1887

Abstract

Infectious myonecrosis virus (IMNV) causes significant economic losses in farmed shrimp, where associated mortality in ponds can reach 70 %. To explore host/pathogen interactions, a next-generation sequencing approach using lymphoid organ tissue from IMNV-infected shrimp was conducted. Preliminary sequence assembly of just the virus showed that there were at least an additional 639 bp at the 5′ terminus and 23 nt at the 3′ terminus as compared with the original description of the IMNV genome (7561 nt). Northern blot and reverse transcription-PCR analysis confirmed the presence of novel sequence at both ends of the genome. Using 5′ RACE, an additional 4 nt were discovered; 3′ RACE confirmed the presence of 22 bp rather than 23 bp of sequence. Based on these data, the IMNV genome is 8226 bp in length. dsRNA was used to trigger RNA interference (RNAi) and suppress expression of the newly revealed genome sections at the 5′ end of the IMNV genome in IMNV-infected . An RNAi trigger targeting a 376 bp length of the 5′ UTR did not improve survival of infected shrimp. In contrast, an RNAi trigger targeting a 381 bp sequence in ORF1 improved survival to 82.2 % as compared with 2.2 % survival in positive control animals. These studies revealed the importance of the new genome sections to produce high-titre infection, and associated disease and mortality, in infected shrimp.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.000137
2015-07-01
2021-10-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/96/7/1821.html?itemId=/content/journal/jgv/10.1099/vir.0.000137&mimeType=html&fmt=ahah

References

  1. Altschul S.F., Gish W., Miller W., Myers E.W., Lipman D.J. (1990). Basic local alignment search toolJ Mol Biol 215403410.[CrossRef] [Google Scholar]
  2. Andrade T.P.D., Srisuvan T., Tang K.F.J., Lightner D.V. (2007). Real-time reverse transcription polymerase chain reaction assay using TaqMan probe for detection and quantification of infectious myonecrosis virus (IMNV)Aquaculture 264915.[CrossRef] [Google Scholar]
  3. Assavalapsakul W., Smith D.R., Panyim S. (2006). Identification and characterization of a Penaeus monodon lymphoid cell-expressed receptor for the yellow head virusJ Virol 80262269.[CrossRef] [Google Scholar]
  4. Brown T., Mackey K., Du T. (2001). Analysis of RNA by Northern and slot-blot hybridizationCurr Protoc Mol Biol. 1012.112.14. [Google Scholar]
  5. Escobedo-Bonilla C.M., Wille M., Alday Sanz V., Sorgeloos P., Pensaert M.B., Nauwynck H.J. (2007). Pathogenesis of a Thai strain of white spot syndrome virus (WSSV) in juvenile, specific pathogen-free Litopenaeus vannamei Dis Aquat Organ 748594.[CrossRef] [Google Scholar]
  6. Garlapati S., Wang C.C. (2005). Structural elements in the 5′-untranslated region of giardiavirus transcript essential for internal ribosome entry site-mediated translation initiationEukaryot Cell 4742754.[CrossRef] [Google Scholar]
  7. Larkin M.A., Blackshields G., Brown N.P., Chenna R., McGettigan P.A., McWilliam H., Valentin F., Wallace I.M., Wilm A., other authors. (2007). Clustal W and Clustal X version 2.0Bioinformatics 2329472948.[CrossRef] [Google Scholar]
  8. Lightner D.V., Redman R.M., Pantoja C.R., Tang K.F., Noble B.L., Schofield P., Mohney L.L., Nunan L.M., Navarro S.A. (2012). Historic emergence, impact and current status of shrimp pathogens in the AmericasJ Invertebr Pathol 110174183.[CrossRef] [Google Scholar]
  9. Liu S., Vijayendran D., Bonning B.C. (2011). Next generation sequencing technologies for insect virus discoveryViruses 318491869.[CrossRef] [Google Scholar]
  10. Loy J.D., Mogler M.A., Loy D.S., Janke B., Kamrud K., Scura E.D., Harris D.L., Bartholomay L.C. (2012). dsRNA provides sequence-dependent protection against infectious myonecrosis virus in Litopenaeus vannamei J Gen Virol 93880888.[CrossRef] [Google Scholar]
  11. Mello M.V., Aragão M.E., Torres-Franklin M.L., Neto J.M., Guedes M.I. (2011). Purification of infectious myonecrosis virus (IMNV) in species of marine shrimp Litopenaeus vannamei in the State of CearáJ Virol Methods 1771014.[CrossRef] [Google Scholar]
  12. Nibert M.L. (2007). ‘2A-like’ and ‘shifty heptamer’ motifs in penaeid shrimp infectious myonecrosis virus, a monosegmented double-stranded RNA virusJ Gen Virol 8813151318.[CrossRef] [Google Scholar]
  13. Pongsomboon S., Wongpanya R., Tang S., Chalorsrikul A., Tassanakajon A. (2008). Abundantly expressed transcripts in the lymphoid organ of the black tiger shrimp, Penaeus monodon, and their implication in immune functionFish Shellfish Immunol 25485493.[CrossRef] [Google Scholar]
  14. Poulos B.T., Lightner D.V. (2006). Detection of infectious myonecrosis virus (IMNV) of penaeid shrimp by reverse-transcriptase polymerase chain reaction (RT-PCR)Dis Aquat Organ 736972.[CrossRef] [Google Scholar]
  15. Poulos B.T., Tang K.F., Pantoja C.R., Bonami J.R., Lightner D.V. (2006). Purification and characterization of infectious myonecrosis virus of penaeid shrimpJ Gen Virol 87987996.[CrossRef] [Google Scholar]
  16. Senapin S., Phewsaiya K., Briggs M., Flegel T.W. (2007). Outbreaks of infectious myonecrosis virus (IMNV) in Indonesia confirmed by genome sequencing and use of an alternative RT-PCR detection methodAquaculture 2663238.[CrossRef] [Google Scholar]
  17. Tang J., Ochoa W.F., Sinkovits R.S., Poulos B.T., Ghabrial S.A., Lightner D.V., Baker T.S., Nibert M.L. (2008). Infectious myonecrosis virus has a totivirus-like, 120-subunit capsid, but with fiber complexes at the fivefold axesProc Natl Acad Sci U S A 1051752617531.[CrossRef] [Google Scholar]
  18. Zerbino D.R., Birney E. (2008). Velvet: algorithms for de novo short read assembly using de Bruijn graphsGenome Res 18821829.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.000137
Loading
/content/journal/jgv/10.1099/vir.0.000137
Loading

Data & Media loading...

Supplements

Supplementary Data



PDF

Most cited this month Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error