Archives of Virology

, Volume 156, Issue 10, pp 1707–1715 | Cite as

Synergistic effects of Cydia pomonella granulovirus GP37 on the infectivity of nucleopolyhedroviruses and the lethality of Bacillus thuringiensis

  • Xiangyang Liu
  • Xiaohe Ma
  • Chengfeng Lei
  • Yuzhou Xiao
  • Zhongxin Zhang
  • Xiulian SunEmail author
Original Article


To initiate an efficient primary infection, it is important for baculovirus virions to penetrate through the peritrophic membrane (PM) of the host insect. It is frequently reported that enhancins of baculoviruses significantly enhance viral infection by degrading the various protein components of PMs. However, not all baculoviruses encode enhancins. GP37s of baculoviruses share high amino acid identity with fusolins, synergistic factors found in entomopoxviruses. In this study, a truncated Cydia pomonella granulovirus GP37 was expressed in Escherichia coli. The expressed GP37 effectively bound to chitin, and binding occurred predominantly within 3 h. GP37 altered the protein profiles of Spodoptera exigua PMs, from which a 50-kDa protein was dissociated. Droplet-feeding bioassays indicated that GP37 significantly enhanced the infectivity of nucleopolyhedroviruses (NPVs) and the lethality of Bacillus thuringiensis (Bt) in S. exigua larvae. This is the first demonstration of the enhancement of NPVs and Bt infection by a baculovirus GP37.


Chitin Potency Ratio Peritrophic Membrane Midgut Epithelial Cell Chitin Powder 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



We appreciate the support from the National Natural Science Foundation of China (30170085), the 973 project (2009CB118903) and the Knowledge Innovation Programs of the Chinese Academy of Sciences (KSCX2-EW-G-16).


