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Expression of Cry1Ac in Transgenic Tobacco Plants Under the Control of a Wound-Inducible Promoter (AoPR1) Isolated from Asparagus officinalis to Control Heliothis virescens and Manduca sexta

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Abstract

Expression of cry1Ac gene from Bacillus thuringiensis (Bt) was evaluated under the control of a wound-inducible AoPR1 promoter from Asparagus officinalis in transgenic tobacco plants. The leaves of transgenic plants were mechanically wounded to evaluate the activity of the AoPR1 promoter in driving the expression of Cry1Ac protein at the wound site. Our results indicate that mechanical wounding of transgenic plants was effective in inducing the expression of Cry1Ac protein. As a result of this induction, the accumulated levels of Cry1Ac protein increased during 6–72 h post-wounding period. The leaves of transgenic tobacco plants were evaluated for resistance against Heliothis virescens and Manduca sexta in insect bioassays in two different ways. The detached tobacco leaves were either fed directly to the insect larvae or they were first mechanically wounded followed by a 72 h post-wounding feeding period. Complete protection of mechanically wounded leaves of transgenic plants was observed within 24 h of the bioassay. The leaves of transgenic plants fed directly (without pre-wounding) to the larvae achieved the same level of protection between 24 and 72 h of the bioassay.

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Acknowledgements

The authors wish to thank Prof. John Draper (University of Wales, Aberystwyth, UK) for critical reading of the manuscript and Leicester University (UK) for permission to use the AoPR1 promoter (Patent No. 6031151) in this research. This study was funded by The Rockefeller Foundation and NSERC grants to IA. SGO gratefully acknowledges a postdoctoral fellowship from The Scientific and Technological Research Council of Turkey.

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Authors and Affiliations

  1. Agricultural Biotechnology Laboratories, Department of Biochemistry Microbiology & Immunology, University of Ottawa, 451 Smyth Rd, Ottawa, ON, K1H 8M5, Canada

    Selma Gulbitti-Onarici, Mohsin Abbas Zaidi, Ibrahim Taga & Illimar Altosaar

  2. The Scientific and Technical Council of Turkey, Genetic Engineering and Biotechnology Institute, P.O. Box 21, 41470, Gebze-Kocaeli, Turkey

    Selma Gulbitti-Onarici

  3. Clinical Biochemistry and Nutrition Unit, Biochemistry Department, Faculty of Sciences, University of Douala, P.O. Box 24157, Douala, Cameroon

    Ibrahim Taga

  4. Department of Field Crops, Faculty of Agriculture, University of Ankara, 06110, Diskapi, Ankara, Turkey

    Sebahattin Ozcan

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  1. Selma Gulbitti-Onarici
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Correspondence to Illimar Altosaar.

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Gulbitti-Onarici, S., Zaidi, M.A., Taga, I. et al. Expression of Cry1Ac in Transgenic Tobacco Plants Under the Control of a Wound-Inducible Promoter (AoPR1) Isolated from Asparagus officinalis to Control Heliothis virescens and Manduca sexta . Mol Biotechnol 42, 341–349 (2009). https://doi.org/10.1007/s12033-009-9168-6

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