Molecular and in Silico Characterization of Achaea janata Granulovirus Granulin Gene

  • Pola Naveen Kumar
  • Yenumula Gerard Prasad
  • Mathyam Prabhakar
  • Arun K. Shanker
  • Divya Bhanu
Original Research Article
First Online:
Received:
Revised:
Accepted:

Abstract

Achaea janata granulovirus (AcjaGV), an insect virus belonging to Baculoviridae, infects semilooper, a widely distributed defoliating pest on castor beans (Ricinus communis L.) and several other plant hosts in India. The propagation and purification of the Hyderabad isolate AcjaGV were performed, granulin gene from this isolate was amplified, cloned and sequenced, and its homology with other known granulin genes was assessed. The 753-bp granulin ORF of AcjaGV encoded for a granulin protein of 250 amino acids with a molecular mass of 29.5 ± 0.7 kDa. This amino acid sequence exhibited significant homology with Spodoptera litura granulovirus (SpliGV) and other GVs infecting insects in the same Noctuidae family of Lepidoptera. Peptide analysis of granulin protein indicated close homology with that of SpliGV. Virtual RFLP patterns from in silico digestions of granulin gene of 18 granuloviruses mapped by 12 restriction enzymes were used for simulated digestions. Implications of the phylogenetic relationships of granulin nucleotide and deduced amino acid sequence are discussed. We have established the sequence identity of granulin gene of AcjaGV and characterized its protein product and the phylogenetic relationship with other known GVs. Our results indicate the presence of unique restriction sites for three restriction enzymes, and this can be used as a tool for identification of AcjaGV from various sources. This is the first report from the Indian subcontinent to describe the complete granulin gene of a GV isolated from A. janata.

Keywords

Baculovirus Granulovirus Achaea janata Semilooper Granulin In silico restriction digestions Phylogenetic studies RFLP 
This is a preview of subscription content, log in to check access.

Notes

Acknowledgments

We thank the AP-Netherlands Biotechnology Programme for Dryland Agriculture for supporting several activities of the work on the granulovirus. The work presented in this paper is a part of Ph. D. work of Mr. P. Naveen Kumar.

