Abstract
Porcine deltacoronavirus (PDCoV) causes severe diarrhea and vomiting in affected piglets. The aim of this study was to establish the basic, in vitro characteristics of the life cycle such as replication kinetics, cellular ultrastructure, virion morphology, and induction of autophagy of PDCoV. Time-course analysis of viral subgenomic and genomic RNA loads and infectious titers indicated that one replication cycle of PDCoV takes 5 to 6 h. Electron microscopy showed that PDCoV infection induced the membrane rearrangements with double-membrane vesicles and large virion-containing vacuoles. The convoluted membranes structures described in alpha- and beta-coronavirus were not observed. PDCoV infection also increased the number of autophagosome-like vesicles in the cytoplasm of cells, and the autophagy response was detected by LC3 I/II and p62 Western blot analysis. For the first time, this study presents the picture of the PDCoV infection cycle, which is crucial to help elucidate the molecular mechanism of deltacoronavirus replication.
Highlights
Porcine deltacoronavirus (PDCoV), which belongs to the genus Deltacoronavirus in the subfamilyCoronavirinae of the family Coronaviridae, order Nidovirales, was first reported in Hong Kong in 2012 [1] and thereafter isolated from pigs in the United States [2] and many Asian countries including China [3,4,5]
Intracellular gRNA load remained low until 5 hpi, followed by a sharp increase at 6 hpi from 2.92 × to 2.07 × copies/106 cells, reaching a final titer of 4.47 × copies/106 cells at 12 hpi (Figure 1B, left panel)
These results indicated that the first generation of progeny was assembled and released from infected cells at about 5 hpi, suggesting that one replication cycle of
Summary
Coronavirinae of the family Coronaviridae, order Nidovirales, was first reported in Hong Kong in 2012 [1] and thereafter isolated from pigs in the United States [2] and many Asian countries including China [3,4,5]. It causes acute diarrhea, vomiting, dehydration and mortality in nursing pigs [6,7]. The genome organization of PDCoV is similar to those of other reported coronaviruses, with the typical gene order 50 -ORF1a/1b-Spike (S)-Envelope (E)-Membrane (M)-NS6-Nucleocapsid (N)/NS7-30.
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