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Multiple insect resistance in transgenic tomato plants over-expressing two families of plant proteinase inhibitors

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Abstract

Protease inhibitors have been proposed as potential defense molecules for increased insect resistance in crop plants. Compensatory over-production of insensitive proteases in the insect, however, has limited suitability of these proteins in plant protection, with very high levels of inhibitor required for increased plant resistance. In this study we have examined whether combined used of two inhibitors is effective to prevent this compensatory response. We show that leaf-specific over-expression of the potato PI-II and carboxypeptidase inhibitors (PCI) results in increased resistance to Heliothis obsoleta and Liriomyza trifolii larvae in homozygote tomato lines expressing high levels (#62;1 the total soluble proteins) of the transgenes. Leaf damage in hemizygous lines for these transformants was, however, more severe than in the controls, thus evidencing a compensation response of the larvae to the lower PI concentrations in these plants. Development of comparable adaptive responses in both insects suggests that insect adaptation does not entail specific recognition of the transgene, but rather represents a general adaptive mechanism triggered in response to the nutritional stress imposed by sub-lethal concentrations of the inhibitors. Combined expression of defense genes with different mechanisms of action rather than combinations of inhibitors may then offer a better strategy in pest management as it should be more effective in overcoming this general adaptive response in the insect.

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References

  • P.B. Bown H.S. Wilkinson J.A. Gatehouse (1998) ArticleTitleMidgut carboxypeptidase from Helicoverpa armigera (Lepidoptera: Noctuidae) larvae: enzyme characterization, cDNA cloning and expression Insect Biochem. Mol. Biol. 28 739–749

    Google Scholar 

  • C.J. Bolter M.A. Jongsma (1995) ArticleTitleColorado potato beetles (Leptinotarsa decemlineata) adapt to proteinase inhibitors induced in potato leaves by methyl jasmonate J. Insect Physiol. 41 1071–1078

    Google Scholar 

  • R.M. Broadway (2000) The response of insects to dietary protease inhibitors D. Michaud (Eds) Recombinant Protease Inhibitors in Plants. Lanoes Bioscience Georgetown 81–90

    Google Scholar 

  • R.M. Broadway (1996) ArticleTitleDietary proteinase inhibitors alter compliment of midgut proteases Arch. Insect Biochem. Physiol. 32 39–53

    Google Scholar 

  • R.M. Broadway S.S. Duffey (1986) ArticleTitlePlant proteinase inhibitors: mechanism of action and effect on the growth and digestive physiology of larval Heliothis zea and Spodoptera exigua J. Insect Physiol. 32 827–833

    Google Scholar 

  • F. Brunelle C. Cloutier D. Michaud (2004) ArticleTitleColorado potato beetles compensate for tomato Cathepsin D inhibitor expressed in transgenic potato Arch. Insect Biochem. Physiol. 55 103–113

    Google Scholar 

  • J.T. Christeller W.A. Laing N.P. Marwick E.P.J. Burgess (1992) ArticleTitleMidgut protease activities in 12 phytophagous lepidopteran larvae: dietary and protease inhibitor interactions Insect Biochem. Mol. Biol. 22 735–746

    Google Scholar 

  • C. Cloutier M. Fournier C. Jean S. Yelle D. Michaud (1999) ArticleTitleGrowth compensation and faster development of Colorado potato beetle (Coleoptera: Chrysomelidae) feeding on potato foliage expressing oryzacystatin I Arch. Insect. Biochem. Physiol. 40 69–79

    Google Scholar 

  • C. Cloutier C. Jean M. Fournier S. Yelle D. Michaud (2000) ArticleTitleAdult Colorado potato beetles, Leptinotarsa decemlineata compensate for nutritional stress on Oryzacystatin I-transgenic potato plants by hypertrophic behavior and over-production of insensitive proteases Arch. Insect Biochem. Physiol. 44 69–81

