Differentiation of two closely related furoviruses using the polymerase chain reaction

• Published in: Phytopathology 1994, Vol.84 (11), p.1366-1369
• Main Authors: Rush, C.M. (Texas Agricultural Experiment Station, Bushland, TX), French, R, Heidel, G.B
• Format: Article
• Language: English
• Subjects: AMPLIFICATION CHAINE POLYMERASE; BETA VULGARIS; Biological and medical sciences; Biotechnology; DIAGNOSTIC; DIAGNOSTICO; Fundamental and applied biological sciences. Psychology; Generalities. Techniques. Transmission, epidemiology, ecology. Antiviral substances, control; Microbiology; PATHOTYPE; PATOTIPOS; Phytopathology. Animal pests. Plant and forest protection; Plant viruses and viroids; PODER PATOGENO; POTYVIRUS; POUVOIR PATHOGENE; REACCION DE CADENAS DE POLIMERASA; Replicative cycle, interference, host-virus relations, pathogenicity, miscellaneous strains; SINTOMAS; SYMPTOME; Virology; VIRUS DE LAS PLANTAS; VIRUS DES VEGETAUX
• ISSN: 0031-949X; 1943-7684
• 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 BSBMVPCR 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