Skip to main content

Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray

VirusDisease volume 26, pages 9–18 (2015)Cite this article

Abstract

White spot syndrome virus (WSSV) continues to be the most devastating viral pathogen infecting penaeid shrimp the world over. The genome of WSSV has been deciphered and characterized from three geographical isolates and significant progress has been made in developing various molecular diagnostic methods to detect the virus. However, the information on host immune gene response to WSSV pathogenesis is limited. Microarray analysis was carried out as an approach to analyse the gene expression in black tiger shrimp Penaeus monodon in response to WSSV infection. Gill tissues collected from the WSSV infected shrimp at 6, 24, 48 h and moribund stage were analysed for differential gene expression. Shrimp cDNAs of 40,059 unique sequences were considered for designing the microarray chip. The Cy3-labeled cRNA derived from healthy and WSSV-infected shrimp was subjected to hybridization with all the DNA spots in the microarray which revealed 8,633 and 11,147 as up- and down-regulated genes respectively at different time intervals post infection. The altered expression of these numerous genes represented diverse functions such as immune response, osmoregulation, apoptosis, nucleic acid binding, energy and metabolism, signal transduction, stress response and molting. The changes in gene expression profiles observed by microarray analysis provides molecular insights and framework of genes which are up- and down-regulated at different time intervals during WSSV infection in shrimp. The microarray data was validated by Real Time analysis of four differentially expressed genes involved in apoptosis (translationally controlled tumor protein, inhibitor of apoptosis protein, ubiquitin conjugated enzyme E2 and caspase) for gene expression levels. The role of apoptosis related genes in WSSV infected shrimp is discussed herein.

This is a preview of subscription content, access via your institution.

Access options

Buy single article

Instant access to the full article PDF.

43,34 €

Price includes VAT (Finland)
Tax calculation will be finalised during checkout.

Fig. 1
Fig. 2

References

  1. Tsai JM, Wang HC, Leu JH, Hsiao HH, Wang AH, Kou H, Lo CF. Genomic and proteomic analysis of thirty-nine structural proteins of shrimp White spot syndrome virus. J Virol. 2004;78:11360–70.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  2. Marks H, Vorst O, van Houwelingen AM, van Hulten MC, Vlak J. Gene-expression profiling of White spot syndrome virus in vivo. J Gen Virol. 2005;86:2081–100.

    CAS  PubMed  Article  Google Scholar 

  3. Liu WJ, Chang YS, Wang CH, Kou GH, Lo CF. Microarray and RT-PCR screening for White spot syndrome virus immediate-early genes in cycloheximide-treated shrimp. Virology. 2005;334:327–41.

    CAS  PubMed  Article  Google Scholar 

  4. Li F, Li M, Ke W, Ji Y, Bian X, Yan X. Identification of the immediate-early genes of White spot syndrome virus. Virology. 2009;385:267–74.

    CAS  PubMed  Article  Google Scholar 

  5. Lan Y, Xu X, Yang F, Zhang X. Transcriptional profile of shrimp White spot syndrome virus (WSSV) genes with DNA microarray. Arch Virol. 2006;151:1723–33.

    CAS  PubMed  Article  Google Scholar 

  6. Wang H, Lin A, Yii D, Chang Y, Kou G, Lo C. DNA microarrays of the White spot syndrome virus genome: genes expressed in the gills of infected shrimp. Mar Biotechnol. 2004;6:S106–11.

    Google Scholar 

  7. Khadijah S, Neo SY, Hossain MS, Miller LD, Mathavan S, Kwang J. Identification of White spot syndrome virus latency-related genes in specific-pathogen-free shrimps by use of a microarray. J Virol. 2003;77:10162–7.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  8. Dhar AK, Dettori A, Roux MM, Klimpel KR, Read B. Identification of differentially expressed genes in shrimp (Penaeus stylirostris) infected with White spot syndrome virus by cDNA microarrays. Arch Virol. 2003;148:2381–96.

    CAS  PubMed  Article  Google Scholar 

  9. Wang B, Li F, Dong B, Zhang X, Zhang C, Xiang J. Discovery of the genes in response to White spot syndrome virus (WSSV) infection in Fenneropenaeus chinensis through cDNA microarray. Mar Biotechnol. 2006;8:491–500.