References

  1. 1.
    Gupta S, Dikshit AK (2010) Biopesticides: an ecofriendly approach for pest control. J Biopest 3(1):186–188Google Scholar
  2. 2.
    Briones ND (2007) Environmental sustainability issues in Philippine agriculture. Asian J Agric Dev 2:67–78Google Scholar
  3. 3.
    Kumar S, Singh A (2014) Biopesticides for integrated crop management: environmental and regulatory aspects. J Biofertil Biopestici 5:e121. doi: 10.4172/2155-6202.1000e121 Google Scholar
  4. 4.
    Ramanujam B, Rangeshwaran R, Sivakumar G, Mohan M, Yandigeri MS (2014) Management of insect pests by microorganisms. Proc Indian Nat Sci Acad 80:455–471CrossRefGoogle Scholar
  5. 5.
    Au S, Wu W, Pante N (2013) Baculovirus nuclear import: open, nuclear pore complex (NPC) sesame. Viruses 5:1885–1900CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Clem RJ, Passarelli AL (2013) Baculoviruses: sophisticated pathogens of insects. PLoS Pathog 9:e1003729CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Shim HJ, Choi JY, Wang Y, Tao XY, Liu Q, Roh JY, Kim JS, Kim WJ, Woo SD et al (2013) Neurobactrus, a novel, highly effective, and environmentally friendly recombinant baculovirus insecticide. Appl Environ Microbiol 79:141–149CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Rohrmann G (2011) Introduction to the baculoviruses, their taxonomy and evolution. In: Bethesda MD (ed) Baculovirus molecular biology, 2nd edn. National Library of Medicine (US), NCBIGoogle Scholar
  9. 9.
    Funk CJ, Braunagel SC, Rohrmann GF (1997) Baculovirus structure. In: Miller LK (ed) The baculoviruses. Plenum Press, New York, pp 7–32CrossRefGoogle Scholar
  10. 10.
    Rohrmann GR (1992) Baculovirus structural proteins. J Gen Virol 73:749–761CrossRefPubMedGoogle Scholar
  11. 11.
    Jehle JA, Lange M, Wang H, Hu Z, Wang Y, Hauschild R (2006) Molecular identification and phylogenetic analysis of baculo-viruses from lepidoptera. Virology 346:180–193CrossRefPubMedGoogle Scholar
  12. 12.
    Zanotto PM, Kessing BD, Maruniak JE (1993) Phylogenetic interrelationships among baculoviruses: evolutionary rates and host associations. J Invertibr Pathol 62:147–164CrossRefGoogle Scholar
  13. 13.
    Vimala Devi PS (1992) Occurrence of granulosis virus and nuclear polyhedrosis virus in castor semilooper Achaea janata Linn. (Lepidoptera: Noctuidae). J Oilseeds Res 9:328–330Google Scholar
  14. 14.
    Singaravelu B, Ramakrishnan N (1998) Characterization of a granulosis virus from the castor semilooper, Achaea janata L. J Invertebr Pathol 71:227–235CrossRefPubMedGoogle Scholar
  15. 15.
    Prasad YG, Srinivas L, Vimala Devi PS (2001) A note of granulovirus infection in Achaea janata Linnaeus. J Oilseeds Res 18:285–286Google Scholar
  16. 16.
    Prasad YG, Prabhakar M, Phanidhara A, Naveen Kumar P (2010) Development of Achaea janata granulosis virus formulation for use a viral biopesticide in the management of semilooper on castor. J Oilseeds Res 27:352–354Google Scholar
  17. 17.
    Naveen Kumar P, Prasad YG, Prabhakar M, Phanidhara A, Venkateswarlu B (2013) Granulovirus of semilooper, Achaea janata (Lepidoptera: Noctuidae): its bioefficacy and safety in mammalian toxicity tests. J Biol Control 27(2):99–104Google Scholar
  18. 18.
    Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Bio Evol 4:406–425Google Scholar
  19. 19.
    Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791CrossRefPubMedGoogle Scholar
  20. 20.
    Zuckerland E, Pauling L (1965) Evolutionary divergence and convergence in proteins. In: Bryson V, Vogel HJ (eds) Evolving genes and proteins. Academic Press, New York, pp 97–166Google Scholar
  21. 21.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599CrossRefPubMedGoogle Scholar
  22. 22.
    Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NYGoogle Scholar
  23. 23.
    Mirza UA, Liu YH, Tang JT, Porter F, Bondoc L (2000) Extraction and characterization of adenovirus proteins from sodium dodecyl sulfate polyacrylamide gel electrophoresis by matrix-assisted laser desorption/ionization mass spectrometry. J Am Soc Mass Spectrom 11:356–361CrossRefPubMedGoogle Scholar
  24. 24.
    Wei W, Davisr RE, Lee IM, Zhao Y (2007) Computer-simulated RFLP analysis of 16S rRNA genes: identification of ten new phytoplasma groups. Int J Syst Evol Microbiol 57:1855–1867CrossRefPubMedGoogle Scholar
  25. 25.
