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Expression of GNA and biting site-restricted cry1Ac in cotton; an efficient attribution to insect pest management strategies

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Abstract

Insect-resistant transgenic cotton has been commercialized for two decades. Most of the introduced cultivars express Bt gene(s) constitutively under the control of 35S promoter in whole-plant tissues. However, there have been other promoters considered by researchers to confine the toxin expression to targeted organ and tissues. We developed a triple-gene construct including GNA, cry1Ac and cp4 epsps genes. We attempted to confine cry1Ac expression to insect biting sites by cloning it to downstream of a wound-inducible promoter isolated from Asparagus officinalis (AoPR1). Moreover, to broaden the range of resistance, GNA was driven by the 35S promoter to target the sap-sucking insects like aphids which impose large losses in cotton production. To select the transformants in selection medium and for glyphosate tolerance, GNA and cry1Ac genes were accompanied with cp4 epsps gene. Two binary vectors harboring desired genes were constructed and utilized in the study (pGTGNAoC1AC and pGTGN35C1AC). Transformation of cultivar GSN-12 was carried out by employing Agrobacterium tumefaciens strain EHA105. Plantlets were primarily screened under glyphosate (N-phosphonomethyl glycine) selection pressure and subsequently subjected to molecular and biotoxicity assays. Introduction of cry1Ac and GNA to cotton plant conferred resistance to Spodoptera littoralis and Aphis gossypii Glover. Restriction of cry1Ac toxin protein to insect biting sites along with a plant lectin attributes significantly to insect pest management strategies.

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Acknowledgements

The PhD. fellowship awarded by The Scientific and Technological Research Council of Turkey (TUBITAK)-BIDEB to Dr. S.D. Khabbazi is deeply appreciated. The authors are grateful to the Leicester University (UK) for giving permission to use AoPR1 promoter for research purposes, Dr. Selma Onarıcı (TÜBİTAK GMBE) for providing pJIT61.cry1Ac plasmid and Prof. Umut Toprak (Department of Crop Protection, Ankara University) for providing S. littoralis larvae.

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Authors and Affiliations

  1. TARBIYOTEK Laboratories, Department of Field Crops, Faculty of Agriculture, Ankara University, Ankara, Turkey

    Saber Delpasand Khabbazi, Dilek Başalma & Sebahattin Özcan

  2. Department of Plant Protection, Faculty of Agriculture, University of Tabriz, Tabriz, Iran

    Afsaneh Delpasand Khabbazi

  3. Central Research Institute for Field Crops, Ministry of Food, Agriculture and Livestock, Ankara, Turkey

    S. Fatih Özcan

  4. Department of Agricultural Genetic Engineering, Faculty of Agricultural Sciences and Technologies, Niğde Ömer Halisdemir University, Niğde, Turkey

    Allah Bakhsh

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  1. Saber Delpasand Khabbazi
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Correspondence to Saber Delpasand Khabbazi or Sebahattin Özcan.

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Khabbazi, S.D., Khabbazi, A.D., Özcan, S.F. et al. Expression of GNA and biting site-restricted cry1Ac in cotton; an efficient attribution to insect pest management strategies. Plant Biotechnol Rep 12, 273–282 (2018). https://doi.org/10.1007/s11816-018-0493-8

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