    Google Scholar 

  • F. De Leo M.A. Bonadé-Bottino L.R. Ceci R. Gallerani L. Jouanin (1998) ArticleTitleOpposite effects on Spodoptera littoralis larvae of high expression level of a trypsin proteinase inhibitor in transgenic plants Plant Physiol. 118 997–1004

    Google Scholar 

  • S.L. Dellaporta J. Wood J.B. Hicks (1983) ArticleTitleA plant DNA minipreparation version II Plant Mol. Biol. Rep. 1 19–21

    Google Scholar 

  • R.G. Donald A.R. Cashmore (1990) ArticleTitleMutation of either G box or I box sequences profoundly affects expression from the Arabidopsis rbcS-1A promoter EMBO J 9 1717–1726 Occurrence Handle1:CAS:528:DyaK3cXlt1Ont7c%3D Occurrence Handle2347304

    CAS  PubMed  Google Scholar 

  • J.A.T. Dow (1992) ArticleTitlepH gradients in lepidopteran midgut J. Exp. Biol. 172 355–375

    Google Scholar 

  • X. Duan X. Li Q. Xue M. Abo-El-Saad D. Xu R. Wu (1996) ArticleTitleTransgenic rice plants harboring an introduced potato proteinase inhibitor II gene are insect resistant Nat. Biotech. 14 494–498

    Google Scholar 

  • T. Fushiki H. Kajiura S.I. Fukuoka K. Kido T. Semba K. Iwai (1989) ArticleTitleEvidence for an intramural mediator in rat pancreatic enzyme secretion: reconstitution of the pancreatic response with dietary protein, trypsin and the monitor peptide J. Nut. 119 622–627

    Google Scholar 

  • Gatehouse, A.M.R., Boulter, D. and Hilder, V.A. 1992. In: A.M.R. Gatehouse, V. A. Hilder, and D. Boulter (Eds.). Plant Genetic Manipulation for Crop Protection. CAB International, pp. 155–181.

  • L.N. Gatehouse J.T. Christeller H.S. Gatehouse X.Y. Zou (2002) ArticleTitleA strong inhibitor of chymotrypsin/eleastase is highly antimetabolic to Helicoverpa armigera larvae New Zealand Plant Protect 55 421–428

    Google Scholar 

  • C. Girard M. LeMétayer B. Zaccomer E. Bartlet I. Williams M. Bonadé-Bottino M.H. Pham-Delegue L. Jouanin (1998) ArticleTitleGrowth stimulation of beetle larvae reared on a transgenic oilseed rape expressing a cysteine protease inhibitor J. Insect Physiol. 44 263–270

    Google Scholar 

  • V.A. Hilder A.M.R. Gatehouse S.E. Sheerman R.F. Barker D. Boulter (1987) ArticleTitleA novel mechanism of insect resistance engineered into tobacco Nature 330 160–163

    Google Scholar 

  • V.A. Hilder A.M.R. Gatehouse D. Boulter (1992) Transgenic plants conferring insect tolerance: protease inhibitor approach S. Kung R. Wu (Eds) Transgenic Plants Academic Press New York, USA 310–338

    Google Scholar 

  • R. Johnson J. Narvaez G. An C.A. Ryan (1989) ArticleTitleExpression of proteinase inhibitors I and II in transgenic tobacco plants: effects on natural defense against Manduca sexta larvae Proc. Natl. Acad. Sci. USA 86 9871–9875

    Google Scholar 

  • K.A. Johnston M.J. Lee C. Brough V.A. Hilder A.M.R. Gatehouse J.A. Gatehouse (1995) ArticleTitleProtease activities in the larval midgut of Heliothis virescens: evidence for trypsin and chymotripsin-like enzymes Insect Biochem. Mol. Biol. 25 375–383

    Google Scholar 

  • M.A. Jongsma P.L. Bakker J. Peters D. Bosch W.J. Stiekema (1995) ArticleTitleAdaptation of Spodoptera exigua larvae to plant proteinase inhibitors by induction of gut proteinase activity insensitive to inhibition Proc. Natl. Acad. Sci. USA 92 8041–8045

