Advertisement

Chinese Journal of Oceanology and Limnology

, Volume 35, Issue 6, pp 1524–1530 | Cite as

Peritrophin-like protein from Litopenaeus vannamei (LvPT) involved in white spot syndrome virus (WSSV) infection in digestive tract challenged with reverse gavage

  • Shijun Xie (谢世筠)
  • Fuhua Li (李富花)
  • Xiaojun Zhang (张晓军)
  • Jiquan Zhang (张继泉)
  • Jianhai Xiang (相建海)
Aquaculture and Fisheries

Abstract

The peritrophic membrane plays an important role in the defense system of the arthropod gut. The digestive tract is considered one of the major tissues targeted by white spot syndrome virus (WSSV) in shrimp. In this study, the nucleotide sequence encoding peritrophin-like protein of Litopenaeus vannamei (LvPT) was amplified from a yeast two-hybrid library of L. vannamei. The epitope peptide of LvPT was predicted with the GenScript OptimumAntigen™ design tool. An anti-LvPT polyclonal antibody was produced and shown to specifically bind a band at ~27 kDa, identified as LvPT. The LvPT protein was expressed and its concentration determined. LvPT dsRNA (4 μg per shrimp) was used to inhibit LvPT expression in shrimp, and a WSSV challenge experiment was then performed with reverse gavage. The pleopods, stomachs, and guts were collected from the shrimp at 0, 24, 48, and 72 h post-infection (hpi). Viral load quantification showed that the levels of WSSV were significantly lower in the pleopods, stomachs, and guts of shrimp after LvPT dsRNA interference than in those of the controls at 48 and 72 hpi. Our results imply that LvPT plays an important role during WSSV infection of the digestive tract.

Keywords

Litopenaeus vannamei digestive tract peritrophin-like protein dsRNA interference binding specificity white spot syndrome virus (WSSV) reverse gavage 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

Notes

Acknowledgment

We sincerely thank the Guangtai Company (Hainan, China) for providing the postlarval shrimp for this study.

