Molecular characterization and In Vitro synthesis of infectious RNA of a Turnip vein-clearing virus isolated from Alliaria petiolata in Hungary

• Published in: PloS one 2019-10, Vol.14 (10), p.e0224398
• Main Authors: Tóth, Tamás, Gyula, Péter, Salamon, Pál, Kis, Szilvia, Sós-Hegedűs, Anita, Szittya, György
• Format: Article
• Language: English
• Subjects: Agricultural biotechnology; Agricultural research; Alliaria petiolata; Arabidopsis; Arabidopsis thaliana; Base sequence; Biology and Life Sciences; Brassicaceae - virology; Chemical synthesis; Chlorosis; Clearing; Cloning; Coat protein; Computer and Information Sciences; Deoxyribonucleic acid; DNA; DNA polymerase; DNA, Complementary - genetics; DNA-directed RNA polymerase; Engineering and Technology; Genes; Genomes; Genomics; Host range; Hungary; Leaves; Methyltransferase; Molecular weight; Movement protein; Nucleotide sequence; Nucleotides; Open reading frames; Phenotypes; Phosphatase; Phylogenetics; Plant viruses; Proteins; Recovery; Replication; Research and Analysis Methods; Ribonucleic acid; RNA; RNA, Viral - biosynthesis; RNA-directed RNA polymerase; Sequence Analysis; Similarity; Tobamovirus; Tobamovirus - genetics; Tobamovirus - isolation & purification; Tobamovirus - metabolism; Transcription; Transcription, Genetic; Transferases; Turnips; Veins (plant anatomy); Viroids; Virus replication; Viruses
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0224398

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.