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Spider Venom Toxin Protects Plants from Insect Attack

Abstract

Many of the toxin proteins, that have been heterogeneously expressed in agricultural crops to provide resistance to insect pests, are too specific or are only mildly effective against the major insect pests. Spider venoms are a complex cocktail of toxins that have evolved specifically to kill insects. Here we show that the ω-ACTX-Hv1a toxin (Hvt), a component of the venom of the Australian funnel web spider (Hadronyche versuta) that is a calcium channel antagonist, retains its biological activity when expressed in a heterologous system. Expressed as a fusion protein in E. coli, the purified toxin fusion immobilized and killed Helicoverpa armigera and Spodoptera littoralis caterpillars when applied topically. Transgenic expression of Hvt in tobacco effectively protected the plants from H. armigera and S. littoralis larvae, with 100% mortality within 48 h. We conclude that the Hvt is an attractive and effective molecule for the transgenic protection of plants from herbivorous insects which should be evaluated further for possible application in agriculture.

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References

  1. A Abdeen A Virgo’s E Olivella J Villanueva X Avile’s R Gabarra et al. (2005) ArticleTitleMultiple insect resistance in transgenic tomato plants over-expressing two families of plant proteinase inhibitors Plant Mol Biol 57 189–202 Occurrence Handle15821877 Occurrence Handle1:CAS:528:DC%2BD2MXjtV2hu7c%3D Occurrence Handle10.1007/s11103-004-6959-9

    PubMed  CAS  Article  Google Scholar 

  2. Atkinson RK, Howden MEH, Tyler MI and Vonarx EJ (1998) Insecticidal toxins derived from funnel web (Atrax or Hadronyche) spiders. US Patent No. 5,763,568, Zeneca Limited, USA.

  3. MM Bradford (1976) ArticleTitleA rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72 248–254 Occurrence Handle942051 Occurrence Handle1:CAS:528:DyaE28XksVehtrY%3D Occurrence Handle10.1016/0003-2697(76)90527-3

    PubMed  CAS  Article  Google Scholar 

  4. JI Fletcher R Smith SI O’Donoghue M Nilges M Connor ME Howden et al. (1997) ArticleTitleThe structure of a novel insecticidal neurotoxin ω-atracotoxin-HV1, from the venom of an Australian funnel web spider Nat Struct Biol 4 559–566 Occurrence Handle9228949 Occurrence Handle1:CAS:528:DyaK2sXksVeksrk%3D Occurrence Handle10.1038/nsb0797-559

    PubMed  CAS  Article  Google Scholar 

  5. E Grishin (1999) ArticleTitlePolypeptide neurotoxins from spider venoms Eur J Biochem 264 276–280 Occurrence Handle10491071 Occurrence Handle1:CAS:528:DyaK1MXlvFWkur4%3D Occurrence Handle10.1046/j.1432-1327.1999.00622.x

    PubMed  CAS  Article  Google Scholar 

  6. F Guerineau P Mullineaux (1993) Plant transformation and expression vectors RRD Croy (Eds) Plant Molecular Biology Labfax Bios Scientific Publishers Oxford, UK 121–147

    Google Scholar 

  7. RP Hellens EA Edwards NR Leyland S Bean PM Mullineaux (2000) ArticleTitlepGreen: a versatile and flexible binary Ti vector for Agrobacterium-mediated plant transformation Plant Mol Biol 42 819–832 Occurrence Handle10890530 Occurrence Handle1:CAS:528:DC%2BD3cXks12jsL0%3D Occurrence Handle10.1023/A:1006496308160

    PubMed  CAS  Article  Google Scholar 

  8. RB Horsch RT Fraley SG Rogers PR Sanders A Lloyd H Hoffman (1984) ArticleTitleInheritance of functional genes in plants Science 223 496–498 Occurrence Handle1:CAS:528:DyaL2cXot12hsg%3D%3D Occurrence Handle17781445

    CAS  PubMed  Google Scholar 

  9. James C (2002) Preview, Global Status of Commercialized Transgenic Crops: 2002. ISAAA Briefs No. 27. ISAAA, Ithaca, New York.

  10. S Lawrence (2005) ArticleTitleAgbio keeps on growing Nat Biotechnol 23 281 Occurrence Handle15765074 Occurrence Handle1:CAS:528:DC%2BD2MXitF2nsLc%3D Occurrence Handle10.1038/nbt0305-281

    PubMed  CAS  Article  Google Scholar 

  11. Y Liu B Tabashnik S Meyer Y Carriere A Bartlett (2001) ArticleTitleGenetics of pink bollworm resistance to Bacillus thuringiensis toxin Cry1Ac J Econ Entomol 94 248–252 Occurrence Handle11233121 Occurrence Handle1:CAS:528:DC%2BD3MXmtVKntrY%3D Occurrence Handle10.1603/0022-0493-94.1.248

    PubMed  CAS  Article  Google Scholar 

  12. L Mehlo D Gahakwa PT Nghia NT Loc T Capell JA Gatehouse et al. (2005) ArticleTitleAn alternate strategy for sustainable pest resistance in genetically enhanced crops Proc Natl Acad Sci (USA) 102 7812–7816 Occurrence Handle1:CAS:528:DC%2BD2MXkvF2jtb8%3D Occurrence Handle10.1073/pnas.0502871102

    CAS  Article  Google Scholar 

  13. S Mouhat B Jouirou A Mosbah M Waard ParticleDe JM Sabatier (2004) ArticleTitleDiversity of folds in animal toxins acting on ion channels Biochem J 378 717–726 Occurrence Handle14674883 Occurrence Handle1:CAS:528:DC%2BD2cXisVyktrY%3D Occurrence Handle10.1042/BJ20031860