    Google Scholar 

  • M.A. Jongsma C. Bolter (1997) ArticleTitleThe adaptation of insects to plant protease inhibitors J. Insect Physiol. 43 885–895

    Google Scholar 

  • L. Jouanin M. Bonadé-Bottino C. Girard G. Morrot M. Giband (1998) ArticleTitleTrangenic plants for insect resistance Plant Sci. 131 1–11

    Google Scholar 

  • A. Katherine (1995) ArticleTitleProtease activities in larva midgut of Helioths virescens: evidence for Trypsin and Chymotrypsin-like enzymes Insect Biochem. Mol. Biol. 25 375–383

    Google Scholar 

  • M. Koornneef M. Jongsma R. Weide P. Zabel J. Hille (1987) Transformation of tomato D.J. Nevins R.A. Jones (Eds) Tomato Biotechnology Alan R. Liss Inc. New York 169–178

    Google Scholar 

  • A. Lecardonnel L. Chauvin L. Jouanin A. Beaujean G. Prévost B. Sangwan-Norreel (1999) ArticleTitleEffects of rice cystatin I expression in transgenic potato on Colorado potato beetle larvae Plant Sci. 140 87–98

    Google Scholar 

  • S.I. Lee S.H. Lee J.C. Koo H.J. Chun C.O. Lim J.H. Mun Y.H. Song M.J. Cho (1999) ArticleTitleSoybean Kunitz trypsin inhibitor (SKTI) confers resistance to the brown planthopper (Nilaparvata lugens Stal) in transgenic rice Mol. Breed. 5 1–9

    Google Scholar 

  • J.C. Leplé M. Bonadé-Bottino S. Augustin G. Pilate V. DumanoisLêTân A. Delplanque D. Cornu L. Jouanin (1995) ArticleTitleToxicity to Chrysomela tremulae (Coleoptera: Crysomelidae) of transgenic poplars expressing a cysteine proteinase inhibitor Mol. Breed. 1 319–328

    Google Scholar 

  • Y.E. Li Z. Zhu Z.X. Chen X. Wu W. Wang S.J. Li (1998) ArticleTitleObtaining transgenic cotton plants with cowpea trypsin inhibitor Acta Gossypii Sinica 10 237–243

    Google Scholar 

  • J. Logemann J. Schell L. Willmitzer (1987) ArticleTitleImproved method for the isolation of RNA from plant tissues Anal Biochem 163 16–20 Occurrence Handle1:CAS:528:DyaL2sXktlKitr0%3D Occurrence Handle2441623

    CAS  PubMed  Google Scholar 

  • M.T. McManus D.W.R. White P.G. McGregor (1994) ArticleTitleAccumulation of a chymotrypsin inhibitor in transgenic tobacco can affect the growth of insect pests Transg. Res. 3 50–58

    Google Scholar 

  • D. Nagadhara S. Ramesh I.C. Pasalu Y. Kondala Rao N.V. Krishnaiah N.P. Sarma D.P. Bown J.A. Gatehouse V.D. Reddy K.V. Rao (2003) ArticleTitleTransgenic indica rice resistant to sap-sucking insects Plant Biotech. J. 1 231–240

    Google Scholar 

  • G.L. Orr J.A. Strickland T.A. Walsh (1994) ArticleTitleInhibition of Diabrotica larval growth by a multicystatin from potato tubers J. Insect Physiol. 40 893–900

    Google Scholar 

  • D. Rivard C. Cloutier D. Michaud (2004) ArticleTitleColorado potato beetles show differential digestive compensatory responses to host plants expressing different sets of defense proteins Arch. Insect Biochem. Physiol. 55 114–123

    Google Scholar 

  • C.A. Ryan (1990) ArticleTitleProteinase inhibitors in plants: genes for improving defenses against insects and pathogens Ann. Rev. Phytopath 28 425–449

