Abstract
A tobamovirus was isolated from leaves of Alliaria petiolata plants, showing vein-clearing, interveinal chlorosis, and moderate deformation. Host range experiments revealed a high similarity of isolate ApH both to ribgrass mosaic viruses and turnip vein-clearing viruses. The complete nucleotide sequence of the viral genome was determined. The genomic RNA is composed of 6312 nucleotides and contains four open reading frames (ORF). ORF1 is 3324 nt-long and encodes a polypeptide of about 125.3 kDa. The ORF1 encoded putative replication protein contains an Alphavirus-like methyltransferase domain. ORF2 is 4806 nt-long and encodes a polypeptide of about 182 kDa. The ORF2 encoded putative replication protein contains an RNA-dependent RNA polymerase, catalytic domain. ORF3 encodes the putative cell-to-cell movement protein with a molecular weight of 30.1 kDa. ORF4 overlaps with ORF3 and encodes the coat protein with a size of 17.5 kDa. Sequence comparisons revealed that the ApH isolate has the highest similarity to turnip vein-clearing viruses and should be considered an isolate of Turnip vein-clearing virus (TVCV). This is the first report on the occurrence of TVCV in Hungary. In vitro transcripts prepared from the full-length cDNA clone of TVCV-ApH were highly infectious and induced typical symptoms characteristic to the original isolate of the virus. Since infectious clones of TVCV-ApH and crTMV (another isolate of TVCV) markedly differed in respect to recovery phenotype in Arabidopsis thaliana, it is feasible to carry out gene exchange or mutational studies to determine viral factors responsible for the symptom recovery phenotype.
Highlights
The genus Tobamovirus represents a group of rod-shaped plant viruses with single-stranded, undivided positive-sense RNA genomes of approximately 6400 nucleotides
Several test plants inoculated by its leaf extract reacted either with local or with local and systemic symptoms characteristic to tobamoviruses
A single lesion subculture of the isolated virus caused symptoms on selected test plants (Fig 1 and Table 1) very similar to those of Ribgrass mosaic virus (RMV), the only tobamovirus known to be pathogenic to cruciferous plants and Solanum betaceum in Hungary
Summary
The genus Tobamovirus (family Virgaviridae) represents a group of rod-shaped (approx. 300– 310 nm long) plant viruses with single-stranded, undivided positive-sense RNA genomes of approximately 6400 nucleotides. We determined the complete nucleotide sequence of the genomic RNA of TVCV isolated from Alliaria petiolata in Hungary and report the preparation of the infectious cDNA clone of the virus. Virions and viral genomic RNA were purified from infected Nicotiana benthamiana plants according to a tobamovirus isolation protocol [16].
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