Thompson Institute for Plant Research, Cornell University

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Cite this Article: Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

Tamborindeguy, C., Gray, S., Jander, G. Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection. J. Vis. Exp. (15), e700, doi:10.3791/700 (2008).

Abstract: Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

Potato loafroll virus (PLRV), from the family Luteoviridae infects solanaceous plants. It is transmitted by aphids, primarily, the green peach aphid. When an uninfected aphid feeds on an infected plant it contracts the virus through the plant phloem. Once ingested, the virus must pass from the insect gut to the hemolymph (the insect blood ) and then must pass through the salivary gland, in order to be transmitted back to a new plant. An aphid may take up different viruses when munching on a plant, however only a small fraction will pass through the gut and salivary gland, the two main barriers for transmission to infect more plants. In the lab, we use physalis plants to study PLRV transmission. In this host, symptoms are characterized by stunting and interveinal chlorosis (yellowing of the leaves between the veins with the veins remaining green). The video that we present demonstrates a method for performing aphid microinjection on insects that do not vector PLVR viruses and tests whether the gut is preventing viral transmission. The video that we present demonstrates a method for performing Aphid microinjection on insects that do not vector PLVR viruses and tests whether the gut or salivary gland is preventing viral transmission.

Protocol: Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

  1. To test if an aphid is able to vector a virus, we feed the aphids on infected tissue for 2 days.
  2. Then, we allow it to feed in a healthy plant for 5 days. The plant is then fumigated and after 3 weeks we start searching for symptoms.
  3. When we find an aphid that cannot vector a virus, we can assess which of the 2 barriers is stopping the virus using a microinjection procedure.
  4. The virus is injected directly into the hemocoel, thereby bypassing the gut transmission barrier.

Please visit Annual Reviews of Plant Biology for more information about this protocol and plant immunity to insect herbivory.

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Disclosures: Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

The authors have nothing to disclose.

Ask the Author: Testing the Physiological Barriers to Viral Transmission in Aphids Using Microinjection

2 Comments

Hi, I have been trying some aphid microinjection recently, and was wondering if anyone had any advice on calibrating the volume that is injected into the aphid? Also, does anyone have any problems with the glass microcapillary blocking after a few injections, would silanising the microcapillary help with this?

1

Reply

Posted by: MiriamNovember 3, 2008, 8:25 AM

Hi,
I'm planning to do micro-injection experiment using white fly as organism of choice. Can you please guide me about the procedure / protocol ? Also what part of white fly should be chosen for micro-injection ? Thanks

Amir Raza

2

Reply

Posted by: Amir R.December 8, 2010, 1:57 PM

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