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Transgenic Expression of Trypsin Inhibitor CMe from Barley in Indica and Japonica Rice, Confers Resistance to the Rice Weevil Sitophilus Oryzae

Transgenic Research volume 12, pages 23–31 (2003)Cite this article

Abstract

Indica and japonica rice (Oryza sativa L.) plants were transformed by particle bombardment with the Itr1 gene encoding the barley trypsin inhibitor BTI-CMe, under the control of its own promoter that confers endosperm specificity, and the maize ubiquitin promoter. From 38 independent transgenic lines of indica (breeding line IR58) and 15 of the japonica (cv Senia) selected, 22 and 11, respectively, expressed the barley inhibitor at detectable levels. The transgene was correctly translated as indicated by western blot analysis with a level of expression in R3 seeds up to 0.31% (IR58) and 0.43% (Senia) of the total extracted protein. The functional integrity of BTI-CMe was confirmed by trypsin activity assays in liquid media and by activity staining gels, performed with seed extracts. The significant reduction of the survival rate of the rice weevil (Sitophilus oryzae, Coleoptera: Curculionidae) reared on homozygous transgenic indica and japonica rice seeds expressing the BTI-CMe, compared to non-transformed controls, and the decrease in the trypsin-like activity of insect crude midgut extracts, confirmed the utility of this proteinase inhibitor gene for the control of important storage pests.

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Author notes

  1. Julio Alfonso-Rubí

    Present address: Centro de Ingeniería Genética y Biotecnología, Apartado 83, CP 60200, Sancti Spiritus, Cuba

Authors and Affiliations

  1. Laboratorio de Bioquímica y Biología Molecular, Departamento de Biotecnología-UPM, E.T.S.Ingenieros Agrónomos, 28040, Madrid, Spain

    Julio Alfonso-Rubí, Pilar Carbonero & Isabel Díaz

  2. Departamento de Biología de Plantas, Centro de Investigaciones Biológicas, CSIC, Velázquez 144, 28006, Madrid, Spain

    Félix Ortego & Pedro Castañera

Authors
  1. Julio Alfonso-Rubí
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  2. Félix Ortego
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  3. Pedro Castañera
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  4. Pilar Carbonero
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  5. Isabel Díaz
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Correspondence to Isabel Díaz.

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Alfonso-Rubí, J., Ortego, F., Castañera, P. et al. Transgenic Expression of Trypsin Inhibitor CMe from Barley in Indica and Japonica Rice, Confers Resistance to the Rice Weevil Sitophilus Oryzae . Transgenic Res 12, 23–31 (2003). https://doi.org/10.1023/A:1022176207180

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  • DOI: https://doi.org/10.1023/A:1022176207180

  • barley trypsin inhibitor
  • Sitophilus oryzae
  • storage pest resistance
  • transgenic rice