    Google Scholar 

  • C.A. Ryan (2000) ArticleTitleThe systemin signaling pathway: differential activation of plant defensive genes Biochim. Biophys. Acta 1477 112–121

    Google Scholar 

  • T.H. Schuler G.M. Poppy B.R. Kerry I. Denholm (1998) ArticleTitleInsect-resistant transgenic plants Trends Biotechnol. 16 168–175

    Google Scholar 

  • A.W. Spannagel D. Guan R.A. Liddle J.R. Reeve G.M. Green (1996) ArticleTitlePurification and characterization of a luminal cholecystokinin-releasing factor (LCRF) from rat intestinal secretion Proc. Natl. Acad. Sci. USA 93 4415–4420

    Google Scholar 

  • J. Stockhaus J. Schell L. Willmitzer (1989) ArticleTitleCorrelation of the expression of the nuclear photosynthetic gene St-LS1 with the presence of chloroplasts EMBO J 8 2445–2451

    Google Scholar 

  • J.G. Turner C. Ellis A. Devoto (2002) ArticleTitleThe jasmonate signal pathway Plant Cell 14 153–164

    Google Scholar 

  • P.E. Urwin K.M. Troth E.I. Zubko H.J. Atkinson (2001) ArticleTitleEffective transgenic resistance to Globodera pallida in potato field trials Mol. Breed. 8 95–101

    Google Scholar 

  • J. Villanueva F. Canals S. Prat D. Ludevid E. Querol F.X. Avilés (1998) ArticleTitleCharacterization of the wound-induced metallocarboxypeptidase inhibitor from potato. cDNA sequence, induction of gene expression, subcellular immunolocalization and potential roles of the C-terminal propeptide FEBS Lett. 440 175–182

    Google Scholar 

  • A. Wu X. Sun Y. Pang K. Tang (2002) ArticleTitleHomozygous transgenic rice lines expressing GNA with enhanced resistance to the rice sap-sucking pest Laodelphax striatellus Plant Breed. 121 93–95

    Google Scholar 

  • D.P. Xu Q.Z. Xue D. McElroy Y. Mawal V.A. Hilder R. Wu (1996) ArticleTitleConstitutive expression of a cowpea trypsin inhibitor gene, CpTi, in transgenic rice plants confers resistance to two major rice insect pests Mol. Breed. 2 167–173

    Google Scholar 

  • K. Zhu-Salzman J.-E. Ahn R.A. Salzman H. Koiwa R.E. Shade S. Balfe (2003) ArticleTitleFusion of a soybean cysteine protease inhibitor and a legume lectin enhaces anti-insect activity synergistically Agric. Forest Entomol. 5 317–323

    Google Scholar 

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Authors and Affiliations

  1. Departmento de Genética Molecular de Plantas, Centro Nacional de Biotecnología-CSIC, Campus Universidad Autónoma, Cantoblanco, 28049, Madrid, Spain

    Ashraf Abdeen, Ariadna Virgós, Elisenda Olivella & Salomé Prat

  2. Institut de Biotecnologia i de Biomedicina, Universitat Autónoma de Barcelona, Bellaterra, 08193, Barcelona, Spain

    Josep Villanueva & Xavier Avilés

  3. Departament d’Entomologia, Centre de Cabrils-IRTA, Carretera de Cabrils s/n, Cabrils, 08348, Barcelona, Spain

    Rosa Gabarra

Authors
  1. Ashraf Abdeen
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  2. Ariadna Virgós
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  3. Elisenda Olivella
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  4. Josep Villanueva
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  5. Xavier Avilés
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  6. Rosa Gabarra
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  7. Salomé Prat
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Correspondence to Salomé Prat.

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Abdeen, A., Virgós, A., Olivella, E. et al. Multiple insect resistance in transgenic tomato plants over-expressing two families of plant proteinase inhibitors. Plant Mol Biol 57, 189–202 (2005). https://doi.org/10.1007/s11103-004-6959-9

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  • DOI: https://doi.org/10.1007/s11103-004-6959-9

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