Detection of petunia vein-clearing virus: model for the detection of DNA viruses in plants with homologous endogenous pararetrovirus sequences.
A number of cases of plant virus sequence integration into host plant genome have been reported. In at least two cases, endogenous pararetrovirus sequences are correlated strongly with subsequent episomal virus infection and there is circumstantial evidence that this also occurs for Petunia vein-clearing virus (PVCV). The detection of viruses is a critical component of plant health and therefore, it is important to have diagnostic procedures that differentiate between the detection of encapsidated viral DNA and homologous sequences in the host genome. PCR-based detection methods targeted at PVCV DNA have been tested and particular attention was paid to design controls that would indicate the existence of host DNA in the reaction. The use of ion-exchange chromatography for the partial purification of plant viruses from other cellular components, including chromosomal DNA, is described. The methods tested for PVCV detection are used to illustrate general principles for the specific detection of virus infections in host plants that carry homologous virus sequences in their genomes.
Harper
Glyn
G
Department of Disease and Stress Biology, John Innes Centre, Colney Lane, NR47UH, Norwich, UK. glyn.harper@bbsrc.ac.uk
Richert-Pöggeler
Katja R
KR
Hohn
Thomas
T
Hull
Roger
R
D002852:Chromatography, Ion Exchange; D004267:DNA Viruses / Q000302:isolation & purification*; D018744:DNA, Plant / Q000032:analysis; D004279:DNA, Viral / Q000032:analysis*; D018745:Genome, Plant; D032306:Petunia / Q000821:virology*; D018515:Plant Leaves / Q000737:chemistry; D010942:Plant Viruses / Q000302:isolation & purification; D010944:Plants / Q000821:virology; D016133:Polymerase Chain Reaction / Q000379:methods; D012190:Retroviridae / Q000737:chemistry* / Q000235:genetics; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D016662:Virus Integration
D018744:DNA, Plant; D004279:DNA, Viral