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Title: Testing the physiological barriers to viral transmission in aphids using microinjection

Author
item TAMBORINDEGUY, CECILIA - Cornell University
item Gray, Stewart
item JANDER, GEORG - Boyce Thompson Institute

Submitted to: Journal of Visualized Experiments
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/30/2008
Publication Date: 5/14/2008
Citation: Tamborindeguy, C., Gray, S.M., Jander, G. 2008. Testing the physiological barriers to viral transmission in aphids using microinjection. Journal of Visualized Experiments. Available: http://www.jove.com/details.php?id=700 doi: 10.3791/700. J Vis Exp. 15.

Interpretive Summary: The viruses that cause barley yellow dwarf disease and potato leafroll virus are economically serious virus disease of small grain crops and potatoes, respectively. They are not controlled by any type of chemical pesticide or agronomic practice and resistance to these viruses is limited and not complete. Understanding how these viruses are transmitted by their insect vectors may identify weaknesses that can be exploited to control this disease. This video and accompanying material describes technologies that are extremely useful for researchers attempting to study the movement of virus through the aphid and attempting to discover points in the aphid that the virus is blocked from further movement. Our lab has used these techniques over the past 20 years to discover the pathway of virus movement and various tissue barriers to virus movement. This has helped develop novel ways to prevent virus transmission between plants.

Technical Abstract: 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.