Identification of Genes Involved in Replication and Movement of Peanut Clump Virus

The genome of peanut clump pecluvirus (PCV) consists of two messenger RNA components which contain, respectively, three and five open reading frames (ORFs). Inoculation of transcripts from full-length cDNA clones derived from the PCV RNAs showed that RNA-1 is able to replicate in the absence of RNA-...

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Bibliographic Details
Published in:Virology (New York, N.Y.) N.Y.), 1998-09, Vol.248 (2), p.312-322
Main Authors: Herzog, E., Hemmer, O., Hauser, S., Meyer, G., Bouzoubaa, S., Fritsch, C.
Format: Article
Language:English
Subjects:
Arachis - virology
Biological Transport
Chenopodium quinoa
DNA, Complementary - analysis
Genes, Reporter
Genes, Viral
Genome, Viral
Glucuronidase - genetics
GUS
Mosaic Viruses - genetics
Mosaic Viruses - pathogenicity
Mosaic Viruses - physiology
Mutagenesis
Nicotiana benthamiana
Peanut clump virus
pecluvirus
plant virus
Protoplasts - virology
RNA replication
RNA, Viral - biosynthesis
RNA, Viral - physiology
Viral Proteins - genetics
Viral Proteins - physiology
virus movement
Virus Replication
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Summary:The genome of peanut clump pecluvirus (PCV) consists of two messenger RNA components which contain, respectively, three and five open reading frames (ORFs). Inoculation of transcripts from full-length cDNA clones derived from the PCV RNAs showed that RNA-1 is able to replicate in the absence of RNA-2 in protoplasts, but both RNAs are necessary for plant infection. To investigate the role of different gene products in viral RNA replication and movement, transcripts from mutant cDNA clones were inoculated to protoplasts and toChenopodium quinoaorNicotiana benthamianaplants, and progeny RNA was detected by Northern blot analysis. The protein P15, encoded by the third ORF of RNA-1, is essential for efficient replication of the viral genome. The three proteins, P51, P14, and P17, of the triple gene block contained in RNA-2 are involved in localized movement of the viral genome, whereas the coat protein (P23) is also required for vascular movement. Insertion of the β-glucuronidase reporter gene (GUS) in place of the P23 or P39 genes (the first and the second genes of RNA-2) allows visualization of the virus infection in inoculated leaves. Although the presence of the GUS gene resulted in a lower accumulation of progeny RNA and, despite instability of the constructin planta,histochemical detection of PCV multiplication was more sensitive than Northern blot detection.
ISSN:0042-6822
1096-0341
DOI:10.1006/viro.1998.9287