  1. 1.
    Barbehenn RV, Martin MM (1995) Peritrophic envelope permeability in herbivorous insects. J Insect Physiol 41:303–311CrossRefGoogle Scholar
  2. 2.
    Bischoff DS, Slavicek JM (1997) Molecular analysis of an enhancin gene in the Lymantria dispar nuclear polyhedrosis virus. J Virol 71:8133–8140PubMedGoogle Scholar
  3. 3.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254PubMedCrossRefGoogle Scholar
  4. 4.
    Cheng XW, Krell PJ, Arif BM (2001) P34.8 (GP37) is not essential for baculovirus replication. J Gen Virol 82:299–305PubMedGoogle Scholar
  5. 5.
    El-Salamouny SA (2007) Observations on the peritrophic membrane of tortricid and noctuid insects and its role in susceptibility and enhancement. J Agric Urban Entomol 24:195–204CrossRefGoogle Scholar
  6. 6.
    Finney DJ (1971) Probit analysis, 3rd edn. Cambridge University Press, Cambridge, LondonGoogle Scholar
  7. 7.
    Gao MY, Li RS, Dai SY, Wu Y, Yi D (2008) Diversity of Bacillus thuringiensis strains from soil in China and their pesticidal activities. Biol Control 44:380–388CrossRefGoogle Scholar
  8. 8.
    Glen DM, Payne CC (1984) Production and field evaluation of codling moth granulosis virus for control of Cydia pomonella in the United Kingdom. Ann Appl Biol 104:87–98CrossRefGoogle Scholar
  9. 9.
    Granados RR, Lawler KA (1981) In vivo pathway of Autographa californica baculovirus invasion and infection. Virology 108:297–308PubMedCrossRefGoogle Scholar
  10. 10.
    Hughes PR, van Beek NAM, Wood HA (1986) A modified droplet feeding method for rapid assay of Bacillus thuringiensis and baculoviruses in noctuid larvae. J Invertebr Pathol 48:187–192CrossRefGoogle Scholar
  11. 11.
    Hukuhara T, Hayakawa T, Wijonarko A (2001) A bacterially produced virus enhancing factor from an entomopoxvirus enhances nucleopolyhedrovirus infection in armyworm larvae. J Invertebr Pathol 78:25–30PubMedCrossRefGoogle Scholar
  12. 12.
    Ivaldi-Sender C (1974) Techniques simples pour élevage permanent de la tordeuse orientale, Grapholita molesta (Lep., Tortricidae), sur milieu artificiel. Ann Zool Ecol Anim 6:337–343Google Scholar
  13. 13.
    Jakubowska AK, Caccia S, Gordon KH, Ferré J, Herrero S (2010) Downregulation of a chitin deacetylase-like protein in response to baculovirus infection and its application for improving baculovirus infectivity. J Virol 84:2547–2555PubMedCrossRefGoogle Scholar
  14. 14.
    Kolbe S, Fischer S, Becirevic A, Hinz P, Schrempf H (1998) The Streptornyces reticuli α-chitin-binding protein CHB2 and its gene. Microbiology+ 144:1291–1297PubMedCrossRefGoogle Scholar
  15. 15.
    Li ZF, Li CB, Yang K, Wang LH, Yin C, Gong YX, Pang Y (2003) Characterization of a chitin-binding protein GP37 of Spodoptera litura multicapsid nucleopolyhedrovirus. Virus Res 96:113–122PubMedCrossRefGoogle Scholar
  16. 16.
    Liu JJ, Carstens EB (1996) Identification, molecular cloning, and transcription analysis of the Choristoneura fumiferana nuclear polyhedrosis virus spindle-like protein gene. Virology 223:396–400PubMedCrossRefGoogle Scholar
  17. 17.
    Luque T, Finch R, Crook N, O’Reilly DR, Winstanley D (2001) The complete sequence of the Cydia pomonella granulovirus genome. J Gen Virol 82:2531–2547PubMedGoogle Scholar
  18. 18.
    Mitsuhashi W, Miyamoto K (2003) Disintegration of the peritrophic membrane of silkworm larvae due to spindles of an entomopoxvirus. J Invertebr Pathol 82:34–40PubMedCrossRefGoogle Scholar
  19. 19.
    Mitsuhashi W, Sato M, Hirai Y (2000) Involvement of spindles of a coleopteran entomopoxvirus (EPV) in infectivity of the EPVs to their host insect. Arch Virol 145:1465–1471PubMedCrossRefGoogle Scholar
  20. 20.
    O’Reilly DR (1997) Auxiliary genes of baculoviruses. In: Miller LK (ed) The Baculoviruses. Plenum Press, New York, pp 267–300Google Scholar
  21. 21.
    Peter W (1992) Peritrophic membranes. Springer, New YorkGoogle Scholar
  22. 22.
    Phanis CG, Miller DP, Cassar SC, Tristem M, Thiem SM, O’Reilly DR (1999) Identification and expression of two baculovirus gp37 genes. J Gen Virol 80:1823–1831PubMedGoogle Scholar
