Detection and Characterization of a Reassortant Mammalian Orthoreovirus Isolated from Bats in Xinjiang, China

Figures

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  Figure 1. Infectivity of the BtMRV in different cell lines. (A) Mock-infected PK-15 cells show a normal appearance with negative detection by IFA. (B–G) CPE induced by BtMRV-QAPpC66 in PK-15, BHK, Vero, Hela, MDBK, and BFK cells with infected cells showing positive detection (bright apple green fluorescence) by IFA (inlet). (H) The replication dynamics of QAPpC11, 15 and 66 in PK-15 at passage 6. (I) The titers of QAPpC66 at passage 3 in five cell lines at 72 h.p.i. (**, p < 0.01; ***, p < 0.001; both by Bonferroni’s multiple comparisons test).

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  Figure 2. Electron microscopy of BtMRV-QAPpC66. (A) Three morphotypes of BtMRV-QAPpC66, i.e., typical virions (open arrow), infectious subvirions (filled triangle) and core particles (filled arrow) can be observed under TEM. (B) Ultrathin section of the infected PK-15 cell shows arrayed virions (arrowed) in cytoplasm.

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  Figure 3. Phylogenetic analyses of the (L1–M3) segments based on nt sequences. The phylogenetic reconstruction was implemented using the maximum likelihood method and the best-fit model with 1000 bootstrap replicates. Each MRV is identified with its serotype, name, host, country, year, and GenBank accession number. Bootstrap value greater than 75 is shown on the node as a triangle with its size indicating the value (shown in panel L2). The three bat MRV isolates in this study are headed with filled stars, and their closest genetic neighbors are indicated with filled circles. The scale bar indicates the genetic distance. The animal cartoons indicate the hosts of the viruses they are next to, and their meanings are shown in Figure 5.

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  Figure 4. Phylogenetic analyses of the (S1–S4) segments based on nt sequences. The phylogeny of S1 indicates that the three isolates belong to MRV serotype 2. The phylogenetic reconstruction was implemented using the maximum likelihood method and applied the best-fit model with 1000 bootstrap replicates. Each MRV strain is identified with its serotype, name, host, country, year, and GenBank accession number. Bootstrap value greater than 75 is shown on the node as a triangle with its size indicating the value (shown in panel S1). The three bat MRV isolates in this study are headed with filled stars, and the closest genetic neighbors are indicated with filled circles. The scale bar indicates the genetic distance. The animal cartoons indicate the hosts of the viruses they are next to, and their meanings are shown in Figure 5.

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  Figure 5. Reassortment analysis of BtMRV-QAPpC66 and other MRVs. (A) Similarity plots and (B) bootscan analyses were performed with the concatenated segments of BtMRV-QAPpC66 as the query and other MRVs, including MRV1/YNSZ/V207/Cattle/CHN/2016, MRV2/SI-MRV07/Human /SLO/2017, MRV3/mew716/Human/SUI/2015, MRV2/WIV4/Bat/CHN/2011, MRV2/OV204/Deer/ USA/2016 and MRV3/MPC/04/Civet/CHN/2004, as potential parental sequences.