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Title: Rapid Detection of Tobacco Rattle Tobravirus in Viruliferous Paratrichodorus allius from Greenhouse and Field Specimens

Author
item RIGA, E - WASHINGTON ST UNIV
item Larsen, Richard
item GUERRA, N - WASHINGTON ST UNIV
item EASTWELL, K - WASHINGTON ST UNIV
item Crosslin, James

Submitted to: Journal of Nematology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 7/9/2008
Publication Date: 1/5/2009
Citation: Riga, E., Larsen, R.C., Guerra, N., Eastwell, K., Crosslin, J. 2009. Rapid Detection of Tobacco Rattle Tobravirus in Viruliferous Paratrichodorus allius from Greenhouse and Field Specimens. Journal of Nematology. 41:60-63.

Interpretive Summary: The stubby root nematode is an important vector in potato crops in the Pacific Northwest. While feeding on potato tubers, it is also capable of transmitting Tobacco ringspot virus that causes the Corky ringspot disease, a disease that renders the tubers unmarketable. Using standard protocols requiring a greenhouse bioassay and serology, several weeks are required to determine whether or not nematodes collected from field samples carry the virus. To significantly reduce the time required to assay soil samples, DNA primers specific to the virus were designed for use in the real-time polymerase chain reaction technique. The technique is highly sensitive and specific, and can detect the virus in as few as five nematodes extracted from field samples. The new assay provides a means of virus detection in a single day.

Technical Abstract: The stubby root nematode, Paratrichodorus allius, is becoming of increasing importance to the potato industry in the Pacific Northwest of USA, because it can vector the Tobacco rattle virus (TRV), which is the causal agent of corky ringspot disease. The current method for determining if the stubby nematodes are viruliferous for TRV takes several weeks, requiring a glasshouse bioassay followed by an ELISA assay. Therefore, a rapid molecular technique has been developed using RT-PCR to identify viruliferous P. allius nematodes in less than 48 hours. A primer pair from the 16 K gene of TRV was used to detect TRV in as few as five adult female stubby nematodes.