Design and application of two novel degenerate primer pairs for the detection and complete genomic characterization of potyviruses.
Two pairs of degenerate primers were designed from sequences within the potyviral CI (CIFor/CIRev) and HC-Pro-coding regions (HPFo/HPRev), and these were shown to be highly specific to members of the genus Potyvirus. Using the CIFor and CIRev primers, three novel potyviruses infecting crop and weed species from Vietnam were detected, namely telosma mosaic virus (TelMV) infecting telosma (Telosma cordata, Asclepiadaceae), peace lily mosaic virus (PeLMV) infecting peace lily (Spathiphyllum patinii, Araceae) and wild tomato mosaic virus (WTMV) infecting wild tomato (Solanum torvum, Solanaceae). The fragments amplified by the two sets of primers enabled additional PCR and complete genomic sequencing of these viruses and a banana bract mosaic virus (BBrMV) isolate from the Philippines. All four viruses shared genomic features typical of potyviruses. Sequence comparisons and phylogenetic analyses indicated that WTMV was most closely related to chilli veinal mottle virus (ChiVMV) and pepper veinal mottle virus (PVMV), while PeLMV, TelMV and BBrMV were related to different extents to members of the bean common mosaic virus (BCMV) subgroup.
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Tropical Crops and Biocommodities Domain, Institute of Health and Biomedical Innovation, Queensland University of Technology, Brisbane, Australia.
Coombs
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Revill
P A
PA
Harding
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RM
Vu
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Dale
J L
JL
D017931:DNA Primers; D004279:DNA, Viral / Q000032:analysis*; D016679:Genome, Viral; D008969:Molecular Sequence Data; D018515:Plant Leaves / Q000821:virology; D016133:Polymerase Chain Reaction; D017800:Potyvirus / Q000145:classification / Q000235:genetics*; D017422:Sequence Analysis, DNA
D017931:DNA Primers; D004279:DNA, Viral