    PubMed  Article  Google Scholar 

  10. Wongpanya R, Aoki T, Hirono I, Yasuike M, Tassanakajon A. Analysis of gene expression in haemocytes of shrimp Penaeus monodon challenged with White spot syndrome virus by cDNA microarray. Science Asia. 2007;33:165–74.

    CAS  Article  Google Scholar 

  11. Robalino J, Almeida JS, McKillen D, Colglazier J, Trent HF III, Chen YA, Peck ME, Browdy CL, Chapman RW, Warr GW, Gross PS. Insights into the immune transcriptome of the shrimp Litopenaeus vannamei: tissue-specific expression profiles and transcriptomic responses to immune challenge. Physiol Genomics. 2007;29:44–56.

    CAS  PubMed  Article  Google Scholar 

  12. Pongsomboon S, Tang S, Boonda S, Aoki T, Hirono I, Tassanakajon A. A cDNA microarray approach for analyzing transcriptional changes in Penaeus monodon after infection by pathogens. Fish Shellfish Immunol. 2011;30:439–46.

    CAS  PubMed  Article  Google Scholar 

  13. Ponprateep S, Tharntada S, Somboonwiwat K, Tassanakajon A. Gene silencing reveals a crucial role for anti-lipopolysaccharide factors from Penaeus monodon in the protection against microbial infections. Fish Shellfish Immunol. 2012;32:26–34.

    CAS  PubMed  Article  Google Scholar 

  14. Kou GH, Peng SE, Chiu YL, Lo CF. Tissue distribution of White spot syndrome virus (WSSV) in shrimp and crabs. In: Flegel TW, editor. Advances in shrimp biotechnology. Bangkok: National Center for Genetic Engineering and Biotechnology; 1998. p. 267–71.

    Google Scholar 

  15. Clavero-Salas A, Sotelo-Mundo RR, Gollas-Galván T, Hernández-López J, Beatriz Peregrino-Uriarte A, Muhlia-Almazán A, Yepiz-Plascencia G. Transcriptome analysis of gills from the white shrimp Litopenaeus vannamei infected with White spot syndrome virus. Fish Shellfish Immunol. 2007;23:459–72.

    CAS  PubMed  Article  Google Scholar 

  16. Tassanakajon A, Somboonwiwat K, Supungul P, Tang S. Discovery of immune molecules and their crucial functions in shrimp immunity. Fish Shellfish Immunol. 2013;34:954–67.

    CAS  PubMed  Article  Google Scholar 

  17. Withyachumnarnkul B, Boonsaeng V, Chomsoong R, Flegel TW, Muangsin S, Nash GL. Seasonal variation in White spot syndrome virus-positive samples in broodstock and post-larvae of Penaeus monodon in Thailand. Dis Aquat Organ. 2003;53:167–71.

    PubMed  Article  Google Scholar 

  18. Lin YR, Hung HC, Leu JH, Wang HC, Kou GH, Lo CF. The role of aldehyde dehydrogenase and hsp70 in suppression of White spot syndrome virus replication at high temperature. J Virol. 2011;85:3517–25.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  19. Woramongkolchai N, Supungul P, Tassanakajon A. The possible role of penaeidin5 from the black tiger shrimp, Penaeus monodon, in protection against viral infection. Dev Comp Immunol. 2011;35:530–6.

    CAS  PubMed  Article  Google Scholar 

  20. Ma FF, Liu QH, Guan GK, Li C, Huang J. Arginine kinase of Litopenaeus vannamei involved in White spot syndrome virus infection. Gene. 2014;539:99–106.

    CAS  PubMed  Article  Google Scholar 

  21. Wang J, Yang K, Zhang X. Characterization of the interaction between arginine kinase and siRNA. Mar Biotechnol. 2013;15:368–74.