  23. 23.
    Rees JS, Jarrett P, Ellar DJ (2009) Peritrophic membrane contribution to Bt Cry d-endotoxin susceptibility in Lepidoptera and the effect of Calcofluor. J Invertebr Pathol 100:139–146PubMedCrossRefGoogle Scholar
  24. 24.
    Regev A, Keller M, Strizhov N, Sneh B, Prudovsky E, Chet I, Ginzberg I, Koncz-kalman Z, Koncz C, Schell J, Zilberstein A (1996) Synergistic activity of a Bacillus thuringiensis d-endotoxin and a bacterial endochitinase against Spodoptera littoralis Larvae. Appl Environ Microb 62:3581–3586Google Scholar
  25. 25.
    Robertson JL, Preisler HK (1992) Pesticide bioassays with arthropods. CRC Press, Baton RougeGoogle Scholar
  26. 26.
    Sambrook J, Russell DW (2001) Molecular cloning: A laboratory manual, 3rd edn. Cold Spring Harbor Laboratory Press, Cold Spring HarborGoogle Scholar
  27. 27.
    Sciocco-Cap A, Parola AD, Goldberg AV, Ghiringhelli PD, Romanowski V (2001) Characterization of a granulovirus isolated from Epinotia aporema Wals. (Lepidoptera: Tortricidae) larvae. Appl Environ Microb 67:3702–3706CrossRefGoogle Scholar
  28. 28.
    Shorey HH, Hale RL (1965) Mass-rearing of the larval of nine noctuid species on a simple artificial medium. J Econ Entomol 58:522–524Google Scholar
  29. 29.
    Smith IRL, Crook NE (1988) Physical maps of the genomes of four variants of Artogeia rapae granulosis virus. J Gen Virol 69:1741–1747CrossRefGoogle Scholar
  30. 30.
    Snedecor GW, Cochran WG (1989) Statistical methods, 8th edn. Iowa State University Press, AmesGoogle Scholar
  31. 31.
    Takatsuka J (2007) Characterization of a nucleopolyhedrovirus of Epinotia granitalis (Lepidoptera: Tortricidae). J Invertebr Pathol 96:265–269PubMedCrossRefGoogle Scholar
  32. 32.
    Tamura K, Dudley J, Nei M, Kumar S (2007) MEGA4: Molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599PubMedCrossRefGoogle Scholar
  33. 33.
    Theilmann DA, Blissart GW, Bonning B, Jehle J, O’Reilly DR, Rohrmann GF, Thiem S, Vlak JM (2005) Baculoviridae. In: Fauquet CM, Mayo MA, Maniloff J, Desselberger U, Ball LA (eds) Virus taxonomy, eighth report of the international committee on taxonomy of viruses. Elsevier Academic Press, London, pp 177–185Google Scholar
  34. 34.
    Thompson JD, Gibson TJ, Plewniak F, Jeanmougin F, Higgins DG (1997) The CLUSTAL_X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Res 25:4876–4882PubMedCrossRefGoogle Scholar
  35. 35.
    Vialard JE, Yuen L, Richardson CD (1990) Identification and characterization of a baculovirus occlusion body glycoprotein which resembles spheroidin, an entomopoxvirus protein. J Virol 64:5804–5811PubMedGoogle Scholar
  36. 36.
    Wang P, Granados RR (1997) An intestinal mucin is the target substrate for a baculovirus enhancin. Proc Natl Acad Sci 94:6977–6982PubMedCrossRefGoogle Scholar
  37. 37.
    Wang P, Granados RR (1997) Molecular cloning and sequencing of a novel invertebrate intestinal mucin cDNA. J Biol Chem 272:16663–16669PubMedCrossRefGoogle Scholar
  38. 38.
    Wang P, Granados RR (2000) Calcofluor disrupts the midgut defense system in insects. Insect Biochem Molec Biol 30:135–143CrossRefGoogle Scholar
  39. 39.
    Wang P, Hammer DA, Granados RR (1994) Interaction of Trichoplusia ni granulosis virus-encoded enhancin with the midgut epithelium and peritrophic membrane of four lepidopteran insects. J Gen Virol 75:1961–1967PubMedCrossRefGoogle Scholar
  40. 40.
    Yang LR, Qiang X, Zhang BQ, Tang MJ, Zhang CX (2009) Characterization of a baculovirus newly isolated from the tea slug moth, Iragoidae fasciata. J Microbiol 47:208–213PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Xiangyang Liu
    • 1
    • 2
  • Xiaohe Ma
    • 1
    • 2
  • Chengfeng Lei
    • 1
  • Yuzhou Xiao
    • 1
  • Zhongxin Zhang
    • 1
  • Xiulian Sun
    • 1
    Email author
  1. 1.Key Laboratory of Agricultural and Environmental Microbiology, Wuhan Institute of VirologyChinese Academy of SciencesWuhanChina
  2. 2.Graduate University of Chinese Academy of SciencesBeijingChina

Personalised recommendations