Use of conserved genomic regions and degenerate primers in a PCR-based assay for the detection of members of the genus Caulimovirus.
The genus Caulimovirus consists of several distinct virus species with a double-stranded DNA genome that infect diverse plant species. A comparative analysis of the sequences of known Caulimovirus species revealed two regions that are conserved in all Caulimovirus species with the exception of Strawberry vein banding virus. Degenerate primers based on these two regions were designed and tested in a polymerase chain reaction-based assay for broad spectrum detection of members of this genus. Cauliflower mosaic virus, Figwort mosaic virus and three distinct caulimoviruses associated with dahlia (Dahlia variabilis) were used to show the utility of this test in detecting diverse caulimoviruses. The primer pair gave an amplicon of expected size (840bp). Amplicons from each virus were cloned and sequenced to verify their identity. The primer pair and the PCR assay provide approach for the broad spectrum detection of several members of the genus Caulimovirus.
Pappu
H R
HR
Department of Plant Pathology, Washington State University, PO Box 646430, Pullman, WA 99164-6430, USA. hrp@wsu.edu
Druffel
K L
KL
D017796:Caulimovirus / Q000235:genetics / Q000302:isolation & purification*; D017124:Conserved Sequence; D017931:DNA Primers / Q000235:genetics*; D004279:DNA, Viral / Q000235:genetics*; D010935:Plant Diseases / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D004279:DNA, Viral