    CAS  PubMed  Article  Google Scholar 

  22. Liu WJ, Chang YS, Wang AHJ, Kou GH, Lo CF. White spot syndrome virus annexes a shrimp STAT to enhance expression of the immediate-early gene ie1. J Virol. 2007;81:1461–71.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  23. Li F, Zhang D, Fujise K. Characterization of fortilin, a novel antiapoptotic protein. J Biol Chem. 2001;276:47542–9.

    CAS  PubMed  Article  Google Scholar 

  24. Bangrak P, Graidist P, Chotigeat W, Hongdara A. Molecular cloning and expression of a mammalian homologue of a translationally controlled tumour protein (TCTP) gene from Penaeus monodon shrimp. J Biotechnol. 1999;108:219–26.

    Article  Google Scholar 

  25. Tonganunt M, Nupan B, Saengsakda M, Suklour S, Wanna W, Senapin Chotigeat W, Phongdara A. The role of Pm–fortilin in protecting shrimp from White spot syndrome virus (WSSV) infection. Fish Shellfish Immunol. 2008;25:633–7.

    CAS  PubMed  Article  Google Scholar 

  26. Graidist P, Fujise K, Wanna W, Sritunyalucksana K, Phongdara A. Establishing a role for shrimp fortilin in preventing cell death. Aquaculture. 2006;255:157–64.

    CAS  Article  Google Scholar 

  27. Flegel TW, Pasharawipas T. Active viral accommodation: a new concept for crustacean response to viral pathogen. In: Flegel TW, editor. Advances in Shrimp Biotechnology. Bangkok: National Centre for Genetic Engineering and Biotechnology; 1998. p. 245–50.

    Google Scholar 

  28. Nupan B, Phongdara A, Saengsakda M, Leu JH, Lo CF. Shrimp Pm-fortilin inhibits the expression of early and late genes of White spot syndrome virus (WSSV) in an insect cell model. Dev Comp Immunol. 2011;35:469–75.

    CAS  PubMed  Article  Google Scholar 

  29. Wu W, Wu B, Ye T, Huang H, Dai C, Yuan J, Wang W. TCTP is a critical factor in shrimp immune response to virus infection. PLoS One. 2013;8(9):e74460.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  30. Thaw P, Baxter NJ, Hounslow AM, Price C, Waltho JP, Craven CJ. Structure of TCTP reveals unexpected relationship with guanine nucleotide-free chaperones. Nat Struct Mol Biol. 2001;8:701–4.

    CAS  Article  Google Scholar 

  31. Wu W, Zong R, Xu J, Zhang X. Antiviral phagocytosis is regulated by a novel Rab-dependent complex in shrimp Penaeus japonicus. J Proteome Res. 2008;7:424–31.

    CAS  PubMed  Article  Google Scholar 

  32. Leu JH, Kuo YC, Kou GH, Lo CF. Molecular cloning and characterization of an inhibitor of apoptosis protein (IAP) from the tiger shrimp, Penaeus monodon. Dev Comp Immunol. 2008;32:121–33.

    CAS  PubMed  Article  Google Scholar 

  33. Leu J, Chen Y, Chen L, Chen K, Huang H, Ho J, Lo C. Litopenaeus vannamei inhibitor of apoptosis protein 1 (LvIAP1) is essential for shrimp survival. Dev Comp Immunol. 2012;38:78–87.

    CAS  PubMed  Article  Google Scholar 

  34. Wang PH, Wan DH, Gu ZH, Qiu W, Chen YG, Weng SP, Yu XQ, He JG. Analysis of expression, cellular localization, and function of three inhibitors of apoptosis (IAPs) from Litopenaeus vannamei during WSSV infection and in regulation of antimicrobial peptide genes (AMPs). PLoS One. 2013;8(8):e72592.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

  35. O’Riordan MX, Bauler LD, Scott FL, Duckett CS. Inhibitor of apoptosis proteins in eukaryotic evolution and development: a model of thematic conservation. Dev Cell. 2008;15:497–508.

    PubMed Central  PubMed  Article  Google Scholar 

  36. Phongdara A, Wanna W, Chotigeat W. Molecular cloning and expression of caspase from white shrimp Penaeus merguiensis. Aquaculture. 2006;252:114–20.