    Bideshi DK, Bigot Y, Federici BA (2000) Molecular characterization and phylogenetic analysis of the Harrisina brillians granulovirus granulin gene. Arch Virol 145:1933–1945CrossRefPubMedGoogle Scholar
  26. 26.
    Oh S, Kim DH, Patnaik BB, Jo YH, Noh MY, Lee HJ, Lee KH, Yoon KH, Kim WJ, Noh JY et al (2013) Molecular and immunohistochemical characterization of granulin gene encoded in Pieris rapae granulovirus genome. J Invertebr Pathol 113:7–17CrossRefPubMedGoogle Scholar
  27. 27.
    Bah A, Bergeron J, Arella M, Lucarotti CJ, Guertin C (1997) Identification and sequence analyses of the granulin gene of Choristoneura fumiferana granulovirus. Arch Virol 142:1577–1584CrossRefPubMedGoogle Scholar
  28. 28.
    Rohrmann G (2011) Structural proteins of baculovirus occlusion bodies and virions. In: Bethesda MD (ed) Baculovirus molecular biology, 2nd edn. National Library of Medicine (US), NCBIGoogle Scholar
  29. 29.
    Federici BA (1997) Baculovirus pathogenesis. In: Miller LK (ed) The baculoviruse. Plenum Press, New York, pp 33–35CrossRefGoogle Scholar
  30. 30.
    Eason JE, Hice RH, Johnson JJ, Federici BA (1998) Effects of substituting granulin or a granulin-polyhedrin chimera for polyhedrin on virion occlusion and polyhedral morphology in Autographa californica multinucleocapsid nuclear polyhedrosis virus. J Virol 72:6237–6243PubMedPubMedCentralGoogle Scholar
  31. 31.
    Liu X, Chen K, Cai K, Yao Q (2008) Determination of protein composition and host-derived proteins of Bombyx mori nucleopolyhedrovirus by 2-dimensional electrophoresis and mass spectrophotometry. Intervirology 51:369–376CrossRefPubMedGoogle Scholar
  32. 32.
    Wang Y, Choi JY, Roh JY, Woo SD, Jin BR, Je YH (2008) Molecular and phylogenetic characterization of Spodoptera litura granulovirus. J Microbiol 46:704–708CrossRefPubMedGoogle Scholar
  33. 33.
    Subramanian S, Rabindra RJ, Palaniswamy S, Sathiah N, Rajasekaran B (2005) Impact of granulovirus infection on susceptibility of Spodoptera litura to insecticides. Biol Control 33:165–172CrossRefGoogle Scholar
  34. 34.
    Mukawa S, Goto C (2008) In vivo characterization of two granuloviruses in larvae of Mythimna separata (Lepidoptera: Noctuidae). J Gen Virol 89:915–921CrossRefPubMedGoogle Scholar
  35. 35.
    Subramanian S, Rabindra RJ, Sithanatham S (2008) Genetic and biological variations among Plutella xylostella granulovirus isolates. Phytoparasitica 36(3):220–230CrossRefGoogle Scholar
  36. 36.
    Payne CC (1986) Insect pathogenic viruses as pest control agents. In: Franz JM (ed) Biological plant and health protection. Fischer Verlag, Stuttgart, pp 183–200Google Scholar
  37. 37.
    Lacey LA, Thomson D, Vincent C, Arthurs SP (2008) Codling moth granulovirus: a comprehensive review. Biocontrol Sci Technol 18:639–663CrossRefGoogle Scholar
  38. 38.
    Wormleaton SL (2000) Molecular and biological studies of fast and slow killing granuloviruses. Dissertation, University of WarwickGoogle Scholar
  39. 39.
    Chakerian R, Rohrmann GF, Nesson MH, Leisy DJ, Beaudreau GS (1985) The nucleotide sequence of the Pieris brassicae granulosis virus Granulin gene. J Gen Virol 66:1263–1269CrossRefPubMedGoogle Scholar
  40. 40.
    Kozlov EA, Rodnin NV, Levitina TL, Gusak NM, Radomskij NF, Palchikovskaya LJ (1992) The amino acid sequence determination of a granulin and polyhedrin from two baculoviruses infecting Agrotis segetum. Virology 189:320–323CrossRefPubMedGoogle Scholar
  41. 41.
    Jehle JA, Backhaus H (1994) The granulin gene region of Cryptophlebia leucotreta granulosis virus: sequence analysis and phylogenetic considerations. J Gen Virol 75:3667–3671CrossRefPubMedGoogle Scholar
  42. 42.
    Hayakawa T, Ko R, Okano K, Seong SI, Goto C, Maeda S (1999) Sequence analysis of the Xestia c-nigrum granulovirus genome. Virology 262:277–297CrossRefPubMedGoogle Scholar
  43. 43.
    Rashidan KK, Nassoury N, Giannopoulos PN, Mauffette Y, Guertin C (2004) Identification, characterization and phylogenic analysis of conserved genes within the odvp-6e/odv-e56 gene region of Choristoneura fumiferana granulovirus. J Biochem Mol Biol 37:206–212PubMedGoogle Scholar

Copyright information

© International Association of Scientists in the Interdisciplinary Areas and Springer-Verlag Berlin Heidelberg 2016

Authors and Affiliations

  1. 1.Division of Crop SciencesICAR - Central Research Institute for Dryland AgricultureHyderabadIndia

Personalised recommendations