VIEW ARTICLE

Techniques

Differentiation of Two Closely Related Furoviruses Using the Polymerase Chain Reaction. C. M. Rush, Texas Agricultural Experiment Station, P.O. Drawer 10, Bushland, TX 79012; R. French(2), and G. B. Heidel(3). (2)USDA-ARS, 406 Plant Science, Lincoln, NE 68583; (3)Texas Agricultural Experiment Station, P.O. Drawer 10, Bushland, TX 79012. Phytopathology 84:1366-1369. Accepted for publication 30 August 1994. Copyright 1994 The American Phytopathological Society. DOI: 10.1094/Phyto-84-1366.

Oligonucleotide primers based on published sequence data for beet necrotic yellow vein virus (BNYVV) were synthesized for use in the reverse transcriptase polymerase chain reaction (RT-PCR) to differentiate beet soilborne mosaic virus (BSBMV) from BNYVV. Primers designed for the 3’ end of BNYVV RNA 1 were effective in PCR amplification of a product of the predicted size, approximately 1,056 bp, from extracts of plants infected by BNYVV. The same primer pair also directed the amplification of a PCR product of approximately 1,000 bp from extracts of plants infected by BSBMV. If extracts from plants infected with BNYVV were mixed with those from plants infected with BSBMV, the primer pair allowed the amplification of only BNYVV. In addition to the slight size difference, the BSBMV product could be distinguished from the BNYVV product by digestion with ThaI, which cleaved the BSBMV product but not the BNYVV product. The BSBMV RT-PCR product was partially sequenced, and primers specific for BSBMV were synthesized. The primers directed the amplification of a PCR product of the predicted size, approximately 691 bp, only with extracts from plants infected by BSBMV. Only one PCR product of the size expected for BSBMV was produced from extracts containing both BSBMV and BNYVV. The BSBMV PCR product obtained with the BSBMV-specific primers could be digested by ThaI. PCR products of similar size were amplified using the BSBMV primers and extracts of several isolates of BSBMV differing in geographic origin and symptom phenotype.