Submitted to: Plant Pathology Journal
Publication Type: Peer reviewed journal
Publication Acceptance Date: 5/4/2006
Publication Date: 2/1/2007
Citation: Gillaspie Jr, A.G., Wang, M.L., Pinnow, D.L., Pittman, R.N. 2007. Polymerase chain reaction for detection of peanut mottle and peanut stripe viruses in arachis hypogaea l. germplasm seedlots. Plant Pathology Journal. 6(1)87-90. Interpretive Summary: A procedure was developed for detection of peanut mottle and peanut stripe viruses in seedlots of peanut. This mehtod involves a nucleic acid methodd called polymerase chain reaction to detect infected seeds using a pooling procedure to check for infected seedlots. Those lots determined to be infected are then tested further by a grid method to determine which seeds are infected. This method is sensitive and more rapid than the currently used serological method. An even faster method combining serology with the nucleic acid method has failed to work consistently. Thisprocedure should speed up the handling of seedlots of international origin that must be tested for virus contamination in order to protect the U.S. peanut crop.
Technical Abstract: Results of peanut mottle and peanut stripe virus detection studies indicated that peanut seedlots were best handled by extracting nucleic acid from samples of seed slices pooled from rows and columns of seeds arranged in a grid. A portion of these RNA samples were then combined (plate pooled sample) and tested via RT-PCR. This approach could detect one infected seed in a 96-seed sample. The samples from the positive seedlots were then tested to determine the location of the virus-infected seeds. A similar approach with IC-RT-PCR tests of samples from rows and columns ground in buffer failed to produce consistent results. Both the RT-PCR and the IC-RT-PCR approaches were compared to the results with DAS-ELISA tests of the individual seeds. The approach of testing plate-pooled samples followed by row and column sample tests by RT-PCR resulted in faster and most reliable testing of seedlots.