    CAS  Article  Google Scholar 

  37. Leu JH, Wang HC, Kou GH, Lo CF. Penaeus monodon caspase is targeted by a White spot syndrome virus anti-apoptosis protein. Dev Comp Immunol. 2008;32:476–86.

    CAS  PubMed  Article  Google Scholar 

  38. Wongprasert K, Sangsuriya P, Phongdara A, Senapin S. Cloning and characterization of a caspase gene from black tiger shrimp (Penaeus monodon)-infected with White spot syndrome virus (WSSV). J Biotechnol. 2007;131:9–19.

    CAS  PubMed  Article  Google Scholar 

  39. Rijiravanich A, Browdy CL, Withyachumnarnkul B. Knocking down caspase-3 by RNAi reduces mortality in Pacific white shrimp Penaeus (Litopenaeus) vannamei challenged with a low dose of white-spot syndrome virus. Fish Shellfish Immunol. 2008;24:308–13.

    CAS  PubMed  Article  Google Scholar 

  40. Wang PH, Wan DH, Chen YG, Weng SP, Yu XQ, He JG. Characterization of four novel caspases from Litopenaeus vannamei (Lvcaspase2-5) and their role in WSSV infection through dsRNA-mediated gene silencing. PLoS One. 2013;8(12):e80418.

    PubMed Central  PubMed  Article  Google Scholar 

  41. Wang L, Zhi B, Wu W, Zhang X. Requirement for shrimp caspase in apoptosis against virus infection. Dev Comp Immunol. 2008;32:706–15.

    CAS  PubMed  Article  Google Scholar 

  42. Shi YG. Mechanisms of caspase activation and inhibition during apoptosis. Mol Cell. 2002;9:459–70.

    CAS  PubMed  Article  Google Scholar 

  43. Deveraux QL, Reed TC. IAP family proteins: suppressors of apoptosis. Genes Dev. 1999;13:239–52.

    CAS  PubMed  Article  Google Scholar 

  44. Yan F, Xia D, Lv S, Qi Y, Xu H. Functional analysis of the orf390 gene of the White spot syndrome virus. Virus Res. 2010;151:39–44.

    CAS  PubMed  Article  Google Scholar 

  45. Tschopp J, Thome M, Hofmann K, Meinl E. The fight of viruses against apoptosis. Curr Opin Genet Dev. 1998;8:82–7.

    CAS  PubMed  Article  Google Scholar 

  46. Huang T, Cui Y, Zhang X. Involvement of viral microRNA in the regulation of antiviral apoptosis in shrimp. J Virol. 2014;88:2544–54.

    PubMed Central  PubMed  Article  Google Scholar 

  47. Chen A, Wang S, Zhao X, Yu X, Wang J. Enzyme E2 from Chinese white shrimp inhibits replication of White spot syndrome virus and ubiquitinates its RING domain proteins. J Virol. 2011;85:8069–79.

    CAS  PubMed Central  PubMed  Article  Google Scholar 

Download references

Acknowledgments

Authors acknowledge Department of Biotechnology, Ministry of Science and Technology, India for funding this study under the project “Molecular studies on sequential pathogenesis of WSSV and defence mechanism in Penaeus monodon”.

Author information

Affiliations

  1. Genetics and Biotechnology Unit, Central Institute of Brackishwater Aquaculture, 75 Santhome High Road, R.A. Puram, Chennai, 600028, India

    M. S. Shekhar, A. Gomathi, G. Gopikrishna & A. G. Ponniah

Authors
  1. M. S. Shekhar
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. A. Gomathi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. G. Gopikrishna
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. A. G. Ponniah
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to M. S. Shekhar.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary material 1 (PDF 5 kb)

Supplementary material 2 (DOC 83 kb)

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Shekhar, M.S., Gomathi, A., Gopikrishna, G. et al. Gene expression profiling in gill tissues of White spot syndrome virus infected black tiger shrimp Penaeus monodon by DNA microarray. VirusDis. 26, 9–18 (2015). https://doi.org/10.1007/s13337-014-0243-7

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13337-014-0243-7

Keywords

  • Microarray
  • WSSV
  • Penaeus monodon