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Spider toxin (Hvt) gene cloned under phloem specific RSs1 and RolC promoters provides resistance against American bollworm (Heliothis armigera)

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Abstract

Spider venoms are neurotoxin proteins that can kill insects. Spider toxin Hvt gene was cloned under two phloem specific RSs1 and RolC promoters, transformed into tobacco plants through Agrobacterium-mediated transformation and tested against Heliothis armigera larvae. Transgenic plants were confirmed through PCR. First instar larvae of H. armigera were released on detached leaves of transformed and non-transformed plants. Insect bioassays showed 93–100% mortality of H. armigera larvae within 72 h on the leaves of transgenic plants while all larvae survived and continued feeding on detached leaves from non-transformed control plants. The Hvt gene expressing under phloem specific RSs1 and RolC promoters could therefore be used for developing H. armigera-resistant, genetically-modified crops.

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References

  • Bajaj S, Mohanty A (2005) Recent advances in rice biotechnology-towards genetically superior transgenic rice. Plant Biotechnol J 3:275–307

    Article  PubMed  CAS  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • Dutta I, Majumder I, Saha P, Ray K, Das S (2005) Constitutive and phloem specific expression of Allium sativum leaf agglutinin (ASAL) to engineer aphid (Lipaphis erysimi) resistance in transgenic Indian mustard (Brassica juncea). Plant Sci 169:996–1007

    Article  CAS  Google Scholar 

  • Foissac X, Loc NT, Christou P, Gatehouse AMR, Gatehouse JA (2000) Resistance to green leafhopper (Nephotettix virescens) and brown planthopper (Nilaparvata lugens) in transgenic rice expressing snowdrop lectin (Galanthus nivalis agglutinin; GNA). J Insect Physiol 46:573–583

    Article  PubMed  CAS  Google Scholar 

  • Hernández-Campuzano B, Suárez R, Lina L, Hernández V, Villegas E, Corzo G, Iturriaga G (2009) Expression of a spider venom peptide in transgenic tobacco confers insect resistance. Toxicon 53:122–128

    Article  PubMed  Google Scholar 

  • Khan SA, Zafar Y, Briddon RW, Malik KA, Mukhtar Z (2006) Spider venom toxin protects plants from insect attack. Transgenic Res 15:349–357

    Article  PubMed  CAS  Google Scholar 

  • King GF, Tedford HW, Maggio F (2002) Structure and function of insecticidal neurotoxins from Australian funnel-web spiders. J Toxicol Toxin Rev 21:359–389

    CAS  Google Scholar 

  • Li Z, Jayasankar S, Gray DJ (2001) Expression of a bifunctional green fluorescent protein (GFP) fusion marker under the control of three constitutive promoters and enhanced derivatives in transgenic grape (Vitis vinifera). Plant Sci 160:877–887

    Article  PubMed  CAS  Google Scholar 

  • Matsuki R, Onodera H, Yamauchi T, Uchimiya H (1989) Tissue specific expression of the RolC promoter of the Ri plasmid in transgenic rice plants. Mol Gen Genet 220:12–16

    Article  CAS  Google Scholar 

  • Mukhtar Z, Khan SA, Zafar Y, Malik KA (2004) Synthetic omega-atracotoxin Hvla gene of funnel web spider (Hadronyche versuta) for development of insect resistant plants. Patent No. 139032, IPO, Pakistan

  • Rao KV, Rathore KS, Hodges TK, Fu X, Stoger E, Sudhakar S, Williams P, Christou P, Bharathi M, Bown DP, Powell KS, Spence J, Gatehouse A, Gatehouse JA (1998) Expression of snowdrop lectin (GNA) in transgenic plants confers resistance to rice brown plant hopper. Plant J 15:469–477

    Article  PubMed  CAS  Google Scholar 

  • Schmuelling T, Schell J, Spena A (1989) Promoters of the rolA, B, and C genes of Agrobacterium rhizogenes are differentially regulated in transgenic plants. Plant Cell 1:665–670

    Article  CAS  Google Scholar 

  • Sharma HC (2001) Cotton bollworm/legume pod borer, Heliothis armigera (Hubner) (Lepidoptera, Noctuidae) biology and management in crop protection Compendium. CAB International, Wallingford, p 72

    Google Scholar 

  • Sharma HC (2005) Heliothis/Helicoverpa management emerging trends and stratigies for future research. Oxford and IBH publishing Co. Pvt. Ltd, New Delhi, p 469

    Google Scholar 

  • Shi Y, Wang MB, Powell KS, Damme EV, Hilder VA, Gatehouse AMR, Boulter D, Gatehouse JA (1994) Use of the rice sucrose synthase-1 promoter to direct phloem-specific expression of β-glucuronidase and snowdrop lectin genes in transgenic tobacco plants. J Exp Bot 45:623–631

    Article  CAS  Google Scholar 

  • Sudhakar D, Fu X, Stoger E, Williams S, Spence J, Brown DP, Bharathi M, Gatehouse JA, Christou P (1998) Expression and immunolocalization of the snowdrop lectin, GNA in transgenic rice plants. Transgenic Res 7:371–378

    Article  PubMed  CAS  Google Scholar 

  • Sugaya S, Hayakawa K, Handa K, Uchimiya H (1989) Cell-specific expression of the RolC gene of the TL-DNA of Ri plasmid in transgenic tobacco plants. Plant Cell Physiol 30:649–653

    CAS  Google Scholar 

  • Wang XH, Connor M, Smith R, Maciejewski MW, Howden MEH, Nicholson GM, Christie MJ, King GF (2000) Discovery and characterization of a family of insecticidal neurotoxins with a rare vicinal disulfide bridge. Nat Struct Mol Biol 7:505–513

    Article  CAS  Google Scholar 

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Acknowledgments

We would like to thank Dr. Tiziana Pandoifini, Italy for providing plasmid vector containing RolC promoter for this study. We acknowledge Dr. Imran Amin and Dr. Muhammad Ashfaq for providing technical support in qPCR and insect bioassays respectively. We are also thankful to Agriculture Department, Government of Sindh, Pakistan for providing PhD study fellowship to Allah Dino Shah.

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Authors and Affiliations

  1. National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box 577, Faisalabad, Pakistan

    Allah Dino Shah, Moddassir Ahmed, Zahid Mukhtar, Sher Afzal Khan, Imran Habib, Zahid Abbas Malik, Shahid Mansoor & Nasir Ahmad Saeed

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  1. Allah Dino Shah
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  2. Moddassir Ahmed
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  3. Zahid Mukhtar
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  4. Sher Afzal Khan
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  5. Imran Habib
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  6. Zahid Abbas Malik
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  7. Shahid Mansoor
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  8. Nasir Ahmad Saeed
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Correspondence to Nasir Ahmad Saeed.

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Shah, A.D., Ahmed, M., Mukhtar, Z. et al. Spider toxin (Hvt) gene cloned under phloem specific RSs1 and RolC promoters provides resistance against American bollworm (Heliothis armigera). Biotechnol Lett 33, 1457–1463 (2011). https://doi.org/10.1007/s10529-011-0572-1

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  • DOI: https://doi.org/10.1007/s10529-011-0572-1

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