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Immunofluorescent Localization of Tobacco Ringspot Nepovirus in the Vector Nematode Xiphinema americanum

September 1998 , Volume 88 , Number  9
Pages  885 - 889

Shouhua Wang and Rose C. Gererich

Department of Plant Pathology, 217 Plant Science Building, University of Arkansas, Fayetteville 72701

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Accepted for publication 26 May 1998.

An indirect immunofluorescent technique was developed to localize tobacco ringspot nepovirus (TRSV) in the vector nematode Xiphinema americanum sensu stricto. A population of this nematode that efficiently transmitted TRSV was given an acquisition access period of 10 days on TRSV-infected cucumber. Treatment of fragments of viruliferous nematodes with a polyclonal antiserum against TRSV followed by fluorescein isothiocyanate-conjugated goat anti-rabbit immunoglobulin G resulted in virus-specific bright fluorescence only in the lumen of the stylet extension and esophagus. Virus-specific fluorescent signals were observed in the virus-retention region of 44% of the nematode fragments examined. The percentage of nematodes labeled with virus-specific fluorescence increased as the acquisition access period increased from 0 to 22 days; the increase paralleled the increase in the transmission efficiency of the nematode population. Visualization of the entire virus-retention region of individual nematodes within a population of vector or nonvector nematodes provides a rapid and simple means of monitoring specific attachment of plant viruses.

Additional keyword: detection .

© 1998 The American Phytopathological Society