Submitted to: Phytopathology
Publication Type: Abstract Only
Publication Acceptance Date: April 1, 2004
Publication Date: July 1, 2004
Citation: Gillaspie Jr, A.G., Wang, M.L., Pittman, R.N., Pinnow, D.L. 2004. Development of a grid rt-pcr method for detection of peanut mottle(pemv) and peanut stripe viruses (pstv) in peanut germ plasm. [abstract]. Phytopathology. 94:534 Technical Abstract: A high throughput method, using three dimensional pooling in a 96-well format, has been developed for detection of PeMV and PStV for peanut seet lots. Samples of seeds and leaves were pooled by rows and columns and extracted by Qiagen miniprep method for total RNA. The plate-pooled sample was a proportional sample from all rows and columns. Plate-pooled samples testing negative by RT-PCR were assumed free of virus requiring no further testing. The pooled-row and column samples were tested when the plate-pooled sample was positive so that virus-infected samples could be located in the grid at the intersect of the row and column. The results from RT-PCR are consistent with the results from individual seed samples tested by DAS-ELISA. Similar results were obtained with leaf tissue. The results obtained by RT-PCR and DAS-ELISA reflect the symptoms observed in the seedlings arising from the tested seeds. The method is sensitive enough to detect and select one infected seed among the 96-plate-pooled samples. The method will be useful for testing newly introduced seed lots.