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Molecular analysis of grass carp reovirus HZ08 genome segments 1–3 and 5–6

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A new grass carp reovirus, assigned HZ08, was isolated from a diseased grass carp case during routine examination in Huzhou City, Zhejiang Province, China. The complete nucleotide sequences of genomic segments S1–S3 and S5–S6 were obtained and comprised 3927, 3870, 3753, 2229, and 2030 bp, respectively. Each segment contained a single open reading frame which encoded putative proteins of 143.6, 143.1, 135.9, 80.5, and 68.4 kDa, respectively. Conserved motifs 5′ (GUAAUUU…UUCAUC) 3′ were found at the ends of each segment. At the amino acid level, HZ08 S1–S3 and S5–S6 showed similarity to the corresponding segments of Aquareovirus. Further phylogenetic analysis using the amino acid sequences of the RNA-dependent RNA polymerases protein encoded by S2 revealed that HZ08 formed a cluster close to the aquareovirus, but was far from the other isolates, which indicated that HZ08 is likely to be a new member of Aquareovirus.

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Funding for this research was provided by the National Special Research Programs for Non-Profit Trades (Agriculture) (No. 200803013), the Earmarked Fund for Modern Agro-industry Technology Research System (nycytx-49-14), and Agricultural Science and Technology Leap Project.

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Correspondence to Shuqin Wu.

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Supplementary Fig. 1

Thin section electron micrograph of CIK cells 5 days after inoculation with new isolate. a: characteristic cytoplasmic inclusion bodies of variable size and shapes in the cytoplasm (bar = 2 μm); b: Virions filling the entire area of the inclusion body towards the late morphogenesis in CIK cells (bar = 100 nm) (TIFF 511 kb)

Supplementary Fig. 2

SDS-PAGE of HZ08 genome (TIFF 323 kb)

Supplementary Fig. 3

Neighbor-joining tree built with the RdRps sequences of HZ08 and other reoviruses. Bootstrap values of 1000 replications are indicated next to the branches. The scale for branch length (0.2 substitutions/site) is shown below the tree. Strains of representative viruses from ten genera of the family Reoviridae and their sequences used: Aquareovirus: Grass carp reovirus 873 (GCRV873) AF260512; Golden shiner reovirus (GSV) NC_005167; Chum salmon reovirus (CSV) AF418295; American grass carp reovirus (AGCRV) NC_010585; Aquareovirus A (Aqua A) NC_007583; Atlantic salmon reovirus TS (TSRV) EF434978; Striped bass reovirus (SBRV) AF450318; Golden ide reovirus (GIRV) AF450323. Orthoreovirus: Mammalian orthoreovirus, subgroup 1, Lang strain (MRV-1) M24734; Jones strain (MRV-2) M31057; Dearing strain (MRV-3) M31058. Orbivirus: African horse sickness virus, serotype 9 (AHSV-9) U94887; Bluetongue virus, serotype 2 (BTV-2) L20508; serotype 10 (BTV-10) X12819; serotype 11 (BTV-11) L20445; serotype 13 (BTV-13) L20446; serotype 17 (BTV-17) L20447. Rotavirus: Rotavirus A, Bovine strain RF (BoRV-A/RF) J04346; bovine strain UK (BoRVA/UK) X55444; Simian strain SA11 (SiRV-A/SA11) AF015955; Simian strain SA11-both (SiRV-A/SA11b) X16830; porcine strain Gottfried (PoRV-A/Go) M32805; Rotavirus B, human/murine rotavirus IDIR (Hu/MuRV-B/IDIR) M97203; Rotavirus C, Porcine Cowden strain (PoRV-C/Co) M74216. Fijivirus: Nilaparvata lugens reovirus, Izumo strain (NLRV-Iz) D49693; Rice black-streaked dwarf virus (RBSDV) AJ294757. Phytoreovirus: Rice dwarf virus, Chinese strain (RDV-Ch) U73201; Rice dwarf virus, strain H (RDV-H) D10222; Rice dwarf virus, strain A (RDV-A) D90198; Oryzavirus: Rice ragged stunt virus, Thai strain (RRSV-Th) U66714. Coltivirus: Colorado tick fever virus strain Florio, (CTFV-Fl) AF133428; European Eyach virus strain Fr578 (EYAV-Fr578) AF282467. Seadornavirus: Banna virus (BAV-In6423) AF133430; Kadipiro virus (KDV-Ja7075) AF133429. Cypovirus: Bombyx mori cypovirus 1 (BmCPV-1) AF323782; Lymantria dispar cypovirus 1 (LdCPV-1) AF389463; Lymantria dispar cypovirus 14 (LdCPV-14) AF389452; Trichoplusia ni cypovirus 15 (TnCPV-15) AF291683 (TIFF 728 kb)

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Zhang, C., Wang, Q., Shi, C. et al. Molecular analysis of grass carp reovirus HZ08 genome segments 1–3 and 5–6. Virus Genes 41, 102–104 (2010).

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