    PubMed  CAS  Article  Google Scholar 

  14. RS Norton PK Pallaghy (1998) ArticleTitleThe cystine knot structure of ion channel toxins and related polypeptides Toxicon 36 1573–1583 Occurrence Handle9792173 Occurrence Handle1:CAS:528:DyaK1cXmsVSqtb0%3D Occurrence Handle10.1016/S0041-0101(98)00149-4

    PubMed  CAS  Article  Google Scholar 

  15. J Sambrook ER Fritsch T Maniatis (1989) Molecular Cloning: a Laboratory Manual Cold Spring Harbor Laboratory Press Woodbury, New York

    Google Scholar 

  16. E Schnepf N Crickmore J Rie ParticleVan D Lereclus J Baum J Feitelson et al. (1998) ArticleTitle Bacillus thuringiensis and its pesticidal crystal proteins Microbiol Mol Biol Rev 62 775–806 Occurrence Handle9729609 Occurrence Handle1:CAS:528:DyaK1cXmtFOju7w%3D

    PubMed  CAS  Google Scholar 

  17. EJ Stewart F Aslund J Beckwith (1998) ArticleTitleDisulfide bond formation in the Escherichia coli cytoplasm: an in vivo role reversal for the thioredoxins EMBO J 17 5543–5550 Occurrence Handle9755155 Occurrence Handle1:CAS:528:DyaK1cXmvVyjsrY%3D Occurrence Handle10.1093/emboj/17.19.5543

    PubMed  CAS  Article  Google Scholar 

  18. FW Studier AH Rosenberg JJ Dunn JW Dubendorff (1990) ArticleTitleUse of T7 RNA polymerase to direct expression of cloned genes Methods Enzymol 185 60–89 Occurrence Handle2199796 Occurrence Handle1:CAS:528:DyaK3cXlsVKltro%3D Occurrence Handle10.1016/0076-6879(90)85008-C

    PubMed  CAS  Article  Google Scholar 

  19. BE Tabashnik T Dennehy MA Sims K Larkin (2002) ArticleTitleControl of resistant Pink Bollworm (Pectinophora gossypiella) by transgenic cotton that produces Bacillus thuringiensis toxin cry2Ab Appl Environ Microbiol 68 3790–3794 Occurrence Handle12147473 Occurrence Handle1:CAS:528:DC%2BD38XmtVaqurw%3D Occurrence Handle10.1128/AEM.68.8.3790-3794.2002

    PubMed  CAS  Article  Google Scholar 

  20. H Tamaoki R Miura M Kusunoki Y Kyogoku Y Kobayashi L Moroder (1998) ArticleTitleFolding motifs induced and stabilized by distinct cystine frameworks Protein Eng 11 649–659 Occurrence Handle9749917 Occurrence Handle1:CAS:528:DyaK1cXmtVWqurg%3D Occurrence Handle10.1093/protein/11.8.649

    PubMed  CAS  Article  Google Scholar 

  21. HW Tedford JI Fletcher GF King (2001) ArticleTitleFunctional significance of the β-hairpin in the insecticidal neurotoxin ω-atracotoxin-Hv1a J Biol Chem 276 26568–26576 Occurrence Handle11313356 Occurrence Handle1:CAS:528:DC%2BD3MXlsVKntLY%3D Occurrence Handle10.1074/jbc.M102199200

    PubMed  CAS  Article  Google Scholar 

  22. HW Tedford N Gilles A Menez CJ Doering GW Zamponi GF King (2004a) ArticleTitleScanning mutagenesis of ω-atracotoxin-Hv1a reveals a spatially restricted epitope that confers selective activity against insect calcium channels J Biol Chem 279 44133–44140 Occurrence Handle1:CAS:528:DC%2BD2cXot1yns7w%3D Occurrence Handle10.1074/jbc.M404006200

    CAS  Article  Google Scholar 

  23. HW Tedford BL Sollod F Maggio GF King (2004b) ArticleTitleAustralian funnel-web spiders: master insecticide chemists Toxicon 43 601–618 Occurrence Handle1:CAS:528:DC%2BD2cXislGjtro%3D Occurrence Handle10.1016/j.toxicon.2004.02.010

    CAS  Article  Google Scholar 

  24. X Wang M Connor D Wilson H Wilson G Nicholson R Smith et al. (2001) ArticleTitleDiscovery and structure of a potent and highly specific blocker of insect calcium channels J Biol Chem 276 40306–40312 Occurrence Handle11522785 Occurrence Handle1:CAS:528:DC%2BD3MXotFGgs7g%3D

    PubMed  CAS  Google Scholar 

  25. JZ Zhao XL Fan XP Shi RM Zhao YL Fan (1997) ArticleTitleGene pyramiding: an effective strategy of resistance management for Helicoverpa armigera and Bacillus thuringiensis Resist Pest Manage Newslett 9 19–21

    Google Scholar 

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Correspondence to Zahid Mukhtar.

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The authors Sher Afzal Khan and Zahid Mukhtar contributed equally to this work.

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Khan, S.A., Zafar, Y., Briddon, R.W. et al. Spider Venom Toxin Protects Plants from Insect Attack. Transgenic Res 15, 349 (2006). https://doi.org/10.1007/s11248-006-0007-2

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Keywords

  • insect
  • Hadronyche versuta
  • resistance
  • spider
  • toxin
  • transgenic