References

  1. Aranguren L F, Tang K F J, Lightner D V. 2010. Quantification of the bacterial agent of necrotizing hepatopancreatitis (NHP-B) by real-time PCR and comparison of survival and NHP load of two shrimp populations. Aquaculture, 307(3–4): 187–192.CrossRefGoogle Scholar
  2. Arts J A J, Taverne-Thiele A J, Savelkoul H F J, Rombout J H W M. 2007. Haemocyte reactions in WSSV immersion infected Penaeus monodon. Fish & Shellfish Immunology, 23(1): 164–170.CrossRefGoogle Scholar
  3. Chou H Y, Huang C Y, Lo C F, Kou G H. 1998. Studies on transmission of white spot syndrome associated baculovirus (WSBV) in Penaeus monodon and P. japonicus via waterborne contact and oral ingestion. Aquaculture, 164(1–4): 263–276.CrossRefGoogle Scholar
  4. Chou H Y, Huang C Y, Wang C H, Chiang H C, Lo C F. 1995. Pathogenicity of a baculovirus infection causing white spot syndrome in cultured penaeid shrimp in Taiwan. Diseases of Aquatic Organisms, 23(3): 165–173.CrossRefGoogle Scholar
  5. Di Leonardo V A, Bonnichon V, Roch P, Parrinello N, Bonami J R. 2005. Comparative WSSV infection routes in the shrimp genera Marsupenaeus and Palaemon. Journal of Fish Diseases, 28(9): 565–569.CrossRefGoogle Scholar
  6. Du Z Q, Lan J F, Weng Y D, Zhao X F, Wang J X. 2013. BAX inhibitor-1 silencing suppresses white spot syndrome virus replication in red swamp crayfish, Procambarus clarkii. Fish & Shellfish Immunology, 35(1): 46–53.CrossRefGoogle Scholar
  7. Eiseman C H, Binnington K C. 1994. The peritrophic membrane: its formation, structure, chemical composition and permeability in relation to vaccination against ectoparasitic arthropods. International Journal for Parasitology, 24(1): 15–26.CrossRefGoogle Scholar
  8. Escobedo-Bonilla C M, Alday-Sanz V, Wille M, Sorgeloos P, Pensaert M B, Nauwynck H J. 2008. A review on the morphology, molecular characterization, morphogenesis and pathogenesis of white spot syndrome virus. Journal of Fish Diseases, 31(1): 1–18.CrossRefGoogle Scholar
  9. Escobedo-Bonilla C M, Wille M, Sanz V A, Sorgeloos P, Pensaert M B, Nauwynck H J. 2005. In vivo titration of white spot syndrome virus (WSSV) in specific pathogenfree Litopenaeus vannamei by intramuscular and oral routes. Diseases of Aquatic Organisms, 66(2): 163–170.CrossRefGoogle Scholar
  10. Lan J F, Li X C, Sun J J, Gong J, Wang X W, Shi X Z, Shi L J, Weng Y D, Zhao X F, Wang J X. 2013. Prohibitin interacts with envelope proteins of white spot syndrome virus and prevents infection in the red swamp crayfish, Procambarus clarkii. Journal of Virology, 87(23): 12756–12765.CrossRefGoogle Scholar
  11. Lee C T, Chen I T, Yang Y T, Ko T P, Huang Y T, Huang J Y, Huang M F, Lin S J, Chen C Y, Lin S S, Lightner D V, Wang H C, Wang A H J, Wang H C, Hor L I, Lo C F. 2015. The opportunistic marine pathogen Vibrio parahaemolyticus becomes virulent by acquiring a plasmid that expresses a deadly toxin. Proceedings of the National Academy of Sciences of the United States of America, 112(34): 10798–10803.CrossRefGoogle Scholar
  12. Livak K J, Schmittgen T D. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2-ΔΔCt method. Methods, 25(4): 402–408.CrossRefGoogle Scholar
  13. Martin G G, Simcox R, Nguyen A, Chilingaryan A. 2006. Peritrophic membrane of the penaeid shrimp Sicyonia ingentis: structure, formation, and permeability. Biological Bulletin, 211(3): 275–285.CrossRefGoogle Scholar
  14. Saleh M C, van Rij R P, Hekele A, Gillis A, Foley E, O’Farrell P H, Andino R. 2006. The endocytic pathway mediates cell entry of dsRNA to induce RNAi silencing. Nature Cell Biology, 8(8): 793–802.CrossRefGoogle Scholar
  15. Sun Y M, Li F H, Chi Y H, Xiang J H. 2013b. Enhanced resistance of marine shrimp Exopalamon carincauda Holthuis to WSSV by injecting live VP28-recombinant bacteria. Acta Oceanologica Sinica, 32(2): 52–58.CrossRefGoogle Scholar
  16. Sun Y M, Li F H, Xiang J H. 2013a. Analysis on the dynamic changes of the amount of WSSV in Chinese shrimp Fenneropenaeus chinensis during infection. Aquaculture, 376–379: 124–132.CrossRefGoogle Scholar
  17. Wang C H, Lo C F, Leu J H, Chou C M, Yeh P Y, Chou H Y, Tung M C, Chang C F, Su M S, Kou G H. 1995. Purification and genomic analysis of baculovirus associated with white spot syndrome (WSBV) of Penaeus monodon. Diseases of Aquatic Organisms, 23(3): 239–242.CrossRefGoogle Scholar
  18. Wang L Y, Li F H, Wang B, Xiang J H. 2012. Structure and partial protein profiles of the peritrophic membrane (PM) from the gut of the shrimp Litopenaeus vannamei. Fish & Shellfish Immunology, 33(6): 1285–1291.CrossRefGoogle Scholar
  19. Wang L Y, Li F H, Wang B, Xiang J H. 2013. A new shrimp peritrophin-like gene from Exopalaemon carinicauda involved in white spot syndrome virus (WSSV) infection. Fish & Shellfish Immunology, 35(3): 840–846.CrossRefGoogle Scholar
  20. Wen R, Li F H, Li S H, Xiang J H. 2014. Function of shrimp STAT during WSSV infection. Fish & Shellfish Immunology, 38(2): 354–360.CrossRefGoogle Scholar
  21. Wen R, Li F H, Sun Z, Li S H, Xiang J H. 2013. Shrimp MyD88 responsive to bacteria and white spot syndrome virus. Fish & Shellfish Immunology, 34(2): 574–581.CrossRefGoogle Scholar
  22. Xie S J, Zhang X J, Zhang J Q, Li F H, Xiang J H. 2015. Envelope proteins of white spot syndrome virus (WSSV) interact with Litopenaeus vannamei peritrophin-like protein (LvPT). PLoS One, 10(12): e0144922.CrossRefGoogle Scholar
  23. Yang H, Li S H, Li F H, Wen R, Xiang J H. 2015. Analysis on the expression and function of syndecan in the Pacific white shrimp Litopenaeus vannamei. Developmental & Comparative Immunology, 51(2): 278–286.CrossRefGoogle Scholar

Copyright information

© Chinese Society for Oceanology and Limnology, Science Press and Springer-Verlag Berlin Heidelberg 2017

Authors and Affiliations

  • Shijun Xie (谢世筠)
    • 1
    • 2
  • Fuhua Li (李富花)
    • 1
    • 3
  • Xiaojun Zhang (张晓军)
    • 1
  • Jiquan Zhang (张继泉)
    • 1
  • Jianhai Xiang (相建海)
    • 1
    • 3
  1. 1.Key Laboratory of Experimental Marine Biology, Institute of OceanologyChinese Academy of SciencesQingdaoChina
  2. 2.University of Chinese Academy of SciencesBeijingChina
  3. 3.Laboratory for Marine Biology and BiotechnologyQingdao National Laboratory for Marine Science and TechnologyQingdaoChina

Personalised recommendations