Isolation and genomic characterization of a classical Muscovy duck reovirus isolated in Zhejiang, China

Abstract

A classical Muscovy reovirus was isolated from a sick Muscovy duck with white necrotic foci in its liver in Zhejiang, China, in 2000. This classical reovirus was propagated in a chicken fibroblast cell line (DF-1) with obvious cytopathic effects. Its genome was 22,967 bp in length, with approximately 51.41% G+C content and 10 dsRNA segments encoding 11 proteins, which formed a 3/3/4 electrophoretic PAGE profile pattern. The length of the genomic segments was similar to those of avian orthoreoviruses (ARV and N-MDRV), ranging from 3959 nt (L1) to 1191nt (S4). All of the segments have the conserved terminal sequences 5'-GCUUUU--UUCAUC-3', and with the exception of the S4 segment, all the genome segments apparently encode one single primary translation product. The genome analysis revealed that the S4 segment of classical MDRV is a bicistronic gene, encoding the overlapping ORFs for p10 and σC but distinct from ARV and N-MDRV/N-GRV, which codes for p10, p18 and σC via the tricistronic S1 segment. A comparative sequence analysis provided evidence indicating extensive sequence divergence between classical MDRV and other avian orthoreoviruses. A phylogenetic analysis based on the RNA-dependent RNA polymerase (RdRp) and the major outer capsid proteins σC was performed. Members of the DRVs in the Avian orthoreovirus species were clustered into two genetic groups (classical MDRV and N-MDRV genotype), and the classical MDRV isolates formed distinct lineages (China and Europe lineages), suggesting that the classical MDRVs isolated in restricted geographical region are evolving by different and independent pathways.

Keywords: Avian orthoreovirus; Classical Muscovy duck reovirus; Genome sequence; Phylogenetic analysis.